Metal organic framework material for targeting bone marrow adipocytes as well as preparation method and application of metal organic framework material

Abstract

The invention discloses a metal organic framework material targeting bone marrow fat cells as well as a preparation method and application of the metal organic framework material, and belongs to the technical field of biomedical materials and tissue engineering. The material comprises: (1) a ZIF-8 core structure used for loading nucleic acid / small molecule drugs; (2) surface modification of lipid droplet targeting peptide: the lipid droplet targeting peptide is combined with a PEDF receptor to realize precise targeting; and (3) anti-miR-188 is loaded inside and is used for inhibiting adipogenic differentiation. The method at least has the following advantages: (1) a bone marrow adipocyte specific targeting MOF material is proposed for the first time, and bone metabolism intervention is converted from a mode of targeting osteoblast or macrophage to targeting adipocyte; (2) the MOF core can quickly respond to a marrow weak acid environment to realize accurate release; (3) the AdipoPep targeting peptide can obviously prolong the residence time of the bone marrow; and (4) adipogenic differentiation is significantly inhibited, osteogenesis microenvironment reconstruction is promoted, and an excellent bone mass recovery effect is shown in an osteoporosis model.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of new bone repair materials and functional nano delivery systems, and particularly relates to a metal organic framework (MOF) material targeting bone marrow adipocytes and a preparation method thereof and application thereof in osteoporosis treatment. BACKGROUND

[0002] Bone marrow adipocytes (BM-Ad) maintain a dynamic balance with bone formation under normal conditions, but under conditions such as osteoporosis, aging, and glucocorticoid application, the number of BM-Ad increases significantly, and the adipokines secreted by BM-Ad inhibit osteogenesis and promote osteoclasts, thereby leading to a decrease in bone mass. Targeted reduction or regulation of BM-Ad has become an important direction for preventing and treating osteoporosis.

[0003] Current intervention methods for bone marrow adipocytes mainly include: (1) systemic administration of miR-188 inhibitors (anti-miR-188), but they cannot be directed to the bone marrow cavity, resulting in large side effects; (2) limited research on adipocyte-targeting nanomaterials, and most of them are limited to white adipose tissue, with low recognition efficiency for bone marrow adipocytes; (3) existing MOF materials have the advantages of being able to load nucleic acid molecules and controllable degradation, but lack bone marrow adipocyte-specific targeting function.

[0004] Therefore, it is of great significance to develop a new MOF material that can precisely target BM-Ad, load nucleic acids, and controllably degrade in the bone marrow microenvironment. SUMMARY

[0005] The present application is based on the deficiencies in the prior art and provides a new MOF material that can precisely target BM-Ad, load nucleic acids, and controllably degrade in the bone marrow microenvironment.

[0006] To achieve the above technical purposes, the present application adopts the following technical solutions: One of the purposes of the present application is to provide a multifunctional metal organic framework material targeting bone marrow adipocytes, comprising: a ZIF-8 metal organic framework core formed by coordination of zinc ions and 2-methyl imidazole, the core encapsulating anti-miR-188 inside; and an AdipoPep peptide modified on the surface of the core through a silane coupling agent layer and an amide bond.

[0007] Further, the silane coupling agent layer is an amino-functional organic layer formed by bonding 3-aminopropyltriethoxysilane (APTES) on the surface of the ZIF-8 core through a hydrolysis condensation reaction.

[0008] Further, the sequence of the AdipoPep peptide is shown in SEQ ID NO. 1.

[0009] Further, the sequence of the anti-miR-188 is shown as SEQ ID NO. 2.

[0010] The second object of the present application is to provide a preparation method of the multifunctional metal organic framework material, comprising the following steps: S1: dissolving anti-miR-188 in RNase-free water to prepare a nucleic acid solution, and dissolving Zn(NO3)2·6H2O and 2-mIM in methanol respectively to prepare Zn solution and 2-mIM solution; S2: mixing the nucleic acid solution with the Zn solution, then quickly adding it into the 2-mIM solution, stirring, centrifuging and washing to obtain ZIF-8@anti-miR-188; S3: dispersing ZIF-8@anti-miR-188 in anhydrous ethanol, then adding APTES dropwise for reaction, and finally centrifuging and washing to obtain NH2-ZIF-8@anti-miR-188; S4: dissolving AdipoPep in MES, adding EDC and NHS for activation, suspending NH2-ZIF-8@anti-miR-188 in PBS, then adding the activated AdipoPep solution into the NH2-ZIF-8@anti-miR-188 suspension for reaction, and finally centrifuging and washing to obtain the multifunctional metal organic framework material, that is, ZIF-8@AdipoPep@anti-miR-188.

[0011] Further, the amount of anti-miR-188 in S1 is 18–22 μg, the amount of RNase-free water is 1.8–2.2 mL, the amount of Zn(NO3)2·6H2O is 0.14–0.16 g, the amount of 2-mIM is 0.75–0.85 g, and the total amount of methanol is 18–22 mL.

[0012] Further, the stirring condition in S2 is magnetic stirring at room temperature for 20–30 min.

[0013] Further, the amount of ZIF-8@anti-miR-188 in S3 is 8–12 mg, the amount of anhydrous ethanol is 18–22 mL, the amount of APTES is 45–55 μL, and the reaction condition is magnetic stirring at room temperature for 1.8–2.2 h.

[0014] Further, the S4 AdipoPep dosage is 0.8-1.2 mg, the MES dosage is 0.8-1.2 mL, the EDC dosage is 8-12 mg, the NHS dosage is 5-7 mg, the activation condition is room temperature oscillation activation for 15-20 min, the NH2-ZIF-8@anti-miR-188 suspension concentration is 4.5-5.5 mg / mL, and the reaction condition is room temperature light shaking reaction for 3-5 h or 3-5℃ overnight.

[0015] A third object of the present application is to provide the use of the multifunctional metal organic framework material and / or the preparation method in the preparation of a product for preventing and treating diseases caused by an increase in bone marrow adipocytes, including osteoporosis, and the product includes a drug.

[0016] Compared with the prior art, the present application has at least the following beneficial effects: The material in the present application includes: ① ZIF-8 core structure: for loading nucleic acids or small molecule drugs; ② surface modification lipid droplet targeting peptide (AdipoPep: LPHSSGHA): can bind to the PEDF receptor on the BM-Ad film, realizing precise targeting; ③ internal loading of miR-188 inhibitors (anti-miR-188): for inhibiting adipogenic differentiation. The present application has the following innovations: (1) the first bone marrow adipocyte specific targeting MOF material is proposed, realizing the mode change of bone metabolism intervention from “targeting osteoblasts or macrophages to targeting adipocytes”; (2) the MOF core can quickly respond to the weak acid environment of bone marrow, realizing precise release; (3) the AdipoPep targeting peptide significantly improves the bone marrow residence time; (4) significantly inhibits adipogenic differentiation, promotes osteogenic microenvironment reconstruction, and shows superior bone mass recovery effect in the osteoporosis model. The present application has good clinical transformation prospects. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 The electron microscope images of ZIF-8, ZIF-8@AdipoPep, and ZIF-8@AdipoPep@anti-miR-188 in Example 2 of the present application.

[0018] Figure 2 The targeting effect of ZIF-8, Dio-ZIF-8, and Dio-ZIF-8@AdipoPep on bone marrow fat in animals in Example 2 of the present application.

[0019] Figure 3 The drug release curve of ZIF-8@anti-miR-188 and ZIF-8@AdipoPep@anti-miR-188 in Example 2 of the present application.

[0020] Figure 4Femoral bone mineral density and marrow fat changes exhibited in each treatment of Example 2 of the present application. DETAILED DESCRIPTION

[0021] The following examples are intended to illustrate the present application but not to limit the scope of the present application. Modifications or substitutions of the method, steps or conditions of the present application, which do not depart from the spirit and essence of the present application, are intended to fall within the scope of the present application. The reagents, products and instruments used in the following examples are commercially available, and the methods used in the examples are consistent with the commonly used methods unless otherwise specified.

[0022] The technical solutions of the present application are further described in detail below in combination with examples.

[0023] Example 1 1. ZIF-8 synthesis procedure 1.1 Reagents (for preparation of about 20 mg ZIF-8) (1) Zn(NO3)2·6H2O: 0.148 g (0.5 mmol).

[0024] (2) 2-methylimidazole (2-mIM): 0.820 g (10 mmol).

[0025] (3) Methanol (or methanol / deionized water mixture): total 20 mL.

[0026] 1.2 Procedure (1) Prepare Zn solution: dissolve Zn(NO3)2·6H2O (0.148 g) in 10 mL methanol, and magnetically stir until clear.

[0027] (2) Prepare 2-mIM solution: dissolve 2-mIM (0.820 g) in another 10 mL methanol, and shake until clear.

[0028] (3) Pour the Zn solution quickly (instantaneously or within 30 s) into the 2-mIM solution, and magnetically stir at room temperature for 20-30 min (suspension or turbidity appears, indicating the formation of ZIF-8).

[0029] (4) Stop stirring for 10 min, centrifuge at 12,000 rpm for 10 min, and discard the supernatant; wash the precipitate with a mixture of methanol / deionized water twice (10 mL each time) to remove unreacted ligands and salts.

[0030] (5) Resuspend the washed precipitate in a small amount of methanol (or RNase-free water, depending on the subsequent use), and freeze-dry or dry at 40°C under vacuum to obtain ZIF-8 powder.

[0031] 2. AdipoPep-modified ZIF-8 (ZIF-8@AdipoPep) synthesis steps 2.1 Reagents (based on 10 mg ZIF-8) (1) ZIF-8 powder: 10 mg.

[0032] (2) Ethanol (absolute): 20 mL.

[0033] (3) APTES (3-aminopropyltriethoxysilane): 50 μL (about 0.22 mmol).

[0034] (4) AdipoPep (carboxyl-terminated, sequence example SEQ ID NO. 1: LPHSSGHA-COOH, Hangzhou Dangang Biotechnology Co., Ltd., 2025-2-6): 1 mg (about 0.6-1.0 μmol, depending on the molecular weight of the peptide).

[0035] (5) EDC: 10 mg (about 0.052 mmol).

[0036] (6) NHS: 6 mg (about 0.052 mmol).

[0037] (7) MES buffer (50 mM, pH 6.0) or PBS (pH 7.4).

[0038] (8) Deionized water / ethanol.

[0039] 2.2 Steps 2.2.1 Surface amination (APTES silanization) (1) Disperse 10 mg ZIF-8 in 20 mL absolute ethanol (ultrasonic assistance for 5 min to assist dispersion).

[0040] (2) Dropwise add APTES (50 μL), room temperature magnetic stirring for 2 h (or 40°C light reflux for 1 h can be accelerated).

[0041] (3) Centrifuge (10,000 rpm, 5 min) to collect the precipitate, wash twice with ethanol and once with deionized water.

[0042] (4) Obtain aminated ZIF-8 (denoted as NH2-ZIF-8), store at 4°C in RNase-free PBS (if RNA loading is required, subsequent operations should be carried out under RNase-free conditions).

[0043] 2.2.2 EDC / NHS activation and coupling of peptides (1) Dissolve AdipoPep (1 mg) in 1 mL MES (50 mM, pH 6.0).

[0044] (2) Add EDC (10 mg) and NHS (6 mg) to the peptide solution, and activate for 15-20 min at room temperature (active NHS ester is generated).

[0045] (3) Meanwhile, suspend NH2-ZIF-8 in 2 mL PBS (pH 7.4) (concentration of about 5 mg / mL).

[0046] (4) Slowly add the activated AdipoPep solution to the NH2-ZIF-8 suspension, and react for 4 h at room temperature or overnight at 4°C with gentle shaking.

[0047] (5) After the reaction, centrifuge (10,000 rpm, 5 min), and wash with PBS for 3 times to remove free peptides, EDC / NHS byproducts.

[0048] (6) Collect the obtained ZIF-8@AdipoPe.

[0049] 3. Preparation of AdipoPep-modified ZIF-8 loaded nucleic acid (ZIF-8@AdipoPep@anti-miR-188) 3.1 Reagents (preparation of 10 mg level drug loading) (1) Zn(NO3)2·6H2O: 0.148 g (0.5 mmol).

[0050] (2) 2-mIM: 0.820 g (10 mmol).

[0051] (3) anti-miR-188 RNase-free solution (anti-miR-188 sequence SEQ ID NO. 2: UGGUACUGGUACAGAGGUG, Beijing Bomeisi Technology Development Co., Ltd., MZIP188-5p-PA-1), final addition amount of 20 μg (adjustable as needed). Note: Because in the NIPO ST.26 version, "T" represents uracil in the RNA sequence and thymine in the DNA sequence, "U" in the above sequence is converted to "T" when making.

[0052] (4) Methanol / RNase-free water: 20 mL (methanol / water mixture can be used for protection of nucleic acids, or water system can be directly adjusted).

[0053] (5) Subsequent APTES, AdipoPep, EDC, NHS are the same as above.

[0054] 3.2 Steps 3.2.1 anti-miR-188 co-encapsulation of ZIF-8 (nucleation co-encapsulation method) (1) Dissolve anti-miR-188 (20 μg) in 2 mL RNase-free water (if necessary, a small amount of RNase-free Tris buffer can be used).

[0055] (2) Dissolve Zn(NO3)2(0.148 g) in 10 mL methanol; slowly add the nucleic acid solution to the Zn solution and mix gently by inverting (be careful of shearing).

[0056] (3) Quickly add the Zn 2+ / nucleic acid mixture to 10 mL 2-mIM solution and stir magnetically at room temperature for 20-30 min. ZIF-8 forms and simultaneously encapsulates the nucleic acid.

[0057] (4) Centrifuge (12,000 rpm, 10 min) and wash 3 times with RNase-free PBS or deionized water to remove unencapsulated nucleic acid (save the eluate for EE determination).

[0058] (5) At this point, ZIF-8@anti-miR-188 (nucleic acid-loaded core) is obtained.

[0059] 3.2.2 Surface amination and AdipoPep conjugation Follow the procedure in the previous section "ZIF-8@AdipoPep" for APTES amination and EDC / NHS conjugation of AdipoPep to ZIF-8@anti-miR-188 to obtain ZIF-8@anti-miR-188.

[0060] 4. Dio encapsulated ZIF-8 (DiO-ZIF-8) synthesis procedure (co-encapsulation DiO) 4.1 Reagents (1) Zn(NO3)2·6H2O: 0.148 g (0.5 mmol).

[0061] (2) 2-mIM: 0.820 g (10 mmol).

[0062] (3) Methanol: 20 mL (in two 10 mL portions).

[0063] (4) DiO solution: DiO 0.2 mg in 200 μL methanol.

[0064] 4.2 Procedure 4.2.1 Preparation of solution A (Zn solution): Dissolve Zn(NO3)2-6H2O (0.148 g) in 10 mL anhydrous methanol, magnetically stir until clear (room temperature).

[0065] 4.2.2 Preparation of solution B (2-mIM + DiO): Dissolve 2-mIM (0.820 g) in another 10 mL anhydrous methanol. Add DiO (0.2 mg, dissolved in 200 μL methanol) into the 2-mIM solution, mix gently by inverting (avoid vigorous shaking to prevent dye aggregation), to disperse DiO evenly in the ligand solution.

[0066] 4.2.3 Nucleation and co-encapsulation: Pour solution A (Zn 2+ ) into solution B (2-mIM + DiO) quickly within 30 s, magnetically stir at room temperature for 20-30 min. The system will change from clear to turbid, indicating the formation of ZIF-8 crystals and the encapsulation / adsorption of DiO into the pores or hydrophobic surface of the crystals during the formation process.

[0067] 4.2.4 Precipitate collection and washing: After the reaction is completed, centrifuge at 12,000 rpm for 10 min. Discard the supernatant, resuspend with 10 mL methanol, sonicate for 1-2 min, and centrifuge to recover; repeat the washing 2 times to remove unbound free DiO and residual reagents.

[0068] 4.2.5 Drying and storage: Freeze-dry (or vacuum dry at 40 °C) the washed precipitate after resuspension in a small amount of methanol to obtain a yellow / light green powder, Dio-ZIF-8. Store in the dark at 4 °C in a dry place.

[0069] 5. Synthesis steps of Dio-encapsulated AdipoPep-modified ZIF-8 (DiO-ZIF-8@AdipoPep) 5.1 Reagents and amounts (based on 10 mg DiO-ZIF-8 as the starting amount) (1) DiO-ZIF-8: 10 mg (dry powder or suspension).

[0070] (2) Anhydrous ethanol: 20 mL (used in portions).

[0071] (3) APTES: 50 μL (about 0.22 mmol).

[0072] (4) AdipoPep (LPHSSGHA-COOH): 1.0 mg (or increase as needed).

[0073] (5) EDC: 10 mg (about 0.052 mmol).

[0074] (6) NHS: 6 mg (about 0.052 mmol).

[0075] (7) MES buffer (50 mM, pH 6.0) or PBS (pH 7.4).

[0076] 5.2 Step 5.2.1 Surface amination (APTES silanization) (1) Disperse 10 mg DiO-ZIF-8 in 20 mL anhydrous ethanol (assist dispersion by ultrasonication for 3-5 min).

[0077] (2) Slowly add APTES (50 μL) at room temperature, magnetically stir for 2 h (or gently reflux at 40 °C for 1 h).

[0078] (3) After reaction, centrifuge (10,000 rpm, 5 min), discard supernatant, wash with ethanol twice and deionized water once to remove unreacted APTES.

[0079] (4) Collect the aminated particles (NH2-DiO-ZIF-8), gently resuspend in PBS (pH 7.4, concentration about 5 mg / mL).

[0080] 5.2.2 EDC / NHS activation of AdipoPep (1) Dissolve AdipoPep (1 mg) in 1 mL MES (50 mM, pH 6.0).

[0081] (2) Add EDC (10 mg) and NHS (6 mg), shake at room temperature for 15-20 min to generate active NHS ester (do not leave for long time to prevent hydrolysis).

[0082] 5.2.3 Coupling reaction (1) Slowly add the activated AdipoPep solution to the NH2-DiO-ZIF-8 suspension (e.g. 2 mL, about 5 mg / mL) at room temperature, gently shake for 4 h or overnight at 4 °C.

[0083] (2) After coupling, centrifuge (10,000 rpm, 5 min), wash with PBS for 3 times to remove free peptide and EDC / NHS byproducts.

[0084] (3) Finally suspend in PBS (or lyophilize) to obtain DiO-ZIF-8@AdipoPep.

[0085] Example 2 1. Material characterization 1.1 TEM shows core-shell structure (1) Experimental procedure ① 10 μL of nanoparticle suspension (1 mg / mL) was dropped on carbon film copper grid.

[0086] ② The grid was left at room temperature for 2 min, and the excess liquid was absorbed by filter paper.

[0087] ③ 2% phosphotungstic acid was added for 1 min.

[0088] ④ After air-drying, the grid was observed under transmission electron microscope (JEOL JEM-2100, voltage 200 kV).

[0089] (2) Results Figure 1 The TEM images showed that the ZIF-8 core had a regular dodecahedron structure with a size of about 500-1000 nm; after modification with the peptide (ZIF-8@AdipoPep), a uniform "gray halo" with a thickness of about 10-15 nm was formed on the surface, confirming that the nanoparticles had a typical core-shell structure; the particle ZIF-8@AdipoPep@anti-miR-188 had a clear edge profile and no obvious aggregation, and the structure was stable.

[0090] 1.2 Application of fluorescence labeling for in vivo tracking in mice (1) Experimental procedure Twelve hours after injection (200 μL) of ZIF-8, Dio-ZIF-8 and Dio-ZIF-8@AdipoPep through the tail vein, the mice (Hubei Beinete Biological Technology Co., Ltd., 002356 (Jackson)) were sacrificed and the heart, liver, spleen, lung, kidney and femur were collected. Then the Vivo FX PRO imaging system was used to detect the distribution of ZIF-8, Dio-ZIF-8 and Dio-ZIF-8@AdipoPep in vivo.

[0091] (2) Experimental results Figure 2 It was shown that 24 hours after injection, the fluorescence intensity of the Dio-ZIF-8@AdipoPep group in the bone region was significantly higher than that of the Dio-ZIF-8 group, while no fluorescence signal was observed in the sham control group injected with ZIF-8. This experiment proved that ZIF-8 modified with AdipoPep could target the bone tissue and avoid damage to liver and kidney function.

[0092] 1.3 Drug release effect (1) Experimental procedure ZIF-8@anti-miR-188 and ZIF-8@AdipoPep@anti-miR-188 were placed in vials containing PBS solution (10.0 mL, 10 mg ZIF per vial) and incubated at room temperature with a constant reciprocating shaking rate of about 100 rpm. Subsequently, the vials were placed in dialysis bags with a molecular weight of 2KD, immersed in a beaker containing PBS solution (1 L), and placed in a constant temperature (37°C) water bath shaker (shaking rate 100 rpm) for reaction. The time was started from 0 hour, and the release medium solution (5 mL) was taken out at the set time points (2, 4, 6, 8, 12, 24, 36 and 48 hours) and supplemented with an equal amount of blank PBS. After the release medium solution taken out was evaporated and dried, acetonitrile (0.5 mL) was added to dissolve it thoroughly, and the supernatant (20 μL) was taken after centrifugation at 15000 rpm, which was detected by liquid chromatography-mass spectrometry to calculate the drug release rate.

[0093] (2) Experimental results Figure 3 The release test results of anti-miR-188 showed that ZIF-8@anti-miR-188 and ZIF-8@AdipoPep@anti-miR-188 could both achieve sustained release of anti-miR-188 within 48 hours, and the introduction of AdipoPep did not have a significant effect on the drug release ability of the material.

[0094] 2. Application of mouse osteoporosis model (1) Experimental animals and feeding conditions Eight-week-old female C57BL / 6J mice (body weight 18-20 g, Hubei Bainter Biotechnology Co., Ltd., 002356 (Jackson)) were randomly divided into three groups: Sham group (sham operation control), OVX group (osteoporosis model group), and OVX + nanoparticle treatment group (ZIF-8@AdipoPep@anti-miR-188). All animals were raised in SPF level animal room, with constant temperature (22±2°C), 12 h light-dark cycle, and free access to food and water.

[0095] (2) OVX operation modeling steps After the mice were anesthetized (1% sodium pentobarbital, 50 mg / kg, intraperitoneal injection), the mice were fixed supine on the operating table, and the skin was prepared and disinfected. Two lateral lumbar incisions were made on the dorsal side of the mouse, the skin and muscle layer were incised, and the bilateral ovaries were exposed; the ovarian blood vessels were ligated with 4# silk and the ovaries were removed; the muscle and skin were sutured in layers; gentamicin was injected to prevent infection.

[0096] (3) Drug administration method The mice in the treatment group were given by tail vein injection: dosage: 10 mg / kg; injection volume: 200 μL (prepared with PBS); administration frequency: twice a week; administration cycle: 4 weeks continuously.

[0097] The control group was injected with the same volume of PBS, ZIF-8, and ZIF-8@AdipoPep.

[0098] (4) Detection method After the treatment, the mice were sacrificed, and the femurs were collected for micro-CT analysis (n=3 per group). The femurs were scanned using a SkyScan 1176 system (parameters: 2400 frames, 5 frames / field, 37 kV, 121 μA), and then fixed with paraformaldehyde overnight. The bone density, bone volume fraction, trabecular number, and bone surface volume ratio were quantified by CTan software. After micro-CT analysis, the femurs were decalcified in 10% ethylenediaminetetraacetic acid solution for 15 days, paraffin-embedded, and sectioned to a thickness of 5–7 μm, and the specimens were stained with hematoxylin-eosin.

[0099] (5) Experimental results The micro-CT analysis results showed that the ZIF-8@AdipoPep@anti-miR-188 group improved the bone density, bone volume fraction, bone surface volume ratio, and trabecular number to varying degrees compared with the ovariectomy group (Fig. 4a, b). Figure 4 The hematoxylin-eosin staining results showed that the fat cell content of the ZIF-8@AdipoPep@anti-miR-188 group was significantly reduced (Fig. 4c, d). Figure 4 c, d). This experiment proved that the final product in the application can indeed improve the bone density of the femur.

[0100] The above-described embodiments are only descriptions of the preferred modes of the application and do not limit the scope of the application. Without departing from the design spirit of the application, various modifications and improvements to the technical solutions of the application made by those of ordinary skill in the art should fall within the protection scope of the claims of the application.

Claims

1. A multifunctional metal-organic framework material targeting bone marrow adipocytes, characterized in that, It comprises: a ZIF-8 metal organic framework core formed by coordination of zinc ions and 2-methyl imidazole, the core encapsulating anti-miR-188 inside; and an AdipoPep peptide modified on the surface of the core through a silane coupling agent layer and an amide bond.

2. The multifunctional metal-organic framework material of claim 1, wherein, The silane coupling agent layer is an amino-functional organic layer formed by hydrolytic condensation reaction of 3-aminopropyl triethoxysilane (APTES) bonded on the surface of the ZIF-8 core.

3. The multifunctional metal-organic framework material of claim 2, wherein, The sequence of the AdipoPep peptide is shown in SEQ ID NO.

1.

4. The multifunctional metal-organic framework material of claim 3, wherein, The sequence of the anti-miR-188 is shown in SEQ ID NO.

2.

5. The method of producing a multifunctional metal-organic framework material according to any one of claims 1 to 4, characterized in that, It comprises the following steps: S1: dissolving anti-miR-188 in RNase-free water to prepare a nucleic acid solution, and dissolving Zn(NO3)2·6H2O and 2-methyl imidazole (2-mIM) in methanol respectively to prepare Zn solution and 2-mIM solution; S2: adding the nucleic acid solution into the Zn solution, mixing well, then quickly adding into the 2-mIM solution, stirring, centrifuging and washing to obtain ZIF-8@anti-miR-188; S3: dispersing ZIF-8@anti-miR-188 in anhydrous ethanol first, then adding APTES dropwise for reaction, and finally centrifuging and washing to obtain NH2-ZIF-8@anti-miR-188; S4: dissolving AdipoPep in MES first, adding EDC and NHS for activation, meanwhile suspending NH2-ZIF-8@anti-miR-188 in PBS, then adding the activated AdipoPep solution into the NH2-ZIF-8@anti-miR-188 suspension for reaction, and finally centrifuging and washing to obtain the multifunctional metal organic framework material, that is, ZIF-8@AdipoPep@anti-miR-188.

6. The preparation method according to claim 5, characterized in that, The amount of anti-miR-188 in S1 is 18–22 μg, the amount of RNase-free water is 1.8–2.2 mL, the amount of Zn(NO3)2·6H2O is 0.14–0.16 g, the amount of 2-mIM is 0.75–0.85 g, and the total amount of methanol is 18–22 mL.

7. The preparation method according to claim 6, characterized in that, The stirring condition in S2 is magnetic stirring at room temperature for 20–30 min.

8. The preparation method according to claim 7, characterized in that, The amount of ZIF-8@anti-miR-188 in S3 is 8–12 mg, the amount of anhydrous ethanol is 18–22 mL, the amount of APTES is 45–55 μL, and the reaction condition is magnetic stirring at room temperature for 1.8–2.2 h.

9. The preparation method according to claim 8, characterized in that, The amount of AdipoPep in S4 is 0.8–1.2 mg, the amount of MES is 0.8–1.2 mL, the amount of EDC is 8–12 mg, the amount of NHS is 5–7 mg, the activation condition is oscillation at room temperature for 15–20 min, the concentration of NH2-ZIF-8@anti-miR-188 suspension is 4.5–5.5 mg / mL, and the reaction condition is light shaking at room temperature for 3–5 h or overnight at 3–5℃.

10. Use of the multifunctional metal-organic framework material according to any one of claims 1 to 4 and / or the production method according to any one of claims 5 to 9 for the production of a product for the prevention and treatment of diseases caused by an increase in bone marrow adipocytes, characterized in that, The disease includes osteoporosis, and the product includes a drug.

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