Use of idebenone for the preparation of a medicament for the treatment of osteosarcoma
By using idebenone preparations to inhibit the proliferation and invasion of osteosarcoma cells, the problems of chemotherapy drug resistance and severe side effects have been solved, thus improving the effectiveness of osteosarcoma treatment and patient survival rates.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
- Filing Date
- 2023-08-29
- Publication Date
- 2026-06-05
AI Technical Summary
Current treatments for osteosarcoma suffer from problems such as chemotherapy drug resistance and significant side effects, as well as insensitivity to radiotherapy, resulting in low patient survival rates and a lack of effective new drug solutions.
Idebenone is used as an antioxidant to inhibit osteosarcoma cell proliferation and invasion, and prevent metastasis. It is prepared into oral or injectable formulations for the treatment of osteosarcoma.
It effectively inhibits the proliferation and invasion of osteosarcoma cells, reduces metastasis, lowers the side effects of chemotherapy drugs, and improves patient survival rates.
Smart Images
Figure CN117159522B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of medicine, and in particular relates to the application of idebenone in the preparation of drugs for treating osteosarcoma. Background Technology
[0002] Osteosarcoma is the most common malignant bone tumor, accounting for about one-third of all malignant bone tumors. It is most common in children and adolescents aged 10-20 years and originates from primitive mesenchymal stem cells. In children, it frequently occurs in the distal femur, proximal tibia, and proximal humerus. Clinically, it presents as a tender soft tissue mass at the metaphysis of long bones. Characteristic radiographic findings include a sunburst peristeal reaction and Codman's triangle. Rapid turnover of damaged bone cells can lead to elevated alkaline phosphatase (ALP) and lactate dehydrogenase (LDH). It is highly malignant, aggressive, and progresses very rapidly, easily metastasizing to the lungs via the bloodstream in the early stages of the tumor, resulting in a high mortality rate and making it a clinically challenging malignant tumor.
[0003] Osteosarcoma has several histological subtypes, including typical osteosarcoma (45%), fibroblastic osteosarcoma (9%), chondroblastic osteosarcoma (27%), degenerative osteosarcoma (17%), capillary osteosarcoma, low-grade central osteosarcoma, and other subtypes (2%). Typical osteosarcoma commonly occurs in the most active epiphyseal growth sites, such as the metaphysis of long tubular bones, most frequently in the distal femur, proximal tibia, and proximal humerus. It also frequently occurs in flat bones such as the pelvis, skull, scapula, ribs, and vertebrae.
[0004] Osteosarcoma often begins as intermittent, dull pain, which quickly transforms into persistent, severe pain. The initial local pain is intermittent and dull, rapidly becoming persistent and severe, especially at night, with significant tenderness. Initially, there is mild local swelling, which expands over time as the tumor grows, potentially forming an eccentric spindle-shaped swelling or mass. Sclerosing osteosarcoma is as hard as stone, while osteolytic osteosarcoma is soft and elastic like rubber. The local skin becomes shiny due to swelling, skin temperature increases, and veins become distended. Osteosarcoma near joints can cause limited joint movement. Local muscle atrophy occurs, leading to limping in the lower limbs. In advanced stages with severe bone destruction, pathological fractures can occur. Systemic symptoms appear early, often including low-grade fever, fatigue, weight loss, anemia, and progressive weakness, eventually leading to cachexia. Lung metastasis is common.
[0005] The pathogenesis of osteosarcoma can be divided into traditional mechanisms and molecular mechanisms. Traditional mechanisms include tumor factor mechanisms, central sensitization, and peripheral sensitization mechanisms. Tumor factor mechanisms mainly include: the gradual compression and damage of peripheral nerves during the growth and proliferation of tumor cells; the release of various inflammatory factors by tumor cells and related immune cells; and local bone tissue hypoxia, decreased pH, and extracellular hypercalcemia, leading to bone cancer pain.
[0006] The molecular mechanisms include osteoclast activation, alterations in ion channels or receptors, changes in signaling pathways, and neurochemicals. The molecular mechanism of osteoclast activation involves the proliferation of osteoclasts, the secretion of related bioactive substances, and the induction of bone destruction leading to bone instability, which in turn causes pain by stimulating mechanoreceptors in the periosteum. The molecular mechanism of altered ion channels or receptors is due to the abundance of ion channels and receptors associated with bone cancer pain, such as N-methyl-D-aspartate (NMDA) receptors, transient receptor potential vanillic acid subtype 1 (TRPV1) receptors, P2X receptors, sodium ion channels, potassium ion channels, and calcium ion channels. Alterations in ion channels and receptors can increase the excitability of sensory neurons and cause abnormal nerve impulse firing, inducing pain sensitization. In bone cancer pain models, ion channel or receptor antagonists can reverse thermal hyperalgesia. The molecular mechanisms of signaling pathway alteration have been discovered through research on signaling pathways. Multiple signaling pathways play important roles in bone cancer pain, such as the Ephrin B-EphB signaling pathway, the MCP-1-ERK signaling pathway, the MEK / ERK signaling pathway, the NF-κB signaling pathway, and the PAR2-NF-κB signaling pathway. In the molecular mechanisms of neurochemicals, tumor cells and various immune cells, such as macrophages, T cells, and neutrophils, can secrete a variety of substances that excite primary afferent nerves, such as tumor necrosis factor (TNF-α), interleukin-1 (IL-1), endothelin-1 (ET-1), prostaglandins (PGE), and nerve growth factor (NGF), as well as various types of growth factors. These substances bind to their corresponding receptors on primary afferent nerves, causing neuronal excitation and transmitting nociceptive information to the central nervous system, thereby inducing pain. Injecting inhibitors or antagonists of related factors and receptors into bone cancer pain models can partially block or reverse hyperalgesia, suggesting that these factors are involved in the formation of bone cancer pain.
[0007] Common treatments for cancer include surgery, radiotherapy, chemotherapy, and immunotherapy. However, because osteosarcoma lacks typical pathological mutations, chromosomal translocations, or gene abnormalities, research into new treatments primarily focuses on increasing the variety of chemotherapy drugs or increasing the dosage of existing standard chemotherapy medications.
[0008] However, current chemotherapy drugs used for osteosarcoma often exhibit drug resistance and significant side effects. The main mechanisms of chemotherapy drug resistance include: methotrexate is transported into cells via reduced folate carriers (RFCs), but studies have reported reduced RFC expression in 65% of pathological biopsies in osteosarcoma, leading to methotrexate resistance; doxorubicin resistance is primarily caused by a glycoprotein (Pgp) encoded by the multidrug resistance gene 1 (MDR1), an AIP-dependent membrane protein that pumps doxorubicin out of tumor cells by consuming ATP; glutathione S-transferase (GST) can transfer glutathione groups to chemotherapy drugs, leading to drug inactivation; cisplatin can induce GSTP1 expression, thereby inhibiting the efficacy of cisplatin.
[0009] The corresponding side effects are as follows: Although methotrexate has a smaller bone marrow suppression effect compared to other chemotherapy drugs, it and its products can deposit in the renal tubules, causing fatal acute renal failure. Methotrexate is almost entirely metabolized by the kidneys, which can lead to a vicious cycle. Doxorubicin has strong cardiotoxicity, and it is dose-dependent. A French study reported that using 420 mg / m²... 2 After a period of treatment with doxorubicin, 39% of the subjects developed myocarditis, while those receiving only 137.5 mg / m² experienced similar outcomes. 2 Only 23% of the treated subjects developed myocarditis; however, the standard dose of doxorubicin chemotherapy is 450 mg / m². 2 Cisplatin can cause damage to renal tubular epithelial cells, leading to decreased glomerular function and charge imbalance (Fanconi syndrome). Studies have reported that 60% of children receiving the standard dose of 500-600 mg / m²... 2 Chemotherapy can lead to a decrease in glomerular filtration rate. Cisplatin's ototoxicity can cause permanent damage, with an incidence rate as high as 78% in osteosarcoma patients. Additionally, up to 90% of patients may develop hypomagnesemia; ifosfamide can cause not only chronic renal insufficiency but also hemorrhagic cystitis. Furthermore, ifosfamide's use as a chemotherapy drug for osteosarcoma is controversial, and its efficacy is uncertain.
[0010] Because osteosarcoma is not sensitive to radiotherapy, it is rarely treated clinically as a single therapy. The current standard clinical treatment for osteosarcoma is a comprehensive approach of neoadjuvant chemotherapy + surgical resection + adjuvant chemotherapy. Even so, the overall survival rate of osteosarcoma has not significantly improved in the past 40 years.
[0011] The main reasons for this are as follows: First, the development of chemotherapy regimens has reached a bottleneck; no better regimens have been explored, and there are still debates regarding drug selection, dosage, administration methods, timing, and requirements. Second, chemotherapy drugs have significant side effects. As cytotoxic drugs, the adverse reactions of chemotherapy drugs can cause considerable damage to patients. The dose-response curves of most chemotherapy drugs show that their cytotoxic effects occur when a certain plasma drug concentration is reached, killing some normally growing cells. For most chemotherapy drugs, toxic reactions such as bone marrow suppression, hair loss, nausea, and vomiting will occur. Furthermore, chemotherapy drugs are difficult to use continuously for extended periods. This limits the application of chemotherapy drugs for osteosarcoma patients with significant toxic side effects. To avoid these side effects, growth factors, protective drugs, and other supportive therapies are used to mitigate the toxicity of chemotherapy drugs, which also increases the economic burden on patients. More importantly, chemotherapy drugs easily lead to drug resistance, which necessitates increasing the dosage of chemotherapy drugs, and even then, the desired therapeutic effect may not be achieved, resulting in a poor prognosis.
[0012] While targeted therapy for osteosarcoma has made some progress, its clinical application remains very limited. This is because, compared to chemotherapy drugs, although the side effects of targeted drugs are significantly fewer, they are still considerable. Secondly, although current research has made progress in understanding the specific molecular mechanisms of targeting osteosarcoma in in vitro models, this has not yet translated into clinical results. One major reason limiting research is likely the need for careful patient selection for targeted therapy. Therefore, research should be conducted based on the presence of specific biomarkers.
[0013] Therefore, developing a new, highly safe drug to treat osteosarcoma patients and thus improve their survival rate is a pressing scientific problem that needs to be solved. Summary of the Invention
[0014] In order to provide a novel drug for treating osteosarcoma patients, the present invention provides the use of idebenone in the preparation of a drug for treating osteosarcoma.
[0015] According to one aspect of the invention, there is provided the use of idebenone or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating osteosarcoma.
[0016] Idebenone is an antioxidant that enhances mitochondrial respiratory activity, glucose utilization, and protects the mitochondrial membrane, thus exerting its antioxidant effect. Therefore, it can be used for ischemic cerebrovascular diseases. Existing literature reports that the main effects of idebenone include: antioxidation, stimulation of neurotrophic factor production, improvement of neurotransmitters, and treatment of skin aging. Therefore, in current technology, idebenone can stimulate the production of neurotrophic factors to treat Alzheimer's disease, and can also improve neurotransmitters in the brain to treat vascular dementia; furthermore, due to its antioxidant properties, idebenone is also commonly used in cosmetics to combat skin aging. The chemical structural formula of idebenone is:
[0017]
[0018] Preferably, the drug is used to inhibit the proliferation of osteosarcoma.
[0019] Preferably, the drug is used to prevent osteosarcoma metastasis or inhibit the growth of micrometastases.
[0020] Preferably, the active ingredient of the drug also includes pharmaceutically acceptable excipients or auxiliary ingredients.
[0021] Preferably, the drug is an oral or injectable formulation; wherein, the oral formulation includes a solid formulation, a liquid formulation, or a suspension formulation.
[0022] The application of idebenone or its pharmaceutically acceptable salt in the preparation of drugs for treating osteosarcoma provided by this invention, through in vitro proliferation experiments (CCK8 assay) and invasion experiments (transwell invasion assay) of osteosarcoma cell lines, with the CCK8 proliferation assay quantifying cell viability and the transwell invasion assay quantifying the number of cells that penetrated; and the construction of nude mouse CDX (cell-derived xenograft) models, PDX (human xenograft) models, and tibial orthotopic / metastatic tumor models, with intraperitoneal injection of idebenone, with the CDX model quantifying tumor volume and weight, the PDX model quantifying tumor volume and weight, and the tibial orthotopic / metastatic tumor model quantifying tumor volume, affected limb weight, and number of lung metastases, fully demonstrates that idebenone can inhibit the proliferation, invasion, and metastasis of osteosarcoma. Attached Figure Description
[0023] Figure 1 The results of the proliferation experiment (CCK8 assay) conducted in three osteosarcoma cell lines with different concentrations of idebenone in Example 1 are shown.
[0024] Labeling explanation: (a) U2OS osteosarcoma cell activity results, (b) indicates activity results of three osteosarcoma cell lines (143B), and (c) indicates activity results of the MG63 osteosarcoma cell line.
[0025] Figure 2Example 2 shows the results of idebenone inhibiting the invasion of osteosarcoma cell lines in vitro;
[0026] Labeling explanations: (a1) and (a2) are microscopic images of the U2OS osteosarcoma cell line after crystal violet staining; (a3) is the statistical result of the number of U2OS osteosarcoma cells in the control group and the idebenone group; (b1) and (b2) are microscopic images of the 143B osteosarcoma cell line after crystal violet staining; (b3) is the statistical result of the number of 143B osteosarcoma cells in the control group and the idebenone group; (c1) and (c2) are microscopic images of the MG63 osteosarcoma cell line after crystal violet staining; (c3) is the statistical result of the number of MG63 osteosarcoma cells in the control group and the idebenone group.
[0027] Figure 3 The results of the CDX model tumor experiment in Example 3;
[0028] Labeling explanation: (a) shows the tumor tissue of the CDX model, with the upper part of the image representing the control group and the lower part representing the idebenone group treated with intraperitoneal injection; (b) shows the statistical results of the tumor tissue volume of the control group and the idebenone group; (c) shows the statistical results of the tumor tissue weight of the control group and the idebenone group.
[0029] Figure 4 The results of the PDX model tumor experiment in Example 4;
[0030] Labeling explanation: (a) shows tumor tissue in nude mouse PDX model, with the upper part of the image representing the control group and the lower part representing the idebenone group administered via intraperitoneal injection; (b) shows the statistical results of tumor tissue volume in the control group and the idebenone group; (c) shows the statistical results of tumor tissue weight in the control group and the idebenone group.
[0031] Figure 5 The results of experiments on the in situ / metastatic tumor model and lungs in Example 5;
[0032] Labeling explanations: (a) shows tumor tissue from the in situ / metastatic tumor model, with the upper image representing the control group and the lower image representing the idebenone group treated with intraperitoneal injection; (b1) and (b2) show lung tumor tissue from the control group and the idebenone group, respectively; (c) shows the statistical results of tumor tissue volume from the control group and the idebenone group; (d) shows the statistical results of tumor tissue weight from the control group and the idebenone group; (e) shows the statistical results of lung metastatic nodules from the control group and the idebenone group. Detailed Implementation
[0033] To enable those skilled in the art to better understand the technical solutions of this invention, the technical solutions of this invention will be clearly and completely described below with reference to the embodiments and accompanying drawings. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this invention.
[0034] Example 1: Idebenone inhibits the proliferation of osteosarcoma cell lines in vitro.
[0035] 1. Experimental Methods
[0036] Using three osteosarcoma cell lines—U2OS, MG63, and 143B—as research subjects, idebenone (catalog number: S2605) purchased from Selleck was diluted with sterile DMSO. Idebenone concentration gradients of 0, 7.5, 15, 30, and 50 nM were set, along with time gradients of 24 h and 48 h. The drug was added to the three cell lines, and cell viability was detected using the CCK8 assay after 24 or 48 h. The specific protocol is shown in the table below.
[0037] Table 1. Experimental cell grouping
[0038]
[0039]
[0040] 2. Experimental Results
[0041] like Figure 1 The experimental results show that different concentrations of idebenone have varying inhibitory effects on the three osteosarcoma cell lines U2OS, 143B, and MG63. However, the results indicate that idebenone has an inhibitory effect on the proliferation of all three osteosarcoma cell lines. Figure 1 *p<0.05, **p<0.01.
[0042] Example 2: Idebenone inhibits the invasion of osteosarcoma cell lines in vitro.
[0043] 1. Experimental Methods
[0044] Three osteosarcoma cell lines, U2OS, MG63, and 143B, were used as research subjects. Idebenone (catalog number: S2605) purchased from Selleck was diluted with sterile DMSO. Corning's matrix gel (catalog number: 356234) was diluted to 0.3 mg / ml with high-glucose DMEM medium and seeded into 8 μm transwell chambers. The three osteosarcoma cell lines were seeded into the transwell chambers, divided into a control group and a 30 nM idebenone group. After 24 h, the cells in the lower chamber were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and photographed under a microscope. The specific protocol is shown in the table below:
[0045] Table 2. Experimental cell grouping
[0046]
[0047] 2. Experimental Results
[0048] like Figure 2 The cell images and cell counts obtained after crystal violet staining under a microscope show that idebenone can inhibit the invasion of three osteosarcoma cell lines, among which... Figure 2 *p<0.05, **p<0.01.
[0049] Example 3: Idebenone inhibits the growth of osteosarcoma in a nude mouse CDX model.
[0050] 1. Experimental Methods
[0051] Idebenone (catalog number: S2605) purchased from Selleck was diluted with sterile solvent (solvent formulation: 5% DMSO + 30% PEG300 + 5% TWEEN80 + 60% sterile water) and administered at a dose of 15 mg / kg based on the weight of nude mice. Ten 4-week-old nude mice were randomly divided into two groups and a CDX model was established. Administration began on day 8 after modeling, with intraperitoneal injections administered for 5 consecutive days each week for a total of 4 weeks. Tumors were then collected from the nude mice, their volume measured, and weighed. The CDX model preparation procedure was as follows: the right axilla of the nude mouse was routinely disinfected, and a 143B cell suspension resuspended in PBS was injected into the right axilla of the nude mouse using a 1 ml syringe, at a dose of 2 x 10⁻⁶ mg / kg. 6 1 cell / 200ul / animal. Specific protocol is shown in the table below:
[0052] Table 3. Grouping of experimental animals
[0053]
[0054] 2. Experimental Results
[0055] like Figure 3The CDX body tissue images and the statistical results of tumor tissue volume and mass shown indicate that idebenone can inhibit the growth of osteosarcoma in the CDX model, with *p<0.05 and **p<0.01.
[0056] Example 4: Idebenone inhibits the growth of osteosarcoma in a nude mouse PDX model.
[0057] 1. Experimental Methods
[0058] Idebenone (catalog number: S2605) purchased from Selleck was diluted with sterile solvent (solvent formulation: 5% DMSO + 30% PEG300 + 5% TWEEN80 + 60% sterile water) and administered at a dose of 15 mg / kg based on the weight of nude mice. Ten 4-week-old nude mice were randomly divided into two groups and a PDX model was established. Administration began on day 15 after modeling, with intraperitoneal injections administered for 5 consecutive days each week for a total of 4 weeks. Afterward, the tumor volume was measured and weighed. The PDX model preparation process was as follows: osteosarcoma tissue obtained during surgery was sent to the laboratory and cut into 2x2 mm tissue blocks. After anesthetizing the nude mice with tribromoethanol, they were placed prone on the operating table, and under routine disinfection and sterile conditions, a 2 mm incision was made on the right side of the mouse's back. The tumor tissue block was inserted subcutaneously using a biopsy needle, and the wound was sutured. Specific procedures are shown in the table below.
[0059] Table 4. Grouping of experimental animals
[0060]
[0061]
[0062] 2. Experimental Results
[0063] like Figure 4 The images of the tumor tissue and the statistical results of the tumor tissue volume and mass shown indicate that idebenone can inhibit the growth of osteosarcoma in the PDX model.
[0064] Example 5: Idebenone inhibits the growth and metastasis of osteosarcoma in a nude mouse tibial orthotopic / metastatic tumor model. 1. Experimental methods
[0065] Idebenone (catalog number: S2605) purchased from Selleck was diluted with sterile solvent (solvent formulation: 5% DMSO + 30% PEG300 + 5% TWEEN80 + 60% sterile water) and administered at a dose of 15 mg / kg based on the weight of nude mice. Ten 4-week-old nude mice were randomly divided into two groups and a tibial orthotopic / metastatic tumor model was established. Administration began on day 15 after modeling, with intraperitoneal injections administered for 5 consecutive weeks for 4 weeks. Afterward, the affected limbs were harvested, and the tumor size and weight of the affected limbs were measured. The preparation procedure for the tibial orthotopic / metastatic tumor model was as follows: After anesthetizing the nude mice with tribromoethanol, they were placed supine on the operating table, and under routine disinfection and sterile conditions, a PBS-resuspended 143B cell suspension was injected into the medullary cavity along the medial side of the left tibial plateau using a 25 μL microsyringe, at a dose of 1 x 10^6 cells / 20 μL / mouse. The specific protocol is shown in the table below.
[0066] Table 5. Grouping of experimental animals
[0067]
[0068] 2. Experimental Results
[0069] like Figure 5 The images shown are of tumor tissue in the affected limbs and lungs of nude mice, along with statistical results on the volume and mass of the tumor tissue. Idebenone can inhibit the growth of osteosarcoma and lung metastasis in the in situ / metastatic tumor model.
[0070] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.
Claims
1. The use of idebenone or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the treatment of osteosarcoma.
2. The use as described in claim 1, characterized in that, The drug is used to inhibit the proliferation of the osteosarcoma.
3. The use as described in claim 1, characterized in that, The drug is used to prevent osteosarcoma metastasis or inhibit the growth of micrometastases.
4. The use as described in any one of claims 1 to 3, characterized in that: The drug also includes pharmaceutically acceptable excipients.
5. The use as described in claim 4, characterized in that: The drug is an oral or injectable formulation; wherein the oral formulation includes a solid or liquid formulation.