Copper-based metal organic framework nano material containing tanshinol as well as preparation method and application of copper-based metal organic framework nano material

Abstract

The invention relates to the technical field of nano medicines, in particular to a tanshinol-containing copper-based metal organic framework nano material and a preparation method and application thereof.According to the prepared tanshinol-containing copper-based metal organic framework nano material, tanshinol is dissolved in a dimethylimidazole aqueous solution, copper ions are rapidly added into the solution, and the tanshinol-containing copper-based metal organic framework nano material is obtained after the tanshinol is dissolved in the dimethylimidazole aqueous solution; the tanshinol-loaded copper-based metal organic framework nano-material is prepared from the copper-based metal organic framework nano-material, the tanshinol-loaded copper-based metal organic framework nano-material has high drug loading rate and acid-sensitive drug release performance, and the defects that a small-molecule drug tanshinol is poor in stability in an articular cavity and is easily removed by synovial fluid can be overcome. Compared with a natural enzyme, the nano-enzyme formed by combining tanshinol and copper-based MOF has the active oxygen scavenging capacity, and the treatment effect on osteoarthritis is jointly improved.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomedicine, and in particular to a copper-based metal organic framework nanomaterial containing danshensu, and a preparation method and application thereof. Background Art

[0002] Osteoarthritis (OA) is a common joint degenerative disease caused by aging, obesity, strain, trauma and congenital joint abnormalities. Currently, an estimated 250 million people worldwide are affected by this disease, which is clinically manifested as progressive joint pain, swelling, stiffness, tenderness, and deformity, seriously affecting the quality of life of patients. OA is characterized by elevated levels of multiple reactive oxygen species (ROS), such as hydroxyl radicals (·OH), hydrogen peroxide (H2O2) and superoxide anions (·O 2- ) leads to its oxidative microenvironment and pathological changes. Overexpression of ROS in inflamed joints leads to degradation of the extracellular cartilage matrix and synovitis. Excessive ROS can be clearly detected in the joint cavity of OA patients, even up to 1mM. Therefore, ROS is considered to be a key marker of OA, and clearing ROS can be used as an effective strategy for OA treatment. In addition, the proliferation of synoviocytes aggravates synovial hypoxia, which destroys the mitochondrial metabolism of synovial macrophages, increases the production of ROS and enhances the inflammatory response. Therefore, the key to effective OA treatment lies in eliminating various types of ROS and alleviating the hypoxic microenvironment.

[0003] Current studies have reported that reactive oxygen scavengers, such as melanin, vitamins, and natural polyphenols show great potential in scavenging ROS and further reducing the severity of OA. Natural antioxidant enzymes, such as superoxide dismutase (SOD), are part of the body's antioxidant system. Studies have shown that direct injection of superoxide dismutase has been shown to be a viable strategy for the treatment of OA. However, natural SOD is easily inactivated and rapidly metabolized in the human body, and natural enzymes cannot remove multiple types of ROS simultaneously. Salvianic acid A (DSS) is an essential water-soluble active ingredient in Danshen and is widely used in the clinical treatment of cardiovascular and cerebrovascular diseases. DSS has multiple pharmacological activities, including antioxidant, anti-inflammatory, antiplatelet, and angiogenic effects. However, the therapeutic application of DSS is severely limited by its poor bioavailability and rapid elimination from the organism, limiting its further application in the clinical treatment of OA. Summary of the invention

[0004] In view of the deficiencies of the above-mentioned prior art, the object of the present invention is to provide a copper-based metal organic framework nanomaterial containing danshensu and a preparation method and application thereof. The object of the present invention is to prolong the retention time of the small molecule drug danshensu in the joint cavity, respond to the acidic microenvironment to quickly release the drug, combine with copper ions to form nanozymes to eliminate reactive oxygen, and improve the therapeutic efficacy of osteoarthritis.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0007] Danshensu is dissolved in a 2-methylimidazole aqueous solution to obtain a mixed solution.

[0008] Soluble copper salt is added into the mixed solution for reaction, and danshensu is loaded on the copper-based metal organic framework during the reaction between 2-methylimidazole and copper ions, and a copper-based metal organic framework nanomaterial containing danshensu is obtained through a one-step reaction.

[0009] The copper-based metal organic framework nanomaterial containing danshensu prepared by the present invention comprises the following steps: dissolving danshensu in a dimethyl imidazole aqueous solution to obtain a mixed solution, adding a soluble copper salt to the mixed solution, and loading danshensu on the copper-based metal organic framework during the reaction of 2-methyl imidazole and copper ions, and forming a copper-based metal organic framework nanomaterial loaded with danshensu through a one-step reaction. The copper-based metal organic framework nanomaterial loaded with danshensu small molecule drug can prolong the retention time of the small molecule drug danshensu in the joint cavity, and the small molecule drug is rapidly released under the acidic microenvironment of arthritis, and is combined with copper ions to form nanoenzymes to remove active oxygen, thereby greatly improving the utilization rate of the drug and the therapeutic effect. In addition, the effective combination of copper metal ions and danshensu small molecule drugs shields the interaction between phenolic hydroxyl groups, resulting in more phenolic hydroxyl groups being exposed, and further promoting the antioxidant effect.

[0010] In a preferred embodiment of the present invention, the mass ratio of danshensu to 2-methylimidazole is 1:1-82.

[0011] In a preferred embodiment of the present invention, the mass ratio of copper ions to 2-methylimidazole in the soluble copper salt is 1:0.5-10.

[0012] In a preferred embodiment of the present invention, the mass concentration of the 2-methylimidazole aqueous solution is 4.1 mg / mL to 82 mg / mL.

[0013] In a preferred embodiment of the present invention, the soluble copper salt is one or more of copper chloride, copper nitrate or copper sulfate and hydrates thereof.

[0014] In a preferred embodiment of the present invention, the reaction temperature is room temperature and the reaction time is 20 min to 60 min.

[0015] Another object of the present invention is to provide a copper-based metal organic framework nanomaterial containing danshensu prepared by any of the preparation methods described above.

[0016] In a preferred embodiment of the present invention, the copper-based metal organic framework nanomaterial containing tanshinone is spherical in shape.

[0017] The third object of the present invention is to provide a use of the copper-based metal organic framework material containing danshensu as described in any of the above items in the preparation of a drug for treating osteoarthritis.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. The copper-based metal organic framework nanomaterial containing danshensu prepared by the present invention is that danshensu is dissolved in a dimethylimidazole aqueous solution to obtain a mixed solution, a soluble copper salt is added to the mixed solution, danshensu is loaded on the copper-based metal organic framework during the reaction of 2-methylimidazole and copper ions, and a copper-based metal organic framework nanomaterial loaded with danshensu is formed by a one-step reaction. The copper-based metal organic framework nanomaterial loaded with danshensu small molecule drug can prolong the retention time of the small molecule drug danshensu in the joint cavity, and the small molecule drug is rapidly released under the acidic microenvironment of arthritis, and the nanoenzyme formed by copper ions is combined to remove active oxygen, which greatly improves the drug utilization rate and therapeutic effect. In addition, the effective combination of copper metal ions and danshensu small molecule drugs shields the interaction between phenolic hydroxyl groups, resulting in more phenolic hydroxyl exposure, further promoting the antioxidant effect.

[0020] 2. The copper-based metal organic framework nanomaterial containing danshensu prepared by the present invention has a high drug loading rate and acid-sensitive drug release performance, which can avoid the shortcomings of small molecule drugs such as danshensu, which have poor stability in the joint cavity and are easily cleared by synovial fluid. Compared with natural enzymes, the nanozymes formed by danshensu combined with copper-based metal organic frameworks have multiple active oxygen scavenging capabilities, which together improve the therapeutic effect of osteoarthritis.

[0021] 3. The present invention can release drugs on demand, respond to the acidic microenvironment of osteoarthritis and quickly release drugs, achieving good anti-inflammatory effects. The copper-based metal organic framework nanomaterial containing danshensu constructed by the present invention can not only effectively remove reactive oxygen species, but also regulate the immune microenvironment of osteoarthritis to exert anti-inflammatory effects, thereby effectively treating osteoarthritis.

[0022] 4. The present invention loads the natural polyphenol drug danshensu into the metal-organic framework nanomaterial to form metal nanoenzymes to remove active oxygen, thereby achieving good osteoarthritis treatment ability. The preparation method of the copper-based metal-organic framework nanomaterial containing danshensu provided by the present invention is simple and rapid, has an ultra-high drug loading capacity of 100%, good antioxidant, anti-inflammatory and antibacterial capabilities, can achieve faster drug release in an acidic environment, and combined with copper ions to exert a good osteoarthritis treatment effect, and has broad application prospects in the field of osteoarthritis treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a schematic diagram of the copper-based metal organic framework nanomaterial containing danshensu prepared by the present invention, wherein 1 is 2-methylimidazole, 2 is danshensu, 3 is copper ion, and 4 is the copper-based metal organic framework nanomaterial containing danshensu.

[0024] Figure 2 This is a morphology diagram of the copper-based metal organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention.

[0025] Figure 3 This is the change in the zeta potential of the copper-based metal organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention.

[0026] Figure 4 These are ultraviolet-visible absorption spectra of the copper-based metal-organic framework nanomaterial containing danshensu, danshensu and the copper-based metal-organic framework nanomaterial prepared in Example 1 of the present invention.

[0027] Figure 5 This is an active oxygen scavenging experiment of the copper-based metal organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0028] The following is a detailed description of the technical solutions in the embodiments of the present invention in combination with the preferred embodiments and the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] It should be noted that all professional terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention. Unless otherwise specifically stated, various raw materials, reagents, instruments and equipment used in the following embodiments of the present invention can be purchased from the market or prepared by existing methods.

[0030] Compared with natural enzymes, nanozymes have multiple antioxidant activities and provide comprehensive antioxidant protection for the treatment of inflammatory diseases, which has attracted the attention of researchers. Recent studies have shown that stable metal-organic framework (MOF) nanomaterials composed of natural herbal derivatives and metal ions can mimic the coordination structure of natural metalloenzymes, with metals as active centers, greatly improving the utilization rate and therapeutic effect of metalloenzymes. With the introduction of active sites, the electron transfer interaction between drugs and metals endows this new agent with superior catalytic activity. Compared with small molecule drugs, MOF nanozymes loaded with small molecule drugs can prolong the retention time of drugs in the body. In addition, small molecule drugs are rapidly released in the acidic microenvironment of arthritis, greatly improving the utilization rate and therapeutic effect of drugs. More importantly, the effective combination of metal ions and small molecule drugs shields the interaction between phenolic hydroxyl groups, resulting in more phenolic hydroxyl exposure, further promoting antioxidant effects.

[0031] Among various metal elements, copper ions have attracted much attention due to their key role in various enzymatic reactions. Increasing evidence shows that copper-based nanoparticles help to enhance electron transfer reactions, thereby shielding the activity of ROS, achieving ROS scavenging function, and ultimately improving the treatment of skin, kidney and liver damage. Importantly, copper ions can also form stable MOFs through bridging ligands as connection points, exerting their synergistic enhancement of antioxidant and anti-inflammatory effects. In summary, copper-based MOF nanomaterials containing tanshinone may be a nanozyme with good reactive oxygen scavenging ability, providing new ideas for the prevention and treatment of various inflammatory diseases.

[0032] Example 1

[0033] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0034] (1) Dissolve 10 mg of danshensu in 10 mL of a 41 mg / mL 2-methylimidazole aqueous solution to obtain a mixed solution.

[0035] (2) 10 mL of 67 mg / mL copper chloride dihydrate was added to the mixed solution, stirred at room temperature for 1 h, centrifuged, and washed to obtain a copper-based metal organic framework nanomaterial containing tanshinone, denoted as SAA@MOF. The formation process is as follows: Figure 1 As shown, 1 is 2-methylimidazole, 2 is danshensu, 3 is copper ion, and 4 is a copper-based metal organic framework nanomaterial containing danshensu. Danshensu is mixed with dimethylimidazole, and then copper ions are added thereto. Danshensu is loaded on the copper-based metal organic framework during the reaction of 2-methylimidazole and copper ions, and the copper-based metal organic framework nanomaterial loaded with danshensu is formed through a one-step reaction.

[0036] Example 2

[0037] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0038] (1) Dissolve 10 mg of danshensu in 10 mL of 8.2 mg / mL 2-methylimidazole aqueous solution to obtain a mixed solution.

[0039] (2) adding 10 mL of 13.4 mg / mL copper chloride dihydrate to the mixed solution, stirring at room temperature for 1 h, centrifuging, and washing to obtain a copper-based metal organic framework nanomaterial containing tanshinone.

[0040] Example 3

[0041] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0042] (1) Dissolve 40 mg of danshensu in 20 mL of a 16.4 mg / mL 2-methylimidazole aqueous solution to obtain a mixed solution.

[0043] (2) Add 20 mL of 26.8 mg / mL copper chloride dihydrate to the mixed solution, stir at room temperature for 1 hour, centrifuge, and wash to obtain a copper-based metal organic framework nanomaterial containing tanshinone.

[0044] Example 4

[0045] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0046] (1) Danshensu was dissolved in a 4.1 mg / mL 2-methylimidazole aqueous solution, wherein the mass ratio of Danshensu to 2-methylimidazole was 1:1, to obtain a mixed solution.

[0047] (2) adding 67 mg / mL of cupric chloride dihydrate to the mixed solution, wherein the mass ratio of copper ions in the cupric chloride dihydrate to 2-methylimidazole is 1:0.5, stirring at room temperature for 20 minutes, centrifuging, and washing to obtain a copper-based metal organic framework nanomaterial containing tanshinone.

[0048] Example 5

[0049] A method for preparing a copper-based metal organic framework nanomaterial containing danshensu comprises the following steps:

[0050] (1) Danshensu was dissolved in 10 mL of 2-methylimidazole aqueous solution, wherein the mass ratio of Danshensu to 2-methylimidazole was 1:82, to obtain a mixed solution.

[0051] (2) adding 67 mg / mL of cupric chloride dihydrate to the mixed solution, wherein the mass ratio of copper ions to 2-methylimidazole in the cupric chloride dihydrate is 1:10, stirring at room temperature for 40 minutes, centrifuging, and washing to obtain a copper-based metal organic framework nanomaterial containing tanshinone.

[0052] Results Analysis

[0053] In order to prove the successful construction of the prepared copper-based metal organic framework nanomaterials containing tanshinone, the morphology of the nanomaterials was observed using a transmission electron microscope. Figure 2 This is a morphology picture of the copper-based metal-organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention. It can be seen that the copper-based metal-organic framework material containing danshensu prepared in the present invention is spherical and relatively uniform in size.

[0054] The Zeta potential of nanoparticles was detected using a multi-angle particle size and high-sensitivity Zeta potential analyzer. Figure 3 The Zeta potential change of the copper-based metal-organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention shows that compared with the positive potential of the copper-based metal-organic framework nanomaterial, after loading danshensu, the Zeta potential of the obtained SAA@MOF becomes negative, indicating that danshensu is successfully loaded. Figure 4 The UV-visible absorption spectrum of the copper-based metal-organic framework nanomaterial containing danshensu prepared in Example 1 of the present invention was analyzed using a UV-visible photometer. Compared with Cu-MOF, SAA@MOF showed an absorption peak similar to that of danshensu, indicating that danshensu was successfully loaded into Cu-MOF.

[0055] Figure 5 In the active oxygen scavenging experiment of the copper-based metal organic framework nanomaterial containing tanshinone prepared in Example 1 of the present invention, the active oxygen detection kit was used to detect the scavenging ability of tanshinone and SAA@MOF on intracellular active oxygen. The results showed that compared with the tanshinone treatment group, SAA@MOF had obvious active oxygen scavenging ability.

[0056] The preparation methods of Examples 2 to 5 of the present invention are the same as those of Example 1, with the only difference being that, when experimental conditions permit, the amount of raw materials and the reaction time are changed, but this does not affect the performance of the obtained copper-based metal organic framework nanomaterial containing tanshinone. Therefore, the nanomaterials prepared in Examples 2 to 5 have similar performance to that of Example 1, and can all prolong the retention time of small molecule drugs in the body, thereby improving drug utilization and therapeutic effects.

[0057] In summary, the copper-based metal organic framework nanomaterial containing danshensu prepared by the present invention dissolves danshensu in a dimethylimidazole aqueous solution, and copper ions are quickly added to the above solution to form a copper-based metal organic framework nanomaterial loaded with danshensu, and the MOF nanozyme loaded with danshensu small molecule drugs can prolong the retention time of the drug in the body. In addition, the small molecule drug is rapidly released under the acidic microenvironment of arthritis, which greatly improves the drug utilization rate and therapeutic effect. More importantly, the effective combination of copper metal ions and danshensu small molecule drugs shields the interaction between phenolic hydroxyl groups, resulting in more phenolic hydroxyl exposure, further promoting the antioxidant effect.

[0058] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the attached claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0059] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing a copper-based metal organic framework nanomaterial containing danshensu, characterized in that: The following steps are involved: dissolving danshensu in a 2-methylimidazole aqueous solution to obtain a mixed solution; Soluble copper salt is added into the mixed solution for reaction to obtain copper-based metal organic framework nanomaterial containing danshensu.

2. The method for preparing the copper-based metal organic framework nanomaterial containing danshensu according to claim 1, characterized in that: The mass ratio of danshensu to 2-methylimidazole is 1:1-82.

3. The method for preparing the copper-based metal organic framework nanomaterial containing danshensu according to claim 1, characterized in that: The mass ratio of copper ions to 2-methylimidazole in the soluble copper salt is 1:0.5-10.

4. The method for preparing the copper-based metal organic framework nanomaterial containing danshensu according to claim 1, characterized in that: The mass concentration of the 2-methylimidazole aqueous solution is 4.1 mg / mL to 82 mg / mL.

5. The method for preparing the copper-based metal organic framework nanomaterial containing danshensu according to claim 1, characterized in that: The soluble copper salt is one or more of copper chloride, copper nitrate or copper sulfate and hydrates thereof.

6. The method for preparing the copper-based metal organic framework nanomaterial containing danshensu according to claim 1, characterized in that: The reaction temperature is room temperature, and the reaction time is 20 min to 60 min.

7. A copper-based metal organic framework nanomaterial containing danshensu prepared by the preparation method according to any one of claims 1 to 6.

8. The copper-based metal organic framework nanomaterial containing danshensu according to claim 7, characterized in that: The copper-based metal organic framework nanomaterial containing danshensu has a spherical shape.

9. Use of the copper-based metal organic framework nanomaterial containing danshensu according to claim 7 in the preparation of a drug for treating osteoarthritis.

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