MMP-9 driven decomposed cobalt-manganese-based nano-cluster as well as preparation method and application of MMP-9 driven decomposed cobalt-manganese-based nano-cluster

Abstract

The preparation method comprises the following steps: dispersing a cobalt salt and a manganese salt in ethylene glycol, stirring until the cobalt salt and the manganese salt are dissolved, carrying out heating reflux, centrifuging, washing and drying, dispersing the obtained cobalt-manganese compound in deionized water, carrying out ultrasonic crushing to obtain a cobalt-manganese-based nano precursor, and carrying out heat treatment to obtain the MMP-9 driven decomposed cobalt-manganese-based nano cluster. And dispersing the cobalt-manganese oxide nanocrystals in a gelatin solution for polymerization to obtain the cobalt-manganese-based nano-cluster. The cobalt-manganese-based nano-cluster has MMP-9 driven decomposition performance, T1-T2 double-weighted enhanced magnetic resonance performance and near-infrared light excited thermal conversion performance, the method is simple, safe, environmentally friendly, low in cost and easy to operate, and the cobalt-manganese-based nano-cluster can serve as a magnetic resonance nano contrast agent to be applied to the field of cancer diagnosis and treatment and has wide application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of multifunctional nanomaterials, and in particular relates to a cobalt-manganese-based nanocluster driven to decompose by MMP-9, and a preparation method and application thereof. Background Art

[0002] Bionanomaterials have attracted increasing attention in the diagnosis and treatment of tumors due to their enhanced tumor tissue targeting and multifunctionality, especially PEGylated liposomal doxorubicin. clinical application and further determine the clinical transformation value of bio-nanomaterials.

[0003] However, targeted drug delivery is a very complex process. Nanodrugs with a particle size of 100-200nm use enhanced permeability and retention (EPR) properties to enter the tumor through the endothelial pores on the blood vessels and are retained in the tumor tissue due to impaired lymphatic filtration. After entering the tumor tissue, the increased interstitial fluid pressure and dense extracellular matrix hinder the diffusion of larger nanodrugs in deep tumors. Nanosystems with transformable particle sizes are expected to maintain a large size in the systemic circulation and transform into a smaller size at the tumor site, thereby achieving efficient targeting and deep penetration of tumor tissues at the same time. Summary of the invention

[0004] Based on this, the main purpose of the present invention is to provide a method for preparing cobalt-manganese-based nanoclusters driven by matrix metalloproteinase MMP-9 (highly expressed in tumor tissues), the required raw materials are easily available, low in price, and the synthesis method is mild.

[0005] Another object of the present invention is to provide a cobalt-manganese-based nanocluster driven to decompose by MMP-9, which is prepared by the preparation method of the cobalt-manganese-based nanocluster, has MMP-9 driven decomposition performance, and has T1-T2 dual-weighted magnetic resonance imaging and photothermal conversion performance.

[0006] Another object of the present invention is to provide the use of the MMP-9 driven decomposition cobalt manganese based nanoclusters in the preparation of T1-T2 dual weighted enhanced magnetic resonance nano contrast agents.

[0007] To achieve the above object, the present invention adopts the following technical solution:

[0008] The present invention provides a method for preparing a cobalt-manganese-based nanocluster driven by MMP-9 decomposition, comprising the following steps:

[0009] (1) dispersing cobalt salt and manganese salt in ethylene glycol, stirring until dissolved, heating to reflux, centrifuging, washing, and drying to obtain a brown cobalt-manganese compound;

[0010] (2) dispersing the cobalt-manganese compound described in step (1) in deionized water and subjecting it to ultrasonic grinding to obtain a cobalt-manganese-based nano-precursor with controllable particle size;

[0011] (3) heat treating the cobalt-manganese-based nano precursor described in step (2) at 150 to 250° C. for 8.0 to 24 hours to obtain cobalt-manganese oxide nanocrystals;

[0012] (4) preparing a clear and transparent gelatin solution, heating it to 60° C., adding the cobalt manganese oxide nanocrystals described in step (3), stirring, centrifuging, and washing to obtain MMP-9 driven decomposition cobalt manganese based nanoclusters.

[0013] Preferably, in step (1), the cobalt salt and manganese salt are selected from one or more of nitrates, acetates or chlorides.

[0014] Preferably, in step (2), the ultrasonic power is 50 to 300 W, and the ultrasonic time is 1 to 60 min. The particle size of the cobalt-manganese-based nano-precursor is regulated according to the ultrasonic power and the ultrasonic time.

[0015] Preferably, in step (3), the atmosphere in which the cobalt-manganese-based nano precursor is heat-treated is air, nitrogen or argon.

[0016] Preferably, in step (4), the mass ratio of cobalt manganese oxide nanocrystals to gelatin is 1:10-300.

[0017] The present invention also provides a cobalt-manganese-based nanocluster decomposed by MMP-9, which is prepared by the preparation method of the cobalt-manganese-based nanocluster decomposed by MMP-9.

[0018] Preferably, the cobalt-manganese-based nanoclusters have MMP-9 driven decomposition performance, and decompose into small-sized nanoparticles with a particle size of 50 to 500 nm in the presence of MMP-9.

[0019] Preferably, the cobalt-manganese-based nanoclusters have T1-T2 dual-weighted magnetic resonance imaging and photothermal conversion performance.

[0020] Preferably, the dispersion of the cobalt-manganese-based nanoclusters in deionized water exhibits concentration-dependent longitudinal and transverse relaxation rates.

[0021] Preferably, the cobalt-manganese-based nanoclusters convert light energy into heat energy under the excitation of 808nm near-infrared light.

[0022] The present invention also provides the use of the MMP-9 driven decomposition cobalt manganese based nano clusters in the preparation of T1-T2 dual weighted enhanced magnetic resonance nano contrast agents.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention proposes a method for preparing a cobalt-manganese-based nanocluster driven to decompose by MMP-9, the required raw materials are easily available and inexpensive, the synthesis method is mild and environmentally friendly, the obtained cobalt-manganese-based nanocluster has MMP-9 driven decomposition performance, and has T1-T2 dual-weighted MR performance and photothermal conversion performance, can realize size changes driven by MMP-9, and can be used as a magnetic resonance nanocontrast agent in the diagnosis and treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : This is the XRD pattern of the cobalt manganese oxide nanocrystals prepared in Example 1.

[0025] Figure 2 TEM images of the cobalt manganese oxide nanocrystals and cobalt manganese-based nanoclusters prepared in Example 1 before and after the action of MMP-9.

[0026] Figure 3 MR properties of the cobalt-manganese-based nanoclusters prepared in Example 1.

[0027] Figure 4 The photothermal conversion performance of the cobalt-manganese-based nanocluster dispersion prepared in Example 1. DETAILED DESCRIPTION

[0028] In order to more fully understand and demonstrate the technical solutions, purposes and advantages of the present invention, the technical effects produced by the present invention are further described in detail and completely in conjunction with the accompanying drawings and specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all. It should be pointed out that for ordinary technicians in this field, other embodiments obtained without departing from the concept of the present invention all belong to the protection scope of the present invention.

[0029] Example 1

[0030] (1) Dispersing 2 mmol of cobalt salt and 1 mmol of manganese salt in ethylene glycol, stirring until dissolved, heating under reflux for 2 h, centrifuging, washing, and drying to obtain a brown cobalt-manganese compound;

[0031] (2) dispersing the cobalt-manganese compound precursor in step (1) in deionized water, and subjecting it to ultrasonic grinding, with an ultrasonic power of 200 W and an ultrasonic time of 40 min, to obtain a cobalt-manganese-based nano-precursor with controllable particle size;

[0032] (3) heat treating the cobalt-manganese-based nano precursor in step (2) at 220° C. for 8.0 h to obtain cobalt-manganese oxide nanocrystals;

[0033] (4) preparing a clear and transparent gelatin solution (20 mg / mL), heating it to 60° C., adding the cobalt manganese oxide nanoparticles (0.2 mg / mL) in step (3), stirring for a period of time, centrifuging and washing, and obtaining the cobalt manganese-based nanoclusters decomposed by MMP-9.

[0034] The XRD pattern of the cobalt manganese oxide nanocrystals prepared in Example 1 is as follows: Figure 1 As shown, the product is cubic cobalt manganese oxide, which corresponds to the standard card (JCPDS 32-0297).

[0035] TEM images of cobalt manganese oxide nanocrystals and cobalt manganese-based nanoclusters are shown in Figure 2. Figure 2 As shown in A and 2B, the cobalt manganese oxide nanocrystals before gelatin coating are particles with a particle size of 18.6 nm, and the size of the cobalt manganese-based nanoclusters after gelatin coating is about 309 nm.

[0036] Example 2

[0037] (1) Dispersing 2 mmol of cobalt salt and 1 mmol of manganese salt in ethylene glycol, stirring until dissolved, heating under reflux for 2 h, centrifuging, washing, and drying to obtain a brown cobalt-manganese compound;

[0038] (2) dispersing the cobalt-manganese compound precursor in step (1) in deionized water and subjecting it to ultrasonic grinding, with an ultrasonic power of 200 W and an ultrasonic time of 60 min, to obtain a cobalt-manganese-based nano-precursor with controllable particle size;

[0039] (3) heat treating the cobalt-manganese-based nano precursor in step (2) at 220° C. for 8.0 h to obtain cobalt-manganese oxide nanocrystals;

[0040] (4) preparing a clear and transparent gelatin solution (20 mg / mL), heating it to 60° C., adding the cobalt manganese oxide nanoparticles (0.2 mg / mL) in step (3), stirring for a period of time, centrifuging and washing, and obtaining the cobalt manganese-based nanoclusters decomposed by MMP-9.

[0041] Example 3

[0042] (1) Dispersing 2 mmol of cobalt salt and 1 mmol of manganese salt in ethylene glycol, stirring until dissolved, heating under reflux for 2 h, centrifuging, washing, and drying to obtain a brown cobalt-manganese compound;

[0043] (2) dispersing the cobalt-manganese compound precursor in step (1) in deionized water and subjecting it to ultrasonic grinding, with an ultrasonic power of 200 W and an ultrasonic time of 40 min, to obtain a cobalt-manganese-based nano-precursor with controllable particle size;

[0044] (3) heat treating the cobalt-manganese-based nano precursor in step (2) at 220° C. for 8.0 h to obtain cobalt-manganese oxide nanocrystals;

[0045] (4) preparing a clear and transparent gelatin solution (20 mg / mL), heating it to 60° C., adding the cobalt manganese oxide nanoparticles (1 mg / mL) in step (3), stirring for a period of time, centrifuging and washing, and obtaining MMP-9 driven decomposition of cobalt manganese based nanoclusters.

[0046] Example 4

[0047] The MMP-9 driven decomposition performance of the cobalt-manganese based nanoclusters obtained in Example 1 was measured.

[0048] The cobalt-manganese-based nanoclusters were dispersed in a solution containing MMP-9, centrifuged and washed after one day, and redispersed in deionized water. The changes in the sample morphology were observed using a transmission electron microscope.

[0049] TEM images of cobalt-manganese-based nanoclusters after the action of MMP-9 are shown in Figure 2 C shows that after the action of MMP-9, the cobalt-manganese-based nanoclusters are decomposed into small particles.

[0050] Example 5

[0051] The in vitro MR properties of the cobalt-manganese-based nanoclusters obtained in Example 1 were measured.

[0052] First, a dispersion of cobalt-manganese-based nanoclusters with gradient concentrations was prepared, and then the relaxation time of cobalt-manganese-based nanoclusters with different concentrations was tested on a magnetic resonance scanner. The results are as follows: Figure 3 As shown, T1, T2 weighted images and relaxation time pseudo-color images of dispersions of different concentrations; the longitudinal relaxation rate and transverse relaxation rate data showed that the cobalt-manganese-based nanoclusters exhibited T1-T2 weighted dual-enhanced MR imaging effect, with the longitudinal relaxation rate being 0.688mM-1s-1 and the transverse relaxation rate being 22.88mM-1s-1.

[0053] Example 6

[0054] The photothermal response performance of the cobalt-manganese-based nanoclusters obtained in Example 1 was measured.

[0055] A dispersion of cobalt-manganese-based nanocluster with gradient concentration was prepared and irradiated with 808 nm near-infrared light (power density: 1.00 W / cm 2 ), real-time recording of temperature changes, data such as Figure 4 As shown, after 10 minutes of illumination, the temperature rise of the solution without cobalt-manganese-based nanoclusters was <5.0°C. The temperature rise gradually increased with the increase of nanocluster concentration, and reached 40°C when the nanocluster concentration reached 1 mg / mL.

[0056] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing cobalt-manganese-based nanoclusters driven by MMP-9 decomposition, characterized in that: The following steps are involved: (1) dispersing a cobalt salt and a manganese salt in ethylene glycol, stirring until dissolved, heating to reflux, centrifuging, washing, and drying to obtain a cobalt-manganese compound; (2) dispersing the cobalt-manganese compound described in step (1) in deionized water and subjecting it to ultrasonic grinding to obtain a cobalt-manganese-based nano-precursor with controllable particle size; (3) heat treating the cobalt-manganese-based nano precursor described in step (2) at 150 to 250° C. for 8.0 to 24 hours to obtain cobalt-manganese oxide nanocrystals; (4) heating the gelatin solution to 50-70° C., adding the cobalt manganese oxide nanocrystals described in step (3), stirring, centrifuging, and washing to obtain cobalt manganese-based nanoclusters decomposed by MMP-9.

2. The method for preparing the cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 1, characterized in that: In step (1), the cobalt salt and manganese salt are selected from one or more of nitrates, acetates or chlorides.

3. The method for preparing the cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 1, characterized in that: In step (2), the ultrasonic power is 50 to 300 W, and the ultrasonic time is 1 to 60 min. The particle size of the cobalt-manganese-based nano precursor is regulated according to the ultrasonic power and the ultrasonic time.

4. The method for preparing the cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 1, characterized in that: In step (3), the atmosphere for heat treatment of the cobalt-manganese-based nano precursor is air, nitrogen or argon.

5. The method for preparing the cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 1, characterized in that: In step (4), the mass ratio of cobalt manganese oxide nanocrystals to gelatin is 1:10-300.

6. A cobalt-manganese-based nanocluster driven by MMP-9 decomposition, characterized in that: The method is used to prepare the cobalt-manganese-based nanoclusters driven to decompose by MMP-9 according to any one of claims 1 to 5.

7. The cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 6, characterized in that: It has the performance of being decomposed by MMP-9, and is decomposed into small-sized nanoparticles with a particle size of 50 to 500 nm in the presence of MMP-9.

8. The cobalt-manganese-based nanoclusters driven by MMP-9 decomposition according to claim 6, characterized in that: The cobalt-manganese-based nanocluster has T1-T2 dual-weighted magnetic resonance imaging and photothermal conversion performance.

9. The cobalt-manganese-based nanoclusters driven to decompose by MMP-9 according to claim 8, characterized in that: The dispersion of the cobalt-manganese-based nanoclusters in deionized water exhibits concentration-dependent longitudinal and transverse relaxation rates; and / or converts light energy into heat energy under 808 nm near-infrared light excitation.

10. Use of the cobalt-manganese-based nanoclusters decomposed by MMP-9 driven by any one of claims 1 to 5 in the preparation of T1-T2 dual-weighted enhanced magnetic resonance nanocontrast agents.

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