Use of a combination of a bmi1 inhibitor and a myc inhibitor for the treatment of cancer

Abstract

The present application belongs to the technical field of biological medicine, and relates to the use of a combination of a BMI1 inhibitor and a Myc inhibitor for treating cancer. After the BMI1 inhibitor and the Myc inhibitor are combined for treating cancer, especially head and neck squamous cell carcinoma (HNSCC), the present application can effectively and continuously eliminate tumor stem cells, thereby inhibiting tumor growth, treating cancer, and preventing tumor recurrence.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to the use of a combination of BMI1 inhibitors and Myc inhibitors for the treatment of cancer. Background Technology

[0002] Head and neck squamous cell carcinoma (HNSCC) originates in the oral cavity, oropharynx, larynx, and hypopharynx, and is a highly aggressive type of tumor. This disease typically occurs in various parts of the oral cavity, including the tongue, floor of the mouth, buccal mucosa, gingiva, and lips. The development of HNSCC is associated with multiple factors, including environmental factors, lifestyle habits, and genetic factors. Because early symptoms of HNSCC are often subtle, patients are frequently diagnosed at an advanced stage, severely impacting their quality of life and prognosis.

[0003] Currently, the main treatment modalities for HNSCC include surgery, radiotherapy, and chemotherapy. However, due to the lack of obvious early symptoms, many patients are already in an advanced stage at the time of diagnosis. For locally advanced HNSCC, concurrent chemoradiotherapy (CRT) is the primary treatment. However, even after CRT treatment, approximately 50% of patients relapse within 3 years, indicating that current treatment modalities have limitations in controlling HNSCC.

[0004] In recent years, targeted therapy and immunotherapy have made significant progress in the treatment of hepatocellular carcinoma (HNSCC). For example, cetuximab, targeting EGFR, has been used in the treatment of HNSCC; immune checkpoint inhibitors (ICIs), such as PD-1 / PD-L1 antibodies, have shown potential in the treatment of HNSCC. However, the response rate of immunotherapy is not 100%, and approximately 70% to 90% of patients with recurrent spontaneous absorptive (RM-HNSCC) may not respond to ICIs; while the efficacy of targeted therapy is limited by tumor heterogeneity and drug resistance. This indicates that further research is needed on therapies for HNSCC to improve treatment outcomes.

[0005] Tumor recurrence, metastasis, and drug resistance are leading causes of death in cancer patients, likely stemming from a subset of cancer cells known as cancer stem cells (CSCs). Therefore, researchers are actively seeking potential targets to eliminate CSCs. Despite recent advancements in treatment, the 5-year survival rate for patients with hepatocellular carcinoma (HNSCC) remains relatively low due to tumor recurrence, metastasis, and drug resistance. Based on the CSC theory, to achieve optimal treatment outcomes, CSCs must be eradicated during continuous treatment of HNSCC.

[0006] Using an in vivo lineage tracing mouse model, recent research has shown that BMI1+ cells are slow-cycle CSCs in head neck squamous cell carcinoma (HNSCC) and are closely related to tumor recurrence, metastasis, and drug resistance. BMI1 (Blymphoma Mo-MLV insertion region 1) is a core member of the multicomb gene family, involved in regulating various fundamental cell biological processes, including cell proliferation, apoptosis, and senescence. Studies have shown that BMI1 is abnormally highly expressed in various human malignancies and is closely related to tumor development, stem cell homeostasis, and poor patient prognosis, making it a potential novel tumor diagnostic biomarker and therapeutic target. The small molecule inhibitor PTC-209 significantly downregulates BMI1 protein expression in HNSCC cells with a concentration- and time-dependent effect, possibly through post-transcriptional inhibition and ubiquitin-proteasome degradation pathways (see: Wang, Q., et al., Pharmacological inhibition of Bmi1 by PTC-209 impaired tumor growth in head neck squamous cell carcinoma. Cancer Cell Int, 2017.17:p.107.). PTC-209 treatment significantly increased ubiquitinated histone 2A in HNSCC cells, inhibiting BMI1 transcription and leading to derepression of its downstream target gene p16 (see: Storti, B., et al., Fluorescence imaging of biochemical relationship between ubiquitinated histone 2A and Polycomb complex protein BMI1. BiophysChem, 2019, 253: p. 106-225.). In vitro intervention with PTC-209 significantly inhibited cell proliferation and exhibited synergistic anticancer effects when combined with cisplatin and 5-FU. Furthermore, PTC-209 induced cell cycle arrest, promoted apoptosis, inhibited cell migration and invasion, reduced cell colony formation rate and tumor spheroid formation, and decreased the proportion of ALDH-positive cell subsets. A heterotopic xenograft model was established by subcutaneously injecting HNSCC cells into nude mice. The effect of intraperitoneal injection of PTC-209 on the growth of the transplanted tumor was observed. The results showed that PTC-209 may significantly inhibit the growth of head and neck squamous cell carcinoma xenografts by downregulating BMI1 expression and inhibiting cell proliferation. Our research shows that the combination therapy of anti-PD1 and cisplatin acts on BMI1 in HNSCC. +CSCs simultaneously inhibited the growth of HNSCC. In contrast, the pharmacological and genetic inhibitory effects of BMI1 eliminated BMI1+CSCs and activated PD1 blockade therapy, thereby inhibiting metastatic HNSCC and preventing HNSCC recurrence. Besides eliminating BMI1... + In addition to CSC, BMI1 inhibition also inhibits CD8 recruitment and activation. + T cells strongly induce an intrinsic immune response in tumor cells. Despite the elimination of BMI1 by PTC-209... + CSCs can limit the growth and metastasis of HNSCC, but this does not lead to tumor regression or eradication.

[0007] V-MYC, the myelocytomatosis viral oncogene homolog, is a proto-oncogene that encodes a nucleophosphoprotein that influences cell cycle progression, apoptosis, and transformation. It also functions as a transcription factor controlling gene expression. Increasing evidence suggests that MYC regulates tumorigenesis through multiple mechanisms and plays a functional role in up to 70% of human cancers. Multiple studies have demonstrated that MYC plays an important role in tumorigenesis and is considered an attractive target for cancer treatment (see: Han, H., et al., Small-Molecule MYC Inhibitors Suppress Tumor Growth and Enhance Immunotherapy. Cancer Cell, 2019, 36(5): p. 483-497e15; Holmes, AG, et al., A MYC inhibitor selectively alters the MYC and MAX cistromes and modulates the epigenomic landscape to regulate target gene expression. Sci Adv, 2022, 8(17): p. eabh3635; Zhang, Y., et al., The m(6)A demethylase ALKBH5-mediated upregulation of DDIT4-AS1 maintains pancreatic cancer stemness and suppresses chemosensitivity by activating the mTOR pathway. Mol Cancer, 2022, 21(1):p.174.). Recently, our research has shown that MYC is associated with lymph node metastasis and poor prognosis in HNSCC. MYCi975 is a small molecule inhibitor of MYC that can disrupt the MYC-MAX dimer, promoting MYC degradation and killing tumor cells. It increases proteasome-mediated MYC degradation by enhancing phosphorylation of MYC at threonine. However, whether MYCi975 is beneficial for the treatment of HNSCC is currently unclear.

[0008] In summary, the treatment of HNSCC faces multiple challenges, including tumor recurrence, metastasis, and drug resistance. Furthermore, treatment-related toxicities and side effects also need to be addressed. For example, while concurrent chemoradiotherapy is effective, its short-term and long-term toxicities remain significant, necessitating the search for new treatment strategies to reduce these side effects. The efficacy of immunotherapy and targeted therapy also requires further improvement. These findings suggest that further research is needed to overcome these challenges. Summary of the Invention

[0009] The purpose of this invention is to provide the use of a combination of BMI1 inhibitors and Myc inhibitors in the preparation of a medicament for treating cancer, so as to effectively and continuously eliminate tumor stem cells after the combined use of BMI1 inhibitors and Myc inhibitors in the treatment of cancer, especially head and neck squamous cell carcinoma (HNSCC), thereby inhibiting tumor growth, treating cancer, and preventing tumor recurrence.

[0010] To achieve this objective, in a basic embodiment, the present invention provides the use of a combination of BMI1 inhibitors and Myc inhibitors for the preparation of a medicament for treating cancer.

[0011] In a preferred embodiment, the present invention provides the use of a combination of a BMI1 inhibitor and a Myc inhibitor for the preparation of a medicament for treating cancer, wherein the BMI1 inhibitor is PTC-209.

[0012] In a preferred embodiment, the present invention provides the use of a combination of a BMI1 inhibitor and a Myc inhibitor for the preparation of a medicament for treating cancer, wherein the Myc inhibitor is MYCi975.

[0013] In a preferred embodiment, the present invention provides the use of a combination of a BMI1 inhibitor and a Myc inhibitor for the preparation of a medicament for treating cancer, wherein the mass ratio of the BMI1 inhibitor to the Myc inhibitor in the combination is 3:5.

[0014] In a preferred embodiment, the present invention provides the use of a combination of BMI1 inhibitors and Myc inhibitors for the preparation of a medicament for treating cancer, wherein the combination is in the form of a composition, preferably a solution.

[0015] In a preferred embodiment, the present invention provides the use of a combination of BMI1 inhibitors and Myc inhibitors for the preparation of a medicament for treating cancer, wherein the combination prepares the medicament for treating cancer by eliminating tumor stem cells.

[0016] In a preferred embodiment, the present invention provides the use of a combination of BMI1 inhibitors and Myc inhibitors in the preparation of a medicament for treating cancer, wherein the cancer is head and neck squamous cell carcinoma.

[0017] In a preferred embodiment, the present invention provides the use of a combination of BMI1 inhibitors and Myc inhibitors in the preparation of a medicament for treating cancer, wherein the cancer is oral squamous cell carcinoma.

[0018] The beneficial effects of this invention are that, when the BMI1 inhibitor and Myc inhibitor are used in combination for the treatment of cancer, especially head and neck squamous cell carcinoma (HNSCC), the invention can effectively and continuously eliminate tumor stem cells, thereby inhibiting tumor growth, treating cancer, and preventing tumor recurrence. Attached Figure Description

[0019] Figure 1 The results of Example 2 are shown below. A is a schematic diagram of time points in PTC209 treatment of PDX#3; B is the tumor volume (n=12) in different treatment groups, ns, with no statistical difference, p>0.05; *p<0.05 and **p<0.01, t test; C is the tumor weight (n=12) in different treatment groups, ns, with no statistical difference, p>0.05, t test; D is the flow cytometry analysis of tumors at different treatment times of PDX#3, where the +DEAB group is the control group to control the interference of background fluorescence in the experiment.

[0020] Figure 2 The results of Example 3 are shown below, where A represents tumor volume in different treatment groups; B represents overall survival in different treatment groups; and C represents median survival time of mice after treatment. The first column indicates the category of inter-group comparisons, the second column shows the statistical analysis results of inter-group comparisons using the log-rank test, * indicates a p-value less than 0.05, and ** indicates a p-value less than 0.01. The values ​​outside the parentheses in the third column represent the median overall survival of the two groups before and after "vs.", separated by "-". The percentages inside the parentheses indicate the percentage increase in overall survival between the group after "vs." and the group before "vs.".

[0021] Figure 3 The results are shown in Example 5, where A is a schematic diagram of the experimental design; B is Bmi1 in a frozen section of a tongue tumor from a transgenic mouse. + Image of tumor stem cells (tomato red fluorescence), white dashed line indicates the boundary between tumor and stromal tissue, scale bar, 50 μm; C is Bmi1 tumor in the tongue of transgenic mice. + Percentage chart of tumor stem cells, ns, no statistically significant difference, p>0.05; **p<0.01. Detailed Implementation

[0022] The following examples further illustrate specific embodiments of the present invention.

[0023] Example 1: Construction of a mouse model of human tumor xenograft in HNSCC (PDX model)

[0024] PDX model (Patient-Derived tumor Xenograft model) refers to a xenograft model in which fresh tumor tissue from a patient is transplanted into an immunodeficient mouse and grows in the environment provided by the mouse (see: Liu, Y., et al., Patient-derived xenograft models in cancer therapy: technologies and applications. Signal Transduct Target Ther, 2023.8(1):p.160.).

[0025] (I) Procedures before tumor tissue transplantation

[0026] The tumor specimens used in this experimental study were obtained from patients with head and neck squamous cell carcinoma who underwent surgery at Peking University School of Stomatology between 2020 and 2022, and were approved by the Biomedical Ethics Committee of Peking University School of Stomatology. The inclusion criteria for patients were: (1) pathological results confirming squamous cell carcinoma;

[0027] (2) The tumor is located on the patient's tongue; (3) The primary cancer and neck dissection were removed without preoperative radiotherapy or chemotherapy; (4) The patient voluntarily signed the informed consent form.

[0028] After removing the tumor tissue during the operation, avoiding the liquefied and necrotic central portion of the tumor, the most representative tumor tissue from the lesion was collected, cut into several small pieces, placed in PBS solution, and stored on ice before being sent to the animal laboratory. In a biosafety cabinet, the collected tumor tissue was placed in cell culture dishes for further processing. Visible liquefied, necrotic, and other non-viable tumor tissues were repeatedly rinsed with PBS and then cut into tissue blocks of approximately 2mm × 2mm × 2mm for subsequent experiments.

[0029] (II) Construction of the PDX model primary (P0)

[0030] In a biosafety cabinet, 6-8 week old female NOD / SCID mice (nude mice, Vitaliva) were anesthetized and fixed with pentobarbital. The skin on the back of the mice was disinfected with 75% alcohol, and the inoculation area was cleaned with a razor and disinfected again. A small incision of about 0.5 cm was made in the inoculation area with autoclaved tissue scissors, and the tissue was sharply dissected with the scissors. 3-4 pieces of prepared tumor tissue were implanted subcutaneously into the inoculation area, and the wound was sutured with a wound clip. The mice after tumor transplantation were housed in cages in a specific pathogen-free (SPF) animal laboratory. The length, width, and weight of the tumor were measured every 3 days using a miniature vernier caliper. The wound clip was removed with a needle holder after one week.

[0031] (III) Succession of PDX Model

[0032] In a biosafety cabinet, mice were anesthetized and the tumor was surgically removed completely. The tumor was then processed according to the primary model construction method and inoculated into the next generation of mice to form the P1 generation PDX model. Passage was performed sequentially according to the original measurement and construction methods.

[0033] Example 2: Animal Experiment for the Treatment of HNSCC (Part 1)

[0034] Twenty-four PDX model mice (PDX#3, each with a tumor volume of approximately 100 cm³) constructed in Example 1 were used. 3 Mice (without ulceration) were randomly divided into two groups of 12 each: a solvent control group (hereinafter referred to as the "control group") and a PTC-209 treatment group (hereinafter referred to as the "PTC-209 group"). Mice in the control group received intraperitoneal injections of 100 μl of PBS on days 1, 4, 8, 11, and 15; mice in the PTC-209 group received intraperitoneal injections of 60 mg / kg of PTC-209 on days 1, 4, 8, 11, and 15. Treatment and observation continued for 36 days. Tumor length (L, longest diameter) and width (W, shortest diameter) of each mouse were measured and recorded every three days using a miniature vernier caliper. Tumor volume was calculated as (L × W). 2 / 2). All mice were sacrificed at 36 days, and tumors were removed and weighed. Experimental methods and results are as follows: Figure 1 A to Figure 1 As shown in Figure C, this indicates that for the PTC-209 group, as long as the drug is administered continuously, the tumor growth of mice will be inhibited, but the tumors will recur rapidly and gradually increase in size after the drug is stopped.

[0035] Cancer stem cells are a small but crucial subpopulation of cells within tumors. They are capable of self-renewal and generating different types of tumor cells, thereby maintaining tumor growth and heterogeneity (see: Batlle, E. and H. Clevers, Cancer stem cells revisited. Nat Med, 2017.23(10):p.1124-1134.).

[0036] Tumor regeneration was rapid after PTC-209 administration was discontinued, prompting the applicant to further explore the dynamic changes of tumor stem cells. To this end, the applicant collected mouse tumor tissue at 0, 1, 18, and 36 days of PTC-209 treatment, minced the tissue, digested it with enzymes at 37°C to obtain a single-cell suspension, and adjusted the concentration of the single-cell suspension to 1×10⁻⁶ cells / mL with PBS. 6 per ml. Subsequently, flow cytometry staining was performed using the ALDEFLUOR assay kit to analyze changes in tumor stem cells (see: Chen, D., et al., Targeting BMI1(+) Cancer Stem Cells Overcomes Chemoresistance and Inhibits Metastases in Squamous Cell Carcinoma. Cell Stem Cell, 2017.20(5):p.621-634e6; Dong, J., et al., Transcriptional super-enhancers control cancerstemness and metastasis genes in squamous cell carcinoma. Nat Commun, 2021.12(1):p.3974; Zhang, W., et al., Targeting KDM4A epigenetically activates tumor-cell-intrinsic immunity by inducing DNA replication stress. Mol Cell, 2021.81(10):p.2148-2165e9; Qin, Z., et al., PVT1 inhibition). Stimulates anti-tumorimmunity, prevents metastasis, and depletes cancer stem cells in squamous cellcarcinoma. Cell Death Dis, 2023.14(3):p.187.). The result is as follows Figure 1 As shown in Figure D, the +DEAB group served as the control group, controlling for background fluorescence interference during the experiment. Experimental results indicate that after one day of PTC-209 administration alone, intratumoral ALDH... + The percentage of cancer stem cells (CSCs) decreased from 1.99% to 1.09%, and within an 18-day treatment cycle, the number of these cancer stem cells decreased to 0.14%. However, the number of cancer stem cells detected in tumors collected 18 days after stopping PTC-209 treatment was 2.33%, indicating a recovery to pre-treatment levels. These results suggest that targeting cancer stem cells with PTC-209 alone is insufficient to eradicate cancer.

[0037] Example 3: Animal Experiments for the Treatment of HNSCC (Part 2)

[0038] Forty-eight PDX model mice (PDX#1, with tumor volumes of approximately 100 cm³) constructed in Example 1 were used. 3 (And without ulceration), and were randomly divided into 4 groups, with 12 animals in each group, namely solvent control group (hereinafter referred to as "control group"), PTC-209 treatment group (hereinafter referred to as "PTC-209 group"), MYCi975 treatment group (hereinafter referred to as "MYCi975 group"), and PTC-209 and MYCi975 combined treatment group (hereinafter referred to as "combined treatment group"). Mice in the control group were intraperitoneally injected with 100 μl of PBS on days 1, 4, 8, 11, and 15; mice in the PTC-209 group were intraperitoneally injected with 60 mg / kg of PTC-209 on days 1, 4, 8, 11, and 15; mice in the MYCi975 group were intraperitoneally injected with 100 mg / kg of MYCi975 on days 1, 4, 8, 11, and 15; and mice in the combined treatment group were intraperitoneally injected with 60 mg / kg of PTC-209 and 100 mg / kg of MYCi975 on days 1, 4, 8, 11, and 15. Observation continued until the mice died. Every three days, the tumor length (L) and width (W) and the weight of each mouse were measured and recorded using a miniature vernier caliper. The tumor volume was calculated as (L × W). 2 / 2). The criteria for mouse death were: (1) natural death; (2) the longest diameter of the mouse tumor reached 15 mm; (3) the mouse weight loss was >20%. The tumors of the dead mice were dissected, removed, and weighed. The tumor volume statistics after 36 days of continuous observation are as follows. Figure 2 As shown in Figure A, this indicates that:

[0039] (1) In the control group, the tumor volume of mice continued to grow rapidly because no drug treatment intervention was performed.

[0040] (2) In the PTC-209 group, tumor growth in mice was inhibited and tumor volume growth slowed down as long as the drug was administered continuously. However, the tumors relapsed rapidly and increased in size quickly after the drug was discontinued. This indicates that only continuous PTC-209 treatment can prevent tumor progression.

[0041] (3) The results of the MYCi975 group were similar to those of the PTC-209 group, indicating that only continuous MYCi975 treatment can prevent tumor progression.

[0042] (4) In the combined treatment group, tumor growth in mice was inhibited during the continuous administration phase, and the tumor volume did not change significantly after drug withdrawal. This indicates that the combined treatment of PTC-209 and MYCi975 is effective in treating HNSCC not only during the administration phase, but also in preventing recurrence of HNSCC after drug withdrawal.

[0043] like Figure 2 As shown in Figure B, the overall survival rate of mice in the combination therapy group was improved compared to mice treated with PTC209 or MYCi975 alone. Figure 2 As shown in Figure C, the median overall survival of the control group, PTC-209 group, MYCi975 group, and combined treatment group were 14 days, 26 days, 27 days, and 48 days, respectively. The results indicate that combined treatment with PTC-209 and MYCi975 can effectively improve the overall survival of mice.

[0044] Example 4: Construction of a 4NQO tumor mouse model

[0045] Choose Bmi1 for 6-8 week old babies CreER Rosa tdTomato Transgenic mice (Jackson Laboratory, USA) were fed drinking water containing 50 μg / ml of 4-nitroquinoline-1-oxide (4NQO) for 16 weeks, followed by 6 weeks of normal drinking water to induce tumor formation and lymph node metastasis, thereby inducing a spontaneous 4NQO tumor mouse model of HNSCC. For details, please refer to: Jia, L., W. Zhang, and CY Wang, BMI1 Inhibition Eliminates Residual Cancer Stem Cells after PD1 Blockade and Activates Antitumor Immunity to Prevent Metastasis and Relapse. Cell Stem Cell, 2020. 27(2): p. 238-253e6.

[0046] Example 5: Animal Experiments for the Treatment of HNSCC (Part 3)

[0047] (I) Animal Experiments

[0048] Forty-eight male mice (6 weeks old, 4NQO water concentration 50 μg / mL, water-fed for 16 weeks) that were used to construct the 4NQO tumor mouse model in Example 4 were randomly divided into four groups of 12 mice each: solvent control group (hereinafter referred to as "control group"), PTC-209 treatment group (hereinafter referred to as "PTC-209 group"), MYCi975 treatment group (hereinafter referred to as "MYCi975 group"), and PTC-209 and MYCi975 combined treatment group (hereinafter referred to as "combined treatment group"). Mice in the control group were intraperitoneally injected with 100 μl of PBS on days 1 and 4 of each week for four consecutive weeks. Mice in the PTC-209 group were intraperitoneally injected with 60 mg / kg of PTC-209 on days 1 and 4 of each week for four consecutive weeks. Mice in the MYCi975 group were intraperitoneally injected with 100 mg / kg of MYCi975 on days 1 and 4 of each week for four consecutive weeks. Mice in the combination treatment group were intraperitoneally injected with 60 mg / kg of PTC-209 and 100 mg / kg of MYCi975 on days 1 and 4 of each week for four consecutive weeks. After each group's administration, mice were given tamoxifen (225 mg / kg, intraperitoneal injection) to induce Bmi1. + Tumor stem cells fluoresce red for visualization in frozen sections. Mice were sacrificed four weeks after administration of tamoxifen.

[0049] (II) Preparation of frozen sections

[0050] After euthanizing the mice, the tongues were collected, photographed, and the size of the tumor lesions was measured. The tongues were fixed overnight in 4 wt% paraformaldehyde. The tongues were then longitudinally cut in half, and one half was randomly selected and placed in PBS solution containing 30 wt% sucrose to reduce tissue water content. After 1-3 days, the tongues were embedded in OCT embedding medium, and stored at -20°C to avoid air bubbles. When using, the processed tongues were removed, a suitable base was selected, and sections were prepared using a cryostat, with each section approximately 5 μm thick. During sectioning, the blade should be parallel to the tongue section to ensure a smooth cut. Each set of sections was labeled and allowed to air dry at room temperature for 20-30 minutes before use, or stored at -20°C for later use.

[0051] (III) Staining and Observation of Frozen Sections

[0052] The staining and observation methods for frozen sections are as follows:

[0053] (1) After removing the slices from the refrigerator, let them warm to room temperature for 30 minutes;

[0054] (2) Immerse the slices in PBS solution and wash them quickly on a shaker for 5 minutes, 3 times;

[0055] (3) Gently shake off the water and circle the tissue with an immunohistochemistry pen to prevent subsequent antibody spillage;

[0056] (4) At room temperature, block nonspecific binding sites with donkey serum for 30 min;

[0057] (5) Gently shake off the blocking solution, immerse in PBS solution and wash quickly on a shaker for 5 minutes, 3 times;

[0058] (6) Add DAPI, stain for 5 min, then wash quickly with PBS solution for 5 min, repeat 3 times;

[0059] (7) Mounting: Wipe the liquid around the tissue dry, drop the fluorescent mounting medium onto the tissue and mount it. Store at 4°C in the dark.

[0060] (8) Take pictures of the stained sections under an optical microscope and count the proportion of positive expression areas (red).

[0061] The test results are as follows Figure 3 As shown, the red fluorescence is Bmi1. + The image shows that the number of tumor stem cells in the PTC-209 and MYCi975 groups was not statistically different from that in the control group after treatment, indicating HNSCC recurrence. In contrast, the combination therapy group showed a sustained inhibitory effect on HNSCC growth and significantly reduced the number of tumor stem cells.

[0062] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims and their equivalents, this invention is also intended to include these modifications and variations. The above embodiments or implementations are merely illustrative examples of this invention, and it can also be implemented in other specific ways or forms without departing from its gist or essential characteristics. Therefore, the described embodiments should be considered illustrative rather than limiting in any respect. The scope of this invention should be defined by the appended claims, and any changes equivalent to the intent and scope of the claims should also be included within the scope of this invention.

Claims

1. Use of a combination of a BMI1 inhibitor, which is PTC-209, and a Myc inhibitor, which is MYC i975, for the manufacture of a medicament for the treatment of cancer, which is head and neck squamous cell carcinoma.

2. Use according to claim 1, characterized in that: The mass ratio of the BMI1 inhibitor to the Myc inhibitor in the combination is 3:

5.

3. Use according to claim 1, characterized in that: The combination is a solution.

4. Use according to claim 1, characterized in that: The combination produces a medicament for the treatment of cancer by eliminating tumor stem cells.

2. Use of a combination of a BMI1 inhibitor, which is PTC-209, and a Myc inhibitor, which is MYC i975, for the manufacture of a medicament for the treatment of cancer, which is head and neck squamous cell carcinoma. The mass ratio of the BMI1 inhibitor to the Myc inhibitor in the combination is 3:

5. The combination is a solution. The combination produces a medicament for the treatment of cancer by eliminating tumor stem cells.

3. Use of a combination of a BMI1 inhibitor, which is PTC-209, and a Myc inhibitor, which is MYC i975, for the manufacture of a medicament for the treatment of cancer, which is head and neck squamous cell carcinoma. The mass ratio of the BMI1 inhibitor to the Myc inhibitor in the combination is 3:

5. The combination is a solution

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