Microarray needle-shaped liquid capsule as well as preparation and application thereof

Abstract

The invention discloses a microarray needle-shaped liquid capsule as well as preparation and application thereof, and belongs to the technical field of biomedical materials. The invention provides a microarray needle-shaped liquid capsule, which is composed of soluble microneedles and a packaging agent, has a hollow structure, can load chemotherapeutic drugs, and is especially suitable for non-water-soluble drugs. According to the design, the problem of insufficient drug loading capacity of a traditional microneedle is effectively solved, the bioavailability of the drug is improved by taking ethanol as a solvent, and the synergistic treatment of chemotherapy and local tumor ablation is realized at the same time. The material not only has excellent mechanical properties and biocompatibility, but also shows significant advantages in treatment of melanoma, reduces skin inflammation reactions, and improves the treatment comfort of patients.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a microarray needle-shaped liquid capsule and a preparation and application thereof. Background Art

[0002] The skin is the largest organ in the human body. Due to its high metastatic and high mortality characteristics, melanoma has become one of the most deadly skin diseases, with its morbidity and mortality rates rising year by year around the world. Nowadays, common methods for treating melanoma include chemotherapy and local ablation therapy. Chemotherapy is a treatment method that uses chemical drugs to kill cancer cells, inhibit the growth and reproduction of cancer cells, and promote the differentiation of cancer cells. It is suitable for most cancer patients. However, when chemotherapy is used alone to treat cancer, the treatment effect may not be sufficient to achieve the desired clinical goals or effects. Commonly used chemotherapy drugs in clinical practice include paclitaxel, doxorubicin and oxaliplatin. Since melanoma is sensitive to paclitaxel, paclitaxel is often used as one of the main drugs for treating melanoma in clinical treatment. Local ablation therapy is a non-invasive treatment method for treating local tumors. It uses physical or chemical methods to directly act on the lesion area to achieve ablation or destruction of the lesion tissue. It has the advantages of minimally invasive and short recovery period, but its treatment failure rate is high. In recent years, common local ablation treatments include percutaneous ethanol injection, which specifically involves injecting anhydrous ethanol into the tumor site to cause cell dehydration and coagulative necrosis in the tumor, thereby killing cancer cells.

[0003] At present, many drug delivery systems such as microcapsules, injectable hydrogels and transdermal patches have been developed to address the problem of how to deliver drugs to tumor sites. Among them, the microneedle patch in the transdermal patch can load the drug into the needle cavity and achieve on-demand drug delivery in a minimally invasive manner. In addition, the microneedle patch can effectively overcome the common shortcomings of other local drug delivery systems, such as low drug penetration and high inflammatory response, and balance the comfort and painlessness of patients as much as possible. Patients can even administer the drug by themselves, providing a promising drug delivery strategy for the treatment of melanoma.

[0004] Many types of microneedle patches have been developed and produced for cancer treatment, including solid microneedles, coated microneedles, hydrogel microneedles, hollow microneedles and soluble microneedles. Solid microneedles are metal or silicon microneedles whose needle tips pierce the skin and create micron-sized channels. Drugs can be delivered directly to the skin layer through these channels. However, solid microneedles may leave metal fragments in the skin, causing erythema and swelling of the skin. Coated microneedles load drugs to the outside of the microneedle tip. When the drug is inserted into the skin, it is quickly delivered. However, too thin a coating will result in a lower drug loading, limiting the efficiency of drug delivery. Hydrogel microneedles are usually prepared from swelling polymers. When the hydrogel microneedles are inserted, the polymers swell into the skin and release the drugs. However, the accuracy of drug delivery is limited. Hollow microneedles are drugs placed in the empty area, and large doses of drugs are loaded and delivered through the device. However, they require external devices to achieve drug delivery. For the combination of drugs and microneedle patches, soluble microneedles are currently the most popular type of microneedle patch. They have low manufacturing costs and are made of materials with good solubility and biocompatibility. When soluble microneedles are inserted into the skin, the polymer will release the drug after dissolving in tissue fluid. However, soluble microneedles also have the disadvantages of poor mechanical properties and limited drug loading.

[0005] Although many studies have been conducted using dissolvable microneedle patches to deliver drugs to cancer sites, there are still some drawbacks:

[0006] (1) The drug loading capacity of soluble microneedles is relatively small.

[0007] (2) There are still few reports on the differences in drug loading capacity of soluble microneedles with different structures.

[0008] (3) Most studies usually use water as a solvent to dissolve chemotherapy drugs. However, the common chemotherapy drugs currently used in clinical practice, such as paclitaxel, are difficult to dissolve in water due to their complex chemical structures, which to a certain extent limits their widespread clinical application.

[0009] (4) For chemotherapy drugs with poor water solubility, most studies use organic solvents such as dimethyl sulfoxide, N-methylpyrrolidone, N,N-dimethylacetamide to dissolve them; or encapsulate poorly water-soluble drugs in liposomes or micelles; or chemically modify the drug structure to make it more soluble in water. However, there are few reports on the use of a solvent that can be taken by the human body, dissolve poorly water-soluble chemotherapy drugs, and treat melanoma. Summary of the invention

[0010] In view of the above defects, the purpose of the present invention is to provide an application of a microarray needle-shaped liquid capsule for treating melanoma after loading drugs. The capsule is composed of soluble microneedles and an encapsulating agent, and has a hollow structure, which can be loaded with a large dose of chemotherapy drugs.

[0011] Specifically, the microarray needle-shaped liquid capsule is equivalent to a portable container loaded with drugs, and there is no substance in the container, so the container can be completely filled with drugs, thereby achieving large-dose delivery of drugs, solving the defect of low drug loading capacity of soluble microneedles.

[0012] The present invention also provides a method for preparing the microarray needle-shaped liquid capsule, which specifically comprises the following steps:

[0013] Step 1: Preparation of soluble microneedles

[0014] 1-1: preparing a high concentration sodium carboxymethyl cellulose solution;

[0015] 1-2: diluting the high concentration sodium carboxymethyl cellulose solution prepared in 1-1 to obtain a low concentration sodium carboxymethyl cellulose solution;

[0016] 1-3: drip a low concentration sodium carboxymethyl cellulose solution into the microneedle mold and remove the bubbles in each needle cavity; then retain the low concentration sodium carboxymethyl cellulose solution in the needle cavity and remove the low concentration sodium carboxymethyl cellulose solution outside the needle cavity; then drip a high concentration sodium carboxymethyl cellulose solution into the microneedle mold and remove the bubbles outside the needle cavity; finally, place the prepared microneedles at room temperature for drying;

[0017] 1-4: The dried microneedles in 1-3 are taken out, and sodium carboxymethyl cellulose forms the shell of the microneedles, and the interior of the microneedles is a hollow structure.

[0018] Furthermore, in step 1, the concentration of the high-concentration sodium carboxymethyl cellulose solution is 0.001-0.03 g / mL, and the concentration of the low-concentration sodium carboxymethyl cellulose solution is 30%-70% of the concentration of the high-concentration sodium carboxymethyl cellulose solution; and the method for removing bubbles includes vacuuming, ultrasound or centrifugation.

[0019] Step 2: Preparation of Encapsulant

[0020] 2-1. Preparation of polydopamine (PDA) nanoparticle dispersion: ammonia water, deionized water and anhydrous ethanol are mixed and stirred until a uniform mixed solution 1 is obtained; then, dopamine hydrochloride is dissolved in deionized water to obtain a dopamine hydrochloride solution; next, the dopamine hydrochloride solution is added to the mixed solution 1 and stirred until a uniform solution is formed to obtain a mixed solution 2; the mixed solution 2 is centrifuged and the lower precipitate is collected to obtain a crude PDA nanoparticle; the crude PDA nanoparticle is washed with anhydrous ethanol and deionized water, and then freeze-dried to obtain PDA nanoparticles; the PDA nanoparticles are stored in a refrigerator at 4°C away from light;

[0021] 2-2. Preparation of silver (Argentum, Ag) modified PDA nanoparticles: dissolve silver nitrate in deionized water to obtain a silver nitrate solution; add ammonia water dropwise to the silver nitrate solution and stir until a clear solution is obtained; add the PDA nanoparticle solution prepared in step 2-1 to the clear solution to obtain a mixed solution 3; stir the mixed solution 3 in the dark and then centrifuge to obtain a crude product of Ag-modified PDA (PDA@Ag) nanoparticles; wash the crude PDA@Ag nanoparticles with anhydrous ethanol and deionized water, and then freeze-dry to obtain PDA@Ag nanoparticles; store the PDA@Ag nanoparticles in the dark at room temperature;

[0022] 2-3. Add chitosan and 2-acrylamido-2-methylpropanesulfonic acid to deionized water and stir until a uniform mixed solution 4 is formed; add acrylamide to the mixed solution 4, and then stir until a uniform and transparent solution is formed to obtain a mixed solution 5; add the PDA@Ag nanoparticles prepared in step 2-2 to the mixed solution 5 and stir until it is uniform to obtain a mixed solution 6; after stirring the mixed solution 6 at room temperature until it is uniform, add a crosslinker and an initiator to the mixed solution 6 and continue stirring at room temperature until it is uniform to obtain a mixed solution 7; finally, pour the mixed solution 7 into a mold for polymerization to obtain an encapsulant.

[0023] Furthermore, in the step 2-1, the mass concentration of ammonia water is 25%-28%; by volume ratio, ammonia water: deionized water: anhydrous ethanol = 1: (40-50): (15-25); the concentration of dopamine hydrochloride solution is 45-55 mg / mL; by volume ratio, dopamine hydrochloride solution: ammonia water = (4.5-5.5): 1; the centrifugal speed is 10000-12000 rpm, and the centrifugal time is 8-10 min.

[0024] Furthermore, in the step 2-2, the mass concentration of ammonia water is 25%-28%, the concentration of silver nitrate solution is 30-35 mg / mL, the concentration of PDA nanoparticles is 8-12 mg / mL, the mass ratio of silver nitrate to PDA nanoparticles is (160-170):1; the centrifugal speed is 10000-12000 rpm.

[0025] Furthermore, in the step 2-3, the mass ratio of chitosan: acrylamide: 2-acrylamido-2-methylpropanesulfonic acid: deionized water: PDA@Ag nanoparticles is (0.25-0.35): 1: (3.5-4.5): (14-16): (0.01-0.02), and the stirring temperature is 45-55°C; the cross-linking agent is N, N'-methylenebisacrylamide, and the initiator is potassium persulfate. The mass ratio of cross-linking agent: initiator is 1: (45-55), and the stirring temperature is 55-65°C.

[0026] Step 3: Assembly of microarray needle-shaped liquid capsules

[0027] The chemotherapy drug is dripped into the soluble microneedles prepared in step 1, and after removing the bubbles, the encapsulant prepared in step 2 is attached to the top of the microneedles to obtain a microarray needle-shaped liquid capsule.

[0028] Further, the chemotherapy drug is paclitaxel;

[0029] Furthermore, paclitaxel is added in the form of paclitaxel ethanol solution, and the concentration of the solution is 2-7 mg / mL.

[0030] The microarray needle-shaped liquid capsule prepared according to the steps of the present invention has excellent mechanical properties, is sufficient to penetrate the skin, and has a small inflammatory reaction after penetrating the skin, and does not affect hair growth. However, when the same dose of the drug is used to treat melanoma by injection, it will produce a great stimulation effect, and the hair will not grow, which shows that the skin inflammatory reaction is strong. Therefore, the microarray needle-shaped liquid capsule can efficiently achieve the delivery of large doses of drugs, which has significant advantages in the treatment of melanoma.

[0031] The present invention has the following beneficial effects:

[0032] 1. The microarray needle-shaped liquid capsule prepared by the present invention not only has mechanical properties sufficient to penetrate the skin, but is also superior to most existing soluble microneedles. When the microarray needle-shaped liquid capsule is inserted into the melanoma site of a mouse, the microneedle will dissolve due to its dissolution characteristics, and can release drugs more quickly in the animal body; its hollow structure can achieve large-dose drug loading, and the microarray needle-shaped liquid capsule overcomes the shortcomings of the current soluble microneedles, which have poor mechanical properties and limited drug loading. At the same time, the present invention also explores the differences in drug loading between different microneedle structures (such as pyramid-shaped and conical microneedle patches).

[0033] 2. The drug selected by the present invention is paclitaxel. Since paclitaxel itself has a complex chemical structure and is insoluble in water, its bioavailability is low, which limits the wide clinical application of paclitaxel to a certain extent. Since paclitaxel has extremely poor solubility in water, researchers usually encapsulate paclitaxel for application. At the same time, paclitaxel can be completely dissolved in ethanol, dimethyl sulfoxide, N-methylpyrrolidone, and N,N-dimethylacetamide by vortexing for 3-5 minutes to form a uniform and stable drug solution.

[0034] 3. The present invention utilizes an ingredient in "alcohol": ethanol, and dissolves paclitaxel in ethanol to solve the problem of utilization rate of poorly water-soluble chemotherapy drugs. Many people "drink" in daily life, which shows that ethanol is a solvent that can be used by the human body under appropriate conditions. And studies have shown that ethanol can cause cell dehydration and coagulative necrosis in the treated tumor and kill cancer cells, thereby achieving local ablation of the tumor. Therefore, the present invention abandons water as a solvent and selects ethanol as a solvent. Ethanol can not only increase the solubility of poorly water-soluble chemotherapy drugs, but also better achieve drug utilization, providing a new use for poorly water-soluble anti-cancer drugs; at the same time, there is no need to use near-infrared lasers, ultrasound and other devices to achieve a strategy of synergistic treatment of melanoma with chemotherapy and percutaneous ethanol injection.

[0035] 4. The drugs that can be loaded by the present invention are not only applicable to paclitaxel, but also applicable to other drugs that are poorly soluble in water but soluble in ethanol.

[0036] 5. The chemotherapeutic drug loaded in the present invention is an ethanol solution of paclitaxel. The strategy of using ethanol as a solvent provides a new solution to the main problem of low bioavailability due to poor water solubility of drugs. In addition, ethanol as a solvent can not only improve the solubility of poorly water-soluble drugs, but also achieve local ablation of tumors, thereby effectively achieving the synergistic treatment of chemotherapy and percutaneous ethanol injection in the simplest way without the aid of any device. Compared with syringe injection in local drug administration treatment, the microarray needle-shaped liquid capsule prepared by the present invention can effectively reduce the inflammatory response to the skin, and demonstrates a new use of drug delivery in the treatment of melanoma.

[0037] 6. The microarray needle-shaped liquid capsule provided by the present invention has good biocompatibility, is harmless to animals or humans, and has the advantage of being biodegradable in vivo. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is the preparation process of the pyramid-shaped microneedles and cone-shaped microneedles prepared in Example 1.

[0039] Figure 2 (a) is a bright field image of the pyramid-shaped microneedle prepared in Example 1; (b) is a bright field image of the conical microneedle prepared in Example 1; (c) is a super-resolution laser confocal microscopy image of the pyramid-shaped microneedle prepared in Example 1; (d) is a super-resolution laser confocal microscopy image of the conical microneedle prepared in Example 1.

[0040] Figure 3 The volume of each pyramid-shaped and cone-shaped needle prepared in Example 1 and the volume of drugs that can be loaded in the pyramid-shaped and cone-shaped microneedle patches.

[0041] Figure 4This is the melanoma tumor treatment effect of each group in Example 4.

[0042] FIG5(a) is an image of skin hair follicles after injection treatment in Example 4; FIG5(b) is an image of skin hair follicles after microarray liquid capsule treatment in Example 4. DETAILED DESCRIPTION

[0043] It should be noted that the following examples will further explain the technical solutions of the present invention, but these examples do not limit the scope of the present invention. Any modification or equivalent substitution of the technical solutions of the present invention, as long as it does not deviate from the basic spirit and scope of the present invention, should be included in the protection scope of the present invention. The devices not particularly specified are all conventional devices in the art, and the technical means in the present invention are all conventional methods well known to those skilled in the art.

[0044] It should be specially noted that the method of the present invention is not limited to soluble materials such as sodium carboxymethyl cellulose, and any soluble material can be used to prepare a microneedle patch with a hollow structure using the method of the present invention.

[0045] Example 1: Preparation of hollow-structured soluble microneedles of sodium carboxymethyl cellulose

[0046] Preparation method of hollow structure soluble microneedles of sodium carboxymethyl cellulose Figure 1 shown.

[0047] Step 1: The specifications of the polydimethylsiloxane (PDMS) microneedle mold are pyramid and cone shapes with a height of 1.5 mm and a diameter of 0.66 mm.

[0048] Step 2: Dissolve each gram of sodium carboxymethyl cellulose in 33 mL of deionized water and stir thoroughly to obtain a high concentration sodium carboxymethyl cellulose solution with a concentration of 0.03 g / mL.

[0049] Step 3, the high concentration sodium carboxymethyl cellulose solution obtained in step 2 is mixed with deionized water at a ratio of 3:7 (V / V), and stirred thoroughly to obtain a low concentration sodium carboxymethyl cellulose solution;

[0050] Step 4. First, drip about 1 mL of low-concentration sodium carboxymethyl cellulose solution into the pyramid and cone-shaped molds respectively, and use vacuum operation to remove the bubbles in each needle cavity; then suck out the low-concentration solution outside the needle cavity, drip about 1 mL of high-concentration sodium carboxymethyl cellulose solution, use a pipette to remove the bubbles, and then place the prepared microneedles at room temperature to dry.

[0051] Step 5: After about 48 hours, the pyramid and cone-shaped microneedles dried in step 4 are taken out. Sodium carboxymethyl cellulose forms the shell of the microneedle, and the interior is a hollow structure.

[0052] Step six, in order to better observe the structure of the microneedle patch, the fluorescent dye fluorescein isothiocyanate is used to label sodium carboxymethyl cellulose in step two.

[0053] The bright field images of the obtained microneedle patches are shown in Figure 2(a) and Figure 2(b). It can be observed that the prepared pyramid-shaped and cone-shaped microneedle patches have uniform and complete morphologies. As shown in Figure 2(c) and Figure 2(d), super-resolution laser confocal microscopy verified that the interiors of the pyramid-shaped and cone-shaped microneedle patches are hollow structures.

[0054] After verifying the hollow structure of the microneedle patch, the present invention explored the dosage of chemotherapy drugs that can be loaded into the microneedle patch. Theoretically, the drug loading capacity of pyramid and cone-shaped microneedles is calculated by their height and bottom diameter. The theoretical internal volume of each needle in the pyramid and cone-shaped microneedle patches is 0.2178 mm 3 and 0.1709mm 3 For the actual amount of drug that can be loaded, Figure 3 As shown in the figure, two types of microneedles with hollow structures were filled by super-resolution laser confocal microscopy, and the actual hollow volume of a single pyramid-shaped microneedle was calculated to be 0.1558 mm 3 The volume of drugs that can be loaded into the pyramid-shaped microneedle patch is 15.58 μL, and the actual hollow volume of a single conical microneedle is 0.1074 mm 3 The volume of the drug that can be loaded into the conical microneedle patch is 10.74 μL. Compared with the theoretical volume of a single needle, the hollow ratio of the pyramidal hollow structure microneedle is 71.55%, and the hollow ratio of the conical hollow structure microneedle is 62.8%.

[0055] Example 2: Preparation of encapsulant

[0056] Step 1, preparation of PDA nanoparticle dispersion: Mix 2 mL of (25%-28%) ammonia solution with 90 mL of deionized water and 40 mL of anhydrous ethanol, and stir at room temperature (i.e., 25°C) for about 1 hour to mix them evenly. Then, dissolve each gram of dopamine hydrochloride in 20 mL of deionized water, slowly pour it into the above mixed solution and stir for about 24 hours to form a uniform and transparent mixed solution 1. Centrifuge the mixed solution 1 at a speed of 10,000-12,000 for 8-10 minutes to collect the lower precipitate to obtain crude PDA nanoparticles. Wash the crude PDA nanoparticles with anhydrous ethanol and deionized water three times respectively to obtain PDA nanoparticles. Finally, freeze-dry the prepared PDA nanoparticles and store them in a refrigerator at 4°C away from light.

[0057] Step 2, preparation of Ag-modified PDA nanoparticles (PDA@Ag): dissolve each gram of silver nitrate in 30 mL of deionized water. Add 25%-28% ammonia water to the above solution and stir until the brown precipitate disappears and the solution becomes clear again. Then add 0.3 mL of the 10 mg / mL PDA nanoparticle solution prepared in step 1 to the clear solution to obtain a mixed solution 2. Stir the mixed solution 2 at 25°C in the dark for about 1 hour, and then centrifuge at a speed of 10000-12000 for 8-10 minutes to obtain a crude Ag-modified PDA (PDA@Ag) nanoparticle. Wash the crude PDA@Ag nanoparticles with anhydrous ethanol and deionized water three times respectively to obtain PDA@Ag nanoparticles. Finally, the prepared PDA@Ag nanoparticles are freeze-dried and stored in the dark at room temperature.

[0058] Step 3, preparation of encapsulant: 0.3g chitosan and 1.0g 2-acrylamido-2-methylpropanesulfonic acid were added to 15mL deionized water, and stirred continuously at 45-55°C for 4h until a uniform mixed solution 3 was formed. 4g acrylamide was added to the mixed solution 3, and then stirred until a uniform transparent solution was formed to obtain a mixed solution 4; 0.015g of PDA@Ag nanoparticles prepared in step 2 was added to the mixed solution 4, and a uniform mixed solution 5 was formed after stirring at room temperature for about 10min. 0.001g of cross-linking agent N,N'-methylenebisacrylamide and 0.05g of initiator potassium persulfate were added to the mixed solution 5 and stirred at room temperature for about 1min to obtain a mixed solution 6. Finally, the mixed solution 6 was poured into a glass mold and polymerized at 55-65°C for about 4h to obtain an encapsulant.

[0059] The encapsulant prepared by the present invention is prepared by adding PDA and Ag to chitosan, 2-acrylamido-2-methylpropanesulfonic acid and acrylamide solution. PDA has attracted much attention due to its excellent adhesion. PDA doped in the encapsulant can increase the adhesion of the encapsulant; Ag is a good antibacterial agent. After the encapsulant is doped with Ag, the encapsulant can have partial antibacterial properties.

[0060] Example 3: Assembly of microarray needle-shaped liquid capsules

[0061] Step 1: dripping the drug into the hollow soluble microneedle prepared in Example 1.

[0062] Step 2: Vacuum to remove air bubbles.

[0063] Step 3: attach the encapsulant prepared in Example 2 onto the soluble microneedles to obtain the microarray needle-shaped liquid capsules of the present invention.

[0064] It should be noted that paclitaxel was added in the form of paclitaxel ethanol solution, and the concentration of the drug solution was 5 mg / mL.

[0065] The purpose of this embodiment is to ensure that the assembled microarray needle-shaped liquid capsule can achieve the encapsulation of paclitaxel and ethanol solution within 7 days, ensure that it does not volatilize, and can better penetrate into the tumor site of mice.

[0066] Example 4: Treatment of melanoma in mice

[0067] The experimental animals were 6-week-old female SPF-grade C57BL / 6 mice, which are inbred strains commonly used in oncology, physiology, immunology, and genetics research. They weighed 18-22g and had a certificate number of No.110322231102976886. The room temperature in the laboratory was 22±2℃, the relative humidity was 40%-60%, and the ventilation was fan-ventilated. The mice were kept in cages, with 5 mice per cage. The cages were cleaned every 3-4 days, and the food and drinking water were changed. The left abdomen of the mice was depilated and inoculated with 1×10 6 The tumors were treated 3 days later, and the concentration of paclitaxel / ethanol in each mouse was 5 mg / kg.

[0068] The control group of this experiment did not receive any treatment except tumor inoculation. The treatment of melanoma was divided into six groups, namely G1: control group, G2: ethanol injection group, G3: microneedle loaded ethanol group, G4: paclitaxel and aqueous solution injection group, G5: paclitaxel and ethanol solution injection group, G6: microneedle loaded paclitaxel and ethanol solution group.

[0069] In step 1, a microarray needle-shaped liquid capsule loaded with solvent ethanol (G3) or ethanol solution of chemotherapy drug paclitaxel (G6) was inserted into the melanoma site of the mouse and maintained for 5 minutes; the injection group was subcutaneously injected into the tumor (G2, G4, G5).

[0070] Step 2: 9 days later, perform the second treatment with the same operation as step 1.

[0071] Step 3: Weigh the body weight and tumor volume of mice every 2 days.

[0072] Table 1 The therapeutic effect of microneedles loaded with paclitaxel / ethanol on mouse melanoma

[0073]

[0074] The experimental data were processed by GraphPadPrism software. The experimental data results are shown in Table 1 and Figure 4 .

[0075] Compared with the control group at the same time: P < 0.05; n = 5

[0076] like Figure 4 The experimental results show that ethanol ablation therapy alone has a partial inhibitory effect on melanoma. When ethanol is used as a solvent to dissolve the anticancer drug paclitaxel, it increases the solubility of paclitaxel. It can also achieve synergistic treatment of melanoma with chemotherapy and local ablation, and has a significant inhibitory effect on melanoma.

[0077] Step 4: In addition, after 14 days of treatment, the tumor volume of mice in the control group had reached the upper limit of ethical regulations (1500mm 3 ). Then, each group of mice was photographed.

[0078] As shown in Figure 5, under the treatment of injection, the treatment site of the group injected with paclitaxel and ethanol solution, i.e., group G5, did not grow hair (Figure 5(a)), and was accompanied by an inflammatory reaction; while under the treatment of microarray needle-shaped liquid capsules, the treatment site of the group loaded with microneedles and paclitaxel and ethanol solution, i.e., group G6, still grew hair (Figure 5(b)). This indicates that compared with injection treatment, the inflammatory reaction of melanoma treatment with microarray needle-shaped liquid capsules is less.

Claims

1. A microarray needle-shaped liquid capsule, It is characterized in that The microarray needle-shaped liquid capsule is composed of soluble microneedles and encapsulating agents, has a hollow structure, and is used for loading chemotherapeutic drugs.

2. The microarray needle-shaped liquid capsule according to claim 1, It is characterized in that The chemotherapy drug is paclitaxel.

3. A method for preparing the microarray needle-shaped liquid capsule according to any one of claims 1 to 2, It is characterized in that The following steps are involved: Step 1: Preparation of soluble microneedles 1-1. Prepare a high concentration sodium carboxymethyl cellulose solution; 1-2, diluting the high concentration sodium carboxymethyl cellulose solution prepared in 1-1 to obtain a low concentration sodium carboxymethyl cellulose solution; 1-3. Drop a low concentration sodium carboxymethyl cellulose solution into the microneedle mold and remove the bubbles in each needle cavity; Then, the low-concentration sodium carboxymethyl cellulose solution in the needle cavity is retained, and the low-concentration sodium carboxymethyl cellulose solution outside the needle cavity is removed; next, a high-concentration sodium carboxymethyl cellulose solution is dripped into the microneedle mold and the bubbles outside the needle cavity are removed; finally, the prepared microneedles are placed at room temperature for drying; 1-4. The dried microneedles in 1-3 are taken out, and sodium carboxymethyl cellulose forms the shell of the microneedles, and the interior of the microneedles is a hollow structure; Step 2: Preparation of Encapsulant 2-1. Preparation of polydopamine nanoparticle dispersion: ammonia water, deionized water and anhydrous ethanol are mixed and stirred until a uniform mixed solution 1 is obtained; then, dopamine hydrochloride is dissolved in deionized water to obtain a dopamine hydrochloride solution; next, the dopamine hydrochloride solution is added to the mixed solution 1 and stirred until a uniform solution is formed to obtain a mixed solution 2; the mixed solution 2 is centrifuged and the lower precipitate is collected to obtain a crude polydopamine nanoparticle; the crude polydopamine nanoparticle is washed with anhydrous ethanol and deionized water, and then freeze-dried to obtain polydopamine nanoparticles; the polydopamine nanoparticles are stored in a refrigerator at 4°C away from light; 2-2. Preparation of silver-modified polydopamine nanoparticles: dissolving silver nitrate in deionized water to obtain a silver nitrate solution; adding ammonia water dropwise to the silver nitrate solution and stirring until a clear solution is obtained; adding the polydopamine nanoparticles prepared in step 2-1 to the clear solution to obtain a mixed solution 3; stirring the mixed solution 3 in the dark and then centrifuging to obtain a crude product of silver-modified polydopamine nanoparticles; washing the crude product of silver-modified polydopamine nanoparticles with anhydrous ethanol and deionized water, and then freeze-drying to obtain silver-modified polydopamine nanoparticles; storing the silver-modified polydopamine nanoparticles in the dark at room temperature; 2-3, chitosan and 2-acrylamido-2-methylpropanesulfonic acid are added to deionized water and stirred until a uniform solution is formed to obtain a mixed solution 4; acrylamide is added to the mixed solution 4 and stirred until a uniform solution is formed to obtain a mixed solution 5; the silver-modified polydopamine nanoparticles prepared in step 2-2 are added to the mixed solution 5 and stirred until a uniform solution is formed to obtain a mixed solution 6; after the mixed solution 6 is stirred at room temperature until it is uniform, a crosslinking agent and an initiator are added to the mixed solution 6 and the mixture is stirred at room temperature until a uniform solution is formed to obtain a mixed solution 7; finally, the mixed solution 7 is poured into a mold for polymerization to obtain an encapsulant; Step 3: Preparation of microarray needle-shaped liquid capsules The chemotherapeutic drug of claim 2 is dripped into the soluble microneedles prepared in step 1, and after removing the bubbles, the encapsulant prepared in step 2 is attached to the top of the microneedles to obtain a microarray needle-shaped liquid capsule.

4. The method for preparing the microarray needle-shaped liquid capsule according to claim 3, It is characterized in that In step 1, the concentration of the high-concentration sodium carboxymethyl cellulose solution is 0.001-0.03 g / mL, and the concentration of the low-concentration sodium carboxymethyl cellulose solution is 30%-70% of the concentration of the high-concentration sodium carboxymethyl cellulose solution; and the method for removing bubbles includes vacuuming, ultrasound or centrifugation.

5. The method for preparing the microarray needle-shaped liquid capsule according to claim 3, It is characterized in that The mass concentration of the ammonia water described in step 2-1 is 25%-28%; by volume ratio, ammonia water: deionized water: anhydrous ethanol = 1: (40-50): (15-25); the concentration of dopamine hydrochloride solution is 45-55 mg / mL; by volume ratio, dopamine hydrochloride solution: ammonia water = (4.5-5.5): 1; the centrifugal speed is 10000-12000 rpm, and the centrifugal time is 8-10 min.

6. The method for preparing the microarray needle-shaped liquid capsule according to claim 3, It is characterized in that In the step 2-2, the mass concentration of ammonia water is 25%-28%, the concentration of silver nitrate solution is 30-35 mg / mL, the concentration of polydopamine nanoparticles is 8-12 mg / mL, the mass ratio of silver nitrate to polydopamine nanoparticles is (160-170):1; the centrifugal speed is 10000-12000 rpm.

7. The method for preparing the microarray needle-shaped liquid capsule according to claim 3, It is characterized in that In the step 2-3, the mass ratio of chitosan: 2-acrylamido-2-methylpropanesulfonic acid: acrylamide: deionized water: silver-modified polydopamine nanoparticles is (0.25-0.35): 1: (3.5-4.5): (14-16): (0.01-0.02), and the stirring temperature is 45-55° C.; the cross-linking agent is N, N'-methylenebisacrylamide, and the initiator is potassium persulfate. The mass ratio of cross-linking agent: initiator is 1: (45-55), and the stirring temperature is 55-65° C.

8. The method for preparing the microarray needle-shaped liquid capsule according to claim 3, It is characterized in that The paclitaxel is added in the form of paclitaxel ethanol solution with a concentration of 2-7 mg / mL.

9. Use of the microarray needle-shaped liquid capsule according to any one of claims 1 to 2 in preparing a drug for treating melanoma.

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