Coenzyme Q10-loaded nano-organic metal framework material Al-MOF@PEG and preparation method thereof

Abstract

This invention belongs to the field of biopolymer materials and nanotechnology, specifically relating to a method for preparing Al-MOF@PEG nanostructured framework material loaded with coenzyme Q10. In this invention, 2,5-dihydroxyterephthalic acid and aluminum (Al) ions are selected to synthesize Al-MOF nanocarriers via a solvothermal method, and coenzyme Q10 is loaded into the pores of the Al-MOF nanocarriers. Subsequently, mPEG5k-NH2, an amino-terminated PEG, is selected and covalently modified onto the Al-MOF nanocarrier surface via a "click chemistry" process between the carboxyl group activated by dicyclohexylcarbodiimide (DCC) on the Al-MOF surface and the amino group at the PEG terminus. The larger pore size of the MOF material increases the drug loading capacity, while the introduction of PEG significantly increases water solubility and prolongs the circulation time of the nanocarrier in the physiological environment, further improving the bioavailability of coenzyme Q10.

Description

Technical Field

[0001] This invention belongs to the field of biopolymer materials and nanotechnology, specifically relating to a method for preparing Al-MOF@PEG nanostructured metal framework material loaded with coenzyme Q10. Background Technology

[0002] Coenzyme Q10 is a natural yellow benzoquinone, a vitamin-like substance commonly found in blood, mitochondria, and cell membranes. It is a natural antioxidant that plays a crucial role in maintaining several biochemical pathways in the human body. It can transfer electrons in the mitochondrial transport chain to synthesize ATP by promoting the transmembrane potential utilized by ATPases, and in its reduced form, it is also a membrane antioxidant, essential for driving the ATP cycle, the body's energy production process. Coenzyme Q10 not only provides sufficient oxygen to protect the heart, scavenge free radicals, and fight oxidation to effectively delay aging, but also keeps cells healthy and produces an anti-fatigue effect. Furthermore, recent studies have shown that coenzyme Q10 also has certain therapeutic effects in cancer prevention and treatment. It also exhibits significant biological activity in chronic heart failure, cardiofacial syndrome, diabetes, cancer, autoimmune diseases, cataracts, asthma, periodontal disease, and thyroid disease. As a fat-soluble antioxidant, coenzyme Q10 is widely used in the cosmetics and healthcare industries; however, its high molecular weight and low water solubility hinder its bioavailability to some extent. Therefore, researchers have developed various technologies to address these issues. Currently, most commercially available coenzyme Q10 formulations are ordinary emulsions or gels, which have poor stability and lack penetration-enhancing and targeted functions, hindering the maximization of coenzyme Q10's efficacy. Therefore, improving the stability of coenzyme Q10, prolonging its effective duration, and promoting effective penetration and targeted release are urgent problems to be solved in its application. Summary of the Invention

[0003] To address the problems existing in the prior art, this invention provides a nano-organic metal framework material carrying coenzyme Q10. The larger pore size of this nanocarrier increases drug loading capacity, and the introduction of PEG significantly increases water solubility and prolongs the circulation time of the nanocarrier in the physiological environment, which will further improve the bioavailability of coenzyme Q10. It also possesses the advantages of nano-drug delivery technology, such as improved drug stability, delayed release, and altered drug distribution in vivo. Another objective of this invention is to provide a method for preparing the aforementioned nanocarrier.

[0004] The first aspect of this invention provides a method for preparing Al-MOF@PEG nanoframework material loaded with coenzyme Q10. 2,5-Dihydroxyterephthalic acid and aluminum (Al) ions are selected, and Al-MOF nanocarriers are synthesized via a solvothermal method. Coenzyme Q10 is then loaded into the pores of the Al-MOF nanocarriers. Subsequently, mPEG5k-NH2, an amino-terminated PEG, is selected and covalently modified onto the Al-MOF nanocarrier surface via a "click chemistry" process between the carboxyl group activated by dicyclohexylcarbodiimide (DCC) on the Al-MOF surface and the amino group at the PEG terminus. The larger pore size of the MOF material increases the drug loading capacity, and the introduction of PEG significantly increases water solubility and prolongs the circulation time of the nanocarrier in the physiological environment, which will further improve the bioavailability of coenzyme Q10.

[0005] Preferably, the preparation method of the coenzyme Q10-loaded metal-organic framework Al-MOF@PEG includes the following steps:

[0006] S1: Preparation of Al-MOF: Nano-Al-MOF was prepared using acetic acid as a modifier. AlCl3·6H2O and 2,5-dihydroxyterephthalic acid were uniformly mixed in DMF. Acetic acid was added, followed by an aqueous ethanol solution, and then pure water was added. The resulting mixture was placed in a 500 mL autoclave and heated to 135 °C for 24 hours. After the reaction was completed, the mixture was cooled to room temperature. The mixture was removed and centrifuged. The supernatant was discarded, and DMF was added and centrifuged three times. The precipitate obtained from the last centrifugation was placed in CH2Cl2 for two days, and fresh CH2Cl2 was continuously replaced to remove unreacted raw materials adhering to the precipitate. After filtration, the nano-carrier Al-MOF was collected by vacuum drying.

[0007] S2: Preparation of Al-MOF nanocarriers carrying coenzyme Q10: Al-MOF nanocarriers were dispersed in pure water, coenzyme Q10 was dissolved in DMF and then added dropwise to pure water and mixed well. The mixture was sonicated for 10 minutes and stirred for 4 hours. Then, the mixture was centrifuged, the supernatant was removed, and the mixture was washed three times with pure water. The solid precipitate obtained by the last centrifugation was freeze-dried to obtain Al-MOF nanocarriers carrying coenzyme Q10.

[0008] S3: Preparation of Al-MOF@PEG nanocarrier carrying coenzyme Q10: Al-MOF was ultrasonically dispersed in CH2Cl2, and then dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine were added; the mixture was stirred at 10℃ for 3 hours, and then CH3O-PEG5k-NH2 was added and reacted at 4℃ for 17 hours; after the reaction was completed, the mixture was removed and centrifuged, the supernatant was discarded, and CH2Cl2 was added and centrifuged and washed three times. Finally, the product, coenzyme Q10-carrying nanoorganic metal framework material Al-MOF@PEG, was collected by vacuum drying.

[0009] Preferably, the mass ratio of AlCl3·6H2O to 2,5-dihydroxyterephthalic acid in S1 is [missing value].

[0010] (4~4.5): 1.

[0011] Preferably, the mass ratio of AlCl3·6H2O to acetic acid in S1 is 1:(17-20).

[0012] Preferably, the amount of DMF used to dissolve AlCl3·6H2O and 2,5-dihydroxyterephthalic acid in S1 is 320-380 mL.

[0013] Preferably, the amount of ethanol aqueous solution and water added in S1 is 20-25 mL.

[0014] Preferably, the mass ratio of the nanocarrier Al-MOF to coenzyme Q10 in S2 is (6-10):1.

[0015] Preferably, the mass ratio of the Al-MOF nanocarrier carrying coenzyme Q10 in S3 to CH3O-PEG5k-NH2 is 1:(1~1.5).

[0016] Preferably, the mass ratio of the coenzyme Q10-carrying nanocarrier Al-MOF, dicyclohexylcarbodiimide (DCC), and 1,6-hexanediamine in S3 is 1:1:(0.8-0.9).

[0017] The second aspect of this invention discloses a method for preparing Al-MOF@PEG nano-organic metal framework material loaded with coenzyme Q10.

[0018] Beneficial effects

[0019] The coenzyme Q10-loaded nano-organic metal framework material of this invention not only possesses high porosity, structural diversity, and controllable function, but also combines the strong drug-carrying capacity of inorganic nanocarriers with the high safety of organic nanocarriers. The introduced PEG structure significantly increases the solubility of coenzyme Q10 in water, which will further improve bioavailability. Simultaneously, nano-drug delivery technology also offers advantages such as improved drug stability, delayed release, and altered drug distribution in vivo. Attached Figure Description

[0020] Figure 1 The infrared (FT-IR) spectrum of Al-MOF;

[0021] Figure 2 Nitrogen adsorption-desorption spectra of Al-MOF@PEG nanocarriers carrying coenzyme Q10;

[0022] Figure 3 Dynamic light scattering (DLS) particle size analysis of Al-MOF@PEG nanocarriers carrying coenzyme Q10.

[0023] Figure 4 The release performance of Al-MOF@PEG nanocarrier carrying coenzyme Q10 in different release media is shown in the figure. Detailed Implementation

[0024] Example 1

[0025] The preparation of Al-MOF@PEG loaded with coenzyme Q10 includes the following steps:

[0026] (1) Preparation of Al-MOF: Nano-Al-MOF was prepared using acetic acid as a modifier. 3.62 g of AlCl3·6H2O and 0.83 g of 2,5-dihydroxyterephthalic acid were mixed into 355 mL of DMF. 60.00 g of acetic acid, 23.50 mL of 95% ethanol aqueous solution, and 23.50 mL of water were added. The resulting mixture was placed in a 500 mL autoclave and heated to 135 °C for 24 hours. After the reaction was complete, the mixture was cooled to room temperature, removed, centrifuged, and the supernatant was discarded. DMF was then added and the mixture was centrifuged and washed three times. The precipitate obtained from the last centrifugation was placed in CH2Cl2 for two days, continuously replaced with fresh CH2Cl2 to remove unreacted raw materials adhering to the precipitate. After filtration, the nano-carrier Al-MOF was collected by vacuum drying.

[0027] (2) Preparation of Al-MOF nanocarrier loaded with coenzyme Q10: 30 mg of Al-MOF nanocarrier was dispersed in pure water, and 5 mg of coenzyme Q10 was dissolved in DMF and then added dropwise to pure water and mixed. The mixture was sonicated for 10 minutes and stirred for 4 hours. Then, the mixture was centrifuged, the supernatant was removed, and the mixture was washed three times with pure water. The solid precipitate obtained by centrifugation was freeze-dried to obtain Al-MOF nanocarrier loaded with coenzyme Q10.

[0028] (3) Preparation of Al-MOF@PEG nanocarrier carrying coenzyme Q10: 100 mg Al-MOF was ultrasonically dispersed in 15 mL CH2Cl2, and then 100 mg dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine were added. The mixture was stirred at 10 °C for 3 hours. 100 mg CH3O-PEG5k-NH2 was added and reacted at 4 °C for 17 hours. After removing the mixed solution, centrifugation was performed, the supernatant was discarded, and CH2Cl2 was added for washing three times. Finally, the product, Al-MOF@PEG nanoorganic metal framework material carrying coenzyme Q10, was collected by vacuum drying.

[0029] Example 2

[0030] Characterization of Al-MOF@PEG nanostructured framework material loaded with coenzyme Q10

[0031] (1) The Al-MOF obtained in reaction S1 of Example 1 was characterized by Fourier transform infrared spectroscopy (FT-IR) (e.g., Figure 1 The KBr tableting method was used. The dried product and KBr were mixed and ground at a mass ratio of 1:100, dried under an infrared lamp, and then compressed into tablets. The tablets were then placed in an instrument for testing using a Nexus infrared spectrometer at 4000-400 cm⁻¹. -1 Test relevant samples within the specified range. 1700cm -1 The peak position of absorption by vibration is 1400 cm⁻¹. -1 and 1590cm -1 The characteristic peak at 1100 cm⁻¹ corresponds to the absorption peak of the stretching vibration of the benzene ring skeleton. -1 It is the stretching vibration of CO, 750cm -1 The absorption peaks at the left and right are Al-O absorption peaks. Infrared spectroscopy analysis further confirms the successful preparation of Al-MOF.

[0032] (2) The structural characteristics of the coenzyme Q10-loaded nanocarrier Al-MOF@PEG were tested using a specific surface area and pore size analyzer. The comprehensive data of specific surface area and porosity calculated based on the model (e.g.) Figure 2The specific surface area and porosity of the samples were measured using a nitrogen adsorption-desorption (BET) instrument at a low temperature (77 K). Before the measurement, approximately 90 mg of the coenzyme Q10-loaded nanocarrier Al-MOF@PEG was weighed and degassed and activated at 120 °C for 12 h to remove residual solvent molecules from the pores. The specific surface area of ​​the coenzyme Q10-loaded nanocarrier Al-MOF@PEG was measured to be 997 m². 2 / g, total pore volume is 0.70m 3 / g, micropore volume is 0.43m 3 / g.

[0033] Example 3

[0034] Performance study of Al-MOF@PEG nanoparticles carrying coenzyme Q10:

[0035] (1) Measurement of nanoparticle size (e.g.) Figure 3 ): The lyophilized powder of Al-MOF@PEG nanoparticles carrying coenzyme Q10 obtained in Example 1 was used to prepare a 0.5 mg / mL nanoparticle solution. After sonication for 10 minutes, impurities were removed by filtration through a 0.45 μm aqueous filter membrane. Subsequently, the solution was placed in a Zeta-Size Nano-ZS 90 dynamic light scattering (DLS) instrument to determine the particle size distribution range and dispersity of the Al-MOF@PEG nanoparticle conjugate. Each group was tested three times.

[0036] (2) Surface morphology characteristics of nanoparticles: A dispersion of Al-MOF@PEG nanocarriers with a concentration of 1 mg / mL coenzyme Q10 was prepared. One drop was added to the surface of a copper mesh carbon film with a pore size of 200 mesh. After about 1 minute, the excess solution on the copper mesh was absorbed with filter paper. After the copper mesh was allowed to dry naturally, the morphology and size of the nanocarriers were observed using a transmission electron microscope (TEM).

[0037] Example 4

[0038] Determination of drug loading in the drug delivery nano-delivery system of this invention:

[0039] It is feasible to prepare an anhydrous ethanol solution of Al-MOF@PEG nanocarriers carrying coenzyme Q10, detect the absorbance at a wavelength of 275 nm using a UV absorbance detector, and measure and calculate the coenzyme Q10 content in the nanocarriers using the Vis-UV method.

[0040] Example 5

[0041] In vitro drug release performance study of Al-MOF@PEG nanoparticles carrying coenzyme Q10:

[0042] The release performance of Al-MOF@PEG nanoparticles carrying coenzyme Q10 in different release media was determined using the UV standard curve method. pH 5.3, pH 6.8, and pH 7.4 were selected as release media to simulate physiological conditions, the extracellular matrix, and the intracellular environment, respectively. The effect of pH on the Al-MOF@PEG system carrying coenzyme Q10 was investigated by measuring the cumulative release of coenzyme Q10 over a certain period. Results are shown below. Figure 4 .

[0043] Example 6

[0044] Coenzyme Q10 skin penetration test

[0045] Coenzyme Q10-loaded nanocarrier Al-MOF@PEG prepared in Example 1 was subjected to a skin permeation test. Preparation of isolated pig ear skin: After euthanizing adult pigs, the skin from the inner side of the auricle was immediately removed, separated from the cartilage, rinsed thoroughly with physiological saline, and the skin integrity was checked. The skin was then placed in physiological saline and stored at 4°C for later use. An in vitro transdermal diffusion experiment of pig ear skin was conducted using a TK-12A transdermal diffusion apparatus (Shanghai Kaikai Technology & Trade Co., Ltd.). The pig ear skin was placed on a Franz diffusion cell, with the stratum corneum facing the supply chamber and the dermis facing the receiving chamber. 3g of sample was accurately weighed into each supply chamber, and the upper opening of the supply chamber was sealed. The receiving chamber was filled with receiving solution. The entire apparatus was placed in a constant temperature water bath at 32±0.1°C, and the electromagnetic stirrer was turned on at a speed of 400 r / min. After the in vitro transdermal assay, ultrathin sections of pig ear skin were prepared using a LEICA 1900 cryostat (LEICA GmbH, Germany). The sections were placed in EP tubes, methanol was added, and the tubes were sealed. The tubes were sonicated three times for 30 min each time, centrifuged at 5000 rpm for 30 min, and filtered through a 0.45 μm filter. The filtrates were then analyzed by high-performance liquid chromatography (HPLC). All experiments were conducted in the dark. The results showed that the prepared Al-MOF@PEG nanocarrier solution carrying CoQ10 achieved a transdermal absorption rate of 25%–35% within 12 h, while the transdermal absorption rate of Al-MOF@PEG nanocarrier solution carrying CoQ10 dissolved directly in 5% TW-80 was less than 8% within 12 h. This indicates that the Al-MOF@PEG nanocarrier significantly improves the transdermal absorption rate of CoQ10, which is beneficial for transdermal drug delivery.

Claims

1. A preparation method of coenzyme Q10-loaded nano-organic metal framework material Al-MOF@PEG, characterized in that, Includes the following steps: (1) Preparation of Al-MOF: Nano-Al-MOF was prepared using acetic acid as a modifier. 3.62g AlCl3·6H2O and 0.83g 2,5-dihydroxyterephthalic acid were mixed into 355mL DMF, 60.00g acetic acid, 23.50mL of 95% ethanol aqueous solution and 23.50mL of water were added. The resulting mixed solution was placed in a 500mL autoclave and heated to 135℃ for 24 hours. After the reaction was completed, the mixed solution was cooled to room temperature. After centrifugation, the supernatant was discarded and DMF was added and centrifuged three times. The precipitate obtained from the last centrifugation was placed in CH2Cl2 for two days and replaced with fresh CH2Cl2 continuously to remove the unreacted raw materials adhering to the precipitate. After filtration, the nano-carrier Al-MOF was collected by vacuum drying. (2) Preparation of Al-MOF nanocarrier loaded with coenzyme Q10: 30 mg of Al-MOF nanocarrier was dispersed in pure water, and 5 mg of coenzyme Q10 was dissolved in DMF and then added dropwise to pure water and mixed. The mixture was sonicated for 10 minutes and stirred for 4 hours. Then, the mixture was centrifuged, the supernatant was removed, and the mixture was washed three times with pure water. The solid precipitate obtained by centrifugation was freeze-dried to obtain Al-MOF nanocarrier loaded with coenzyme Q10. (3) Preparation of Al-MOF@PEG nanocarrier carrying coenzyme Q10: 100 mg Al-MOF was ultrasonically dispersed in 15 mL CH2Cl2, and then 100 mg dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine were added. The mixture was stirred at 10 °C for 3 hours, and then 100 mg CH3O-PEG5k-NH2 was added and reacted at 4 °C for 17 hours. After removing the mixed solution, centrifugation was performed, the supernatant was discarded, and CH2Cl2 was added and centrifuged and washed three times. Finally, the product, Al-MOF@PEG nanoorganic metal framework material carrying coenzyme Q10, was collected by vacuum drying.

2. A method for preparing a coenzyme Q10-loaded metal-organic framework Al-MOF@PEG according to claim 1.

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