Application of cardiac glycoside medicine or pharmaceutically acceptable salt or derivative thereof in preparation of products for inhibition, alleviation, adjuvant therapy or treatment of cancer and medicine

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of cardiac glycoside medicines or pharmaceutically acceptable salts or derivatives thereof in preparation of products for inhibition, relief, adjuvant therapy or treatment of cancers and medicines. The invention provides application of cardiac glycoside drugs or pharmaceutically acceptable salts or derivatives of the cardiac glycoside drugs in preparation of products for inhibition, alleviation, adjuvant therapy or treatment of cancers and the drugs, and the cardiac glycoside drugs comprise deacetylated chaenoside, deacetylated chaenoside, deacetylated chaenoside, deacetylated chaenoside, deacetylated chaenoside, deacetylated chaenoside, deacetylated chaenoside and deacetylated chaenoside. According to the application disclosed by the invention, the effects of inhibiting, relieving, assisting in treating or treating cancers and potential mechanisms of the Deslanoside are researched in SH-SY5Y and SK-N-SH neuroblastoma cell lines, and results show that the Deslanoside can be used for inhibiting proliferation of SH-SY5Y cells and SK-N-SH cells, promoting apoptosis and respectively inducing a cell cycle to stop at a G0 / G1 phase; and G1 / S and G2 / M periods.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of a cardiac glycoside or its pharmaceutically acceptable salt or derivative in the preparation of products for inhibiting, alleviating, adjuvant treatment or treating cancer, and to the use of a drug. Background Technology

[0002] Neuroblastoma (NB) is the most common extracranial solid tumor in children, with approximately 90% of cases diagnosed before the age of 10. This tumor originates from neural crest precursor cells in the developing sympathetic nervous system, commonly found in sympathetic ganglia and the adrenal glands. NB accounts for 15% of all childhood cancer deaths. Clinical treatment outcomes are often hampered by delayed diagnosis, high recurrence rates, and strong metastatic potential. Therefore, there is an urgent need to develop effective treatment strategies to meet the pressing need for NB treatment.

[0003] Cardiac glycosides are a class of plant-derived compounds with over two centuries of history of use in the treatment of heart failure and atrial arrhythmias. Deslanoside is an important member of the cardiac glycoside family. Deslanoside is a metabolite of digoxin, and it works by inhibiting Na+... + / K + -ATPase increases intracellular calcium ion concentration, thereby enhancing myocardial contractility and improving symptoms in patients with heart failure. Recent epidemiological studies have unexpectedly found that cardiac glycosides can significantly reduce mortality in cancer patients. No reports exist in the current technology regarding the anti-cancer effects of deslanoside. Summary of the Invention

[0004] Therefore, the technical problem to be solved by the present invention is to overcome the defect of poor NB treatment effect in the prior art, thereby providing an application and medicine of cardiac glycosides or their pharmaceutically acceptable salts or derivatives in the preparation of products for inhibiting, alleviating, adjuvant treatment or treating cancer, wherein the cardiac glycoside deslanoside has good inhibitory, alleviating, adjuvant treatment or treatment effect on cancer.

[0005] Therefore, the present invention provides the following technical solution: This invention provides the use of cardiac glycosides or their pharmaceutically acceptable salts or derivatives in the preparation of products for inhibiting, alleviating, adjuvant treatment or treating cancer.

[0006] Preferably, the cardiac glycoside includes deslanoside.

[0007] Preferably, the inhibition, relief, adjuvant treatment or treatment of cancer includes the inhibition, relief, adjuvant treatment or treatment of neuroblastoma.

[0008] Preferably, the inhibition, relief, adjuvant therapy, or treatment of neuroblastoma includes at least one of the following: 1) Regulate the MAPK and / or mTORC1 signaling pathways to inhibit the proliferation of neuroblastoma cells; 2) Induces apoptosis and / or necrosis of neuroblastoma cells; 3) Inducing cell cycle arrest in neuroblastoma cells; the cell cycle arrest includes one or more of G0 / G1 phase arrest, G1 / S phase arrest and G2 / M phase arrest.

[0009] This invention demonstrates through in vitro experiments that Deslanoside can significantly inhibit the proliferation of neuroblastoma cell lines (SH-SY5Y and SK-N-SH), induce apoptosis, and cause cell cycle arrest. Specifically: Inhibition of cell proliferation: Deslanoside inhibited the proliferation of SH-SY5Y and SK-N-SH cells in a dose-dependent manner, IC50 50 The concentrations were 2.26 μM (24 h) and 23.29 μM (48 h), respectively.

[0010] Induction of apoptosis: Annexin V / PI double staining, Caspase-3 activity assay, and mitochondrial membrane potential assay confirmed that Deslanoside can activate the mitochondrial-mediated endogenous apoptosis pathway and the Caspase-dependent exogenous pathway, thereby inducing apoptosis.

[0011] Induction of cell cycle arrest: Deslanoside can induce G0 / G1 phase arrest in SH-SY5Y cells, G1 / S and G2 / M phase arrest in SK-N-SH cells, and downregulate the expression of Cyclin D1 and Cyclin B1 proteins.

[0012] Transcriptomic mechanism: RNA-seq analysis showed that Deslanoside regulates the expression of multiple genes related to cell cycle, apoptosis, MAPK, mTORC1 signaling pathway and cellular senescence.

[0013] Preferably, the product includes a drug.

[0014] This invention provides a medicament for inhibiting, alleviating, adjuvant treatment or treatment of cancer, comprising a cardiac glycoside or a pharmaceutically acceptable salt or derivative thereof and a pharmaceutically acceptable carrier or excipient; optionally, the cardiac glycoside comprises deslanoside.

[0015] Preferably, the dosage form of the drug includes liquid, semi-solid, or solid dosage forms.

[0016] Preferably, the dosage form of the drug includes one or more of the following: injection, oral, sustained-release, and targeted formulation.

[0017] Preferably, the oral preparation includes one or more of tablets, granules, and capsules; the injectable preparation includes an injection solution.

[0018] Preferably, the excipients include any one or a combination of at least two of the following: carrier, diluent, excipient, filler, binder, wetting agent, disintegrant, emulsifier, cosolvent, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent, or buffer.

[0019] The technical solution of this invention has the following advantages: 1. The application of deslanoside or its pharmaceutically acceptable salts or derivatives provided by this invention in the preparation of products for inhibiting, alleviating, adjuvant therapy, or treating cancer. This invention investigated the inhibitory, alleviating, adjuvant therapy, or therapeutic effects of deslanoside on cancer and its potential mechanisms in SH-SY5Y and SK-N-SH neuroblastoma cell lines. Results showed that deslanoside inhibited the proliferation of SH-SY5Y cells, promoted apoptosis, and induced cell cycle arrest at the G0 / G1 phase; while in SK-N-SH cells, it induced cell cycle arrest at the G1 / S and G2 / M phases. Deslanoside induces apoptosis in neuroblastoma cells through intrinsic mitochondrial pathways and extrinsic caspase-dependent pathways. Furthermore, RNA sequencing analysis revealed that deslanoside regulates the expression of multiple genes and biological pathways, including cell cycle regulation, apoptosis, and the MAPK and mTORC1 signaling pathways. Further REACTOME and WIKIPATHWAYS analyses showed its enrichment in aging-related pathways, providing new insights into the mechanisms of deslanoside's inhibitory, alleviating, adjuvant therapy, or therapeutic effects on cancer. These findings suggest that Deslanoside possesses inhibitory, alleviating, adjuvant, or therapeutic properties in neuroblastoma cells, and support further repositioning of cardiac glycosides as potential therapeutic agents for neuroblastoma.

[0020] 2. The present invention provides a drug for inhibiting, alleviating, adjuvant treatment or treatment of cancer, which can inhibit the proliferation of neuroblastoma cells, induce apoptosis and / or necrosis of neuroblastoma cells, and induce cell cycle arrest in neuroblastoma cells.

[0021] The purpose of this invention is to provide new uses for Deslanoside in the preparation of drugs for inhibiting, alleviating, adjuvant treatment or treating neuroblastoma, and to provide new drug candidates for the treatment of neuroblastoma.

[0022] Deslanoside, also known as deslanoside C or deslanoside D, is a deacetylated derivative of deslanoside C and belongs to the class of fast-acting cardiac glycosides. It is a white crystalline powder, slightly soluble in methanol, and practically insoluble in water or chloroform. This drug works by inhibiting the Na+ ionization of myocardial cell membranes. + -K + ATPase enhances myocardial contractility (positive inotropic effect), improves hemodynamics, and slows heart rate (negative chronotropic effect). Clinically, it is mainly used for the rapid treatment of acute heart failure, atrial fibrillation, and flutter. This invention is the first to discover that Deslanoside has significant inhibitory, alleviating, adjuvant, or therapeutic activity against neuroblastoma, providing a new direction for the "drug repurposing" of cardiac glycosides and holding promise for adjuvant therapy or new drug development in high-risk neuroblastoma patients. Attached Figure Description

[0023] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0024] Figure 1 The inhibitory effect of deslanoside on the proliferation of SH-SY5Y and SK-N-SH cells and its IC50 value. 50 Measurement results; Figure 2 Flow cytometry analysis results of Deslanoside-induced cell cycle arrest in SH-SY5Y and SK-N-SH cells; Figure 3 Annexin V / PI double staining, Caspase-3 activity and mitochondrial membrane potential detection results for Deslanoside-induced apoptosis in SH-SY5Y and SK-N-SH cells; Figure 4 Western blot analysis results of MAPK / ERK, AKT, and mTORC1 signaling pathway-related proteins in SH-SY5Y and SK-N-SH cells after Deslanoside treatment; Figure 5 A diagram illustrating the mechanism of action of Deslanoside on cancer cells based on transcriptome analysis; Figure 6 A diagram showing that Deslanoside does not inhibit the proliferation of the non-cancer cell line HT22. Detailed Implementation

[0025] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0026] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0027] Example 1 1. Experimental Materials The human neuroblastoma SH-SY5Y cell line (Expasy: CVCL_0019) was provided by Professor Wang Yun of the Institute of Neuroscience, Peking University.

[0028] The SK-N-SH cell line was purchased from Procell Biotechnology Co., Ltd. (catalog number: CL-0214).

[0029] The HT22 mouse hippocampal neuron cell line was purchased from Servicebio (catalog number: STCC20011P-1) (see "https: / / www.servicebio.cn / goodsdetail?id=12632&specificationId=725&specificationGroupId").

[0030] Deslanoside (CAS No.: 17598-65-1) was purchased from MedChemExpress (Shanghai, China) and dissolved in DMSO according to the instructions to prepare stock solutions of 10 mmol / L and 80 mmol / L.

[0031] 2. Cell Culture and Drug Treatment SH-SY5Y and HT22 cells were cultured in DMEM medium (Gibco, USA) containing 10% fetal bovine serum (Gibco, USA) and 1% penicillin / streptomycin (Gibco, USA) under a humidified environment of 37°C and 5% CO2. After reaching 70%-80% confluence, cells were passaged using 2.5% trypsin (Gibco, USA) and seeded at a density of 250,000 cells per well in 24-well plates. The medium was changed every two days.

[0032] SK-N-SH cells were cultured in MEM medium (Pricella, China) containing 10% fetal bovine serum and 1% penicillin / streptomycin, under a humidified environment of 37°C and 5% CO2. After reaching 70%-80% confluence, cells were passaged using 2.5% trypsin (Gibco, USA) and seeded at a density of 250,000 cells per well in 24-well plates. The medium was changed every two days.

[0033] The SH-SY5Y cell assay consisted of six groups: a blank control group (no drug), a solvent control group (1‰ DMSO), and four Deslanoside treatment groups (final concentrations of 1.25 μM, 2.5 μM, 5 μM, and 10 μM, respectively). The SK-N-SH cell assay also consisted of six groups: a blank control group (no drug), a solvent control group (1‰ DMSO), and four Deslanoside treatment groups (final concentrations of 10 μM, 20 μM, 40 μM, and 80 μM, respectively).

[0034] After processing, the 24-well plate was placed in a zenCELL Owl 24-channel microscope (innoME GmbH, Germany) and observed continuously in an incubator for about 48 hours, taking pictures every hour.

[0035] The results showed that deslanoside selectively induced apoptosis in neuroblastoma cells in a dose-dependent manner. In SH-SY5Y cells, significant cell death was observed after 8 hours of treatment. Figure 1 The cell death rate was significantly higher after 24 hours of treatment with higher concentrations than after 8 hours of treatment. Figure 1 (A). This experiment involved hourly imaging, with the 24-hour half-maximal inhibitory concentration (IC50) determined. 50 The value was 2.255 ± 0.73 μM (95% confidence interval: 1.43–3.08 μM). Figure 1 China A and Figure 1 (C)

[0036] Significant cell death was observed in SK-N-SH cells after 48 hours of treatment. Figure 1 The cell death rate after 48 hours of treatment with higher concentrations was significantly higher than that after 24 hours of treatment. Figure 1 (Middle B). 48h IC 50 The value was 23.29 ± 6.12 μM (95% confidence interval: 12.39–34.18 μM). Figure 1 B, Figure 1 (C)

[0037] 3. Cell proliferation detection To further clarify the effect of deslanoside on cell proliferation, an EdU incorporation assay was used to detect DNA synthesis. The BeyoClick EdU Cell Proliferation Detection Kit (Beyotime, China) was used to assess cell proliferation. The specific procedure was as follows: cells were inoculated at 4 × 10⁶ cells per well. 5 Cells were seeded at a density of [number] cells per cell in 20 mm glass-bottomed culture dishes. SH-SY5Y cells were treated with a final concentration of 2.5 μM Deslanoside for 24 h, and SK-N-SH cells were treated with a final concentration of 20 μM Deslanoside for 48 h. Then, 20 μM EdU was added, and the cells were incubated at 37°C for 2 h. After fixation and permeabilization with 4% paraformaldehyde, the cells were stained with EdU Azide Alexa Fluor 488. The cells were observed and photographed using a laser confocal microscope, and the images were analyzed using ImageJ software. Results are shown below. Figure 1 As shown in Figure D, after 24 h of treatment with 2.5 μM Deslanoside, the EdU fluorescence intensity of SH-SY5Y cells was significantly lower than that of DMSO and the control group (EdU positivity rate: 2.5 μM vs DMSO = 21.20% ± 11.72% vs 92.94% ± 17.98%). p <0.0001, n =10; 2.5μM vs control = 21.20%±11.72% vs 93.34%±19.26%, p <0.0001, n =10)( Figure 1 (D). EdU is 5-ethynyl-2'-deoxyuridine.

[0038] Similarly, after SK-N-SH cells were treated with 20 μM Deslanoside for 48 h, the EdU fluorescence intensity was also significantly reduced (EdU positivity rate: 20 μM vs DMSO = 13.65% ± 5.25% vs 80.64% ± 12.89%, p < 0.0001). n =10; 20 μM vs control = 13.65% ± 5.25% vs 81.26% ± 12.38%, p < 0.0001, n =10)( Figure 1 (D).

[0039] It is known that Deslanoside inhibits the proliferation of SH-SY5Y and SK-N-SH cells in a dose-dependent manner.

[0040] 4. Cell cycle analysis Cells were distributed at a rate of 4 × 10⁶ cells per well. 5Cells were seeded at a density of [number] cells per well in 6-well plates. SH-SY5Y cells were treated with 2.5 μM Deslanoside for 24 h, and SK-N-SH cells were treated with 20 μM Deslanoside for 48 h. After collection, cells were fixed with 70% ethanol for 24 h, stained with PI, and cell cycle distribution was analyzed using an Attune NXT flow cytometer (Invitrogen). Controls included: blank control group (no drug, only culture medium and solvent) and control group (1‰ DMSO).

[0041] The results showed that after 24 h of treatment with 2.5 μM Deslanoside, the proportion of SH-SY5Y cells in G0 / G1 phase significantly increased, while the proportion in S phase significantly decreased (G0 / G1 phase: 2.5 μM vs DMSO = 47.46% ± 7.38% vs 42.46% ± 5.89%). p =0.0373, n =8; 2.5μM vs control = 47.46%±7.38% vs 39.21%±6.05%, p =0.0014, n =8; S phase: 2.5μM vs DMSO = 32.34%±9.93% vs 44.12%±8.21%, p =0.0733, n =8; 2.5μM vs control = 32.34%±9.93% vs 44.48%±5.89%, p =0.0005, n =8)( Figure 2 (A)

[0042] In SK-N-SH cells, treatment with 20 μM Deslanoside for 48 h significantly increased the proportions in both G0 and G1 phases (G0 / G1 phase: 20 μM vs control = 54.72% ± 6.87% vs 44.02% ± 1.86%, p = 0.0140, n = 5). Unlike SH-SY5Y cells, SK-N-SH cells also showed a significantly increased proportion in the G2 / M phase (G2 / M phase: 20 μM vs control = 13.52% ± 5.52% vs 5.60% ± 0.48%, p = 0.0175, n = 5). Figure 2 (B)

[0043] Western blot analysis further validated the expression changes of cell cycle-related proteins Cyclin B1 and Cyclin D1. The results showed that Deslanoside significantly inhibited the expression of these two proteins. Figure 2 C and Figure 2The presence of D indicates that it exerts inhibitory, alleviating, adjuvant, or therapeutic effects on tumors by interfering with G1 / S and G2 / M phase regulation.

[0044] 5. Apoptosis detection Apoptosis was assessed using a Caspase-3 activity assay kit (Beyotime, China). The specific procedure was as follows: 4 × 10⁴ cells per well. 5 Cells were seeded at a density of 100 cells per glass-bottomed culture dish. SH-SY5Y cells were treated with 2.5 μM Deslanoside for 24 h, and SK-N-SH cells were treated with 20 μM Deslanoside for 48 h.

[0045] Control: Blank control group: No drug, only culture medium.

[0046] DMSO: Solvent control group: 1‰ DMSO.

[0047] Add Annexin V-mCherry, GreenNuc™ Caspase-3 substrate and Hoechst 33342 staining solution, incubate in the dark and observe under a fluorescence microscope.

[0048] In addition, changes in mitochondrial membrane potential were detected using the Mito-Tracker Red CMXRos and Annexin V-FITC double staining kits, and the Annexin V-FITC / PI double staining kit was used to distinguish between apoptotic and necrotic cells.

[0049] Fluorescence microscopy revealed that SH-SY5Y cells treated with 2.5 μM Deslanoside showed a significant increase in the proportion of apoptotic cells (Annexin V positivity rate: 2.5 μM vs DMSO = 17.07% ± 5.61% vs 4.41% ± 2.34%). p <0.0001, n =15; 2.5μM vs control = 17.07%±5.61% vs 4.34%±2.65%, p <0.0001, n =15)( Figure 3 (A)

[0050] Treatment of SK-N-SH cells with 20 μM Deslanoside significantly increased the proportion of apoptotic cells (Annexin V positivity rate: 20 μM vs DMSO = 13.80% ± 9.43% vs 6.25% ± 2.66%). p =0.0315, n=12; 20μM vs control = 13.80%±9.43% vs 6.25%±2.75%, p =0.0319, n =12)( Figure 3 China A and Figure 3 (C)

[0051] Mitochondrial membrane potential assays showed that after Deslanoside treatment, the fluorescence intensity of Mito-Tracker Red CMXRos in SH-SY5Y and SK-N-SH cells was significantly reduced, indicating a decrease in mitochondrial membrane potential, suggesting that it induces apoptosis through the mitochondrial pathway. Figure 3 B, Figure 3 (C)

[0052] Furthermore, PI staining showed an increased proportion of late-stage apoptotic / necrotic cells in the Deslanoside treatment group. Caspase-3 activity assays revealed that Deslanoside significantly increased caspase-3 activity in both cell types, suggesting that it induces apoptosis via a caspase-dependent pathway. Figure 3 (D).

[0053] 6. Western blot analysis Cells were distributed at a rate of 4 × 10⁶ cells per well. 5 Cells were seeded at a density of [number] cells per well in 6-well plates. SH-SY5Y cells were treated with 2.5 μM Deslanoside for 24 h, and SK-N-SH cells were treated with 20 μM Deslanoside for 48 h. DMSO treatment groups included a blank control group (no drug, only culture medium) and a solvent control group (1‰ DMSO). After treatment, cells were collected, and total protein was extracted using RIPA lysis buffer. Protein concentration was determined by the BCA method. The protein concentration range at loading was 20–30 μg. After separation by 10% SDS-PAGE, the protein was transferred to a membrane and incubated overnight at 4°C with primary antibody (e.g., p-ERK, ERK, p-AKT, AKT, Cyclin B1, Cyclin D1, etc.). Secondary antibody incubation was performed at room temperature for 1 h. ECL imaging was performed, and band quantification was performed using ImageJ software.

[0054] According to previous literature, the PI3K / Akt / mTOR and RAS-MAPK pathways play key roles in NB cell proliferation. Western blot results showed that after 24 h of treatment with 2.5 μM Deslanoside, the p-ERK / ERK ratio of SH-SY5Y cells was significantly increased (p-ERK / ERK: 2.5 μM vs DMSO = 62.81% ± 38.03% vs 46.45% ± 32.80%). p=0.0256, n =9; 2.5μM vs control = 62.81%±38.03% vs 41.59%±22.95%, p =0.0495, n =9)( Figure 4 The levels of p-AKT and p-S6 did not change significantly. In SK-N-SH cells, after treatment with 20 μM Deslanoside for 48 h, the levels of p-ERK, p-AKT, and p-S6 did not change significantly.

[0055] 7. RNA sequencing and bioinformatics analysis RNA was extracted from SH-SY5Y cells treated with 10 μM Deslanoside for 4 days in the DMSO control group and sent to the Chigene Translational Medicine Research Center for RNA sequencing. Differentially expressed genes (DEGs) were screened using the DEGseq R package, with the criteria being an adjusted p-value <0.05 and |log2 fold change| ≥1. GO and KEGG pathway enrichment analysis was performed using the DAVID database, and volcano plots and GO analysis plots were generated using the ggplot2 and GOplot packages in R. Figure 5 ).according to Figure 5 It can be seen that a total of 451 genes meet these DEG criteria, including 248 upregulated genes and 203 downregulated genes. Figure 5 (A). To further explore the biological significance of these DEGs, we performed gene ontology (GO) analysis, classifying them according to their involvement in biological pathways, cellular components, and molecular functions. GO enrichment analysis identified 37 significantly enriched differentially expressed genes (p<0.05) and their main biological pathways. Figure 5 B, Figure 5 (C). These pathways mainly include cell cycle regulation, apoptosis, G1 / S transition in mitotic cell cycle, negative regulation of cell adhesion, negative regulation of cell proliferation, and regulation of MAPK and mTORC1 signaling pathways. Furthermore, GO analysis of cellular components showed that, compared to the SH-SY5Y control group, these differentially expressed genes were significantly localized in the nucleus, nucleoplasm, cytoplasm, and mitochondria.

[0056] 8. Deslanoside does not inhibit cell proliferation in non-cancer cell lines. To verify whether deslanoside also has cytotoxicity on non-cancer cell lines, cell experiments were conducted using HT22 mouse hippocampal neurons (a recognized primary neuron-like model). The HT22 cell experiments were divided into six groups: a control group (no drug), a solvent control group (1‰ DMSO), and four deslanoside treatment groups (final concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM, respectively).

[0057] After treatment, the 24-well plates were placed in a zenCELL Owl 24-channel microscope (innoME GmbH, Germany) and continuously observed in an incubator for approximately 48 hours, with images taken every hour. The results showed that deslanoside at a concentration of 20 μM (48 h treatment) did not induce significant cell death in HT22 cells, supporting its safety profile in non-cancerous neuronal cells. Figure 6 ).

[0058] In summary, this invention evaluated the inhibitory, alleviating, adjuvant, or therapeutic activities of deslanoside in human SH-SY5Y and SK-N-SH neuroblastoma cell lines. The study found that deslanoside effectively inhibited the proliferation of cancer cell lines (SH-SY5Y and SK-N-SH), promoted apoptosis, and induced cell cycle arrest, while having no effect on the non-cancer cell line (HT22). RNA sequencing analysis further revealed that differentially expressed genes were significantly enriched in cell cycle regulation, apoptosis, and signaling pathways such as MAPK and mTORC1, providing new insights into the mechanisms by which deslanoside inhibits, alleviates, adjuvants, or treats cancer.

[0059] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. The use of cardiac glycosides or their pharmaceutically acceptable salts or derivatives in the preparation of products for the inhibition, relief, adjuvant treatment or treatment of cancer.

2. The application according to claim 1, characterized in that, The cardiac glycosides mentioned include deslanoside.

3. The application according to claim 1, characterized in that, The inhibition, relief, adjuvant therapy, or treatment of cancer includes the inhibition, relief, adjuvant therapy, or treatment of neuroblastoma.

4. The application according to claim 3, characterized in that, The inhibition, relief, adjuvant therapy, or treatment of neuroblastoma includes at least one of the following: 1) Regulate the MAPK and / or mTORC1 signaling pathways to inhibit the proliferation of neuroblastoma cells; 2) Induces apoptosis and / or necrosis of neuroblastoma cells; 3) Inducing cell cycle arrest in neuroblastoma cells; the cell cycle arrest includes one or more of G0 / G1 phase arrest, G1 / S phase arrest and G2 / M phase arrest.

5. The application according to claim 1, characterized in that, The products include pharmaceuticals.

6. A drug for inhibiting, alleviating, adjuvantly treating, or treating cancer, characterized in that, Includes the cardiac glycosides as described in any one of claims 1-5, or pharmaceutically acceptable salts or derivatives thereof, and pharmaceutically acceptable carriers or excipients; optionally, the cardiac glycosides include deslanoside.

7. The drug according to claim 6, characterized in that, The dosage form of the drug includes liquid, semi-solid, or solid dosage forms.

8. The drug according to claim 6, characterized in that, The dosage form of the drug includes one or more of the following: injection, oral, sustained-release, and targeted formulation.

9. The medicament according to claim 8, characterized in that, The oral preparations include one or more of tablets, granules, and capsules; the injectable preparations include injection solutions.

10. The medicament according to any one of claims 5-9, characterized in that, The excipients include any one or a combination of at least two of the following: carrier, diluent, excipient, filler, binder, wetting agent, disintegrant, emulsifier, cosolvent, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent, or buffer.

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