Coenzyme q10-loaded nano-organic metal framework material, and preparation method and application thereof

Abstract

The application discloses a coenzyme Q10-loaded nano organic metal framework material and a preparation method and application thereof, and relates to the field of biological macromolecular materials and nanotechnology. The preparation method comprises the following steps: (1) preparation of Al-MOF; (2) preparation of the coenzyme Q10-loaded nano carrier Al-MOF; and (3) preparation of the coenzyme Q10-loaded nano carrier Al-MOF@PEG. In the application, 2,5-dihydroxyterephthalic acid and aluminum (Al) ions are selected, Al-MOF nano carriers are synthesized through a solvothermal method, coenzyme Q10 is loaded into the interspace of the Al-MOF nano carriers, then mPEG5k-NH2, i.e. a kind of PEG with an amino group, is selected, and the carboxyl on the surface of the Al-MOF is activated by dicyclohexyl carbodiimide (DCC), and the amino at the end of the PEG is covalently modified to the surface of the nano carrier Al-MOF through “click chemistry” between the carboxyl and the amino. The nano carrier Al-MOF has a larger pore size, the drug loading capacity is increased, the water solubility is significantly increased after the introduction of the PEG, the circulation time of the nano carrier in a physiological environment is prolonged, and the bioavailability of the coenzyme Q10 is further improved.

Description

Technical Field

[0001] This invention relates to the field of biopolymer materials and nanotechnology, specifically to a coenzyme Q10-loaded nanoorganic metal framework material, its preparation method, and its application. 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. 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, thus effectively delaying aging, but also keeps cells healthy and produces an anti-fatigue effect. Furthermore, recent research indicates that coenzyme Q10 also has certain therapeutic effects in cancer prevention and treatment. It has also shown significant biological activity in chronic heart failure, cardiofacial syndrome, diabetes, cancer, autoimmune diseases, cataracts, asthma, periodontal disease, and thyroid disease.

[0003] Coenzyme Q10, a fat-soluble antioxidant, 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 improve 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, thus hindering the maximization of coenzyme Q10's efficacy.

[0004] Therefore, how to improve the stability of coenzyme Q10, prolong its effective action time, and promote effective penetration and targeted release are urgent problems to be solved in current applications. Summary of the Invention

[0005] This invention provides a coenzyme Q10-loaded nano-organic metal framework material, its preparation method, and its applications, aiming to solve the problems existing in the background art. The coenzyme Q10-loaded nano-organic metal framework material provided by this invention utilizes the larger pore size of the nanocarrier to increase drug loading capacity. Simultaneously, 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. It also incorporates nano-drug delivery technology, improving drug stability, delaying release, and altering drug distribution in vivo.

[0006] This invention synthesizes Al-MOF nanocarriers via a solvothermal method using 2,5-dihydroxyterephthalic acid and aluminum (Al) ions. Coenzyme Q10 is then loaded into the pores of the Al-MOF nanocarriers. Subsequently, mPEG5k-NH2, an amino-containing PEG, is selected and covalently modified onto the Al-MOF nanocarrier surface through a "click chemistry" process between the carboxyl group activated by dicyclohexylcarbodiimide (DCC) on the Al-MOF surface and the amino group at the end of the PEG. This process results in a larger pore size, increasing the drug loading capacity. Simultaneously, 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.

[0007] To achieve the above-mentioned technical objectives, the present invention mainly adopts the following technical solutions:

[0008] In a first aspect, the present invention discloses a method for preparing a nano-organic metal framework material loaded with coenzyme Q10, comprising the following steps:

[0009] (1) Preparation of Al-MOF

[0010] AlCl3·6H2O and 2,5-dihydroxyterephthalic acid were uniformly mixed in DMF, then acetic acid was added, followed by the dropwise addition of ethanol and pure water. The mixture was placed in a high-temperature and high-pressure reactor for reaction. After the reaction was completed, the reaction solution was cooled to room temperature, centrifuged and the supernatant was discarded. The solution was washed, and then CH2Cl2 was used to remove the residual chemical substances in the washing product. After filtration, the product was vacuum dried to obtain the nanocarrier Al-MOF.

[0011] (2) Preparation of Al-MOF nanocarriers carrying coenzyme Q10

[0012] The nanocarrier Al-MOF prepared in step (1) was dispersed in pure water. Coenzyme Q10 was dissolved in DMF and then added dropwise to pure water and mixed. After ultrasonic treatment, the mixture was stirred to obtain a mixture. The mixture was then centrifuged, the supernatant was discarded, washed, and freeze-dried to obtain the nanocarrier Al-MOF carrying coenzyme Q10.

[0013] (3) Preparation of Al-MOF@PEG nanocarriers carrying coenzyme Q10

[0014] The Al-MOF nanocarrier carrying coenzyme Q10 prepared in step (2) was ultrasonically dispersed in CH2Cl2, and then dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine were added. After stirring at 10°C for an appropriate time, CH3O-PEG5k-NH2 was added, and the reaction was continued at 4°C to obtain a reaction solution. The reaction solution was centrifuged, the supernatant was discarded, washed, and vacuum dried to obtain the Al-MOF@PEG nanocarrier carrying coenzyme Q10.

[0015] In a preferred embodiment of the present invention, in step (1), the mass ratio of AlCl3·6H2O to 2,5-dihydroxyterephthalic acid is (4-4.5):1.

[0016] In a preferred embodiment of the present invention, in step (1), the method for removing residual chemical substances from the washing product by CH2Cl2 is as follows: the product obtained after the last centrifugal washing is placed in CH2Cl2 for two days and continuously replaced with fresh CH2Cl2 to remove residual chemical substances.

[0017] In a preferred embodiment of the present invention, in step (1), the mass ratio of AlCl3·6H2O to acetic acid is 1:(16-20).

[0018] In a preferred embodiment of the present invention, in step (1), the volume ratio of DMF to ethanol and pure water is (12-17):1:1, and 1gAlCl3·6H2O is dissolved in 0.8-1ml DMF.

[0019] In a preferred embodiment of the present invention, in step (2), the mass ratio of the nanocarrier Al-MOF to coenzyme Q10 is (6-10):1.

[0020] In a preferred embodiment of the present invention, in step (3), the mass ratio of the coenzyme Q10-loaded nanocarrier Al-MOF, dicyclohexylcarbodiimide (DCC), 1,6-hexanediamine and CH3O-PEG5k-NH2 is 1:1:(0.8-0.9):(1-1.5).

[0021] In a second aspect, the present invention discloses an Al-MOF@PEG nanostructured metal framework material carrying coenzyme Q10, prepared by the preparation method described in the first aspect.

[0022] Thirdly, this invention discloses the application of Al-MOF@PEG, a nano-organic metal framework material carrying coenzyme Q10 as described in the second aspect, in the preparation of pharmaceuticals, cosmetics, or health products.

[0023] In a preferred embodiment of the present invention, the dosage form of the medicine, cosmetic or health product is a patch.

[0024] The coenzyme Q10-loaded nano-organic metal framework material provided by this invention not only has the characteristics of high porosity, diverse structure and controllable function, but also combines the strong drug loading capacity of inorganic nanocarriers with the high safety of organic nanocarriers.

[0025] This invention significantly increases the solubility of coenzyme Q10 in water by introducing a PEG structure, thereby further improving its bioavailability.

[0026] The coenzyme Q10-loaded nano-organic metal framework material provided by this invention has the advantages of nano-drug delivery technology, which can improve drug stability, delay release, and change the distribution of drugs in the body. Attached Figure Description

[0027] Figure 1 The infrared (FT-IR) spectrum of Al-MOF provided in the embodiments of the present invention;

[0028] Figure 2 Nitrogen adsorption-desorption spectra of Al-MOF@PEG provided in embodiments of the present invention;

[0029] Figure 3 The image shows the dynamic light scattering DLS particle size analysis of Al-MOF@PEG nanocarriers provided in this embodiment of the invention.

[0030] Figure 4 Release performance diagram of Al-MOF@PEG carrying coenzyme Q10 in different release media provided in the embodiments of the present invention. Detailed Implementation

[0031] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0032] Example 1

[0033] The preparation of Al-MOF@PEG loaded with coenzyme Q10 includes the following steps:

[0034] (1) Preparation of Al-MOF: Nano-Al-MOF was prepared by using acetic acid as a modulator.

[0035] 3.62 g of AlCl3·6H2O and 0.83 g of 2,5-dihydroxyterephthalic acid were mixed in 355 mL of LDM. 60.00 g of acetic acid, 23.50 mL of ethanol, and 23.50 mL of water were added. The resulting mixture was placed under a pressure of 1 x 10⁻⁶ m³ / s. 5 The reaction mixture was placed in a 500ml autoclave at approximately 135℃ and heated for 24 hours. After the reaction was complete, the reaction solution was allowed to cool to room temperature. The mixture was then removed, centrifuged, and the supernatant was discarded. DMF was added, and the mixture was washed three times by centrifugation. The precipitate obtained from the last centrifugation and washing was placed in CH2Cl2 for two days, continuously replaced with fresh CH2Cl2 to remove residual chemicals. After filtration, the Al-MOF nanocarrier was collected by vacuum drying.

[0036] (2) Preparation of Al-MOF nanocarriers carrying coenzyme Q10

[0037] Take 30 mg of the nanocarrier Al-MOF obtained in step (1) and disperse it in pure water. Then, dissolve 5 mg of coenzyme Q10 in DMF and add it dropwise to pure water and mix well. Sonicate for 10 minutes and stir the mixture for 4 hours. Then, centrifuge the mixture, remove the supernatant and wash it three times with pure water. Freeze-dry the solid precipitate obtained by the last centrifugation to obtain the coenzyme Q10-loaded nanocarrier Al-MOF.

[0038] (3) Preparation of Al-MOF@PEG nanocarriers carrying coenzyme Q10

[0039] 100 mg of the coenzyme Q10-loaded Al-MOF nanocarrier obtained in step (2) was ultrasonically dispersed in 15 mL of CH2Cl2, followed by the addition of 100 mg of dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine. The mixture was stirred at 10 °C for 3 hours. 100 mg of CH3O-PEG5k-NH2 was added, and the mixture was reacted at 4 °C for 17 hours. The mixture was removed, centrifuged, the supernatant was discarded, and the mixture was washed three times with CH2Cl2. Finally, the product was collected by vacuum drying, yielding the coenzyme Q10-loaded Al-MOF@PEG nanocarrier.

[0040] Example 2

[0041] Characterization of Al-MOF@PEG nanocarriers carrying coenzyme Q10

[0042] (1) The reaction product Al-MOF obtained in step (1) of Example 1 was characterized by Fourier transform infrared spectroscopy (FT-IR) as follows: The dried product and KBr were mixed and ground at a ratio of 1:100 using the KBr pelleting method, dried under an infrared lamp, and then pelleted. The pellets were then placed in an instrument for testing, using a Nexus infrared spectrometer at 4000-400 cm⁻¹. -1 Test relevant samples within the scope.

[0043] The results are as follows Figure 1 As shown, 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.

[0044] (2) The structural characteristics of the coenzyme Q10 nanocarrier Al-MOF@PEG were tested using a specific surface area and pore size analyzer as follows: The specific surface area and porosity of the sample were tested using a nitrogen adsorption-desorption (BET) analyzer at low temperature (77K). Before the measurement, approximately 90 mg of the sample was weighed and degassed and activated at 120 °C for 12 h to remove residual solvent molecules from the pores.

[0045] The results are as follows Figure 2 As shown, the combined data of specific surface area and porosity directly calculated by the computer of the testing instrument indicate that the specific surface area of ​​the coenzyme Q10-loaded nanocarrier Al-MOF@PEG is 997 m². 2 / g, total pore volume is 0.70m 3 / g, micropore volume is 0.43m 3 / g.

[0046] Example 3

[0047] Performance Study of Al-MOF@PEG Nanocarriers Loaded with Coenzyme Q10

[0048] (1) The particle size of the nanocarrier was determined by the following method: The freeze-dried powder of Al-MOF@PEG nanoparticles carrying coenzyme Q10 prepared in Example 1 was prepared into a solution with a concentration of 0.5 mg / mL nanoparticles. After sonication for 10 minutes, impurities were removed by a 0.45 μm aqueous filter membrane. Then, the solution was placed in a Zeta-Size Nano-ZS 90 dynamic light scattering instrument (DLS) to determine the particle size distribution range and dispersion of the Al-MOF@PEG conjugate carrying coenzyme Q10. Each group was tested three times.

[0049] The results are as follows Figure 3 As shown, by Figure 3 It can be seen that the average particle size of the Al-MOF@PEG conjugate carrying coenzyme Q10 is 240 nm and the dispersion is 0.201.

[0050] (2) The surface morphology of the nanocarrier was determined using the following method: A dispersion of Al-MOF@PEG nanocarrier 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, excess solution was absorbed from the copper mesh using filter paper. After the copper mesh dried naturally, the morphology and size of the nanocarrier were observed using a transmission electron microscope (TEM). It was found that the crystals were distributed in a strip-like pattern.

[0051] Example 4

[0052] Determination of drug loading capacity in the drug delivery nano-delivery system of the present invention

[0053] The sample was prepared into anhydrous ethanol solution, and the absorbance was detected at a wavelength of 275 nm using a UV absorbance detector. The coenzyme Q10 content in the nanocarrier was measured and calculated using the Vis-UV method. The drug loading content (DLC) was calculated according to the following formula and the result was 30.18%, indicating that the coenzyme Q10-loaded nanocarrier Al-MOF@PEG prepared in this invention has a good effect on loading coenzyme Q10.

[0054]

[0055] In the formula, W t For the initial amount of Coenzyme Q10, W s The value represents the residual coenzyme Q10 in the supernatant after drug loading, and W represents the mass of the coenzyme Q10-loaded nanocarrier.

[0056] Example 5

[0057] In vitro drug release performance study of Al-MOF@PEG nanocarrier carrying coenzyme Q10:

[0058] The release performance of coenzyme Q10-loaded nanoparticles in different release media was determined using the ultraviolet standard curve method. Release media at pH 5.3, pH 6.8, and pH 7.4 were selected to simulate physiological conditions, the extracellular matrix, and the intracellular environment, respectively. The effect of pH on the coenzyme Q10-loaded nanosystem was investigated by measuring the cumulative release of coenzyme Q10 over a certain period.

[0059] The results are as follows Figure 4 As shown, by Figure 4 It is known that the PEG-modified Al-MOF@PEG nanocarrier carrying coenzyme Q10 has the characteristics of delayed drug release, improved water solubility, and is more likely to release coenzyme Q10 in a weakly acidic skin environment.

[0060] Example 6

[0061] Coenzyme Q10 skin penetration test

[0062] Experimental group: The Al-MOF@PEG nanocarrier carrying coenzyme Q10 prepared in Example 1 was dissolved in 5% TW-80.

[0063] Control group: Al-MOF solution carrying coenzyme Q10 was dissolved in an equal volume of 5% TW-80 as the experimental group.

[0064] The coenzyme Q10-loaded solutions prepared in the experimental and control groups were subjected to a skin penetration test, as follows:

[0065] 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 its integrity was checked. The skin was then stored in physiological saline at 4℃ for later use. An in vitro transdermal diffusion experiment 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℃, and the electromagnetic stirrer was turned on at a speed of 400r / 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, the tubes were sealed, and the samples were sonicated three times for 30 min each time. After centrifugation at 5000 rpm for 30 min, the sections were filtered through a 0.45 μm filter membrane, and the filtrates were analyzed by high-performance liquid chromatography (HPLC). All experiments were performed under light-protected conditions.

[0066] The results showed that the Al-MOF@PEG solution carrying CoQ10 in the experimental group achieved a transdermal absorption rate of 25%–35% within 12 hours, while the Al-MOF solution carrying CoQ10 dissolved in 5% TW-80 used directly in the control group had a transdermal absorption rate of less than 8% within 12 hours. This indicates that Al-MOF@PEG carrying CoQ10 significantly improved the transdermal absorption rate of CoQ10 and effectively enhanced its water solubility, which is beneficial for transdermal drug delivery.

[0067] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. The application of Al-MOF@PEG nanostructured metal framework material loaded with coenzyme Q10 in the preparation of patch dosage forms of pharmaceuticals, cosmetics or health products, characterized in that, The preparation method of the coenzyme Q10-loaded nano-organic metal framework material includes the following steps: (1) Preparation of Al-MOF AlCl3·6H2O and 2,5-dihydroxyterephthalic acid were uniformly mixed in DMF, then acetic acid was added, followed by the dropwise addition of ethanol and pure water. The mixture was placed in a high-temperature and high-pressure reactor for reaction. After the reaction was completed, the reaction solution was cooled to room temperature, centrifuged and the supernatant was discarded. The solution was washed, and then CH2Cl2 was used to remove the residual chemical substances in the washing product. After filtration, the product was vacuum dried to obtain the nanocarrier Al-MOF. (2) Preparation of Al-MOF nanocarriers carrying coenzyme Q10 The nanocarrier Al-MOF prepared in step (1) was dispersed in pure water. Coenzyme Q10 was dissolved in DMF and then added dropwise to pure water and mixed. After ultrasonic treatment, the mixture was stirred to obtain a mixture. The mixture was then centrifuged, the supernatant was discarded, washed, and freeze-dried to obtain the nanocarrier Al-MOF carrying coenzyme Q10. (3) Preparation of Al-MOF@PEG nanocarriers carrying coenzyme Q10 The Al-MOF nanocarrier carrying coenzyme Q10 prepared in step (2) was ultrasonically dispersed in CH2Cl2, and then dicyclohexylcarbodiimide (DCC) and 1,6-hexanediamine were added. After stirring at 10°C for an appropriate time, CH3O-PEG5k-NH2 was added, and the reaction was continued at 4°C to obtain a reaction solution. The reaction solution was centrifuged, the supernatant was discarded, washed, and vacuum dried to obtain the Al-MOF@PEG nanocarrier carrying coenzyme Q10.

2. The application according to claim 1, characterized in that, In step (1), the mass ratio of AlCl3·6H2O to 2,5-dihydroxyterephthalic acid is (4-4.5):

1.

3. The application according to claim 1, characterized in that, In step (1), the method for removing residual chemicals from the washing product using CH2Cl2 is as follows: the product obtained after the last centrifugal washing is placed in CH2Cl2 for two days and continuously replaced with fresh CH2Cl2 to remove residual chemicals.

4. The application according to claim 1, characterized in that, In step (1), the mass ratio of AlCl3·6H2O to acetic acid is 1:(16-20).

5. The application according to claim 1, characterized in that, In step (1), the volume ratio of DMF to ethanol and pure water is (12-17):1:1, and 1gAlCl3·6H2O is dissolved in 0.8-1ml DMF.

6. The application according to claim 1, characterized in that, In step (2), the mass ratio of the nanocarrier Al-MOF to coenzyme Q10 is (6-10):

1.

7. The application according to claim 1, characterized in that, In step (3), the mass ratio of the coenzyme Q10-loaded nanocarrier Al-MOF, dicyclohexylcarbodiimide (DCC), 1,6-hexanediamine and CH3O-PEG5k-NH2 is 1:1:(0.8-0.9):(1-1.5).

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