Application of c-Myc G4 targeting small molecule medicine in preparation of medicine for treating multiple myeloma

Abstract

The invention provides application of a c-Myc G4 targeting small molecule drug in preparation of a drug for treating multiple myeloma, and belongs to the technical field of multiple myeloma. The pharmaceutical composition for treating multiple myeloma provided by the invention takes a compound shown as a formula Y502-2304 or a pharmaceutically acceptable salt thereof as an active ingredient or a main active ingredient, and is supplemented with a pharmaceutically acceptable carrier. In multiple myeloma, the compound Y502-2304 regulates apoptosis and cycle progress of cells by stabilizing the c-Myc G4 structure, promotes death of myeloma cells, can be used as a drug for treating multiple myeloma, and provides a new thought for research and development of drugs for resisting multiple myeloma. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of multiple myeloma, and particularly relates to a small molecule drug targeting c-Myc G4, namely compound Y502-2304 or a pharmaceutically acceptable salt thereof, and its application in the preparation of a drug for treating multiple myeloma by stabilizing c-Myc G4, inhibiting the expression of c-Myc, and regulating the expression levels of related genes at the mRNA and protein levels. Background Art

[0002] Cancer, characterized by uncontrolled cell growth, remains a major global health challenge and one of the leading causes of global mortality. Despite significant progress in our understanding of its molecular mechanisms, the incidence of cancer is still rising due to population aging and lifestyle changes. Current treatment methods, including surgery, chemotherapy, and radiotherapy, aim to eliminate cancer cells; however, they often come with significant side effects and limited efficacy. Small molecule targeted therapy, as a promising alternative, offers specific treatment while reducing systemic toxicity. However, there is an urgent need to develop more molecular targets for targeted therapy to address the complexity and heterogeneity of cancer. This highlights the necessity of continuous research in this field.

[0003] Aberrant expression of the c-Myc oncogene plays a key role in the development of many types of cancer. Designing drugs targeting the expression of c-Myc protein is challenging due to its inherent instability and lack of well-defined binding sites. Within the nuclease hypersensitive element III 1 (NHEIII 1) region upstream of the P1 promoter of the c-Myc gene, there is a guanine-rich DNA strand that can form a unique secondary structure called G-quadruplex (G4), which is thought to play a role in inhibiting transcriptional elements. In this context, targeting G-quadruplex has become an effective anti-cancer therapeutic strategy.

[0004] To date, a variety of potential c-Myc G4 stabilizers have been reported. The macrocyclic compound TMPyP4 exhibits tumor inhibitory effects but has poor selectivity for the G4 structure. Pidnarulex and APTO-253 have both entered their respective Phase I clinical trials and shown good tolerability. The tetraene derivative IZCZ-3 also shows certain inhibitory effects on human cervical squamous cell carcinoma in vivo. QN-1 and IZTZ-1 show significant anti-cancer activity both in vitro and in vivo and have a greater preference for the c-Myc G4 structure. Certain isaindigotone derivatives can downregulate c-Myc transcription by affecting the interaction between G4 and proteins. However, only a portion of the ligands show effects. Therefore, there is a need to further study ligands that can specifically target different G4 structures and evaluate their therapeutic potential. Summary of the Invention

[0005] Based on the prior art, the object of the present invention is to provide a small molecule drug targeting c-Myc G4, namely the use of compound Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating multiple myeloma.

[0006] The second object of the present invention is to provide the use of the above-mentioned compound Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a c-Myc G4 stabilizer drug.

[0007] The third object of the present invention is to provide the use of the above-mentioned compound Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a drug for inhibiting the expression of c-Myc and regulating the mRNA and protein expression levels of related genes.

[0008] The technical solution of the present invention is as follows:

[0009] Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating multiple myeloma,

[0010]

[0011] When the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for treating multiple myeloma, the drug can be made into a liquid preparation, a solid preparation or a semi-solid preparation. Further, based on the technical solution provided by the present invention, the drug can be made into an injection, a tablet, a capsule or an oral liquid. Pharmaceutically acceptable excipients play a key role in the entire development process of pharmaceutical formulations. In many formulations, excipients account for the majority, and thus to a large extent, the properties of the excipients determine the properties of the formulations. Excellent excipients can enhance the stability of the main drug and extend the shelf life of the pharmaceutical; can regulate the release rate of the main drug in vivo and in vitro; can change the absorption of the drug in the body and increase the bioavailability. The specific type of excipients is not limited.

[0012] In a preferred embodiment, the present invention provides a pharmaceutical composition for treating multiple myeloma, which contains the compound represented by formula Y502-2304 mentioned in the present invention or a pharmaceutically acceptable salt thereof as the active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier. Further, the pharmaceutical composition can be formulated into a liquid preparation, a solid preparation or a semi-solid preparation. Furthermore, the pharmaceutical composition can be formulated into an injection, a tablet, a capsule or an oral liquid. Pharmaceutically acceptable excipients play a crucial role in the entire development process of pharmaceutical formulation technology. In many formulations, excipients account for the majority, so to a large extent, the properties of excipients determine the properties of the formulation. Excellent excipients can enhance the stability of the main drug and extend the shelf life of the pharmaceutical; can regulate the release rate of the main drug in vivo and in vitro; can change the absorption of the drug in vivo and increase the bioavailability, and no specific type of excipient is limited.

[0013] Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a c-Myc G4 stabilizer drug,

[0014]

[0015] When the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof is used in the preparation of a c-Myc G4 stabilizer drug, the drug can be formulated into a liquid preparation, a solid preparation or a semi-solid preparation. Further, based on the technical solution provided by the present invention, the drug can be formulated into an injection, a tablet, a capsule or an oral liquid. Pharmaceutically acceptable excipients play a crucial role in the entire development process of pharmaceutical formulation technology. In many formulations, excipients account for the majority, so to a large extent, the properties of excipients determine the properties of the formulation. Excellent excipients can enhance the stability of the main drug and extend the shelf life of the pharmaceutical; can regulate the release rate of the main drug in vivo and in vitro; can change the absorption of the drug in vivo and increase the bioavailability, and no specific type of excipient is limited.

[0016] In a preferred embodiment, the present invention provides a pharmaceutical composition that can be used as a c-Myc G4 stabilizer, which contains the compound represented by formula Y502-2304 mentioned in the present invention or a pharmaceutically acceptable salt thereof as the active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier. Further, the pharmaceutical composition can be formulated into a liquid preparation, a solid preparation or a semi-solid preparation. Furthermore, based on the technical solution provided by the present invention, the drug can be formulated into an injection, a tablet, a capsule or an oral liquid.

[0017] Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a drug for inhibiting the expression of c-Myc and regulating the mRNA and protein expression levels of related genes,

[0018]

[0019] The use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a drug for inhibiting the expression of c-Myc and regulating the expression levels of the mRNA and protein of related genes. The drug can be formulated into a liquid preparation, a solid preparation or a semi-solid preparation. Further, based on the technical solution provided by the present invention, the drug can be formulated into an injection, a tablet, a capsule or an oral liquid. Pharmaceutically acceptable excipients play a key role in the entire development process of pharmaceutical formulations. In many formulations, excipients account for the majority, and thus to a large extent, the properties of the excipients determine the properties of the formulations. Excellent excipients can enhance the stability of the main drug and extend the shelf life of the pharmaceutical; can regulate the release rate of the main drug in vivo and in vitro; can change the absorption of the drug in vivo and increase the bioavailability. The specific type of excipient is not limited.

[0020] In a preferred embodiment, the present invention provides a pharmaceutical composition for inhibiting the expression of c-Myc and regulating the expression levels of the mRNA and protein of related genes. It contains the compound represented by formula Y502-2304 mentioned in the present invention or a pharmaceutically acceptable salt thereof as the active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier. Further, the pharmaceutical composition can be formulated into a liquid preparation, a solid preparation or a semi-solid preparation. Further, based on the technical solution provided by the present invention, the drug can be formulated into an injection, a tablet, a capsule or an oral liquid.

[0021] Through a number of experiments such as FRET experiments, immunofluorescence, qPCR, Western blot, CCK-8, flow cytometry and DNA damage detection, etc., this application verified the stabilizing effect of compound Y502-2304 on the c-Myc G4 structure, successfully inhibited the transcription and translation of the c-Myc gene, and significantly reduced the c-Myc protein level. In addition, Y502-2304 also inhibited cell proliferation, induced apoptosis, and caused DNA damage, indicating its potential anti-cancer effect. It can be used as a drug for the treatment of multiple myeloma, providing a new idea for the research and development of anti-multiple myeloma drugs.

[0022] The advantages of adopting the technical solution of the present invention are as follows:

[0023] The present invention confirmed the important role of compound Y502-2304 in anti-multiple myeloma. Experiments showed that compound Y502-2304 can target c-Myc G4 to promote the apoptosis of multiple myeloma cells and the death of myeloma cells. It can be used as a drug for the treatment of multiple myeloma, providing a new idea for the research and development of anti-multiple myeloma drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Y502-2304 has anti-multiple myeloma activity;

[0025] Figure 2 Y502-2304 induces apoptosis in multiple myeloma cells and shows concentration dependence;

[0026] Figure 3 Y502-2304 can regulate the cell cycle of multiple myeloma cells and arrest it at the G2 / M phase;

[0027] Figure 4 Y502-2304 can activate the apoptosis pathway of multiple myeloma cells;

[0028] Figure 5 Y502-2304 can specifically reduce the expression level of c-Myc mRNA;

[0029] Figure 6 Y502-2304 can reduce the expression level of c-Myc protein;

[0030] Figure 7 Y502-2304 can stabilize the G4 structure;

[0031] Figure 8 Y502-2304 can induce DNA damage in multiple myeloma cells;

[0032] Figure 9 Y502-2304 can significantly increase the protein expression of γH2AX in multiple myeloma cells;

[0033] Figure 10 Y502-2304 can specifically stabilize the c-Myc G4 structure; wherein, Figure 10 A has no significant effect on the stability of Telomeric G4 by Y502-2304; Figure 10 B can stabilize the c-Myc G4 structure by Y502-2304. Specific embodiments

[0034] In order to more clearly understand the technical solution of the present invention, the present invention is further described by the following examples, but these examples do not constitute any limitation to the present invention. Specific embodiments

[0036] 1. Detection of the anti-tumor activity of Y502-2304 by CCK-8 assay

[0037] Experimental materials:

[0038] (1) Cells

[0039] The cell lines were from the cell bank of the Medical College of Anhui University of Science and Technology. The human cell lines RPMI 8226 and U266 were cultured in RPMI-1640 medium (KeyGEN, Shanghai, China) supplemented with 10% fetal bovine serum (FBS; Zeta Life, USA). All cells were maintained in a humidified atmosphere of 37 °C and 5% CO2. The cell lines were identified by short tandem repeat (STR) analysis and mycoplasma contamination was regularly detected.

[0040] (2) CCK-8 kit

[0041] Experimental procedures:

[0042] (1) Cell counting: Cells were digested routinely with trypsin. After adding complete medium to terminate digestion, they were transferred to a centrifuge tube. Suspended cells could be directly collected and centrifuged at 1000 rpm for minutes; the supernatant was aspirated and discarded, and an appropriate amount of medium was added to resuspend the cells. The cell concentration was calculated using the cell counting method;

[0043] (2) Cell seeding: 5000 cells per well, with a volume of 100 μL, and 3 replicates were set for each group. Calculate the required cell volume and the volume of complete medium needed to make up, and after thoroughly pipetting and mixing, inoculate into a 96-well plate and culture in a constant temperature cell incubator;

[0044] (3) CCK-8 assay: 48 hours after cell administration, 10 μL of CCK-8 reagent was added to each well and cultured for another 2 hours. Subsequently, the absorbance values of each well were measured at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and a growth curve was plotted.

[0045] Experimental results:

[0046] The study found that the IC50 values of Y502-2304 for the RPMI 8226 and U226 cell lines were 2.87 μM and 6.00 μM respectively ( Figure 1 ), indicating that Y502-2304 has significant cytotoxicity against these two typical MM cell lines.

[0047] 2. Cell apoptosis cycle experiment

[0048] Experimental procedures

[0049] Culture RPMI 8226 and U266 cells in an appropriate medium to ensure that the cells are in the logarithmic growth phase. Divide the cells into four groups and treat them as the control group (no drug treatment), 2 μM of Y502-2304, 5 μM of Y502-2304, and 10 μM of Y502-2304 for 24 hours. Collect the suspended cells, wash them with PBS, and then use the Annexin V-FITC / PI double staining kit. Transfer them to a flow cytometry tube and perform detection on the instrument.

[0050] Experimental results:

[0051] We investigated the effects of Y502-2304 at different concentrations on the apoptosis of RPMI 8226 and U266 multiple myeloma cells. The results showed that at concentrations of 5 μM and 10 μM, Y502-2304 significantly promoted the apoptosis of RPMI 8226 and U266 cancer cells ( Figure 2 ). At the same time, Y502-2304 was able to regulate the cell cycle of multiple myeloma cells and arrest them in the G2 / M phase ( Figure 3 ). Moreover, the results of the Western blot experiment of apoptosis-related proteins showed that Y502-2304 might induce the apoptosis of RPMI 8226 and U226 cells by activating the apoptosis pathway, increasing the expression of the apoptosis markers cleaved-caspase 3 and BAX, and simultaneously inhibiting the expression of the anti-apoptotic protein Bcl-2 ( Figure 4 ).

[0052] 3. Y502-2304 inhibits the mRNA expression level of c-Myc.

[0053] Experimental procedures

[0054] (1) Extract the RNA of RPMI 8226 and U266 cells after Y502-2304 treatment for 48 h: Collect the cells, wash the cells three times with an appropriate amount of PBS, then add an appropriate amount of Trizol and centrifuge at 12,000 r / min. Aspirate the upper chloroform layer and add isopropanol. Let it stand in a 4°C refrigerator for 20 minutes and then centrifuge at 12,000 r / min. After washing three times with 70% ethanol, measure the purity and concentration.

[0055] (2) Use a reverse transcription kit to reverse transcribe the RNA into a cDNA template and dilute it to 10 times the original concentration with deionized water for later use.

[0056] (3) Transfer the prepared qPCR reaction system into an eight-strip tube, ensuring that the volume of each well is consistent. Then, tighten the tube caps to prevent the reaction solution from leaking during centrifugation. Place the eight-strip tube in a palm centrifuge for a short centrifugation to ensure that the reaction system is thoroughly mixed. After centrifugation, gently tap the bottom of the tube to concentrate the liquid inside, and then centrifuge again to ensure that any bubbles generated during the pipetting process move to the tube mouth and are removed.

[0057] (4) After the PCR reaction is completed, collect the CT values of each sample. Analyze these data using appropriate statistical methods and controls to calculate and compare the relative expression levels of the target gene in different samples.

[0058] Experimental results:

[0059] We investigated the effect of Y502-2304 on the expression of c-Myc gene in RPMI 8226 and U266 cells. The treatments were divided into a control group (no drug treatment), 2 μM of Y502-2304, 5 μM of Y502-2304, and 10 μM of Y502-2304. The results showed that Y502-2304 inhibited the c-Myc mRNA level in a concentration-dependent manner in both multiple myeloma cell lines ( Figure 5 ). In addition, we also evaluated the effect of Y502-2304 on the expression of cancer-related genes (Bcl-2, c-Kit, k-RAS, h-RAS, and Telomeric) in RPMI 8226 and U266 cells. It was found that Y502-2304 had no significant effect on other genes. These results indicate that Y502-2304 can selectively inhibit the expression of the c-Myc gene.

[0060] 4. Y502-2304 downregulates the protein expression level

[0061] Western blot experiment

[0062] Experimental procedure:

[0063] (1) Cell protein extraction: Centrifuge to collect cells, wash them 3 times with PBS buffer, and discard the supernatant. Prepare a protein lysate according to the ratio of RIPA:SPMF:phosphatase inhibitor = 100:1:2, add it to the cells, shake with a vortex oscillator for 10 s, and lyse on ice for 30 min, shaking for 10 s every 10 min. Pre-cool a 4°C centrifuge, after lysis on ice, centrifuge at 12,000 r / min at 4°C for 20 min, and collect the supernatant.

[0064] (2) Measuring protein concentration by BCA method: According to the kit instructions, prepare the BCA working solution with A solution:B solution = 50:1. Add 2 μl of protein supernatant, 18 μl of PBS buffer, and 200 μl of BCA working solution into a 96-well plate, incubate in the dark for 30 min, measure the absorbance at 462 nm, and calculate the protein concentration. After mixing the protein supernatant with the loading buffer and boiling for 15 min, store it at -20 °C.

[0065] (3) Prepare 10% separating gel and 5% stacking gel.

[0066] (4) Protein loading: Take out the protein, load it after boiling.

[0067] (5) Electrophoresis: Run electrophoresis at 80 V until the Marker boundary is obvious, then adjust to 120 V and run electrophoresis until the bottom of the separating gel.

[0068] (6) Wet transfer: Cut a PVDF membrane of appropriate size according to the separation distance of the Marker and the molecular weight of the target protein, mark it for standby. Place the SDS-PAGE gel in the electrotransfer buffer, moisten the sandwich clip, place the gel, place the PVDF membrane, remove the bubbles, fix it, add the electrotransfer buffer into the electrotransfer tank, and perform electrotransfer at 100 V on ice. The time is set according to the molecular weight. Blocking: After electrotransfer, take out the PVDF membrane, place it in TBST containing 5% skim milk powder, and gently shake it on a shaker for 2 h.

[0069] (7) Membrane washing: After blocking, take out the PVDF membrane, place it in TBST, and quickly shake it on a shaker for 5 min × 3 times.

[0070] (8) Primary antibody incubation: Blot the PVDF membrane dry, put it into the primary antibody, and incubate it overnight on a shaker at 4 °C.

[0071] (9) Membrane washing: After primary antibody incubation, wash it with TBST for 5 min × 3 times.

[0072] (10) Secondary antibody incubation: Place the PVDF membrane in the secondary antibody, gently shake it on a shaker, and incubate it at room temperature for 1 h. Membrane washing: Take out the PVDF membrane and wash it with TBST for 5 min × 3 times.

[0073] (11) ECL development: Prepare the ECL developing solution and develop it with the instrument.

[0074] Experimental results:

[0075] Western blot analysis showed that Y502-2304 effectively downregulated the expression of c-Myc in a concentration-dependent manner in the RMPI 8226 and U266 cell lines ( Figure 6)。This downregulation was mutually verified with the experimental results of inhibiting the mRNA expression level of the gene, further confirming that Y502-2304 has a stabilizing effect on the G-quadruplex (G4) structure in the promoter region of the c-Myc gene.

[0076] 5. Immunofluorescence experiment

[0077] (1) Collect suspension cells, remove the culture medium by centrifugation, and wash twice with PBS to remove impurities.

[0078] (2) Fix the cells with 4% paraformaldehyde (PFA), incubate at room temperature for 20 minutes, and then wash three times with PBS to remove the residual fixative. Subsequently, permeabilize the cells with a PBS solution containing 0.1% Triton X-100, incubate at room temperature for 10 minutes, and wash three times again with PBS.

[0079] (3) To reduce non-specific binding, block the cells with a PBS solution containing 1% bovine serum albumin (BSA) for 30 minutes to 1 hour.

[0080] (4) After blocking, add the diluted primary antibody to the cell suspension, incubate overnight at 4°C or for 1-2 hours at room temperature, and then wash three times with PBS to remove the unbound primary antibody.

[0081] (5) Subsequently, add the diluted fluorescently labeled secondary antibody, incubate at room temperature for 1 hour in the dark, and after incubation, wash three times with PBS to remove the excess secondary antibody.

[0082] (6) Then stain the cell nuclei with DAPI, incubate at room temperature for 5-10 minutes, and wash twice with PBS.

[0083] (7) Finally, drop the stained cell suspension onto a glass slide, allow some liquid to evaporate naturally, add an anti-fluorescence quenching mounting medium, cover with a coverslip for mounting. Observe and record the cell fluorescence signals through a fluorescence microscope or a laser confocal microscope.

[0084] Experimental results:

[0085] The BG4 immunofluorescence experiment showed that Y502-2304 could stabilize the G4 structure in the cell nucleus ( Figure 7 ). The fluorescence experiment of the DNA damage marker γH2AX showed that Y502-2304 could induce DNA damage in cells ( Figure 8 ), and the level of γH2AX protein in multiple myeloma cells increased significantly after treatment with Y502-2304 ( Figure 9 ).

[0086] 6. FRET experiment confirmed that Y502-2304 could stabilize the c-Myc G4 structure

[0087] Experimental materials:

[0088] (1) Tris-HCl buffer (pH 7.4)

[0089] (2) KCL solution (1M)

[0090] (3) Primers (ordered from Sangon Biotech)

[0091] Experimental procedures:

[0092] (1) Fluorescently labeled Telomeric and c-Myc G-quadruplex (G4) structure oligonucleotides were provided by Sangon Biotech. The oligonucleotides were labeled with FAM (carboxyfluorescein) at the 5' end and TAMRA (6-carboxytetramethylrhodamine) at the 3' end.

[0093] (2) Single-stranded DNA oligonucleotides were annealed to form G-quadruplex structures: The oligonucleotides were heated to 95°C for 5 minutes and then slowly cooled to room temperature, and incubated at 4°C for 24 hours in Tris-HCl buffer (pH 7.4) containing 50 mM KCl.

[0094] (3) Performed on a ThermoFisher QuantStudio 3 real-time PCR detection system. Fluorescently labeled G4 DNA (0.5 μM) was mixed with Y502-2304 at different molar ratios in Tris-HCl buffer (pH 7.4) with a total reaction volume of 20 μL. The melting temperature (Tm) of the G4 structure in the presence of Y502-2304 was determined by recording the fluorescence values as a function of temperature. The samples were heated from 25°C to 95°C at a heating rate of 1°C / min

[0095] Experimental results:

[0096] The FRET melting curve of c-Myc shifted significantly towards higher temperatures with increasing concentrations of Y502-2304, indicating that these G4 structures were strongly stabilized. For Telomeric G4, the ΔTm value showed no significant change, indicating that Y502-2304 had little stabilizing effect on the Telomeric G4 structure ( Figure 10 ). In summary, Y502-2304 selectively stabilizes the c-Myc G4 structure.

[0097] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: it is still possible to modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating multiple myeloma, 2. The application according to claim 1, wherein The medicament is made into a liquid preparation, a solid preparation or a semi-solid preparation.

3. The application according to claim 2, characterized in that, The medicament is made into an injection, a tablet, a capsule or an oral liquid.

4. A pharmaceutical composition for treating multiple myeloma, which uses the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof as the active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier.

5. Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for stabilizing c-Myc G4, 6. The application according to claim 5, characterized in that, The medicament is made into a liquid preparation, a solid preparation or a semi-solid preparation.

7. The application according to claim 6, wherein The medicament is made into an injection, a tablet, a capsule or an oral liquid.

8. Use of the compound represented by formula Y502-2304 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for inhibiting the expression of c-Myc and regulating the expression levels of the mRNA and protein of related genes, 9. The application according to claim 8, wherein The medicament is made into a liquid preparation, a solid preparation or a semi-solid preparation.

10. The application according to claim 9, characterized in that, The medicament is made into an injection, a tablet, a capsule or an oral liquid.

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