Pharmaceutical compositions and use in the manufacture of a medicament for the treatment of EGFRi-induced skin toxicity

Abstract

The application discloses a pharmaceutical composition, which comprises chlorogenic acid, ferulic acid and a zeolitic imidazolate framework material, the mass ratio of the chlorogenic acid to the ferulic acid is 4:1-1:4, and the zeolitic imidazolate framework material is a Zn-based zeolitic imidazolate framework material. In the composition, the chlorogenic acid and the ferulic acid are compatible to synergistically regulate the proliferation, migration and apoptosis of keratinocytes, and restore the physiological function imbalance of human skin keratinocytes caused by EGFRIs. The ZIF can efficiently co-carry the chlorogenic acid and the ferulic acid, the nanometer size particle size is beneficial to the penetration of the drug into the skin, the hydroxyl groups on the surface of the ZIF can form hydrogen bonds with ceramides in the skin to retain the drug in the lesion site. The composition preparation has good moisturizing performance, is simple to use, and can effectively treat skin toxicity reactions such as papulopustular skin rash, dry and itchy skin, periungual inflammation, abnormal hair regeneration and the like caused by EGFRIs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a pharmaceutical composition for treating skin toxicity caused by EGFRIs, its preparation method, and its application in the preparation of therapeutic drugs for EGFRI-induced skin toxicity. Background Technology

[0002] In cancer treatment, epidermal growth factor receptor inhibitors (EGFR inhibitors) are widely used clinically to treat cancer patients with EGFR activating mutations, especially non-small cell lung cancer, breast cancer, and colorectal cancer. However, with the use of EGFRIs, patients have experienced various adverse reactions, among which skin adverse reactions (including papulopustular rashes, dry and itchy skin, paronychia, and abnormal hair regrowth) are the most common, with an incidence rate as high as 79-88%. Early skin toxicity severely impacts patients' daily lives and emotions. If toxic reactions are not treated promptly and effectively, they will gradually worsen with continued EGFRI use. Severe skin toxicity may force a reduction in the dosage of EGFRIs or even discontinuation of the drug, leading to treatment interruption and exacerbating cancer progression.

[0003] Currently, expert consensus on treating adverse reactions to EGFRIs recommends topical antibiotics, steroids, and emollients, or adjunctive therapy with oral antibiotics and immunomodulators; however, clinical efficacy is unsatisfactory, with recurrent episodes and long-term use leading to side effects and secondary harm to patients' physical and mental well-being. Therefore, there is an urgent need to develop drug formulations with proven efficacy against EGFRI-induced skin toxicity for clinical application.

[0004] The applicant discovered through research (application number 202510176114X) that a drug composition composed of chlorogenic acid, ferulic acid and zinc salt has a good effect on treating adverse reactions of EGFRIs. However, the drug composition has significant differences in solubility and transdermal properties and poor stability. Simply administering the drug composition cannot achieve long-term retention in the basal layer of the skin, resulting in low drug utilization and the risk of affecting the anti-tumor efficacy of EGFRIs. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the primary objective of this invention is to provide a pharmaceutical composition for treating skin toxicity caused by EGFRIs, a method for preparing the same, and its application in the preparation of therapeutic drugs for EGFRI-induced skin toxicity. This pharmaceutical composition uses a zeolite imidazole framework (ZIF) as a carrier material and utilizes the zinc element contained in the ZIF as a component of the therapeutic agent. This not only eliminates the need for preservatives, reducing the introduction of risky substances and thus lowering the risk of skin allergies, making it safer, easier to carry and store, but also prolongs the drug's retention time in the skin, achieving better therapeutic effects.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: A pharmaceutical composition comprising chlorogenic acid, ferulic acid, and a zeolite imidazole framework (ZIF) material, wherein the mass ratio of chlorogenic acid to ferulic acid is 4:1 to 1:4, preferably 2:1 to 1:2, and most preferably 1.5:1. The zeolite imidazole framework material is a Zn-based zeolite imidazole framework material.

[0007] Preferably, the zeolite imidazole framework (ZIF) material is selected from ZIF91, ZIF90, or ZIF8. ZIF91 with hydroxyl groups on its surface is preferred.

[0008] Preferably, the total amount of chlorogenic acid and ferulic acid is in a mass ratio of 3:1 to 1:3 to the mass of Zn in the zeolite imidazole framework (ZIF) material, more preferably 2:1 to 1:2, and most preferably 4:3.

[0009] Preferably, the ZIF91 is prepared by the following steps: (1) The zinc salt and the organic ligand were reacted in a mixed solution containing trioctylamine for 1-24 h to obtain the intermediate product; (2) The intermediate product is reacted in a methanol solution of sodium borohydride to obtain ZIF91.

[0010] Preferably, the zinc salt zinc acetate or zinc nitrate mentioned in step (1) is used, more preferably ZnCH3(COO)2·2H2O.

[0011] Preferably, in the mixed solution described in step (1), the volume fraction of trioctylamine is 4-8%.

[0012] Preferably, the mass ratio of the zinc salt to the organic ligand in step (1) is (0.15~0.25):(0.1~0.2).

[0013] Preferably, the organic ligand in step (1) is imidazole-2-formaldehyde.

[0014] Preferably, the reaction temperature in step (2) is 55~65℃ and the reaction time is 10~12 h.

[0015] Preferably, the concentration of sodium borohydride in step (2) is 10~20 μg / mL.

[0016] Preferably, the pharmaceutical composition further includes a matrix material. When the matrix material is a gel matrix and water, the pharmaceutical composition is a gelling agent; When the matrix material is a spinning matrix, the pharmaceutical composition is a nanofiber membrane dressing.

[0017] Preferably, the gel matrix is ​​one or a mixture of several of hyaluronic acid, carbomer, sodium alginate, xanthan gum or cellulose derivatives, with hyaluronic acid being the most preferred.

[0018] Preferably, the molecular weight of the hyaluronic acid is 10 k to 120 w, and most preferably 10 k.

[0019] Preferably, in the gelling agent, the mass fraction of the gel matrix is ​​1-12%, more preferably 6-10%, and most preferably 8%.

[0020] Preferably, the total amount of chlorogenic acid, ferulic acid and zeolite imidazole framework (ZIF) material in the gelling agent is 0.5-2.0% by mass.

[0021] Preferably, the spinning matrix is ​​one or a mixture of several of hyaluronic acid, gelatin, silk fibroin, collagen, polyethylene glycol or polylactic acid, preferably a mixture of hyaluronic acid and gelatin, and more preferably, the mass ratio of hyaluronic acid to gelatin is 1:(4.8~7.2).

[0022] More preferably, in the nanofiber membrane dressing, the mass fraction of the spinning matrix material is 85-95%.

[0023] A second objective of this invention is to provide a method for preparing the pharmaceutical composition, comprising the following steps: Chlorogenic acid, ferulic acid, and zeolite imidazole framework (ZIF) materials were mixed in a methanol solution in the dark, followed by solid-liquid separation to obtain drug particles.

[0024] Preferably, the mixing is a stirring mixture.

[0025] Preferably, the mixing time is 6 to 24 hours, more preferably 10 to 12 hours.

[0026] Preferably, the preparation method further includes the steps of solid-liquid separation followed by washing and freeze-drying.

[0027] Preferably, the solid-liquid separation method is centrifugation.

[0028] Preferably, when the pharmaceutical composition is a gel, the preparation method further includes the following step A: Step A: Mix the gel matrix and water to make the gel matrix swell, then add the remaining components and mix evenly to obtain the gel agent.

[0029] Preferably, the swelling or uniform mixing step is carried out under stirring conditions, with a stirring speed of 1000~2000 rpm and a stirring time of 0.5~5 h. More preferably, it is 1400~1600 rpm for 2~4 h.

[0030] Preferably, when the pharmaceutical composition is a nanofiber membrane dressing, the preparation method further includes the following step B: Step B: The spinning matrix and the drug particles are made into a spinning solution, and then the spinning solution is electrospinned to obtain the nanofiber membrane dressing.

[0031] Preferably, the spinning solution is prepared by the following method: The spinning matrix is ​​swollen in a spinning solvent, and then the remaining components are added and mixed evenly to obtain the spinning solution.

[0032] Preferably, the spinning solvent is any one or a mixture of hexafluoroisopropanol, dichloromethane, chloroform, formic acid, or water, preferably a mixture of hexafluoroisopropanol and water, more preferably a mixture of hexafluoroisopropanol and water in a volume ratio of 6:4 to 8:2, and even more preferably a mixture of hexafluoroisopropanol and water in a volume ratio of 7:3.

[0033] Preferably, when the spinning matrix is ​​a mixture of hyaluronic acid and gelatin, the mass concentration of hyaluronic acid in the spinning solution is 2.5-3.5%, and the mass concentration of gelatin is 12-18%. More preferably, the mass concentration of hyaluronic acid in the spinning solution is 2.8-3.2%, and the mass concentration of gelatin is 14-16%.

[0034] Preferably, the electrospinning conditions are a voltage of 20~28 kV, a feed speed of 0.5~1.5 mL / h, and a receiving distance of 12~18 cm.

[0035] Preferably, the voltage is 24~26 kV; the propulsion speed is 0.8~1.2 mL / h; and the receiving distance is 14~16 cm.

[0036] A third objective of this invention is to provide the use of the pharmaceutical composition in the preparation of a therapeutic agent for EGFRI-induced skin toxicity.

[0037] Preferably, the EGFRIs include erlotinib, imatinib, erlotinib, sorafenib, rituximab, trastuzumab, cetuximab, bevacizumab, or panitumumab.

[0038] The skin toxicities of the EGFRIs include papulopustular rashes, dry and itchy skin, desquamation, periungual inflammation, or abnormal hair regrowth.

[0039] The beneficial effects of the pharmaceutical compositions, formulations, and applications described in this invention are manifested in the following aspects: ZIF's nanoscale size facilitates the penetration of chlorogenic acid and ferulic acid into the skin, while its unique drug-carrying capacity allows for the control of their loading ratio, enabling optimal delivery to the disease site. The abundant hydroxyl groups on the ZIF-91 surface interact with ceramides via hydrogen bonding, increasing retention time and forming a subcutaneous drug reservoir. This effectively addresses the issue of drugs being delivered only to the basal layer of the skin, reducing the risk of impacting the anti-tumor effects of EGFRIs. Furthermore, ZIF-91 compensates for zinc deficiency in the skin caused by EGFRIs, thus aiding in the treatment of EGFRI-induced skin toxicity.

[0040] This invention discovers that ferulic acid and chlorogenic acid can reverse the inhibitory effect of EGFRIs on the activity of human skin keratinocytes (HaCa-T cells), reduce EGFRI-induced HaCa-T cell apoptosis, restore the ability of EGFRIs to inhibit HaCa-T cell migration, and effectively treat EGFRI-induced papulopustular rashes, dry and itchy skin, desquamation, periungual inflammation, abnormal hair regeneration, and other skin toxicities, with an efficacy rate of up to 90%, providing a pharmaceutical composition for the clinical treatment of such skin toxicities.

[0041] This invention preferentially utilizes ZIF91 nanoparticle drug delivery carriers, which have relatively uniform particle size, around 65 nm. This nanoscale size facilitates the penetration of chlorogenic acid and ferulic acid into the skin. The carrier can co-load chlorogenic acid and ferulic acid, which have significant differences in solubility and transdermal properties. By controlling the amount and ratio of ferulic acid and chlorogenic acid added, the optimal drug loading ratio of ferulic acid and chlorogenic acid in ZIF91 can be achieved at 1:1.5, with a total drug loading of approximately 25%. Its surface hydroxyl groups interact with ceramides in the skin through hydrogen bonding, retaining the drug at the target region where EGFRIs induce skin toxicity, prolonging the drug's retention time in the skin. This increases the retention of ferulic acid and chlorogenic acid in the skin by 3 times and 13.5 times, respectively, improving drug utilization and reducing dosage. Furthermore, the Zn element in the ZIF91 carrier has a regulatory effect on many physiological processes such as keratinocyte proliferation, apoptosis, and migration, and can synergistically exert the therapeutic effect of chlorogenic acid and ferulic acid on skin toxicity.

[0042] The gel of this invention has excellent moisturizing properties, helping the skin retain moisture, hydrating dry skin, and relieving symptoms of dry and itchy skin caused by EGFRIs. The gel formulation of the pharmaceutical composition is effective, easy to apply, simple to use, and has good patient compliance, thus meeting clinical needs well.

[0043] This invention prepares drug-loaded ZIF91 nanoparticles into a nanofiber membrane dressing, forming a "solid membrane" that requires no preservatives, reducing the risk of skin allergies and making it safer, easier to carry, and easier to store. The nanofiber membrane immediately permeates into a film upon contact with water, adhering more closely to the skin. When hyaluronic acid is used as the material for the nanofiber membrane, its moisturizing effect is even better, helping to relieve dry skin. It also has good adhesion, making it less likely to stain clothing, providing more options to meet patient needs. Attached Figure Description

[0044] Figure 1 The particle size distribution and zeta potential diagram of ZIF91 obtained in Example 1 are shown.

[0045] Figure 2 The image shown is a SEM image of ZIF91 obtained in Example 1.

[0046] Figure 3 These are inverted microscope images of the scratches at 0 h and 24 h in Example 4.

[0047] Figure 4 The scratch healing rate is shown for each group in Example 4.

[0048] Figure 5 This is a comparison chart of the amount of ferulic acid retained and permeated in the skin in each group in Example 5 (n=3, X±SD).

[0049] Figure 6 This is a comparison chart of the amount of chlorogenic acid retained and permeated in the skin in each group in Example 5 (n=3, X±SD).

[0050] Figure 7 This is a comparison of DSC images of skin at different temperatures in each group in Example 6.

[0051] Figure 8 This is a graph showing the reduction of skin toxicity symptoms in each model mouse in Example 7.

[0052] Figure 9 Figure 7 shows a comparison of the body weights of the model mice in each group (n=10, X±SD, *). P <0.05,** P <0.01).

[0053] Figure 10 A comparison of spleen indices in different groups of model mice in Example 7 (n=10, X±SD, *P <0.05,** P <0.01).

[0054] Figure 11 A comparison of liver indices in different groups of model mice in Example 7 (n=10, X±SD, * P <0.05,** P <0.01).

[0055] Figure 12 This is a comparison chart of the thymus index of each group of model mice in Example 7.

[0056] Figure 13 This is a comparison chart of skin water content in each group of model mice in Example 7.

[0057] Figure 14 The results of HE and Oil Red staining of the skin of model mice in each group after treatment in Example 7 are shown (the scale bar in the figure is 100 μm). Detailed Implementation

[0058] The sources of cells, animals, drugs, reagents, and instruments involved in the examples are as follows: Experimental cells: HaCa-T cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0059] Laboratory animals: SPF-grade BALB / c female mice, purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd., Laboratory Animal License No. (SCXK(Su)2022-0009), housed at the Laboratory Animal Center of Jiangsu Academy of Traditional Chinese Medicine under light / dark (12 h / 12 ​​h) and temperature (22±3)℃, with free access to food and water, Laboratory Animal Use License (SYXK(Su)2016-0018).

[0060] Experimental pig skin: Bama fragrant pig skin (Linxi County Jingde Agricultural Products Sales Co., Ltd.).

[0061] Drugs and reagents: Chlorogenic acid (HPLC≥98%, Sichuan Pusi Biotechnology Co., Ltd.); Ferulic acid (HPLC≥98%, Sichuan Pusi Biotechnology Co., Ltd.); Tween 80 (Biofroxx); Erlotinib hydrochloride (Aladdin); Zn(CH3COO)2·2H2O, Imidazole-2-formaldehyde, Dimethylformamide, Trioctylamine (Ron Reagent); Methanol, Ethanol, Chloromethane, Chloroform, Formic acid (Xilong Scientific); DMEM medium (Gibco); Fetal bovine serum (Deary Tech); CCK8 (Tongren Chemical); DMSO (Ron Reagent); (Xilong Scientific); Gelatin, Carbomer, Sodium alginate, Xanthan gum, Carboxyethyl cellulose, Silk fibroin, Collagen, Polyethylene glycol, Polylactic acid, Hexafluoroisopropanol (Maclean).

[0062] Instruments: Mutiskan Go microplate reader, ST16R centrifuge, 311 CO2 incubator (Thermo Fisher); clean bench (Suantai Air Technology Co., Ltd.); cell counter (IC1000, Countstar); TriStar II 3020 surface area and porosity analyzer (Micromeritics); ZEN 3600 particle size analyzer (Malvin Instruments Co., Ltd.); MS205DU 0.0001 g electronic balance (Mettler Toledo); XMTD-8222 high-temperature oven (Jinghong Instruments); OS20-Pro mechanical stirrer (SCILogex); ACQUITY ARC high-performance liquid chromatograph (Waters, USA); freeze dryer (Shanghai Jingyan Technology Co., Ltd.); electrospinning machine (high voltage power supply (Dalian Jieman Technology Co., Ltd.), micro-propellant pump (Yanhang Power Technology Co., Ltd.)).

[0063] Unless otherwise specified, all other sources are commercial.

[0064] Example 1: Preparation and physicochemical characterization of chlorogenic acid-ferulic acid nanoparticles Formula 1: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0065] Formula 2: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 carrier. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 carrier, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF90 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF90.

[0066] Formula 3: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-methyl in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF8 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF8 support, and 10 mL of methanol. Dissolve chlorogenic acid and ferulic acid in methanol, add the ZIF8 support, stir in the dark for 12 h, centrifuge at 5500 rpm for 10 min, collect the precipitate, wash the precipitate with an appropriate amount of methanol, and freeze-dry for 12 h to obtain a ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), i.e., CA&FA-ZIF8.

[0067] Formula 4: Dissolve 0.17 g of Zn(NO3)2·6H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 carrier. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 carrier, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF90 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF90.

[0068] Formula 5: Dissolve 0.15 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF90 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0069] Formula 6: Dissolve 0.2 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF90 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0070] Prescription 7: Dissolve 0.25 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF90 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0071] Formula 8: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.10 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0072] Prescription 9: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.20 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0073] Formula 10: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.4 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0074] Formula 11: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.8 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0075] Prescription 12: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 6 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0076] Prescription 13: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 24 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0077] Formula 14: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 10 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0078] Prescription 15: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 20 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF90 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0079] Formula 16: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.1% chlorogenic acid, 0.1% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0080] Prescription 17: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.1% chlorogenic acid, 0.2% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0081] Formula 18: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.3% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0082] Prescription 19: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.4% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0083] Formula 20: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.5% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0084] Prescription 21: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.6% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 12 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0085] Prescription 22: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.4% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 12 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 6 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0086] Prescription 23: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.4% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 10 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 6 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0087] Prescription 24: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 support. Place the ZIF90 support in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 support. Take 0.2% chlorogenic acid, 0.4% ferulic acid, 0.1% ZIF91 support, and 10 mL of methanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 24 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 6 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0088] Prescription 25: Dissolve 0.17 g of ZnCH3(COO)2·2H2O and 0.15 g of imidazole-2-formaldehyde in 10 mL of DMF. Stir magnetically for 5 min. Add 0.6 mL of trioctylamine to the reaction solution and continue stirring for 12 h. Centrifuge the reaction mixture at 12000 rpm for 0.5 h, discard the supernatant, wash the precipitate three times with ethanol, centrifuge at 5500 rpm for 10 min, collect the precipitate, and freeze-dry for 12 h to obtain the ZIF90 carrier. Place the ZIF90 carrier in a methanol solution containing 15 μg / mL sodium borohydride, stir in the dark for 24 h, and freeze-dry for 12 h to obtain the ZIF91 carrier. Take 0.2% chlorogenic acid, 0.4% ferulic acid, 0.1% ZIF91 carrier, and 10 mL of ethanol. Chlorogenic acid and ferulic acid were dissolved in methanol, and ZIF91 carrier was added. After stirring in the dark for 24 h, the mixture was centrifuged at 5500 rpm for 10 min and the precipitate was collected. The precipitate was washed with an appropriate amount of methanol and freeze-dried for 6 h to obtain ZIF co-loaded with chlorogenic acid (CA) and ferulic acid (FA), namely CA&FA-ZIF91.

[0089] Physicochemical property characterization: Determination of drug loading: Take the supernatant (M2) after drug loading from Formula 22, centrifuge at 12000 rpm for 10 min, filter through a 0.22 μm filter membrane, collect the filtrate, and inject for analysis. Take the chlorogenic acid and ferulic acid reference standard stock solutions and prepare a series of reference standard solutions with mass concentrations of 0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, 4.0 μg / mL, and 8.0 μg / mL using methanol, and process them in the same way as the samples described above. Determine the chlorogenic acid and ferulic acid reference standard solutions and the test solution according to the chromatographic conditions, record the peak areas, and plot a standard curve with the concentration of chlorogenic acid and ferulic acid reference standard solutions as the abscissa (X) and the peak area (Y) as the ordinate. Calculate the content of chlorogenic acid and ferulic acid using the standard curve. The chromatographic column was an Agilent (4.6 mm × 250 mm, 5 μm), the mobile phase was acetonitrile and 0.1% phosphate solution, gradient elution was used, the flow rate was 1.0 mL / min, the column temperature was 35 ℃, the injection volume was 10 μL, the detection wavelength was 327 nm, and the elution gradient is shown in Table 1.

[0090] 0→9 13→16 87→84 9→10 16→25 84→75 10→16 25→45 75→55 The drug loading rate (%) is calculated using the following formula: (M1 - M2) / M3 × 100% Where M1 is the amount of drug administered, M2 is the amount of drug remaining in the supernatant, and M3 is the mass of CA / FA-ZIF91.

[0091] The loadings of chlorogenic acid and ferulic acid were approximately 14.95% and 10.56%, respectively, with a total drug loading of approximately 25%. The particle size distribution, zeta potential plot, and SEM image of the ZIF-91 nanoformulation are shown below. Figure 1 , 2 As shown, the particle size is relatively uniform, around 65 nm, and the zeta potential is around -17 mV.

[0092] Example 2: Study on the preparation process of gelling agent (CA&FA-ZIF91 Gel) The fractions used in the prescription of this embodiment are mass fractions, and the CA&FA-ZIF91 used is prepared from prescription 24 of Example 1.

[0093] Prescription 1: Take 6% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 hours, mix evenly, and you will get the product.

[0094] Prescription 2: Take 0.8% of Carbomer 940, add an appropriate amount of pure water to make it completely swollen, add 0.15% CA&FA-ZIF91 to the Carbomer 940 matrix, add sodium hydroxide to adjust the pH value to 6.0, add pure water to 10 g, continue stirring at 1500 rpm for 5 hours, mix evenly, and the product is ready.

[0095] Prescription 3: Take 2% xanthan gum, add an appropriate amount of pure water to make it fully swollen, add 0.15% CA&FA-ZIF91 to the xanthan gum matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 h, mix evenly, and the product is ready.

[0096] Prescription 4: Take 2% sodium alginate, add an appropriate amount of pure water to make it completely swollen, add 0.15% CA&FA-ZIF91 to the sodium alginate matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 h, mix evenly, and the product is ready.

[0097] Prescription 5: Take 2% CMC-Na, add an appropriate amount of pure water to make it completely swollen, add 0.15% CA&FA-ZIF91 to the CMC-Na matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 h, mix evenly, and the product is ready.

[0098] Prescription 6: Take 2% hyaluronic acid (40k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 hours, mix evenly, and you will get the product.

[0099] Prescription 7: Take 1.5% hyaluronic acid (80k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 hours, mix evenly, and you will get the product.

[0100] Prescription 8: Take 0.8% hyaluronic acid (120w), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 hours, mix evenly, and you will get the product.

[0101] Prescription 9: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1000 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1000 rpm for 1 hour, mix evenly, and you will get the product.

[0102] Prescription 10: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 1 hour, mix evenly, and you will get the product.

[0103] Prescription 11: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 2000 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 2000 rpm for 2 hours, mix evenly, and you will get the product.

[0104] Prescription 12: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 0.5 h to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 0.5 h, mix evenly, and you will get the product.

[0105] Prescription 13: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 1 h to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 1 h, mix evenly, and you will get the product.

[0106] Prescription 14: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 3 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 3 hours, mix evenly, and you have the product.

[0107] Prescription 15: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 5 h to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 5 h, mix evenly, and you will get the product.

[0108] Prescription 16: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 1.5 hours, mix evenly, and you will get the product.

[0109] Prescription 17: Take 8% hyaluronic acid (10k), add an appropriate amount of pure water, stir at 1500 rpm for 2 hours to make it completely swollen, add 0.15% CA&FA-ZIF91 to the hyaluronic acid matrix, add pure water to 10 g, continue stirring at 1500 rpm for 1 hour, mix evenly, and you will get the product.

[0110] Example 3: Preparation of nanofiber membrane dressing (HA@CA&FA-ZIF91) The fractions used in the prescription of this embodiment are mass fractions, and the CA&FA-ZIF91 used is prepared from prescription 24 of Example 1.

[0111] Formula 1: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0112] Formula 2: Using 10 mL of 60% formic acid as a solvent, prepare a 1% polyethylene glycol solution and magnetically stir at room temperature for 24 h; then add 16% type I collagen peptide COP and 0.10% CA&FA-ZIF91, and magnetically stir at room temperature to obtain a homogeneous COP / PEG spinning solution. A uniform electrospinning solution is obtained; it is then prepared using an electrospinner under the conditions of 25 kV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0113] Formula 3: Add 40% PVP and 0.10% CA&FA-ZIF91 to 6 mL of anhydrous ethanol, and magnetically stir at room temperature for 8 h until completely dissolved. Then, ultrasonically degas for 20 min to prepare a homogeneous PVP shell spinning solution containing the drug composition. Separately, weigh an appropriate amount of PLA and dissolve it in an appropriate amount of chloroform-acetone (volume ratio 3:1) to prepare a 6% mixed solution. After magnetically stirring at room temperature for 15 h, ultrasonically degas for 20 min to obtain a PLA core-shell emulsion spinning solution. Aspirate the core-shell spinning solution into two 10 mL syringes, attach them to a microinjection pump, and connect the two interfaces of a coaxial needle. Prepare the solution using an electrospinning apparatus under the conditions of 25 kV voltage, a feed rate of 0.8 mL / h, and a receiving distance of 15 cm.

[0114] Formula 4: Add 2.5% hyaluronic acid and 14% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 6:4) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0115] Formula 5: Add 2.5% hyaluronic acid and 16% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0116] Formula 6: Add 2.5% hyaluronic acid and 18% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 8:2) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0117] Formula 7: Add 3.0% hyaluronic acid and 14% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0118] Formula 8: Add 3.0% hyaluronic acid and 16% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 8:2) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0119] Formula 9: Add 3.0% hyaluronic acid and 18% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 6:4) to swell, then add 0.10% green CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 25 Kv, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0120] Formula 10: Add 3.5% hyaluronic acid and 14% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 8:2) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0121] Formula 11: Add 3.5% hyaluronic acid and 16% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 6:4) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0122] Formula 12: Add 3.5% hyaluronic acid and 18% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0123] Formula 13: Add 2.8% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0124] Formula 14: Add 3.2% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0125] Formula 15: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.05% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0126] Formula 16: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.15% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0127] Formula 17: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.20% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 25 Kv, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0128] Formula 18: Add 3.0% hyaluronic acid and 12% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 25 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0129] Formula 19: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 20 KV, feed speed 0.5 mL / h, and receiving distance 12 cm.

[0130] Formula 20: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 20 KV, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0131] Formula 21: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 20 KV, feed speed 1.5 mL / h, and receiving distance 18 cm.

[0132] Formula 22: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 24 KV voltage, feed speed 0.5 mL / h, and receiving distance 15 cm.

[0133] Formula 23: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 24 Kv, feed speed 1.0 mL / h, and receiving distance 18 cm.

[0134] Formula 24: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 24 KV voltage, feed speed 1.5 mL / h, and receiving distance 12 cm.

[0135] Formula 25: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 28 Kv, feed speed 0.5 mL / h, and receiving distance 18 cm.

[0136] Formula 26: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 28 Kv, feed speed 1.0 mL / h, and receiving distance 12 cm.

[0137] Formula 27: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 28 Kv, feed speed 1.5 mL / h, and receiving distance 15 cm.

[0138] Formula 28: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 25 Kv, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0139] Formula 29: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 26 KV, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0140] Formula 30: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare the solution using an electrospinning apparatus under the conditions of 24 KV voltage, feed speed 0.8 mL / h, and receiving distance 15 cm.

[0141] Formula 31: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 24 Kv, feed speed 1.2 mL / h, and receiving distance 15 cm.

[0142] Formula 32: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 24 Kv, feed speed 1.0 mL / h, and receiving distance 14 cm.

[0143] Formula 33: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 24 KV, feed speed 1.0 mL / h, and receiving distance 16 cm.

[0144] Formula 34: Add 3.0% hyaluronic acid and 15% gelatin to 10 mL of a mixed solution of hexafluoroisopropanol and water (volume ratio 7:3) to swell, then add 0.10% CA&FA-ZIF91 to the above solution and mix well to obtain a uniform electrospinning solution; prepare it using an electrospinning apparatus under the conditions of voltage 25 Kv, feed speed 1.0 mL / h, and receiving distance 15 cm.

[0145] Example 4: Effects of HA@CA&FA-ZIF91 on HaCa-T cell migration HaCa-T cells in logarithmic growth phase were harvested, digested, and the cell density was adjusted to 4 × 10⁻⁶. 5 Cells / mL: Add the above cell suspension to 6-well plates, 2 mL per well, and incubate at 37°C in a 5% CO2 incubator. Observe the cells until they reach a confluence state, then perform a scratch test. Using a 10 μL sterile pipette tip, scratch the cells perpendicular to the pre-defined horizontal lines, ensuring the pipette tip is perpendicular to the 6-well plate and the scratch width is consistent. After scratching, slowly aspirate the culture medium with the pipette, add an appropriate amount of PBS, and wash the cells three times to remove cell debris. Discard PBS, add complete culture medium to each well, and add erlotinib hydrochloride (16 μM, model group), erlotinib hydrochloride (16 μM) and CA&FA (mass ratio 1.5:1, CA&FA group), erlotinib hydrochloride (16 μM) and HA@ZIF91 (HA@ZIF91 group, without CA&FA, prepared according to the method of formulation 24 in Example 1 and formulation 1 in Example 3), and erlotinib hydrochloride (16 μM) and HA@CA&FA-ZIF91 (HA@CA&FA-ZIF91 group, formulation 1 in Example 3). The amounts added to the CA and FA groups are kept consistent (CA: 0.02 μg / mL, FA: 0.014 μg / mL). Each concentration is repeated in triplicate. Add an equal volume of complete culture medium as a blank group. At different time points (0, 24 h) after scratching, photograph the 6-well plate under an inverted microscope to observe the degree of scratch healing before and after modeling and compared with the model group. Calculate the healing rate according to the following formula: Healing rate = (width of 0 h - width of 24 h) / width of 0 h × 100%.

[0146] The results are as follows Figure 3 , 4 As shown, compared with the model group, the CA&FA, HA@ZIF91, and HA@CA&FA-ZIF91 groups significantly restored the ability of HaCa-T cells to migrate. This indicates that HA@CA&FA-ZIF91 has the effect of restoring the ability of erlotinib hydrochloride to inhibit HaCa-T cell migration.

[0147] Example 5: Retention of CA&FA, CA&FA-ZIF91 and HA@CA&FA-ZIF91 in the skin A Franz vertical diffusion cell was used, with an effective diffusion area of ​​3.14 cm². 2 The receiving pool volume was 8 mL. Instrument settings were: rotation speed 400 r / min, temperature 32℃. Using pig back skin as a model, hair was physically removed to ensure the integrity of the stratum corneum and prevent surface damage. The skin was stored at -80℃, thawed at room temperature before the experiment, cleaned with physiological saline, and cut into 3 cm squares. The receiving solution was PBS buffer (pH 7.4) containing 1% polyethylene glycol-400 and 0.1% gentamicin (to prevent bacterial fermentation of the skin from affecting drug penetration and retention). 1 mL of CA&FA 20% ethanol solution (CA&FA group), CA&FA-ZIF91 solution (Formula 24 from Example 1 dispersed in an appropriate amount of water), and HA@CA&FA-ZIF91 dressing (Formula 1 from Example 3 applied to pig back skin and sprayed with an appropriate amount of water to allow permeation) were added to the supply pool. All tested formulations contained 1 mg of ferulic acid and 1.5 mg / mL of chlorogenic acid. After 24 hours, 2 mL of sample was taken from the receiving pool to detect the skin penetration of the drug.

[0148] Skin retention: After 24 h, skin was removed from the diffusion cell and the residual drug on the pigskin surface was washed with PBS buffer (pH 7.4) containing 1% (w / %) polyethylene glycol-400. The skin was dried, cut into small pieces with scissors, ground, and extracted with 1 mL of methanol by sonication. The resulting mixture was centrifuged at 12000 r / min for 15 min, then filtered through a 0.22 μm filter membrane, and the filtrate was analyzed by HPLC. The method for determining the CA&FA content was the same as in Example 4.

[0149] The results are as follows Figure 5 , 6 As shown, the retention amounts of ferulic acid in the skin of CA&FA solution, CA&FA-ZIF91, and HA@CA&FA-ZIF91 were 22.11±0.96, 58.8±6.03, and 60.12±2.29 μg / cm³, respectively. 2 There were no significant differences between the groups. The retention amounts of chlorogenic acid in the skin from CA&FA solution, CA&FA-ZIF91, and HA@CA&FA-ZIF91 were 13.33±0.56, 93.90±4.26, and 95.51±4.15 μg / cm³, respectively. 2 Compared with the CA&FA group, the retention of chlorogenic acid in the skin was significantly increased in CA&FA-ZIF91 and HA@CA&FA-ZIF91. This indicates that ZIF91 can significantly increase drug retention in the skin.

[0150] Example 6: The interaction between CA&FA-ZIF91 and ceramides in the skin A Franz vertical diffusion cell was used, with an effective diffusion area of ​​3.14 cm². 2 The receiving pool volume was 8 mL. Instrument settings were: rotation speed 400 r / min, temperature 32 ℃. Using pig back skin as a model, hair was physically removed to ensure the integrity of the stratum corneum and prevent surface damage. The skin was stored at -80 ℃, thawed at room temperature before the experiment, cleaned with physiological saline, and cut into 3 cm squares. The release medium was PBS buffer (pH 7.4) containing 1% (w / %) polyethylene glycol-400 and 0.1% gentamicin (to prevent bacterial fermentation of the skin from affecting drug penetration and retention). A blank group, a ZIF91 group (without CA&FA, prepared according to the method of Formulation 24 in Example 1, dispersed with an appropriate amount of water), a CA&FA group (in 20% ethanol solution), and a CA&FA-ZIF91 group (Formulation 24 in Example 1, dispersed with an appropriate amount of water) were set up. All test formulations contained 1 mg / mL ferulic acid and 1.5 mg / mL chlorogenic acid. After 24 hours, the skin was removed, the administration site was cut off, rinsed with physiological saline, and lyophilized. A suitable amount of skin sample was taken and analyzed by DSC to determine the interaction between CA&FA-ZIF91 and ceramides in the skin. Detection conditions: heating rate 10 ℃ / min, heating range 30–200 ℃, nitrogen flow rate 50 mL / min.

[0151] The results are as follows Figure 7 As shown, the DSC pattern of pig back skin treated with CA&FA showed similar wavenumber changes to the control group. The phase transition temperature and enthalpy change of pig back skin treated with ZIF91 and CA&FA-ZIF91 were significantly increased, indicating that ZIF91 forms hydrogen bonds with ceramides in the skin, allowing the drug to remain in the skin and facilitating its therapeutic effect at the target areas where EGFRIs induce skin toxicity.

[0152] Example 7: Efficacy evaluation of the pharmaceutical composition for treating dermal toxicity induced by EGFRIs BALB / c mice (approximately 20 g) were randomly divided into two groups according to body weight after acclimatization for 3 days. The mice were randomly assigned to a control group (n=10) and an erlotinib hydrochloride group (n=50). The day before administration, the BALB / c mice underwent hair removal on their backs and were administered erlotinib hydrochloride solution by gavage at a dose of 150 mg / kg (0.015 mL / g) once daily until skin toxicity appeared (approximately 48 days).

[0153] After establishing the animal model of skin toxicity induced by EGFRIs, the rats in the erlotinib hydrochloride group were further divided into four groups: erlotinib hydrochloride group (n=10), erlotinib hydrochloride + drug solution group (n=10), erlotinib hydrochloride + blank matrix group (n=10), erlotinib hydrochloride + drug composition nanofiber membrane dressing group (n=10), and erlotinib hydrochloride + drug composition gel group (n=10) (CA&FA-ZIF91 Gel). All groups of mice continued to be administered erlotinib hydrochloride 150 mg / kg by gavage once daily. In addition, the drug treatment groups received topical application of the corresponding preparations (obtained using the same methods as in Example 5) at a dose of 125 mg / kg, while the model group received topical application of physiological saline once daily; this treatment continued for 14 consecutive days.

[0154] After the animal experiment, the skin toxicity of each group of mice was observed and the severity of skin toxicity was scored. Figure 8 ), and statistically analyzed the weight change curves of mice in each group ( Figure 9 The thymus, spleen, and liver of mice in different groups were weighed, organ indices were calculated, and differences between groups were compared. Figures 10-12 The water content of a portion of the skin on the back of a mouse was measured. Figure 13 The efficacy of the drug combination was observed by examining skin sections stained with HE and Oil Red. Figure 14 ).

[0155] The results are shown in Tables 2-4: The severity of dry, scaly, red, swollen, herpes, and rashes on the mouse skin indicates that the model group mice experienced hair loss around the eyes, neck, and back, and dry, scaly, cracked, and herpes-like skin around the mouth and limbs, resulting in decreased activity. The CA&FA group, HA@ZIF91 group, HA@CA&FA-ZIF91 group, and CA&FA-ZIF91 Gel group all showed varying degrees of therapeutic effect on skin toxicity. The HA@CA&FA-ZIF91 group and CA&FA-ZIF91 Gel group showed better efficacy, significantly reducing the incidence of redness, swelling, herpes, and rashes on the mouse skin. Hair regrowth resumed, symptoms significantly disappeared, and the treatment of skin toxicity was effective, with an efficacy rate of 90%.

[0156] Blank group 10 10 - - - - Model group 10 - - 7 3 100% CA&FA 10 5 4 1 - 50% HA@ZIF91 10 1 5 3 1 90% HA@CA&FA-ZIF91 10 9 1 - - 10% CA&FAZIF91 Gel 10 9 1 - - 10%

[0157] Blank group 10 10 - - - - Model group 10 5 2 3 - 50% CA&FA 10 8 1 1 - 20% HA@ZIF91 10 6 3 1 - 40% HA@CA&FA-ZIF91 10 10 - - - - CA&FA-ZIF91 Gel 10 10 - - - -

[0158] Blank group 10 10 - - - - Model group 10 - 2 5 3 100% CA&FA 10 8 2 - - 20% HA@ZIF91 10 6 4 - - 40% HA@CA&FA-ZIF91 10 10 - - - - CA&FA-ZIF91 Gel 10 10 - - - - During treatment, the body weight of mice in the model group did not change significantly, while the body weight of mice in the CA&FA, HA@ZIF91, HA@CA&FA-ZIF91, and CA&FA-ZIF91 Gel groups increased, indicating that the drug improved the quality of life of the mice. The liver and spleen indices of the model group mice were significantly increased, and the thymus index was decreased, indicating that erlotinib hydrochloride severely damaged the immune system of the mice. Treatment with the CA&FA, HA@ZIF91, HA@CA&FA-ZIF91, and CA&FA-ZIF91 Gel groups resulted in varying degrees of recovery, indicating that the drug can reverse and protect against organ damage caused by erlotinib hydrochloride, with the HA@CA&FA-ZIF91 and CA&FA-ZIF91 Gel groups showing more significant effects.

[0159] Analysis of skin hydration results showed that, compared with the control group, the hydration content of mice in the model group was significantly reduced, indicating that erlotinib hydrochloride severely damaged the skin barrier of mice and affected their moisturizing function. After administration of CA&FA, HA@ZIF91, HA@CA&FA-ZIF91, and CA&FA-ZIF91 Gel, the skin hydration content increased significantly, indicating that topical application of the drug composition gel can have a certain repairing effect on the skin barrier.

[0160] HE staining results showed that, compared with the control group, the model group mice exhibited characteristics of skin inflammation, namely parakeratosis (retention of stratum corneum cell nuclei), significant thickening of the stratum corneum, and altered skin structure. Treatment with CA&FA, HA@ZIF91, HA@CA&FA-ZIF91, and CA&FA-ZIF91 Gel improved the skin toxicity of the mice, with the HA@CA&FA-ZIF91 and CA&FA-ZIF91 Gel groups showing the most significant improvement. Oil Red staining results showed that, compared with the control group, the number of inhibited and swollen follicles in the sebaceous glands was significantly reduced in the model group. Compared with the model group, the number of swollen follicles in the skin of mice treated with CA&FA, HA@ZIF91, HA@CA&FA-ZIF91, and CA&FA-ZIF91 Gel was significantly increased, indicating that the drug combination can effectively restore the inhibition of sebaceous gland secretion by erlotinib hydrochloride.

Claims

1. A pharmaceutical composition, characterized in that, The material comprises chlorogenic acid, ferulic acid, and zeolite imidazole framework, wherein the mass ratio of chlorogenic acid to ferulic acid is 4:1 to 1:

4. The zeolite imidazole framework material is a Zn-based zeolite imidazole framework material.

2. The pharmaceutical composition according to claim 1, characterized in that, The mass ratio of chlorogenic acid to ferulic acid is 2:1 to 1:

2.

3. The pharmaceutical composition according to claim 2, characterized in that, The mass ratio of chlorogenic acid to ferulic acid is 1.5:

1.

4. The pharmaceutical composition according to any one of claims 1-3, characterized in that, The zeolite imidazole framework material is selected from ZIF91, ZIF90 or ZIF8.

5. The pharmaceutical composition according to any one of claims 1-3, characterized in that, The total amount of chlorogenic acid and ferulic acid is in a mass ratio of 3:1 to 1:3 to the Zn element in the zeolite imidazole framework material.

6. The pharmaceutical composition according to claim 5, characterized in that, The total amount of chlorogenic acid and ferulic acid is in a mass ratio of 2:1 to 1:2 to the Zn element in the zeolite imidazole framework material.

7. The pharmaceutical composition according to claim 6, characterized in that, The total amount of chlorogenic acid and ferulic acid is in a mass ratio of 4:3 to the Zn element in the zeolite imidazole framework material.

8. The pharmaceutical composition according to any one of claims 1-3, characterized in that, The pharmaceutical composition also includes a matrix material. When the matrix material is a gel matrix and water, the pharmaceutical composition is a gelling agent; When the matrix material is a spinning matrix, the pharmaceutical composition is a nanofiber membrane dressing.

9. The pharmaceutical composition according to claim 8, characterized in that, The gel matrix is ​​one or a mixture of several of the following: hyaluronic acid, carbomer, sodium alginate, or xanthan gum; The spinning matrix is ​​one or a mixture of several of the following: hyaluronic acid, gelatin, silk fibroin, collagen, polyethylene glycol, or polylactic acid.

10. The pharmaceutical composition according to claim 9, characterized in that, The spinning matrix is ​​a mixture of hyaluronic acid and gelatin.

11. The pharmaceutical composition according to claim 10, characterized in that, The mass ratio of hyaluronic acid to gelatin is 1:(4.8~7.2).

12. The pharmaceutical composition according to claim 8, characterized in that, In the gelling agent, the mass fraction of the gel matrix is ​​1-12%; the total mass fraction of the chlorogenic acid, ferulic acid, and zeolite imidazole framework material in the gelling agent is 0.5-2.0%. In the nanofiber membrane dressing, the mass fraction of the spinning matrix is ​​85-95%.

13. The pharmaceutical composition according to claim 12, characterized in that, In the gelling agent, the mass fraction of the gel matrix is ​​6-10%.

14. The pharmaceutical composition according to claim 13, characterized in that, In the gelling agent, the mass fraction of the gel matrix is ​​8%.

15. A method for preparing the pharmaceutical composition according to any one of claims 1-14, characterized in that, Includes the following steps: Chlorogenic acid, ferulic acid, and zeolite imidazole framework material are mixed in a methanol or ethanol solution in the dark, followed by solid-liquid separation to obtain drug particles.

16. The preparation method according to claim 15, characterized in that, When the pharmaceutical composition is a gel, the preparation method further includes step A; when the pharmaceutical composition is a nanofiber membrane dressing, the preparation method further includes step B. Step A: Mix the gel matrix and water to make the gel matrix swell, then add the drug particles and mix evenly to obtain the gel; Step B: Prepare a spinning solution by combining the spinning matrix and the drug particles, and then use electrospinning to obtain the nanofiber membrane dressing.

17. The preparation method according to claim 16, characterized in that, The electrospinning conditions are: voltage 20~28kV, feed speed 0.5~1.5 mL / h, and receiving distance 12~18 cm.

18. The preparation method according to claim 17, characterized in that, The electrospinning conditions are: voltage 24~26kV, feed speed 0.8~1.2 mL / h, and receiving distance 14~16 cm.

19. The preparation method according to claim 16, characterized in that, The spinning solution is prepared by the following method: The spinning matrix is ​​swollen in a spinning solvent, and then the drug particles are added and mixed evenly to obtain the spinning solution. The spinning solvent is any one or a mixture of hexafluoroisopropanol, dichloromethane, trichloromethane, formic acid, or water.

20. The preparation method according to claim 19, characterized in that, The spinning solvent is a mixture of hexafluoroisopropanol and water.

21. The preparation method according to claim 20, characterized in that, The spinning solvent is a mixture of hexafluoroisopropanol and water in a volume ratio of 6:4 to 8:

2.

22. The preparation method according to claim 21, characterized in that, The spinning solvent is a mixture of hexafluoroisopropanol and water in a volume ratio of 7:

3.

23. The preparation method according to any one of claims 19-22, characterized in that, The spinning matrix is ​​a mixture of hyaluronic acid and gelatin, and the mass concentration of hyaluronic acid in the spinning solution is 2.5-3.5%, and the mass concentration of gelatin is 12-18%.

24. The preparation method according to claim 23, characterized in that, The spinning solution contains 2.8-3.2% hyaluronic acid and 14-16% gelatin by mass.

25. Use of the pharmaceutical composition of any one of claims 1-14 in the preparation of a therapeutic remedy for erlotinib-induced skin toxicity.

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