A drug-loaded microneedle patch for treating psoriasis and a preparation method thereof

Abstract

The present invention proposes a drug-loaded microneedle patch for the treatment of psoriasis and a preparation method thereof, and belongs to the field of medical technology. Zr-ferrocene-MOF material is prepared, the surface is modified by tannic acid, and the compound coupled with glycine modified by acryloyl chloride and adalimumab and secukinumab is coupled, and then copolymerization is initiated, and the prepolymer liquid is poured into the PDMS microneedle mold, ultrasonically treated, illuminated, demoulded, and dried to obtain a drug-loaded microneedle patch for the treatment of psoriasis. The microneedle patch prepared by the present invention has high mechanical strength, anti-fatigue, certain photothermal antibacterial and anti-inflammatory effects, and can release drugs adalimumab and secukinumab in a controlled and sustained release. At the same time, it has a long attachment time on the skin, is firmly attached, and is more conducive to releasing drugs, thereby having an excellent therapeutic effect, and has minimal side effects, high patient compliance, and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a drug-loaded microneedle patch for treating psoriasis and a preparation method thereof. Background Art

[0002] Psoriasis is a chronic inflammatory skin disease with the main clinical manifestations of erythema and scaling. It can occur all over the body, but is more common on the scalp and extensor side of the limbs.

[0003] Although the treatment of psoriasis has made rapid progress after the promotion of biological agents and small molecule targeted drugs, there are still many patients in clinical practice who have some stubborn skin lesions that are difficult to eliminate, and thus cannot achieve a PASI 90% clearance rate.

[0004] Chinese invention patent CN109431967B discloses a soluble microneedle for treating psoriatic arthritis, but its 24-hour drug skin retention is still low, thus affecting the treatment effect. Chinese invention patent application CN101879336A discloses a microneedle array based on a flexible substrate for subcutaneous drug injection and its preparation method, wherein metal, silicon, and glass are used to prepare microneedles, and silica gel is used to make a flexible backing. The biocompatibility of the material is poor, and it is difficult to avoid the microneedles from breaking in the skin and being difficult to metabolize out of the body, causing potential dangers. Summary of the invention

[0005] The purpose of the present invention is to propose a drug-loaded microneedle patch for the treatment of psoriasis and a preparation method thereof, which has high mechanical strength, anti-fatigue, certain photothermal antibacterial and anti-inflammatory effects, and can release the drugs adalimumab and secukinumab in a sustained and controlled manner. At the same time, it can be attached to the skin for a long time and firmly, which is more conducive to the release of drugs, thereby having excellent therapeutic effects, and extremely small side effects, high patient compliance, and broad application prospects.

[0006] The technical solution of the present invention is achieved in this way:

[0007] The invention provides a method for preparing a drug-loaded microneedle patch for treating psoriasis. The method comprises the following steps: preparing a Zr-ferrocene-MOF material, modifying the surface of the material with tannic acid, coupling the material with a composite of glycine modified with acryloyl chloride and adalimumab and secukinumab, initiating copolymerization, pouring a prepolymer solution into a PDMS microneedle mold, ultrasonically treating the material, irradiating the material with light, demolding the material, and drying the material to obtain the drug-loaded microneedle patch for treating psoriasis.

[0008] As a further improvement of the present invention, the following steps are included:

[0009] S1. dissolving zirconium chloride, 1,1'-ferrocenecarboxylic acid and acetic acid in a DMF solution, performing a hydrothermal reaction, centrifuging, washing and drying to obtain a Zr-ferrocene-MOF material;

[0010] S2. Adding Zr-ferrocene-MOF material to water, adding tannic acid and a catalyst, heating and stirring to react, centrifuging, washing, and drying to obtain a modified Zr-ferrocene-MOF material;

[0011] S3. Add glycine to the alkali solution, add the THF solution of acryloyl chloride under ice-water bath condition, stir to react, remove the solvent under reduced pressure, dissolve the alkali solution, precipitate the acid solution, filter, wash, dry, add the solid to water, add NHS and EDC, stir to activate, add adalimumab and secukinumab, stir to react, dialyze, and freeze-dry to obtain a monoclonal antibody complex;

[0012] S4. Add the modified Zr-ferrocene-MOF material to water, add NHS and EDC, stir to activate, add the monoclonal antibody complex, add the initiator, stir to react, and prepare a prepolymer solution;

[0013] S5. Pour the prepolymer solution into the PDMS microneedle mold, perform ultrasonic treatment, irradiate with light, demold, and dry to prepare a drug-loaded microneedle patch for the treatment of psoriasis.

[0014] As a further improvement of the present invention, the molar ratio of zirconium chloride, 1,1'-ferrocenecarboxylic acid and acetic acid in step S1 is 1:1:45-55, the temperature of the hydrothermal reaction is 150-160°C, and the time is 10-12h.

[0015] As a further improvement of the present invention, the mass ratio of the Zr-ferrocene-MOF material, tannic acid and catalyst in step S2 is 10:2-3:0.1-0.2, the temperature of the heating and stirring reaction is 40-50°C, and the time is 3-5h; the catalyst is a Tris-HCl solution with a pH of 8.5-9.5.

[0016] As a further improvement of the present invention, the alkali solution in step S3 is a 0.5-1.5 mol / L NaOH or KOH solution, the acid solution is a 1-2 mol / L HCl solution, the mass ratio of glycine and acryloyl chloride is 10:12-15, the mass ratio of the solid, EDC, NHS, adalimumab, and secukinumab is 10:3-5:2-4:7-12:10-14, the stirring activation time is 20-30 min, the dialysis bag pore size used in the dialysis is 500-1000 kDa, and the time is 24-36 h.

[0017] As a further improvement of the present invention, the mass ratio of the modified Zr-ferrocene-MOF material, NHS, EDC, monoclonal antibody complex, and initiator in step S4 is 12-15:4-6:3-5:15-20:0.1-0.15, and the initiator is photoinitiator I2959 The stirring activation time is 20-30 minutes, and the stirring reaction time is 0.5-1 hour.

[0018] As a further improvement of the present invention, the PDMS microneedle mold in step S5 is a quadrangular pyramid, with a needle length of 300-400μm, a bottom diameter of 280μm×280μm, a needle tip distance of 800-1000μm, a quantity array of 14×14, a microneedle patch size of 15mm×15mm, and a groove depth of 1.5-2.5mm; the power of the ultrasonic treatment is 1000-1500W, the time is 10-20min, and the illumination is ultraviolet light curing, and the time is 10-15min.

[0019] As a further improvement of the present invention, the present invention specifically comprises the following steps:

[0020] S1. Dissolve 1 molar equivalent of zirconium chloride, 1 molar equivalent of 1,1'-ferrocenecarboxylic acid and 45-55 molar equivalents of acetic acid in a DMF solution, heat to 150-160°C, perform hydrothermal reaction for 10-12h, centrifuge, wash and dry to obtain a Zr-ferrocene-MOF material;

[0021] S2. Add 10 parts by weight of Zr-ferrocene-MOF material to water, add 2-3 parts by weight of tannic acid and 0.1-0.2 parts by weight of catalyst, heat to 40-50°C, stir the reaction for 3-5h, centrifuge, wash, and dry to obtain a modified Zr-ferrocene-MOF material;

[0022] The catalyst is a Tris-HCl solution with a pH of 8.5-9.5;

[0023] S3. Add 10 parts by weight of glycine to a 0.5-1.5 mol / L NaOH or KOH solution, add 12-15 parts by weight of a THF solution of acryloyl chloride under ice-water bath conditions, stir and react for 3-5 hours, remove the solvent under reduced pressure, dissolve in a 0.5-1.5 mol / L NaOH or KOH solution, precipitate in a 1-2 mol / L HCl solution, filter, wash, and dry, add 10 parts by weight of the obtained solid to water, add 3-5 parts by weight of NHS and 2-4 parts by weight of EDC, stir and activate for 20-30 minutes, add 7-12 parts by weight of adalimumab and 10-14 parts by weight of secukinumab, stir and react for 2-4 hours, dialyze for 24-36 hours using a dialysis bag with a pore size of 500-1000 kDa, and freeze-dry to obtain a monoclonal antibody complex;

[0024] S4. Add 12-15 parts by weight of modified Zr-ferrocene-MOF material to water, add 4-6 parts by weight of NHS and 3-5 parts by weight of EDC, stir and activate for 20-30 minutes, add 15-20 parts by weight of monoclonal antibody complex, add 0.1-0.15 parts by weight of photoinitiator I 2959 , stirring and reacting for 0.5-1h to obtain a prepolymer solution;

[0025] S5. Pour the prepolymer into the PDMS microneedle mold, ultrasonically treat at 1000-1500W for 10-20min, UV-curing for 10-15minn, demolding, and drying to obtain a drug-loaded microneedle patch for the treatment of psoriasis;

[0026] The PDMS microneedle mold is a quadrangular pyramid with a needle length of 300-400 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 800-1000 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 1.5-2.5 mm.

[0027] The present invention further protects a drug-loaded microneedle patch for treating psoriasis prepared by the above preparation method.

[0028] The present invention further protects the use of the above-mentioned drug-loaded microneedle patch for treating psoriasis in the preparation of a drug for treating psoriasis.

[0029] The present invention has the following beneficial effects:

[0030] Secukinumab is an interleukin-17 (IL-17) inhibitor approved for treatment. Secukinumab is a high-affinity fully human anti-IL-17 monoclonal antibody that can improve inflammatory responses in patients with few adverse reactions. Adalimumab (ADA) is a fully humanized anti-TNF monoclonal antibody that is administered subcutaneously. The synergistic effect of the two is widely used in dermatology to treat arthritis psoriasis, plaque psoriasis, etc., bringing complete cure to many patients, especially those who have not responded to traditional treatments. Compared with single medication, it greatly reduces the dosage and side effects.

[0031] The present invention uses Zr-ferrocene-MOF material as the matrix material, which not only has good biocompatibility, but also can convert light energy into heat energy under near-infrared light irradiation to generate local heat, causing irreversible cell damage in a local treatment mode, and has the advantages of remote controllability, low toxicity and fewer side effects. At the same time, the prepared Zr-ferrocene-MOF material has the advantages of large specific surface area, large load capacity and greatly improved mechanical strength.

[0032] The surface of Zr-ferrocene-MOF material is modified by coating with tannic acid. Tannic acid is a naturally occurring polyphenol compound, which contains a large amount of catechol and benzenetriol in its molecular structure. The hydroxyl groups on the catechol groups can form strong hydrogen bonds, and the benzene rings on the groups can form π-π electronic interactions with other benzene rings, and can form strong hydrogen bonds, metal coordination bonds, π-π interactions, etc. with the groups on the surface of the material, and adhere to the surface of the substrate through non-covalent bonds. The introduction of tannic acid into the hydrogel can effectively enhance its adhesion, mechanical strength and fatigue resistance, and can also enhance the photothermal sterilization and anti-inflammatory effects with Zr-ferrocene-MOF materials.

[0033] The present invention modifies glycine with acryloyl chloride to obtain a monomer, which is coupled with adalimumab and secukinumab to form a complex with strong hydrophilicity and good biocompatibility. The complex is then coupled to a modified Zr-ferrocene-MOF material to achieve a sustained release effect.

[0034] The drug-loaded microneedle patch for treating psoriasis prepared by the present invention has high mechanical strength, is anti-fatigue, has certain photothermal antibacterial and anti-inflammatory effects, can release drugs adalimumab and secukinumab in a sustained and controlled manner, and at the same time, can be attached to the skin for a long time and firmly, which is more conducive to the release of drugs, thereby having an excellent therapeutic effect, and having extremely small side effects, high patient compliance, and having broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0036] Figure 1 This is a comparison of the back skin lesion pictures of each group of mice in Test Example 1 of the present invention. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] Example 1

[0039] This embodiment provides a method for preparing a drug-loaded microneedle patch for treating psoriasis, which specifically comprises the following steps:

[0040] S1. 1 mmol zirconium chloride, 1 mmol 1,1'-ferrocenecarboxylic acid and 45 mmol acetic acid were dissolved in 50 mL DMF solution, heated to 150°C, hydrothermally reacted for 10 h, centrifuged, washed and dried to obtain Zr-ferrocene-MOF material;

[0041] S2. Add 10 mg of Zr-ferrocene-MOF material to 50 mL of water, add 2 mg of tannic acid and 0.1 mg of catalyst, heat to 40 ° C, stir and react for 3 h, centrifuge, wash, and dry to obtain a modified Zr-ferrocene-MOF material;

[0042] The catalyst is a Tris-HCl solution with a pH of 8.5;

[0043] S3. Add 10 mg of glycine to 50 mL of 0.5 mol / L NaOH solution, add 10 mL of THF solution containing 12 mg of acryloyl chloride under ice-water bath conditions, stir and react for 3 h, remove the solvent under reduced pressure, dissolve with 20 mL of 0.5 mol / L NaOH solution, add 50 mL of 1 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid to water, add 3 mg of NHS and 2 mg of EDC, stir and activate for 20 min, add 7 mg of adalimumab and 10 mg of secukinumab, stir and react for 2 h, dialyze with a dialysis bag with a pore size of 500 kDa for 24 h, and freeze-dry to obtain a monoclonal antibody complex;

[0044] S4. Add 12 mg of modified Zr-ferrocene-MOF material to 50 mL of water, add 4 mg of NHS and 3 mg of EDC, stir and activate for 20 min, add 15 mg of monoclonal antibody complex, add 0.1 mg of photoinitiator I 2959 , stirring and reacting for 0.5h to obtain a prepolymer solution;

[0045] S5. The prepolymer solution was poured into the PDMS microneedle mold, ultrasonically treated at 1000W for 10min, UV-cured for 10min, demolded, and dried to obtain a drug-loaded microneedle patch for the treatment of psoriasis;

[0046] The PDMS microneedle mold is a quadrangular pyramid with a needle length of 350 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 900 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 2 mm.

[0047] Example 2

[0048] This embodiment provides a method for preparing a drug-loaded microneedle patch for treating psoriasis, which specifically comprises the following steps:

[0049] S1. Dissolve 1 mmol of zirconium chloride, 1 mmol of 1,1'-ferrocenecarboxylic acid and 55 mmol of acetic acid in 50 mL of DMF solution, heat to 160°C, perform hydrothermal reaction for 12 h, centrifuge, wash and dry to obtain Zr-ferrocene-MOF material;

[0050] S2. Add 10 mg of Zr-ferrocene-MOF material to 50 mL of water, add 3 mg of tannic acid and 0.2 mg of catalyst, heat to 50 ° C, stir and react for 5 h, centrifuge, wash, and dry to obtain a modified Zr-ferrocene-MOF material;

[0051] The catalyst is a Tris-HCl solution with a pH of 9.5;

[0052] S3. Add 10 mg of glycine to 50 mL of 1.5 mol / L KOH solution, add 10 mL of THF solution containing 15 mg of acryloyl chloride under ice-water bath conditions, stir and react for 5 h, remove the solvent under reduced pressure, dissolve with 20 mL of 1.5 mol / L KOH solution, add 50 mL of 2 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid to water, add 5 mg of NHS and 4 mg of EDC, stir and activate for 30 min, add 12 mg of adalimumab and 14 mg of secukinumab, stir and react for 4 h, dialyze with a dialysis bag with a pore size of 500 kDa for 36 h, and freeze-dry to obtain a monoclonal antibody complex;

[0053] S4. Add 15 mg of modified Zr-ferrocene-MOF material to 50 mL of water, add 6 mg of NHS and 5 mg of EDC, stir and activate for 30 min, add 20 mg of monoclonal antibody complex, add 0.15 mg of photoinitiator I 2959 , stirring and reacting for 1 hour to obtain a prepolymer solution;

[0054] S5. The prepolymer solution was poured into the PDMS microneedle mold, ultrasonically treated at 1500W for 20min, UV-cured for 15minn, demolded, and dried to obtain a drug-loaded microneedle patch for the treatment of psoriasis;

[0055] The PDMS microneedle mold is a quadrangular pyramid with a needle length of 350 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 900 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 2 mm.

[0056] Example 3

[0057] This embodiment provides a method for preparing a drug-loaded microneedle patch for treating psoriasis, which specifically comprises the following steps:

[0058] S1. Dissolve 1 mmol of zirconium chloride, 1 mmol of 1,1'-ferrocenecarboxylic acid and 50 mmol of acetic acid in 50 mL of DMF solution, heat to 155°C, perform hydrothermal reaction for 11 h, centrifuge, wash and dry to obtain Zr-ferrocene-MOF material;

[0059] S2. Add 10 mg of Zr-ferrocene-MOF material to 50 mL of water, add 2.5 mg of tannic acid and 0.15 mg of catalyst, heat to 45 ° C, stir and react for 4 h, centrifuge, wash, and dry to obtain a modified Zr-ferrocene-MOF material;

[0060] The catalyst is a Tris-HCl solution with a pH of 9;

[0061] S3. Add 10 mg of glycine to 50 mL of 1 mol / L NaOH solution, add 10 mL of 13 mg of acryloyl chloride in THF solution under ice-water bath conditions, stir and react for 4 h, remove the solvent under reduced pressure, dissolve with 20 mL of 1 mol / L NaOH solution, add 50 mL of 1.5 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid to water, add 4 mg of NHS and 3 mg of EDC, stir and activate for 25 min, add 10 mg of adalimumab and 12 mg of secukinumab, stir and react for 3 h, dialyze for 30 h using a dialysis bag with a pore size of 500 kDa, and freeze-dry to obtain a monoclonal antibody complex;

[0062] S4. Add 13 mg of modified Zr-ferrocene-MOF material to 50 mL of water, add 5 mg of NHS and 4 mg of EDC, stir and activate for 25 min, add 17 mg of monoclonal antibody complex, add 0.12 mg of photoinitiator I 2959 , stirring and reacting for 1 hour to obtain a prepolymer solution;

[0063] S5. The prepolymer solution was poured into the PDMS microneedle mold, ultrasonically treated at 1200W for 15min, UV-cured for 12minn, demolded, and dried to obtain a drug-loaded microneedle patch for the treatment of psoriasis;

[0064] The PDMS microneedle mold is a quadrangular pyramid with a needle length of 350 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 900 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 2 mm.

[0065] Comparative Example 1

[0066] Compared with Example 3, the difference is that adalimumab is not added in step S3.

[0067] The details are as follows:

[0068] S3. Add 10 mg of glycine to 50 mL of 1 mol / L NaOH solution, add 10 mL of 13 mg of acryloyl chloride in THF solution under ice-water bath conditions, stir and react for 4 hours, remove the solvent under reduced pressure, dissolve with 20 mL of 1 mol / L NaOH solution, add 50 mL of 1.5 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid into water, add 4 mg of NHS and 3 mg of EDC, stir and activate for 25 minutes, add 22 mg of secukinumab, stir and react for 3 hours, dialyze using a dialysis bag with a pore size of 500 kDa for 30 hours, and freeze-dry to obtain a monoclonal antibody complex.

[0069] Comparative Example 2

[0070] Compared with Example 3, the difference is that secukinumab is not added in step S3.

[0071] The details are as follows:

[0072] S3. Add 10 mg of glycine to 50 mL of 1 mol / L NaOH solution, add 10 mL of 13 mg of acryloyl chloride in THF solution under ice-water bath conditions, stir and react for 4 h, remove the solvent under reduced pressure, dissolve with 20 mL of 1 mol / L NaOH solution, add 50 mL of 1.5 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid into water, add 4 mg of NHS and 3 mg of EDC, stir and activate for 25 min, add 22 mg of adalimumab, stir and react for 3 h, dialyze using a dialysis bag with a pore size of 500 kDa for 30 h, and freeze-dry to obtain a monoclonal antibody complex.

[0073] Comparative Example 3

[0074] Compared with Example 3, the difference is that secukinumab and adalimumab are not added in step S3.

[0075] The details are as follows:

[0076] S3. Add 10 mg of glycine to 50 mL of 1 mol / L NaOH solution, add 10 mL of 13 mg of acryloyl chloride in THF solution under ice-water bath conditions, stir and react for 4 hours, remove the solvent under reduced pressure, dissolve with 20 mL of 1 mol / L NaOH solution, add 50 mL of 1.5 mol / L HCl solution to precipitate, filter, wash, and dry to obtain a complex.

[0077] Comparative Example 4

[0078] Compared with Example 3, the difference is that step S2 is not performed.

[0079] The details are as follows:

[0080] S1. Dissolve 1 mmol of zirconium chloride, 1 mmol of 1,1'-ferrocenecarboxylic acid and 50 mmol of acetic acid in 50 mL of DMF solution, heat to 155°C, perform hydrothermal reaction for 11 h, centrifuge, wash and dry to obtain Zr-ferrocene-MOF material;

[0081] S2. Add 10 mg of glycine to 50 mL of 1 mol / L NaOH solution, add 10 mL of 13 mg of acryloyl chloride in THF solution under ice-water bath conditions, stir and react for 4 h, remove the solvent under reduced pressure, dissolve with 20 mL of 1 mol / L NaOH solution, add 50 mL of 1.5 mol / L HCl solution to precipitate, filter, wash, and dry, add 10 mg of the obtained solid to water, add 4 mg of NHS and 3 mg of EDC, stir and activate for 25 min, add 10 mg of adalimumab and 12 mg of secukinumab, stir and react for 3 h, dialyze for 30 h using a dialysis bag with a pore size of 500 kDa, and freeze-dry to obtain a monoclonal antibody complex;

[0082] S3. Add 13 mg Zr-ferrocene-MOF material to 50 mL water, add 5 mg NHS and 4 mg EDC, stir and activate for 25 min, add 17 mg monoclonal antibody complex, add 0.12 mg photoinitiator I 2959 , stirring and reacting for 1 hour to obtain a prepolymer solution;

[0083] S4. The prepolymer solution was poured into the PDMS microneedle mold, ultrasonically treated at 1200W for 15min, UV-cured for 12minn, demolded, and dried to obtain a drug-loaded microneedle patch for the treatment of psoriasis;

[0084] The PDMS microneedle mold is a quadrangular pyramid with a needle length of 350 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 900 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 2 mm.

[0085] Comparative Example 5

[0086] Compared with Example 3, the difference is that in step S3, the monoclonal antibody is not coupled with acryloyl chloride-modified glycine.

[0087] The details are as follows:

[0088] S3. Mix 10 mg of adalimumab and 12 mg of secukinumab for 10 minutes to prepare a monoclonal antibody mixture.

[0089] Test Example 1

[0090] BALB / c mice were selected, 2×3 cm of back hair was removed, and they were randomly divided into a normal control group (vaseline group), a model control group (IMQ group), Example 1-3 groups, and Comparative Example 1-5 groups, with 10 mice in each group. Establishment of psoriasis mouse model: Imiquimod (IMQ) induction method.

[0091] Dosage:

[0092] ① Vaseline group: 62.5 mg / day of vaseline was applied to the hair-depleted areas on the backs of mice for 7 consecutive days.

[0093] ②IMQ group: IMQ cream 62.5 mg / day was applied to the hair-depleted area on the back of mice for 7 consecutive days.

[0094] ③ Groups of Example 1-3 and Comparative Example 1-5: Same as the IMQ group, 6 hours after the application of IMQ cream on D4-D7, the corresponding drug-loaded microneedle patch for the treatment of psoriasis was applied, and far-infrared irradiation was performed for 1 hour every day.

[0095] Observation indicators:

[0096] ① Check the back of the mice every day to see if there are characteristic changes of psoriasis such as erythema, skin hypertrophy, and scaling, and record the PASI score. The scoring criteria are shown in Table 1.

[0097] Table 1 Scoring method for skin lesion severity in animal models of psoriasis

[0098]

[0099] ②Relative expression levels of inflammatory factors (TNF-α, IL-17, IL-22, IL-23) mRNA in mouse back skin lesions.

[0100] The results are as follows:

[0101] On the 8th day, the back skin lesions of mice in each group were as follows Figure 1 As can be seen from the figure, the drug-loaded microneedle patch for the treatment of psoriasis prepared in Examples 1-3 can effectively improve the symptoms of the IMQ-induced psoriasis mouse model.

[0102] The PASI scores for evaluation of erythema, skin hypertrophy, and scaling on day 8 are shown in Table 2 .

[0103] Table 2

[0104]

[0105]

[0106] As can be seen from the above table, the drug-loaded microneedle patches for the treatment of psoriasis prepared in Examples 1-3 of the present invention can effectively reduce the PASI scores of erythema, skin hypertrophy and scaling in IMQ-induced psoriasis mice.

[0107] The relative expression levels of mRNA of inflammatory factors (TNF-α, IL-17, IL-22, IL-23) in the back lesions of mice on the 8th day are shown in Table 3.

[0108] Table 3

[0109]

[0110] Note: * compared with the normal control group, P < 0.05; # compared with the model group, P < 0.05.

[0111] It can be seen from the above table that the drug-loaded microneedle patches for the treatment of psoriasis prepared in Examples 1-3 of the present invention can effectively reduce the relative expression levels of mRNA of inflammatory factors (TNF-α, IL-17, IL-22, IL-23) in the back lesions of psoriasis mice induced by IMQ.

[0112] Test Example 2

[0113] Take 10 μL of Escherichia coli (ATCC25922) solution (10 6 cfu / mL) were dropped onto the drug-loaded microneedle patch for the treatment of psoriasis prepared in Examples 1-3 or Comparative Examples 4-5, and irradiated with 808 nm near-infrared light for 10 min (2.6 W / cm 2 ) and then cultured in a 37°C incubator. After 4 hours, 900 μL LB liquid medium was added to dilute the bacterial solution and plated, with 3 parallel samples in each group. The number of colonies was recorded after 24 hours of culture. The control group was co-cultured with 100 μL phosphate (PBS) buffer and 100 μL bacterial solution.

[0114] Antibacterial rate (%) = (1-N 样品 / N 对照 )×100%

[0115] The results are shown in Table 4.

[0116] Table 4

[0117] Group Antibacterial rate (%) Example 1 98.9 Example 2 99.2 Example 3 99.6 Comparative Example 4 88.9 Comparative Example 5 93.4

[0118] It can be seen from the above table that the antibacterial rate of the drug-loaded microneedle patches for the treatment of psoriasis prepared in Examples 1-3 of the present invention is relatively high.

[0119] Test Example 3

[0120] Take 4 mL of fresh blood, collect the red blood cells at the bottom by centrifugation, wash with PBS buffer until the supernatant is clear, and finally prepare the red blood cells with PBS buffer into a 2% solution by volume. Take 100 μL of each of the drug-loaded microneedle patch for the treatment of psoriasis prepared in Example 1-3 or Comparative Example 1-5 (prepared into a 1 mg / mL solution with PBS buffer), 2 wt% of Triton and PBS buffer solution and mix with 100 μL of red blood cells, wherein 2 wt% of Triton and PBS buffer solution are the positive control group and the negative control group, respectively, and then incubate in an oven at 37°C for 3 hours. After incubation, centrifuge, aspirate 100 μL of supernatant into a 96-well plate, and test the optical density (OD) value at 576 nm. Hemolysis rate (R H ) is calculated as follows:

[0121] Hemolysis rate R H =(OD 样品 -OD 阴性 ) / (OD 阳性 -OD 阴性 )×100%

[0122] The results are shown in Table 5.

[0123] Table 5

[0124] Group Hemolysis rate (%) Example 1 0.32 Example 2 0.29 Example 3 0.27 Comparative Example 1 0.55 Comparative Example 2 0.59 Comparative Example 3 0.62 Comparative Example 4 0.48 Comparative Example 5 0.50

[0125] As can be seen from the above table, the hemolysis rate of the drug-loaded microneedle patches for the treatment of psoriasis prepared in Examples 1-3 of the present invention is relatively low, meeting the hemolysis standard of biomedical materials (less than 5%).

[0126] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a drug-loaded microneedle patch for the treatment of psoriasis, characterized in that: Zr-ferrocene-MOF material was prepared, the surface of which was modified with tannic acid and coupled with a complex of glycine modified with acryloyl chloride and adalimumab and secukinumab, and then copolymerization was initiated. The prepolymer solution was poured into a PDMS microneedle mold, subjected to ultrasonic treatment, illumination, demolding, and drying to obtain a drug-loaded microneedle patch for the treatment of psoriasis.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. dissolving zirconium chloride, 1,1'-ferrocenecarboxylic acid and acetic acid in a DMF solution, performing a hydrothermal reaction, centrifuging, washing and drying to obtain a Zr-ferrocene-MOF material; S2. Adding Zr-ferrocene-MOF material to water, adding tannic acid and a catalyst, heating and stirring to react, centrifuging, washing, and drying to obtain a modified Zr-ferrocene-MOF material; S3. Add glycine to the alkali solution, add the THF solution of acryloyl chloride under ice-water bath condition, stir to react, remove the solvent under reduced pressure, dissolve the alkali solution, precipitate the acid solution, filter, wash, dry, add the solid to water, add NHS and EDC, stir to activate, add adalimumab and secukinumab, stir to react, dialyze, and freeze-dry to obtain a monoclonal antibody complex; S4. Add the modified Zr-ferrocene-MOF material to water, add NHS and EDC, stir to activate, add the monoclonal antibody complex, add the initiator, stir to react, and prepare a prepolymer solution; S5. Pour the prepolymer solution into the PDMS microneedle mold, perform ultrasonic treatment, irradiate with light, demold, and dry to prepare a drug-loaded microneedle patch for the treatment of psoriasis.

3. The preparation method according to claim 2, characterized in that: In step S1, the molar ratio of zirconium chloride, 1,1'-ferrocenecarboxylic acid and acetic acid is 1:1:45-55, the temperature of the hydrothermal reaction is 150-160° C., and the time is 10-12 hours.

4. The preparation method according to claim 2, characterized in that: In step S2, the mass ratio of the Zr-ferrocene-MOF material, tannic acid and catalyst is 10:2-3:0.1-0.2, the temperature of the heating and stirring reaction is 40-50°C, and the time is 3-5h; the catalyst is a Tris-HCl solution with a pH of 8.5-9.

5.

5. The preparation method according to claim 2, characterized in that: In step S3, the alkali solution is a 0.5-1.5 mol / L NaOH or KOH solution, the acid solution is a 1-2 mol / L HCl solution, the mass ratio of glycine and acryloyl chloride is 10:12-15, the mass ratio of the solid, EDC, NHS, adalimumab, and secukinumab is 10:3-5:2-4:7-12:10-14, the stirring activation time is 20-30 min, the dialysis bag pore size used in the dialysis is 500-1000 kDa, and the time is 24-36 h.

6. The preparation method according to claim 2, characterized in that: The mass ratio of the modified Zr-ferrocene-MOF material, NHS, EDC, monoclonal antibody complex, and initiator in step S4 is 12-15:4-6:3-5:15-20:0.1-0.15, and the initiator is photoinitiator I 2959 The stirring activation time is 20-30 minutes, and the stirring reaction time is 0.5-1 hour.

7. The preparation method according to claim 2, characterized in that: The PDMS microneedle mold in step S5 is a quadrangular pyramid, with a needle length of 300-400 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 800-1000 μm, a number array of 14×14, a microneedle patch size of 15 mm×15 mm, and a groove depth of 1.5-2.5 mm; the power of the ultrasonic treatment is 1000-1500 W, the time is 10-20 min, and the illumination is ultraviolet light curing, and the time is 10-15 min.

8. The preparation method according to claim 2, characterized in that: The specific steps include: S1. Dissolve 1 molar equivalent of zirconium chloride, 1 molar equivalent of 1,1'-ferrocenecarboxylic acid and 45-55 molar equivalents of acetic acid in a DMF solution, heat to 150-160°C, perform hydrothermal reaction for 10-12h, centrifuge, wash and dry to obtain a Zr-ferrocene-MOF material; S2. Add 10 parts by weight of Zr-ferrocene-MOF material to water, add 2-3 parts by weight of tannic acid and 0.1-0.2 parts by weight of catalyst, heat to 40-50°C, stir the reaction for 3-5h, centrifuge, wash, and dry to obtain a modified Zr-ferrocene-MOF material; The catalyst is a Tris-HCl solution with a pH of 8.5-9.5; S3. Add 10 parts by weight of glycine to a 0.5-1.5 mol / L NaOH or KOH solution, add 12-15 parts by weight of a THF solution of acryloyl chloride under ice-water bath conditions, stir and react for 3-5 hours, remove the solvent under reduced pressure, dissolve in a 0.5-1.5 mol / L NaOH or KOH solution, precipitate in a 1-2 mol / L HCl solution, filter, wash, and dry, add 10 parts by weight of the obtained solid to water, add 3-5 parts by weight of NHS and 2-4 parts by weight of EDC, stir and activate for 20-30 minutes, add 7-12 parts by weight of adalimumab and 10-14 parts by weight of secukinumab, stir and react for 2-4 hours, dialyze for 24-36 hours using a dialysis bag with a pore size of 500-1000 kDa, and freeze-dry to obtain a monoclonal antibody complex; S4. Add 12-15 parts by weight of modified Zr-ferrocene-MOF material to water, add 4-6 parts by weight of NHS and 3-5 parts by weight of EDC, stir and activate for 20-30 minutes, add 15-20 parts by weight of monoclonal antibody complex, add 0.1-0.15 parts by weight of photoinitiator I 2959 , stirring and reacting for 0.5-1h to obtain a prepolymer solution; S5. Pour the prepolymer into the PDMS microneedle mold, ultrasonically treat at 1000-1500W for 10-20min, UV-curing for 10-15minn, demolding, and drying to obtain a drug-loaded microneedle patch for the treatment of psoriasis; The PDMS microneedle mold is a quadrangular pyramid with a needle length of 300-400 μm, a bottom diameter of 280 μm×280 μm, a needle tip distance of 800-1000 μm, a number array of 14×14 needles, a microneedle patch size of 15 mm×15 mm, and a groove depth of 1.5-2.5 mm.

9. A drug-loaded microneedle patch for treating psoriasis prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the drug-loaded microneedle patch for treating psoriasis as claimed in claim 9 in the preparation of a medicine for treating psoriasis.

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