Preparation method and application of self-assembled bortezomib nanodrug

Abstract

The application discloses a preparation method and application of self-assembled bortezomib nanodrugs, and the preparation method comprises the following steps: dissolving bortezomib and a targeting peptide cRGD in an organic solvent, dissolving ZnCl2 in ultrapure water, then mixing the two, magnetically heating and stirring, obtaining a nanodrug suspension, removing supernatant by centrifugation, and then freeze-drying to obtain bortezomib nanodrugs. The preparation method is a carrier-free nanodrug delivery system, has good biosafety, and improves the targeted effect of the nanodrug on tumor cells.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a self-assembled bortezomib nanomedicine, its preparation method, and its application. Background Technology

[0002] Bortezomib is a synthetic borate dipeptide compound and the first proteasome inhibitor clinically approved for the treatment of multiple myeloma. It primarily works by inhibiting the NF-κB signaling pathway in tumor cells, thereby downregulating the expression of genes targeting anti-apoptotic targets and ultimately inducing tumor cell death. However, bortezomib has several drawbacks in clinical treatment, such as causing peripheral neuropathy, thrombocytopenia, neutropenia, nausea, diarrhea, and fatigue. Furthermore, it suffers from drug resistance, poor stability, and limited efficacy against solid tumors, restricting its further clinical application.

[0003] Nanomedicine delivery systems represent the application of nanotechnology in the pharmaceutical field. They enable the packaging and delivery of drugs to their sites of action via nanoscale carriers, exhibiting highly efficient drug delivery. While carrier-based nanomedicine delivery utilizes physical encapsulation to deliver drugs, it still faces numerous scientific challenges, such as low drug loading capacity, complex fabrication processes, and potential carrier toxicity. Furthermore, the fabrication processes of some carrier-based nanomedicines are quite complex, hindering their clinical translation. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preparing and applying self-assembled bortezomib nanomedicines, which is a carrier-free nanomedicine delivery system with good biosafety and improved targeting effect of nanomedicines on tumor cells.

[0005] This invention employs the following technical solution: a method for preparing self-assembled bortezomib nanomedicine, the preparation method comprising:

[0006] Bortezomib and the targeting peptide cRGD were dissolved in an organic solvent, and ZnCl2 was dissolved in ultrapure water. The two were then mixed and magnetically heated and stirred to obtain a nanodrug suspension. The supernatant was removed by centrifugation, and then lyophilized to obtain bortezomib nanodrug.

[0007] Furthermore, the aforementioned bortezomib and Zn 2+ The molar ratio of substances is 3:1, 1:1, or 1:3.

[0008] Furthermore, the organic solvent is DMSO, and the ratio of the organic solvent to ultrapure water is 1:1, 1:2, or 1:4.

[0009] Furthermore, the mass ratio of the targeting peptide cRGD to BTZ is 1:1, 1:2, or 1:10.

[0010] Furthermore, the conditions for magnetic heating and stirring are as follows: reaction temperature 37°C, stirrer speed 1100 rpm.

[0011] This invention also discloses the application of the self-assembled bortezomib nanomedicine prepared by the above-mentioned method in the preparation of drugs for treating subcutaneous tumors and bone tumors.

[0012] The beneficial effects of this invention are: (1) Bortezomib nanomedicine is synthesized using a polypeptide self-assembly method, which is a carrier-free nanomedicine delivery system. It is simple to prepare, has high drug loading efficiency, high biosafety, and the nanomedicine can also target tumor accumulation. (2) The introduction of cRGD further improves the targeting effect of the nanomedicine on tumor cells. (3) Compared with monomeric drugs, bortezomib nanomedicine has a stronger inhibitory effect on tumor cells and lower toxic side effects on normal cells. (4) In in vitro experiments, bortezomib nanomedicine has a stronger inhibitory effect on subcutaneous tumors and bone tumors in mice, and no obvious organ damage, proving that the nanomedicine has good tumor treatment ability and biocompatibility. (5) Subcutaneous tumor experiments have shown that bortezomib nanomedicine has a certain therapeutic effect on solid tumors, providing support for the clinical application of bortezomib in the treatment of solid tumors. Attached Figure Description

[0013] Figure 1 For two types of cells with BTZ and Zn 2+ Cell viability graphs after 48 hours of incubation with BTZNDs synthesized in different amounts of different substances at different concentrations.

[0014] Figure 2 The graph shows the cell viability of two cell types after incubating BTZNDs synthesized under different pH conditions at different concentrations for 48 hours.

[0015] Figure 3 The cell viability graph shows the cell viability of two cell types after incubating with BTZNDs at different concentrations for 48 hours.

[0016] Figure 4 For BTZ and Zn 2+ The killing effect of BTZNDs synthesized under a molar ratio of 1:3 and BTZNDs-cRGD synthesized under different mass ratios of BTZ and cRGD on MDA-MB-231 cells is shown in the figure.

[0017] Figure 5 a and b are high-resolution transmission electron microscopy (TEM) images and HAADF images of bortezomib nanodrug BTZNDs-cRGD, respectively, along with their corresponding EDS elemental mapping images.

[0018] Figure 6The surface charge of bortezomib nanomedicines BTZNDs and BTZNDs-cRGD.

[0019] Figure 7 In the figure, a and b are the XPS full spectrum and XPS spectrum of bortezomib nanodrug BTZNDs-cRGD and B1s, respectively.

[0020] Figure 8 The in vitro drug release curves are for bortezomib nanomedicines BTZNDs and BTZNDs-cRGD.

[0021] Figure 9 The cell survival rates of the monomeric drug bortezomib, the bortezomib nanodrug BTZNDs, and BTZNDs-cRGD against tumor cells MDA-MB-231 and normal cells L929 were measured.

[0022] Figure 10 Hemolysis analysis of BTZNDs-cRGD at different concentrations.

[0023] Figure 11 The study compared the therapeutic effects of bortezomib (a single drug), bortezomib nanomedicine, and a control group on subcutaneous tumors in mice.

[0024] Figure 12 In the figure, a and b represent the changes in tumor volume and mouse weight during the treatment of subcutaneous tumors in mice, respectively, for the monomeric drug bortezomib, the bortezomib nanomedicine, and the control group.

[0025] Figure 13 Micro-CT images of the forelimb bones of mice with bone tumors, including the monomeric drug bortezomib, bortezomib nanomedicine, and the control group, as well as 3D modeling images of some cancellous bone.

[0026] Figure 14 Micro-CT images of the hind limb bones of mice with bone tumors, showing the effects of the monomeric drug bortezomib, bortezomib nanomedicine, and the control group. Detailed Implementation

[0027] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0028] This invention discloses a method for preparing self-assembled bortezomib nanomedicines, the method comprising:

[0029] Bortezomib (BTZ) and the targeting peptide cRGD were dissolved in an organic solvent, and ZnCl2 was dissolved in ultrapure water. The two were then mixed, and the pH of the mixture was adjusted. The reaction was carried out under magnetic heating and stirring at a temperature of 37°C. After the reaction was completed, the mixture was centrifuged and freeze-dried to obtain bortezomib nanomedicine.

[0030] The organic solvent mentioned above is DMSO; the bortezomib and Zn 2+ The molar ratio of substances is 3:1, 1:1, or 1:3.

[0031] The ratio of organic solvent to ultrapure water is 1:1, 1:2, or 1:4, and the pH of the mixture is 7, 8, 9, or 10.

[0032] The mass ratio of the targeting peptide cRGD to BTZ is 1:1, 1:2, or 1:10.

[0033] The conditions for magnetic heating and stirring are as follows: reaction temperature 37℃, stirrer speed 1100rpm, and reaction time 30min.

[0034] This invention also discloses the application of a self-assembled bortezomib nanomedicine in the preparation of drugs for treating subcutaneous tumors and bone tumors.

[0035] Example 1

[0036] This embodiment relates to a method for preparing self-assembled bortezomib nanomedicines (BTZNDs), the preparation method of which is as follows:

[0037] Weigh 10 mg of bortezomib (BTZ) and dissolve it in 500 μL of DMSO. Weigh 10.62 mg of ZnCl2 and dissolve it in 1 mL of ultrapure water. Mix the two solutions in a glass reaction flask. BTZ and Zn... 2+ The molar ratio of the two components was 1:3, and the pH of the solution was adjusted to 7 using 1M NaOH solution. At room temperature, a magnetic stir bar was added to the reaction flask, and the flask was placed on a magnetic stirrer. The reaction was carried out at 37°C and 1100 rpm for 30 min. After the reaction, the reaction solution was transferred to a 2 mL centrifuge tube and centrifuged at 14,000 rpm for 10 min. The supernatant was discarded, and the precipitate was resuspended in 1 mL of ultrapure water. The centrifugation process was repeated, and the supernatant was discarded. After resuspending the precipitate in 1 mL of ultrapure water, it was freeze-dried to obtain bortezomib nanomedicine BTZNDs. BTZ and Zn 2+ The reaction steps for molar ratios of 1:1 and 3:1 are the same as above.

[0038] The killing ability of BTZNDs and BTZ against MDA-MB-231 cells and their cytotoxicity against L929 cells were investigated using the CCK-8 assay. MDA-MB-231 and L929 cells were treated with BTZ and Zn. 2+ Cell viability was assessed after incubation of BTZNDs synthesized at different concentrations (50, 100, and 200 nM) for 48 hours at varying molar ratios. Results showed that BTZ and Zn... 2+ BTZNDs synthesized under a molar ratio of 1:3 exhibit stronger cytotoxicity against tumor cells, such as... Figure 1As shown in Figure a, it is also less toxic to normal cells, such as Figure 1 As shown in b. Therefore, BTZ and Zn are finally determined. 2+ The optimal molar ratio is 1:3.

[0039] Example 2

[0040] The difference between this embodiment and Example 1 lies in the pH value of the reaction. 10 mg of BTZ was weighed and dissolved in 500 μL of DMSO, and 10.62 mg of ZnCl2 was weighed and dissolved in 1 mL of ultrapure water. The two were mixed in a glass reaction flask. 2+ The molar ratio of the two components was 1:3, and the pH of the solution was adjusted to 8 using 1M NaOH solution. At room temperature, a magnetic stir bar was added to the reaction flask, and the flask was placed on a magnetic stirrer. The reaction was carried out at 37°C and 1100 rpm for 30 min. After the reaction, the reaction solution was transferred to a 2 mL centrifuge tube, centrifuged at 14,000 rpm for 10 min, and the supernatant was discarded. The precipitate was then resuspended in 1 mL of ultrapure water, and the centrifugation process was repeated, discarding the supernatant each time. After resuspending the precipitate in 1 mL of ultrapure water, it was freeze-dried to obtain bortezomib nanomedicines (BTZNDs). The reaction steps for pH values ​​of 9 and 10 were the same.

[0041] MDA-MB-231 and L929 cells were incubated with BTZNDs synthesized under different pH conditions at different concentrations (100 and 200 nM) for 48 hours, and cell viability was then assessed. The results showed that BTZNDs had no killing effect on tumor cells at pH 10. BTZNDs synthesized at pH 9 showed weak toxicity to normal cells, but the killing effect on tumor cells was also not ideal. Comparing pH 7 and pH 8, BTZNDs synthesized at pH 7 showed the best killing effect on tumor cells. Figure 2 As shown in Figure a, it also has relatively weak toxicity to normal cells, such as Figure 2 As shown in b. Therefore, the final pH of the reaction solution was determined to be 7.

[0042] Example 3

[0043] The difference between this embodiment and Example 1 lies in the volume ratio of the organic solvent DMSO and ultrapure water. 10 mg of BTZ was weighed and dissolved in 500 μL of DMSO, and 10.62 mg of ZnCl2 was weighed and dissolved in 500 μL of ultrapure water. The two were mixed in a glass reaction flask. 2+The molar ratio of the active ingredients was 1:3, and the pH of the solution was adjusted to 7 using 1M NaOH solution. At room temperature, a magnetic stir bar was added to the reaction flask, and the flask was placed on a magnetic stirrer. The reaction was carried out at 37°C and 1100 rpm for 30 min. After the reaction, the reaction solution was transferred to a 2 mL centrifuge tube, centrifuged at 14,000 rpm for 10 min, and the supernatant was discarded. The precipitate was then resuspended in 1 mL of ultrapure water, and the centrifugation process was repeated, discarding the supernatant each time. After resuspending the precipitate in 1 mL of ultrapure water, it was freeze-dried to obtain bortezomib nanomedicines (BTZNDs). The reaction procedure for organic solvent DMSO and ultrapure water in a volume ratio of 1:4 was the same.

[0044] BTZNDs synthesized with different volume ratios of DMSO and H2O at various concentrations (50, 100, and 200 nM) were incubated with MDA-MB-231 and L929 cells for 48 hours, and cell viability was then assessed. The results showed that BTZNDs synthesized at a DMSO to H2O volume ratio of 1:2 achieved a tumor cell killing effect of approximately 30% at a relatively low concentration (50 nM). Figure 3 As shown in Figure a, and at this concentration, it also exhibits relatively weak toxicity to normal cells, such as Figure 3 As shown in Figure b. Therefore, the final volume ratio of DMSO to H2O was chosen to be 1:2.

[0045] Example 4

[0046] This embodiment relates to a method for preparing a self-assembled bortezomib nanomedicine BTZNDs-cRGD.

[0047] The difference between this embodiment and Example 1 lies in the amount of cRGD added. 10 mg of BTZ was weighed and dissolved in 500 μL of DMSO, and then 10 mg of cRGD was added until completely dissolved. 10.62 mg of ZnCl2 was weighed and dissolved in 1 mL of ultrapure water, and the two were mixed in a glass reaction flask. BTZ and Zn... 2+ The molar ratio of BTZ to cRGD was 1:3, and the mass ratio of BTZ to cRGD was 1:1. The pH of the solution was adjusted to 7 using 1M NaOH solution. At room temperature, a magnetic stir bar was added to the reaction flask, and the flask was placed on a magnetic stirrer. The reaction was carried out at 37°C and 1100 rpm for 30 min. After the reaction, the reaction solution was transferred to a 2 mL centrifuge tube and centrifuged at 14,000 rpm for 10 min. The supernatant was discarded, and the precipitate was resuspended in 1 mL of ultrapure water. The centrifugation process was repeated, and the supernatant was discarded. After resuspending the precipitate in 1 mL of ultrapure water, it was freeze-dried to obtain the bortezomib nanodrug BTZNDs-cRGD. The reaction steps for BTZ to cRGD with a mass ratio of 1:2 and 1:10 are the same.

[0048] BTZ and Zn in Example 1 were detected by CCK-8 assay.2+ The killing effects of BTZNDs synthesized under a molar ratio of 1:3 and BTZNDs-cRGD synthesized under different BTZ and cRGD mass ratios on MDA-MB-231 cells were investigated. The results are as follows: Figure 4 As shown, BTZNDs-cRGD exhibits stronger tumor cell killing activity than BTZNDs, indicating that cRGD was successfully introduced into bortezomib nanomedicine and achieved tumor cell targeting. When 1 mg of cRGD was introduced (BTZ to cRGD mass ratio of 1:10), the tumor cell killing activity was even stronger, demonstrating that a small amount of cRGD is sufficient for bortezomib nanomedicine to achieve tumor cell targeting. Therefore, it was ultimately determined that introducing 1 mg of cRGD into the BTZNDs reaction system would participate in the synthesis of BTZNDs-cRGD.

[0049] Example 5

[0050] This embodiment relates to the morphology and elemental analysis of the self-assembled bortezomib nanomedicine BTZNDs-cRGD.

[0051] The morphology of the self-assembled bortezomib nanomedicine was characterized using transmission electron microscopy. The bortezomib nanomedicine BTZNDs-cRGD prepared in Example 4 at a BTZ:cRGD mass ratio of 1:10 was resuspended in ultrapure water and uniformly dispersed by sonication. After dilution by a certain factor, 10 μL was dropped onto a 100-mesh copper grid and allowed to evaporate overnight at room temperature. The morphological characteristics and corresponding elemental energy dispersive spectroscopy (EDS) images of the nanomedicine were observed under a transmission electron microscope.

[0052] Experimental results are as follows Figure 5 As shown in the transmission electron microscopy (TEM) image, BTZNDs-cRGD exhibits a nanorod-like morphology. Further elemental analysis of BTZNDs-cRGD using TEM revealed the following: Figure 1 As shown in b, the HAADF image and the corresponding EDS elemental mapping image show that the nanomedicine contains Zn and the characteristic elements N and O of the peptide, proving that the peptide drug BTZ contains Zn. 2+ Self-assembly was successfully performed.

[0053] Example 6

[0054] This embodiment relates to BTZ and Zn in Embodiment 1. 2+ Surface charge characterization of bortezomib nanomedicine BTZNDs prepared under a molar ratio of 1:3 and bortezomib nanomedicine BTZNDs-cRGD prepared under a mass ratio of 1:10 for BTZ and cRGD in Example 4.

[0055] The surface zeta potential of bortezomib nanomedicines was determined using a zeta potential meter. Bortezomib nanomedicines BTZNDs and BTZNDs-cRGD were resuspended in ultrapure water and sonicated to achieve uniform dispersion. After dilution by a certain factor, 0.8 mL of the solution was added to the sample cell of the zeta potential meter. The sample cell was placed in the sample slot of the instrument, and the instrument parameters were set before measuring the zeta potential. Each sample was measured three times.

[0056] Experimental results are as follows Figure 6 As shown in the figure, the surface charge of BTZNDs is 16.5 mV. Since the pH of the reaction solution is 7, bortezomib molecules are positively charged under these conditions, therefore BTZNDs are positively charged. The surface charge of BTZNDs-cRGD is 20.0 mV. Since cRGD is also positively charged under pH 7 conditions, the higher surface charge of BTZNDs-cRGD compared to BTZNDs proves the successful introduction of cRGD.

[0057] Example 7

[0058] This embodiment relates to the XPS characterization of the bortezomib nanomedicine BTZNDs-cRGD prepared under the condition that the mass ratio of BTZ to cRGD is 1:10 as in Example 4.

[0059] Experimental results are as follows Figure 7 As shown, Zn, B, O, C, and N elements were all detected, proving that BTZ and Zn... 2+ Successful self-assembly.

[0060] Example 8

[0061] This embodiment relates to BTZ and Zn in Embodiment 1. 2+ In vitro drug release studies were conducted on bortezomib nanomedicine BTZNDs prepared under a molar ratio of 1:3 and bortezomib nanomedicine BTZNDs-cRGD prepared under a mass ratio of 1:10 for BTZ and cRGD in Example 4.

[0062] Weigh 5 mg of lyophilized BTZNDs and BTZNDs-cRGD and resuspend them in 1 mL of PBS. Place them in a dialysis bag (3500 Da) and seal it. Immerse the dialysis bag containing the sample in a glass reaction flask containing 20 mL of PBS. Add a magnetic stir bar to the reaction flask and place it on a constant temperature water bath with a magnetic stirrer at 37 °C and 200 rpm for dialysis. Take 1 mL of solution from the reaction flask at different time points and use a UV spectrophotometer to determine the release of BTZ at 270 nm.

[0063] Experimental results are as follows Figure 8As shown, the maximum drug release of BTZNDs can reach 58%, while the maximum drug release of BTZNDs-cRGD can reach 78%. This may be because BTZNDs contain only BTZ and Zn. 2+ The structure of BTZNDs-cRGD is relatively compact, while BTZNDs-cRGD also contains cyclic peptide cRGD, making its structure more loosely distributed. Therefore, BTZ is more easily released from BTZNDs-cRGD. The results show that the self-assembled bortezomib nanomedicine prepared in Example 1 has good drug release performance.

[0064] Example 9

[0065] This embodiment involves the monomeric drug bortezomib, BTZ and Zn from Example 1. 2+ The in vitro killing effect of bortezomib nanomedicine BTZNDs prepared under the condition of a molar ratio of 1:3 and the bortezomib nanomedicine BTZNDs-cRGD prepared under the condition of a mass ratio of 1:10 of BTZ and cRGD in Example 4 on tumor cells MDA-MB-231 and toxic effects on normal cells L929 were investigated.

[0066] Experimental results are as follows Figure 9 As shown in the figure, tumor cells MDA-MB-231 and normal cells L929 were incubated with different concentrations of BTZ, BTZNDs, and BTZNDs-cRGD for 48 hours, and cell viability was then assessed. The results showed that at concentrations of 10, 50, 100, and 200 nM, BTZNDs-cRGD exhibited better cell-killing effects on MDA-MB-231 cells than the monomeric drug BTZ, demonstrating that the introduction of cRGD allows bortezomib nanomedicine to better target tumor cells, achieving better therapeutic effects. Furthermore, BTZNDs-cRGD showed significantly lower toxicity to L929 cells than the monomeric drug BTZ. These results indicate that compared to the monomeric drug, the bortezomib nanomedicine BTZNDs-cRGD has a stronger inhibitory effect on tumor cells and lower toxicity to normal cells.

[0067] Example 10

[0068] This embodiment relates to the hemolysis analysis of the bortezomib nanomedicine BTZNDs-cRGD prepared under the condition that the mass ratio of BTZ and cRGD is 1:10 as in Example 4.

[0069] Mouse blood was incubated with different concentrations (50, 100, 200, 500, 1000 μg / mL) of BTZNDs-cRGD solution at 37°C for 45 min. PBS was used as a negative control, and deionized water as a positive control. The experimental results are as follows: Figure 10As shown, incubation of different concentrations of BTZNDs-cRGD with blood did not induce significant hemolysis, while the positive control group showed very obvious hemolysis. Calculations showed that the hemolysis rate of BTZNDs-cRGD was less than 2%, indicating that BTZNDs-cRGD has good biocompatibility.

[0070] Example 11

[0071] This embodiment involves the monomeric drug bortezomib, composed of BTZ and Zn from Example 1. 2+ The therapeutic effects and related parameter changes of bortezomib nanomedicine BTZNDs prepared under the condition of a molar ratio of 1:3 and BTZ to cRGD prepared under the condition of a mass ratio of 1:10 in Example 4 on subcutaneous tumors in mice were investigated.

[0072] Tumor-bearing mice were randomly divided into 7 groups of 4 mice each: PBS control group, 0.5 mg / kg BTZ group, 1.0 mg / kg BTZ group, 0.5 mg / kg BTZNDs group, 1.0 mg / kg BTZNDs group, 0.5 mg / kg BTZNDs-cRGD group, and 1.0 mg / kg BTZNDs-cRGD group. Mice were treated with the drugs via tail vein injection every two days, repeated three times.

[0073] The longest and shortest diameters of the mouse tumors were measured every two days to calculate the tumor volume, and the mouse's weight changes were recorded at the same time.

[0074] like Figure 11 As shown, compared with the control group, the tumor size of mice in the BTZNDs-cRGD group was significantly smaller, indicating that bortezomib nanomedicine is more effective than the monomeric drug bortezomib in treating subcutaneous tumors in mice. The relative tumor volume change curves for each group of mice are shown in the figure. Figure 12 As shown in Figure a, the treatment results indicate that the tumors in the PBS group grew the fastest. Low-dose BTZ and BTZNDs showed little inhibitory effect on the tumors, while high-dose BTZ and BTZNDs both achieved some tumor-suppressing effect. The high-dose BTZNDs-cRGD showed the most significant inhibitory effect on mouse tumors, indicating that bortezomib nanomedicine has a good inhibitory effect on subcutaneous tumors in mice. The relative body weight change curves of each group of mice are shown in Figure a. Figure 12 As shown in Figure b, the mice in the high-dose BTZ group experienced a weight loss of over 30%, and two mice died during the treatment period, indicating that the high-dose BTZ had significant toxicity to the mice. In contrast, the weight of mice in other experimental groups did not change significantly, suggesting that the bortezomib nanomedicine reduced the toxicity of the monomeric drug bortezomib and exhibited good biocompatibility in vivo.

[0075] Example 12

[0076] This embodiment involves the monomeric drug bortezomib, BTZ and Zn from Example 1. 2+ The therapeutic effects and related parameter changes of bortezomib nanomedicine BTZNDs prepared under the condition of a molar ratio of 1:3 and BTZ to cRGD prepared under the condition of a mass ratio of 1:10 in Example 4 on mouse bone tumors were investigated.

[0077] Tumor-bearing mice were randomly divided into four groups: PBS control group, 0.5 mg / kg BTZ group, 0.5 mg / kg BTZNDs group, and 0.5 mg / kg BTZNDs-cRGD group. Mice were treated with the drugs via tail vein injection every two days, repeated three times. The survival status of the mice was observed daily during the treatment period.

[0078] After treatment, the forelimb and hindlimb bones of the mice were removed, and the muscle tissue on the bone surface was cleaned off. The bones were then fixed in a 4% paraformaldehyde solution. A micro-computed tomography (MCT) scanner was used to scan the forelimb and hindlimb bones to observe bone destruction. Furthermore, 3D models of some cancellous bone from the mice were created using the Micro-CT software to further observe the bone condition.

[0079] Figure 13 Micro-CT images of the forelimb bone of mice with bone tumors, including the monomeric drug bortezomib, bortezomib nanomedicine, and the control group, as well as 3D modeling images of some cancellous bone, are presented. Figure 14 Micro-CT images of the hind limb bones of mice with bone tumors were obtained from the single drug bortezomib, bortezomib nanomedicine, and a control group. Image analysis showed that the untreated PBS group mice had more severe bone destruction, while the BTZ and BTZNDs treatment groups showed less bone destruction compared to the control group. The BTZNDs-cRGD group showed the least bone destruction. Therefore, BTZNDs-cRGD showed the best therapeutic effect on mice with bone tumors.

Claims

1. A method for preparing a self-assembled bortezomib nanomedicine, characterized in that, The preparation method includes: Bortezomib and the targeting peptide cRGD were dissolved in an organic solvent, and ZnCl2 was dissolved in ultrapure water. The two were then mixed and magnetically heated and stirred to obtain a nanodrug suspension. The supernatant was removed by centrifugation and then lyophilized to obtain bortezomib nanodrug. The mass ratio of the targeting peptide cRGD to BTZ is 1:1, 1:2, or 1:

10.

2. The method for preparing a self-assembled bortezomib nanomedicine as described in claim 1, characterized in that, The substance of the above bortezomib and Zn 2+ The amount of substance ratio is 3:1, 1:1 or 1:

3.

3. The method for preparing a self-assembled bortezomib nanomedicine as described in claim 1, characterized in that, The organic solvent is DMSO, and the ratio of the organic solvent to ultrapure water is 1:1, 1:2, or 1:

4.

4. The method for preparing a self-assembled bortezomib nanomedicine as described in claim 1, characterized in that, The conditions for magnetic heating and stirring are as follows: reaction temperature 37℃, stirrer speed 1100rpm.

5. The self-assembled bortezomib nanomedicine prepared by the method for preparing a self-assembled bortezomib nanomedicine according to any one of claims 1-4, and its application in the preparation of drugs for treating subcutaneous tumors and bone tumors.

Images