A method for preparing a metal carrier loaded with deaminotyrosine and its application

Abstract

This invention discloses a method for preparing a metal carrier loaded with deaminotyrosine and its application. The preparation method involves first synthesizing Zr-MOFs with specific particle size and structure using ZrOCl2•8H2O, TCPP, and benzoic acid in specific proportions and concentrations, and then reacting them with a certain concentration of deaminotyrosine. DAT@Zr-MOFs are prepared in two steps. The prepared DAT@Zr-MOFs have a high loading of deaminotyrosine and good stability, effectively overcoming the problems of enzymatic hydrolysis and oxidation of deaminotyrosine. They can be used for external application to treat wounds and promote tissue repair.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to a method for preparing and applying a metal carrier loaded with deaminotyrosine. Background Technology

[0002] Deaminotyrosine (DAT), also known as p-hydroxyphenylpropionic acid, is a metabolite derived from gut microbiota. Studies have shown that DAT possesses antioxidant, anti-inflammatory, and lipid metabolism-regulating effects, demonstrating promising applications in reducing inflammatory responses, controlling infection, anti-aging, antioxidant therapy, and neuronal protection. However, because DAT is easily degraded by enzymes or oxidized in the physiological environment, its free molecules are rapidly metabolized in vivo, making it difficult to achieve effective concentrations.

[0003] Metal-organic frameworks (MOFs) are porous materials formed by the self-assembly of metal ions / clusters and organic ligands. They possess characteristics such as high specific surface area, tunable pore size, and surface functionalization, and are widely used in drug delivery, catalysis, and gas adsorption. The structural flexibility of MOFs allows for the tuning of their properties through the design of ligands and metal nodes. Among them, Zr... 4+ Zr-based metal-organic frameworks (Zr-MOFs) are a class of metals based on zirconium ions (Zr). 4+ Zr-MOFs are porous materials with metal nodes and organic ligands as connecting units. Due to their ultra-high stability, structural diversity, and functional tunability, they have become a research hotspot in the field of MOFs. The high bond energy of the Zr-O bond in Zr-MOFs (approximately 800 kJ / mol) endows the materials with excellent chemical stability, making them resistant to decomposition in body fluids (pH 4-8), blood, and high-temperature environments, thus preventing premature drug leakage. Compared to liposomes or polymer micelles, Zr-MOFs exhibit strong resistance to degradation by proteases, nucleases, and other biological enzymes, making them suitable for long-term in vivo circulation. Furthermore, Zr-MOFs have a specific surface area of ​​up to 3000 m² / g, and their microporous / mesoporous structure (0.5-3 nm) can physically adsorb or chemically bond drug molecules, achieving drug loading capacities as high as 30%-50% (by weight). Simultaneously, Zr... 4+ It is a biologically inert metal, and its metabolic products (such as ZrO2) are non-toxic and can be slowly excreted through the kidneys, making it relatively safe for long-term use.

[0004] To overcome the problem that deaminotyrosine is easily enzymatically hydrolyzed and oxidized, our laboratory has explored a metal carrier capable of loading deaminotyrosine by combining Zr-MOFs with deaminotyrosine. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the present invention aims to provide a method for preparing and applying a metal carrier loaded with deaminotyrosine, so as to solve the problems mentioned in the background art.

[0006] The technical solution adopted by the present invention to achieve the above objectives is as follows:

[0007] A method for preparing a metal support loaded with deaminotyrosine includes the following steps:

[0008] S1. Mix ZrOCl2•8H2O solution, TCPP solution, and benzoic acid solution thoroughly, then reflux at 90℃ for 5 hours. After the reaction is complete, the product Zr-MOFs is obtained.

[0009] S2. Mix deaminotyrosine with the Zr-MOFs obtained in step S1 and reflux at 90°C for 5 hours. After the reaction is complete, dialyze through deionized water 3-4 times to obtain the product DAT@Zr-MOF.

[0010] Preferably, the mass concentration ratio of ZrOCl2•8H2O, TCPP and benzoic acid is 6:1-1.5:28.

[0011] Preferably, the mass concentration of ZrOCl2•8H2O is not less than 15 mg / mL.

[0012] Preferably, the mass concentration of deaminotyrosine is not less than 10 mg / mL.

[0013] The present invention describes a method for preparing a metal carrier loaded with deaminotyrosine. First, Zr-MOFs with specific particle sizes and structures are synthesized using ZrOCl2•8H2O, TCPP, and benzoic acid in specific proportions and concentrations. Then, these are reacted with a certain concentration of deaminotyrosine to prepare DAT@Zr-MOFs in a one-step process. The prepared DAT@Zr-MOFs have a high loading capacity of deaminotyrosine, good stability, and can effectively overcome the problems of enzymatic hydrolysis and oxidation of deaminotyrosine.

[0014] Preferably, the present invention also provides the application of the metal carrier of deaminotyrosine prepared above in the preparation of a drug for external application to treat wounds and promote tissue repair.

[0015] The present invention has the following beneficial effects:

[0016] 1. The metal carrier loaded with deaminotyrosine prepared by the present invention is a spherical nanoparticle. The deaminotyrosine loaded is uniformly distributed and has good stability. Moreover, the preparation method is simple and the conditions are mild.

[0017] 2. The metal carrier loaded with deaminotyrosine prepared by this invention has low cytotoxicity, is not easily decomposed in body fluids, blood and high temperature environments, and will not cause premature drug leakage.

[0018] 3. The carrier prepared by this invention can be used to prepare topical medications loaded with deaminotyrosine, enabling them to treat wounds and rapidly promote tissue repair; Attached Figure Description

[0019] Figure 1 This is a flowchart of the preparation of a metal framework loaded with deaminotyrosine;

[0020] Figure 2 This is a transmission electron microscope image of the prepared metal framework loaded with deaminotyrosine (scale bar: 200 nm).

[0021] Figure 3 This invention demonstrates how the metal framework loaded with deaminotyrosine promotes wound healing in mice. The top image shows HE staining of mouse skin wounds; the bottom image shows Masson staining of mouse skin wounds. Detailed Implementation

[0022] To further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, and not for limiting the scope of the claims. Example

[0023] The preparation of the metal framework M1 loaded with deaminotyrosine includes the following steps (such as...) Figure 1 ):

[0024] S1. Mix ZrOCl2•8H2O solution (2.5 mL, 15 mg / mL, dissolved in DMF), TCPP solution (5 mL, 2.5 mg / mL, dissolved in DMF), and benzoic acid (5 mL, 70 mg / mL, dissolved in DMF) thoroughly. Reflux the reaction mixture at 90 °C for 5 h. After the reaction is complete, the product Zr-MOFs is obtained.

[0025] S2. The above reaction product and deaminotyrosine solution (DAT, 3.75 mL, 10 mg / mL, dissolved in DMF) are mixed and refluxed at 90 °C for 5 h. After the reaction is complete, the mixture is dialyzed through deionized water 3-4 times to obtain product M1. Example

[0026] The preparation of the metal framework M2 loaded with deaminotyrosine includes the following steps:

[0027] S1. Mix ZrOCl2•8H2O solution (2.5 mL, 15 mg / mL, dissolved in DMF), TCPP solution (5 mL, 2.5 mg / mL, dissolved in DMF), and benzoic acid (5 mL, 70 mg / mL, dissolved in DMF) thoroughly. Reflux the above reaction solution at 90 °C for 5 h. After the reaction is complete, the product Zr-MOFs is obtained.

[0028] S2. The above reaction product and deaminotyrosine solution (DAT, 3.75 mL, 12 mg / mL, dissolved in DMF) are mixed and refluxed at 90 °C for 5 h. After the reaction is complete, the mixture is dialyzed through deionized water 3-4 times to obtain product M2. Example

[0029] The preparation of the metal framework M3 loaded with deaminotyrosine includes the following steps:

[0030] S1. Mix ZrOCl2•8H2O solution (2.5 mL, 15 mg / mL, dissolved in DMF), TCPP solution (5 mL, 3.75 mg / mL, dissolved in DMF), and benzoic acid (5 mL, 70 mg / mL, dissolved in DMF) thoroughly. Reflux the reaction mixture at 90 °C for 5 h. After the reaction is complete, the product Zr-MOFs is obtained.

[0031] S2. The above reaction product and deaminotyrosine solution (DAT, 3.75 mL, 12 mg / mL, dissolved in DMF) are mixed and refluxed at 90 °C for 5 h. After the reaction is complete, the mixture is dialyzed through deionized water 3-4 times to obtain product M3. Example

[0032] The particle size distribution of M1-M3 obtained in Examples 1-3 was characterized by transmission electron microscopy (TEM), ultraviolet-visible absorption spectroscopy, infrared spectroscopy, and dynamic light scattering. The results showed that all M1-M3 particles were uniformly distributed spherical nanoparticles, with the smallest particle size of the prepared M2 product being 143.8 nm. Figure 2 As shown. Example

[0033] The M2 product prepared in Example 2 was formulated into a topical application for treating trauma, and its ability to promote tissue repair was tested.

[0034] S1. Mix M2 with poloxamer 407 (1 mL, 20%, dissolved in DAT@Zr-MOF aqueous solution) and dissolve in a shaker at 4°C for 8 hours to obtain the product DAT@Zr-MOF hydrogel.

[0035] S2. Take 6-8 week old C57 mice, anesthetize them with 1% pentobarbital at a dose of 50 mg / kg, remove hair from the back skin and disinfect with iodine, and then use ophthalmic scissors to make a circular skin wound with a diameter of 9 mm.

[0036] S3. After disinfecting again with povidone-iodine, add 80 μL of uLDAT@Zr-MOF hydrogel. After the hydrogel solidifies, wrap the wound with gauze.

[0037] S4 and 14 days later, the skin on the back of the mice was fixed in 4% paraformaldehyde, and the healing of the mouse skin wound was examined by paraffin embedding, sectioning, HE staining, and Masson staining.

[0038] S5, by Figure 3 As can be seen, the metal framework DAT@Zr-MOF hydrogel loaded with deaminotyrosine can effectively promote wound healing on the skin surface of mice.

Claims

1. The application of a metal carrier loaded with deaminotyrosine in the preparation of a drug for external application to treat wounds and promote tissue repair, characterized in that, The method for preparing the metal support loaded with deaminotyrosine includes the following steps: S1. ZrOCl2•8H2O solution, TCPP solution and benzoic acid solution were mixed and heated under reflux at 90℃ to obtain Zr-MOFs; The mass concentration ratio of ZrOCl2•8H2O, TCPP and benzoic acid is 6:1:28; The mass concentration of ZrOCl2•8H2O is 15 mg / mL; S2. Deaminotyrosine at a mass concentration of 12 mg / mL was mixed with Zr-MOFs and refluxed at 90 °C. After the reaction was complete, the mixture was dialyzed through deionized water 3-4 times to obtain a metal carrier loaded with deaminotyrosine with a particle size of 143.8 nm.

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