Application of vanadium pentoxide nanoparticle NIP-FOS or NIP-PVP in preparation of medicine for treating pancreatic cancer
Through targeted binding of vanadium pentoxide nanoparticles to HSP60, inhibiting M2 macrophage polarization, the problem that the existing technology is difficult to inhibit the early development of pancreatic cancer is solved, effective treatment of pancreatic cancer is achieved, and patient survival rate is improved.
Patent Information
- Application Number
- CN202510452533.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-11
- Publication Date
- 2025-06-24
AI Technical Summary
The prior art is difficult to effectively inhibit the early development of pancreatic cancer, resulting in most patients who have progressed to middle and late stages when they are diagnosed. The effectiveness of surgical treatment is limited, and the survival rate of patients is low.
Vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP are used to inhibit the expression level of HSP60 and polarization of M2 macrophages by targeting the binding of heat shock protein 60 (HSP60).
Vanadium pentoxide nanoparticles significantly inhibit the proliferation and invasion of pancreatic cancer cells, have good therapeutic effects, have significant therapeutic effects on the pancreatic precancerous lesion model (PanIN), and improve the patient's survival rate.
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Abstract
Description
Technical Field
[0001] The present invention relates to the application of vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP in the preparation of drugs for treating pancreatic cancer, belonging to the field of new drug research and development. Background Art
[0002] Pancreatic cancer is a digestive system tumor with extremely high malignancy, and its incidence and mortality are relatively high. Up to now, pancreatic cancer has become one of the four major causes of cancer death globally. Among them, pancreatic ductal adenocarcinoma is the most common type of pancreatic cancer, accounting for about 90% of all pancreatic malignancies. This disease lacks specific early symptoms and effective screening methods, resulting in more than 85% of pancreatic cancer patients progressing to the middle and advanced stages when they are clearly diagnosed. Surgery is the main method for treating pancreatic cancer, but only 15 - 20% of patients have the opportunity to undergo radical surgery, and the 5-year survival rate of patients is less than 9%. In the early development process of PDAC, the transition from acinar ductal metaplasia (ADM) to pancreatic intraepithelial neoplasia (PanIN) is a key link. If pancreatic cancer is diagnosed and treated early, the survival rate of patients can be increased to 30% - 60%. Therefore, there is an urgent need to find new and effective drugs to inhibit the development of pancreatic cancer.
[0003] Heat shock protein 60 (HSP60) is an important member of the heat shock protein family. In a variety of tumor cells, HSP60 shows a high expression state, playing a key promoting role in the proliferation and survival of tumor cells. Currently, there are studies showing that HSP60 may affect the polarization of M2 macrophages in the tumor microenvironment. When HSP60 released by tumor cells is recognized by macrophages, a series of intracellular signaling pathways will be activated. HSP60 binds to Toll-like receptor 4 (TLR4) on the surface of macrophages, and then activates the myeloid differentiation factor 88 (MyD88)-dependent signaling pathway, leading to the activation of nuclear factor κB (NF-κB), promoting the polarization of macrophages into the M2 type, thereby promoting the proliferation, migration and invasion of tumor cells. Therefore, HSP60 and M2 macrophages can be used as new therapeutic targets for pancreatic cancer. Vanadium is a new type of metal anti-cancer drug. Numerous studies have shown that vanadium compounds have the ability to induce apoptosis of cancer cells, mainly through the mitochondrial pathway and the death receptor pathway. In the present invention, through a large number of studies, it has been confirmed that vanadium pentoxide nanoparticles can target and bind to HSP60 to play a role, inhibit the expression level of HSP60, and thus inhibit M2 macrophage polarization.
[0004] Therefore, treating pancreatic cancer by targeting HSP60 with vanadium pentoxide nanoparticles to inhibit M2 macrophage polarization has good academic value and clinical significance. Summary of the Invention
[0005] Vanadium pentoxide nanoparticles, as metal nanoparticles, are widely used in therapeutic applications due to their multivalent nature, ability to participate in various redox processes, and being key components of various enzymes, electron transfer proteins, etc. They have excellent biocompatibility, high chemical and biological stability, and biodegradability. Our previous results showed that vanadium pentoxide nanoparticles had a significant killing effect on pancreatic cancer cells and had a good therapeutic effect on the pancreatic in situ cancer model. Therefore, in this study, NIP-FOS or NIP-PVP was used for the treatment of PanIN, providing a scientific experimental basis for the application of vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP in the treatment of early pancreatic cancer.
[0006] The technical solution of the present invention:
[0007] Application of vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP in the preparation of a drug for treating pancreatic cancer.
[0008] Furthermore, the preparation method of the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP is as follows: Mix V2O5 powder, citrate, hydrogen peroxide, and fructooligosaccharide (FOS) or polyvinylpyrrolidone (PVP), and vigorously stir at room temperature to form a suspension; add sodium borohydride, and stir at room temperature to synthesize a black colloid; add propanol with the same volume as the black colloid, centrifuge, and collect the centrifuged precipitate; wash the precipitate with a mixed washing solution of deionized water and propanol, centrifuge, and collect the precipitate for vacuum drying to obtain vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP.
[0009] Furthermore, the mass concentrations of V2O5, citrate, fructooligosaccharide or polyvinylpyrrolidone in the suspension are 0.3% - 0.8%, 1% - 1.4%, 0.2% - 0.8% respectively; the mass ratio of hydrogen peroxide to V2O5 is 7 - 11:400; the mass concentration of sodium borohydride in the black colloid is 0.2% - 0.8%; the volume ratio of the mixed washing solution of water and propanol is 1 - 1.5:1.
[0010] Furthermore, the citrate is sodium citrate or potassium citrate, used as an etching agent; fructooligosaccharide and polyvinylpyrrolidone are used as surfactants; sodium borohydride is used as a reducing agent.
[0011] Furthermore, the administration dose of the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP is: intravenous injection three times a week, with a dose of 5 mg / kg.
[0012] Furthermore, the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP can target and bind to HSP60, thereby inhibiting the expression level of HSP60 and the polarization level of M2 macrophages, and further treating pancreatic cancer.
[0013] Further, in the above technical solution, the pre - pancreatic cancer lesion model uses a genetically engineered mouse primary pancreatic cancer model, which simulates early - stage pancreatic cancer patients in clinical practice.
[0014] Further, in the above technical solution, vanadium pentoxide nanoparticles have a good therapeutic effect on PanIN.
[0015] Further, the vanadium pentoxide nanoparticles can target - bind to recombinant human HSP60 protein.
[0016] Further, in the above technical solution, vanadium pentoxide nanoparticles can inhibit the oligomerization of HSP60 protein.
[0017] Further, in the above technical solution, vanadium pentoxide nanoparticles down - regulate the expression levels of HSP60 and the M2 macrophage surface marker CD206 in pancreatic cancer tissues.
[0018] Advantages of the present invention: The present invention uses a pre - pancreatic cancer lesion model to prove that vanadium pentoxide nanoparticles have a good therapeutic effect on PanIN; uses BLI molecular interaction to prove that vanadium pentoxide nanoparticles have a target - binding effect on HSP60; and proves that vanadium pentoxide nanoparticles can inhibit the oligomerization and expression level of HSP60 in the state of pancreatic cancer, thereby inhibiting M2 macrophage polarization. The present invention provides a basis for using vanadium pentoxide nanoparticles as a new drug for treating PanIN. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] Figure 1 Therapeutic effects of vanadium pentoxide nanoparticles NIP - FOS and NIP - PVP on PanIN.
[0020] Among them, A is H&E staining of mouse pancreatic tissue. B is Masson staining of mouse pancreatic tissue. C is Ki67 immunohistochemical staining of mouse pancreatic tissue.
[0021] Figure 2 Targeting effects of vanadium pentoxide nanoparticles NIP - FOS and NIP - PVP on HSP60 protein.
[0022] Among them, A BLI detects the binding ability of NIP - FOS and NIP - PVP to recombinant human HSP60 protein. B Oligomerization of HSP60 in mouse pancreatic tissue. C mRNA level of HSP60 in mouse pancreatic tissue. Compared with the model group, *P < 0.05, **p < 0.01, ***p < 0.001. D Immunohistochemical staining of HSP60 in mouse pancreatic tissue. E Immunofluorescence staining of HSP60 in mouse pancreatic tissue.
[0023] Figure 3 Vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP inhibit M2 macrophage polarization.
[0024] A Protein expression level of CD206 in mouse pancreatic tissue. B Immunofluorescence staining results of M2 macrophages in mouse pancreatic tissue. Specific implementation manners
[0025] The following non-limiting examples can enable those of ordinary skill in the art to more comprehensively understand the present invention, but do not limit the present invention in any way.
[0026] 1. Experimental protocol:
[0027] (1) In vivo experiment:
[0028] Construction of a KC (LSL-KrasG12D, Pdx1-Cre) mouse pre-cancerous lesion model of pancreatic cancer: Select 30 male KC mice about 6 weeks old and randomly divide them into the following 4 groups: control group, model group, NIP-FOS administration group, and NIP-PVP administration group. Dissolve tamoxifen in corn oil and inject it subcutaneously into the mice (200 mg / kg). One week later, inject caerulein (100 μg / kg) intraperitoneally into the mice 3 times a day, every other day, for 3 days a week, for a total of 8 weeks. At the same time, the control group was injected with an equal amount of normal saline in the same manner. In addition to caerulein given to the model group, the drug containing vanadium pentoxide nanoparticles NIP-FOS or NIP–PVP was injected into the mice through the tail vein (5 mg / kg) twice a week. After 8 weeks of model establishment, all the mice were euthanized, and their serum and pancreatic tissue samples were collected and stored in a -80 °C refrigerator for later use. At the same time, a part of the pancreatic tissue was fixed in 4% paraformaldehyde solution and made into tissue sections by paraffin embedding for later use. H&E staining, Masson staining, and Ki67 immunohistochemical staining were used to evaluate the degree of damage to the pancreatic tissue; BLI molecular interaction was used to judge the binding ability of NIP-FOS and NIP-PVP to HSP60 protein; WB was used to detect the effect of NIP-FOS and NIP-PVP on HSP60 oligomerization; qPCR, immunohistochemistry, and immunofluorescence were used to detect the expression level of HSP60 in the pancreatic tissue; WB, F4 / 80, and CD206 immunofluorescence were used to detect macrophage recruitment and M2 polarization in the pancreatic tissue.
[0029] (2) Binding experiment
[0030] First, pre-wet the SA sensor in 200 μL of PBST for 10 min. Then, add 200 μL each of 10 μg / mL biotinylated HSP60 protein solution, 1 μg / mL VIP-FOS or VIP-PVP solution, and PBST buffer into a black 96-well plate. First, immerse the well-equilibrated SA sensor into the solution containing biotinylated HSP60 protein and bind for 10 min, so that the biotinylated HSP60 protein is immobilized on the surface of the SA sensor through the specific binding of biotin and streptavidin. Place the sensor into the PBST buffer and perform baseline measurement on the BLI detection system for 60 s. Then, immerse the sensor into the VIP-FOS or VIP-PVP solution respectively, and record the signal changes during the binding process of the sensor and vanadyl ions. The binding time is 180 s. After the binding reaction is completed, transfer the sensor back into the PBST buffer, monitor the dissociation process of vanadyl ions from the immobilized HSP60 protein, and record the signal change over time. The dissociation time is 180 s. Use the data analysis software supporting the Octet RH16 detection system to analyze the signal data of the binding and dissociation processes.
[0031] (3) The preparation method of the vanadium pentoxide nanoparticles in the present invention is as follows:
[0032] (1) Mix V2O5 powder (0.2 g of V2O5 powder is added to 15 mL of water), sodium citrate (0.4 g of sodium citrate is added to 10 mL of water), H2O2 (mass concentration is 3%, 150 μL), and fructooligosaccharide (FOS) or polyvinylpyrrolidone (PVP) (0.2 g of fructooligosaccharide or polyvinylpyrrolidone is added to 10 mL of water), and stir vigorously at room temperature (rotation speed 1500 r / min) to prepare a suspension.
[0033] (2) Add NaBH4 (0.24 g of NaBH4 is added to 10 mL of water) to the suspension, stir at room temperature for 3 hours, and then stop the reaction to synthesize a black colloid.
[0034] (3) Add propanol with the same volume as the black colloid, centrifuge at 10000 rpm for 8 min, and collect the centrifuged precipitate.
[0035] (4) Prepare a mixed washing solution of deionized water and propanol with a volume ratio of 1:1, wash the precipitate, centrifuge at 10000 rpm for 5 min, collect the precipitate and perform vacuum drying to obtain vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP.
[0036] 2. Experimental results:
[0037] 2.1 Vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP have good therapeutic effects on PanIN
[0038] To study the therapeutic effects of NIP-FOS and NIP-PVP on pancreatic intraepithelial neoplasia (PanIN), a pre-cancerous lesion of pancreatic cancer, we observed pancreatic tissue sections of each group by H&E staining. In the model group, the PanIN lesion tissues showed abnormal hyperplasia of ductal epithelial cells, with enlarged and deeply stained cell nuclei, dysregulated nuclear-cytoplasmic ratio, and varying degrees of distortion and deformation of the ductal structure, presenting typical pre-cancerous lesion characteristics. In the two nano-material administration groups, the tissue morphology was significantly improved. The degree of epithelial cell hyperplasia was significantly reduced, the morphology and size of the cell nuclei were closer to normal cells, and the distortion and deformation of the ductal structure were also alleviated to a certain extent, indicating that these two nano-materials, NIP-FOS and NIP-PVP, have good therapeutic effects on PanIN lesions ( Figure 1 as shown in A of Figure 1 ). In the PanIN lesion tissues of the model group, a large number of randomly distributed collagen fibers were visible, suggesting a high degree of tissue fibrosis, which is related to tissue remodeling caused by pre-cancerous lesions. In the two nano-material administration groups, the content of collagen fibers decreased, indicating that the fibrosis process of the tissue was effectively inhibited ( Figure 1 as shown in B of
[0039] 2.2 Targeting effect of vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP on HSP60
[0040] To further explore the interaction between NIP-FOS, NIP-PVP and HSP60, this study used biolayer interferometry (BLI) to conduct molecular interaction experiments. During the experiment, recombinant human HSP60 protein was immobilized on the surface of the BLI biosensor, and then the two vanadium pentoxide nano-materials, NIP-FOS and NIP-PVP, were introduced into the reaction system. The BLI device was used to collect and analyze the interference signals on the surface of the biosensor in real time. The experimental results showed that when the NIP-FOS and NIP-PVP nano-materials contacted the immobilized recombinant human HSP60 protein, significant changes occurred in the BLI signals, indicating specific binding between the two ( Figure 2A) in it. Under normal physiological conditions, HSP60 usually exists in the form of monomers with specific structures and functions within cells. In cancer, due to the cells being in a continuous stress state and the abnormal activation of multiple signaling pathways, the oligomerization of HSP60 tends to occur more frequently and significantly. We detected the oligomerization of HSP60 in PanIN tissues, and the results showed that in the model group, obvious oligomerization bands of HSP60 appeared, while in the NIP-FOS and NIP-PVP nanomaterial administration groups, the degree of oligomerization of HSP60 was significantly reduced ( Figure 2 B) in it. In the molecular level detection, compared with the model group, the mRNA level of HSP60 in the NIP-FOS and NIP-PVP administration groups was significantly down-regulated ( Figure 2 C) in it. The immunohistochemistry and immunofluorescence results were consistent with the above, and the protein expression level of HSP60 in the pancreatic tissues of the two administration groups was also significantly reduced ( Figure 2 D-E) in it. These results further revealed that NIP-FOS and NIP-PVP inhibited the development of pancreatic cancer by targeting HSP60.
[0041] 2.3 Vanadium pentoxide nanoparticles NIP-FOS and NIP-PVP inhibit M2 macrophage polarization
[0042] To further clarify the mechanism of action of NIP-FOS and NIP-PVP in the treatment of pancreatic cancer, the protein expression level of the M2 macrophage marker CD206 was detected. Compared with the model group, the level of CD206 in the two administration groups was significantly reduced ( Figure 3 A) in it. The immunofluorescence staining results showed that the number of M2 macrophages double-positive for F4 / 80 and CD206 was also significantly reduced ( Figure 3 B) in it. These results indicate that the vanadium pentoxide nanoparticles NIP-PVP and NIP-FOS can effectively inhibit M2 macrophage polarization, providing an important basis for further understanding their mechanism of action in inhibiting the development of pancreatic cancer.
Claims
1. Application of vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP in the preparation of drugs for treating pancreatic cancer.
2. The use according to claim 1, characterized in that: The preparation method of the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP is as follows: V2O5 powder, citrate, hydrogen peroxide, and oligofructose or polyvinyl pyrrolidone are mixed, and the mixture is stirred vigorously at room temperature to form a suspension; sodium borohydride is added, and the mixture is stirred at room temperature to synthesize a black colloid; propanol with the same volume as the black colloid is added, centrifuged, and the centrifugal precipitate is collected; the precipitate is washed with a mixed washing liquid of deionized water and propanol, centrifuged, and the precipitate is collected and vacuum dried to obtain the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP.
3. The use according to claim 2, characterized in that: The mass concentrations of V2O5, citrate, oligofructose or polyvinyl pyrrolidone in the suspension are 0.3%-0.8%, 1%-1.4% and 0.2%-0.8% respectively; the mass ratio of hydrogen peroxide to V2O5 is 7-11:400; the mass concentration of sodium borohydride in the black colloid is 0.2%-0.8%; and the volume ratio of the mixed washing liquid of water and propanol is 1-1.5:
1.
4. The use according to claim 2, characterized in that: The citrate is sodium citrate or potassium citrate, which is used as an etchant; oligofructose and polyvinyl pyrrolidone are used as surfactants; and sodium borohydride is used as a reducing agent.
5. The use according to claim 1, 2, 3 or 4, characterized in that: The dosage of the vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP is: intravenous injection three times a week, with a dosage of 5 mg / kg.
6. The use according to claim 1, 2, 3 or 4, characterized in that: The vanadium pentoxide nanoparticles NIP-FOS or NIP-PVP can target and bind to HSP60, thereby inhibiting the expression level of HSP60 and the polarization level of M2 macrophages, thereby treating pancreatic cancer.