Use of an acetyltransferase naa10 inhibitor in the manufacture of a medicament for treating lymphoma

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to an application of an acetyltransferase NAA10 inhibitor in a drug for treating lymphoma. More particularly, the application relates to an application of the acetyltransferase NAA10 inhibitor in a drug for treating diffuse large B-cell lymphoma. The application discloses that the acetyltransferase NAA10 is a key factor for regulating the transcriptional activity of a proto-oncogene Myc in the diffuse large B-cell lymphoma. Through in-depth bioinformatics analysis and experimental verification, it is proved that the NAA10 can enhance the transcriptional activity of Myc, and promote abnormal proliferation and clonal formation of the diffuse large B-cell lymphoma cells. In addition, through high-throughput screening, a natural small molecule compound myricetin is screened and verified as an effective NAA10 inhibitor, can effectively inhibit self-acetylation of the NAA10, block the transcriptional activation of Myc by the NAA10, and inhibit proliferation of the diffuse large B-cell lymphoma cells, thereby providing a promising lead compound for subsequent drug research and development, and accelerating the development process of potential therapeutic drugs.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of an acetyltransferase NAA10 inhibitor in the treatment of lymphoma. Background Technology

[0002] Lymphoma is one of the most common aggressive hematologic malignancies in adults, especially diffuse large B-cell lymphoma, with approximately 150,000 new cases worldwide each year, accounting for about 30% of non-Hodgkin's lymphoma cases. Currently, the standard treatment for diffuse large B-cell lymphoma is R-CHOP (anti-CD20 antibody combined with cyclophosphamide, doxorubicin, vincristine, and steroids). This regimen has been used for over 20 years, but the long-term overall survival (OS) rate is only about 60%. For patients with early relapse or refractory disease, the prognosis is even worse; even with high-dose chemotherapy and autologous stem cell transplantation, the median OS is only 6.3 months, and the 3-year event-free survival rate is as low as 20%. In recent years, the application of immunotherapies such as chimeric antigen receptor T-cell therapy (CAR-T) has improved the survival rate of some relapsed / refractory patients, but most patients ultimately succumb to disease progression. Therefore, exploring new therapeutic targets and biomarkers closely related to poor patient prognosis is of significant clinical importance.

[0003] From a transcriptional regulation perspective, Myc, as a key transcription factor, has been confirmed to participate in the development and progression of diffuse large B-cell lymphoma (DBL). However, due to its disordered structure and lack of a well-defined small molecule binding pocket, it has become an "untreatable" target. Therefore, screening for key molecules that can regulate Myc transcriptional activity is of great significance for inhibiting the progression of DBL. NAA10 (N-α-acetyltransferase 10), as a novel cancer biomarker, has been shown to be closely related to the development and progression of various tumors. In terms of regulatory mechanisms, there is a reciprocal regulatory relationship between NAA10 and Myc. On the one hand, Myc can affect the expression of NAA10 through transcriptional regulation mechanisms; on the other hand, NAA10, through its acetyltransferase activity and interaction with PAI1, regulates the invasive ability of cells, thereby affecting the biological function of Myc. However, whether NAA10 plays a role in DBL in conjunction with Myc still requires further in-depth research. This will not only help to elucidate the pathogenesis of DBL but may also provide important evidence for the development of new therapeutic strategies. Summary of the Invention

[0004] To address the aforementioned shortcomings, this invention provides the application of an acetyltransferase NAA10 inhibitor in the treatment of lymphoma.

[0005] This invention, through extensive bioinformatics analysis, revealed elevated Myc transcriptional activity in patients with diffuse large B-cell lymphoma (DBL) exhibiting high Myc expression. Furthermore, it was found that in patients with similar Myc expression levels, the expression levels of downstream Myc genes were further activated, suggesting the potential existence of key factors regulating Myc transcriptional activity. Further bioinformatics data analysis, combined with cell and animal experiments, revealed for the first time that the acetyltransferase NAA10 is a key factor regulating Myc transcriptional activity in DBL and plays an important role in the development and progression of DBL. High expression of NAA10 in DBL patient tissues was significantly associated with enhanced Myc transcriptional activity and poor patient prognosis.

[0006] At the molecular level, firefly luciferin reporter gene assays confirmed that overexpression of NAA10 promotes Myc transcriptional activation. RNA-seq experiments comprehensively analyzed gene expression changes induced by NAA10 silencing, confirming that NAA10 is a key factor promoting Myc transcriptional activity. A cell model overexpressing NAA10 was constructed, demonstrating that overexpression significantly promotes the proliferation and colony formation of diffuse large B-cell lymphoma cells. Two shRNAs specifically targeting NAA10 were designed, confirming that silencing NAA10 significantly inhibits the proliferation and colony formation of diffuse large B-cell lymphoma cells, thereby alleviating the malignant progression of diffuse large B-cell lymphoma. Lentiviral-packaged shCtrl and shNAA10 were used to silence NAA10 expression in mice, confirming that silencing NAA10 inhibits the in vivo malignant progression of diffuse large B-cell lymphoma. Subsequently, a NAA10 inhibitor screening model was constructed. High-throughput screening identified and validated that the natural small molecule compound myricetin can act as a specific NAA10 inhibitor in diffuse large B-cell lymphoma. A NAA10 protein purification plasmid was constructed, and by purifying the target NAA10 protein to simulate its mechanism of action, it was confirmed that Myricetin can inhibit the autoacetylation of NAA10, and this inhibition may be achieved by regulating NAA10 function, thereby inhibiting the progression of diffuse large B-cell lymphoma. The effect of Myricetin on Myc transcriptional activity was detected using a luciferase reporter gene assay, confirming that Myricetin inhibits the activation of Myc transcriptional activity by NAA10. Based on the diffuse large B-cell lymphoma cell line U2932 and the lymphoma cell line OCI-LY10, which highly expresses NAA10, it was confirmed that Myricetin can inhibit the proliferation of tumor cells and suppress the malignant progression of diffuse large B-cell lymphoma.

[0007] These results indicate that the activity of acetyltransferase NAA10 has a close regulatory effect on the growth of diffuse large B-cell lymphoma cells. It not only provides new ideas for antitumor targets and targeted drugs for the treatment of diffuse large B-cell lymphoma, but also provides new molecular indicators for judging the prognosis of diffuse large B-cell lymphoma.

[0008] The first aspect of the present invention provides the use of an acetyltransferase NAA10 inhibitor in the preparation of a drug for treating lymphoma.

[0009] Furthermore, the lymphoma is non-Hodgkin lymphoma.

[0010] Furthermore, the lymphoma is a diffuse large B-cell lymphoma.

[0011] Furthermore, the NAA10 inhibitor can reduce intracellular Myc transcriptional activity.

[0012] Furthermore, the NAA10 inhibitor is selected from natural small molecule compounds.

[0013] Furthermore, the natural small molecule compound is a flavonoid compound.

[0014] Furthermore, the flavonoid compound is myricetin.

[0015] Furthermore, the myricetin exists in its pharmaceutically acceptable salt, ester, prodrug, or isomer form.

[0016] A second aspect of the invention provides a pharmaceutical composition comprising a therapeutically effective amount of an acetyltransferase NAA10 inhibitor, and one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0017] Furthermore, the NAA10 inhibitor is myricetin or a pharmaceutically acceptable salt, ester, prodrug, or isomer thereof.

[0018] The beneficial effects of this invention are: This invention identifies and validates a novel therapeutic target and its key role in diffuse large B-cell lymphoma (DBL): For the first time, this invention reveals that the acetyltransferase NAA10 is a key factor regulating the transcriptional activity of the proto-oncogene Myc in DBL. Through in-depth bioinformatics analysis and experimental validation, it was demonstrated that NAA10 can enhance the transcriptional activity of Myc, promoting the abnormal proliferation and colony formation of DBL cells. High expression of NAA10 in DBL tissues is significantly associated with tumor progression and poor patient prognosis, establishing NAA10 as a potential therapeutic target for DBL.

[0019] This invention provides an effective strategy and potential drug for inhibiting the acetyltransferase NAA10: Experiments have demonstrated that inhibiting NAA10 significantly reduces Myc transcriptional activity in diffuse large B-cell lymphoma, effectively suppressing tumor cell proliferation and colony formation in vitro. In particular, in a mouse xenograft model, silencing NAA10 significantly delayed tumor progression and prolonged animal survival, validating the therapeutic efficacy of targeting NAA10 in vivo. Furthermore, high-throughput screening identified and validated myricetin, a natural small molecule compound, as an effective NAA10 inhibitor. Myricetin inhibits NAA10 autoacetylation and suppresses the proliferation of diffuse large B-cell lymphoma cells by blocking its transcriptional activation of Myc, providing a promising lead compound for subsequent drug development and accelerating the development of potential therapeutics. Attached Figure Description

[0020] Figure 1 A heatmap showing the expression levels of Myc downstream target genes in patients with diffuse large B-cell lymphoma who express Myc at low and high levels, as shown in the TCGA database.

[0021] Figure 2 This is a statistical graph showing the regulation of Myc activity by NAA10 in diffuse large B-cell lymphoma tissues, based on statistics from the TCGA database. Figure 2 A is a Venn diagram of differentially expressed genes that affect Myc transcriptional activity and tumor / normal tissue expression, diffuse large B-cell lymphoma-specific high expression, and CRISPR sensitivity during the development of diffuse large B-cell lymphoma. Figure 2 B Display Figure 2 Ranking of the impact of 30 differentially expressed genes selected by A on the five-year survival rate of patients with diffuse large B-cell lymphoma.

[0022] Figure 3 This is a statistical graph based on the TCGA database showing that high NAA10 expression leads to poor prognosis in patients with diffuse large B-cell lymphoma (validated using the GSE58445 dataset).

[0023] Figure 4 This is a statistical graph showing the effects of NAA10 on the expression of downstream target genes of Myc and poor prognosis, based on data from the TCGA database. Figure 4 A is a cluster heatmap of the activation of Myc transcription by acetyltransferase NAA10 in a dataset of patients with diffuse large B-cell lymphoma; Figure 4 B represents the survival analysis curves for patients with diffuse large B-cell lymphoma, specifically those with high / low NAA10 and Myc expression.

[0024] Figure 5 This is a result showing that overexpression of NAA10 can significantly activate Myc transcriptional activity. Figure 5Figure A shows the results of NAA10 and Myc overexpression in the Western-Blot assay for luciferase reporter gene detection. Figure 5 Figure B shows the results of the firefly luciferase reporter gene assay for the effect of NAA10 on Myc transcriptional activity.

[0025] Figure 6 This is the result of silencing NAA10 to inhibit Myc transcriptional activity. Figure 6 A is the enrichment result graph of the HALLMARK dataset. Figure 6 B is the enrichment result graph of the Myc downstream dataset analyzed by GSEA.

[0026] Figure 7 This is the result of the tumor-promoting effect of NAA10 overexpression on BaF3 in normal B cells. Figure 7 Figure A shows the results of NAA10 overexpression in BaF3 detected by RT-qPCR; Figure 7 B shows the cell counting results; Figure 7 C represents the result of cell clonal formation.

[0027] Figure 8 This is the result of the in vitro silencing effect of NAA10 on the anti-tumor activity of diffuse large B-cell lymphoma. Figure 8 A is the result of proliferation detection after silencing NAA10; Figure 8 B is a graph showing the results of cloning ability testing after silencing NAA10.

[0028] Figure 9 This is the result of the in vivo silencing of NAA10 on the anti-cancer effect of diffuse large B-cell lymphoma. Figure 9 A is a fluorescence imaging result of tumors in mice; Figure 9 B is a graph showing the results of mouse weight and survival time.

[0029] Figure 10 The figure shows the results of high-throughput screening of the NAA10-specific inhibitor Myricetin from a library of natural small molecule compounds.

[0030] Figure 11 The image shows the Western-Blot results of the inhibitory effect of Myricetin on NAA10 autoacetylation.

[0031] Figure 12 This is the result of Myricetin inhibiting the ability of NAA10 to activate Myc transcriptional activity. Figure 12 Figure A shows the results of the Western-Blot study on the overexpression of NAA10 and Myc in the luciferase reporter gene detection experiment. Figure 12 Figure B shows the results of firefly luciferase reporter gene assays, which showed that the compound Myricetin significantly reduced the activation of NAA10 on Myc transcriptional activity.

[0032] Figure 13 This is the result of Myricetin inhibiting the promoting effect of NAA10 on diffuse large B-cell lymphoma cells. Figure 13 Figure A shows the results of CCK-8 testing on the inhibitory effect of Myricetin. Figure 13 Figure B shows the results of Myricetin dose-dependent inhibition of the proliferation ability of NAA10-overexpressing cells. Detailed Implementation

[0033] The following detailed description, in conjunction with embodiments, illustrates the application of an acetyltransferase NAA10 inhibitor in the treatment of lymphoma. For the sake of simplicity, this document does not exhaustively list all alternative technical features and embodiments included in this invention. Therefore, those skilled in the art should understand that any technical feature and embodiment within this embodiment does not limit the scope of protection of this invention. The scope of protection includes all alternative technical features and embodiments adopted by those skilled in the art without inventive effort. Specifically, any embodiment obtained by replacing any technical feature in this invention or by combining any two or more technical features provided by this invention should be within the scope of protection of this invention.

[0034] Example Example 1 Example 1: Based on the TCGA tumor database, it was confirmed that a key factor regulating Myc transcriptional activity exists during the progression of diffuse large B-cell lymphoma. Myc is a crucial factor driving the development and progression of tumors such as diffuse large B-cell lymphoma. High activation of Myc can regulate the malignant progression of diffuse large B-cell lymphoma by modulating cell proliferation, apoptosis, and metabolic pathways. However, due to its status as a "drug-incompatible" target, its upstream and downstream regulatory mechanisms have attracted considerable attention. The upstream regulatory mechanisms targeting Myc transcriptional activity remain unclear in current technologies, and effective intervention strategies are lacking, necessitating further research to identify key targets.

[0035] In this example, patients with diffuse large B-cell lymphoma from the TCGA database were divided into those with high Myc expression and those with low Myc expression using the cbioportal website. Downstream genes of Myc from the HALLMARK-Myc-TARGET dataset provided by the GSEA website were analyzed for activation using the Lianchuan Bio Cloud Platform to rank the patients. The patients were then categorized as follows: Group A: Low Myc expression, low Myc transcriptional activity; Group B: Low Myc expression, high Myc transcriptional activity; Group C: High Myc expression, low Myc transcriptional activity; Group D: High Myc expression, high Myc transcriptional activity. The heatmap results of the expression levels of downstream target genes of Myc are shown below. Figure 1As shown, Myc transcriptional activity was further increased in patients with high Myc expression, and in some patients with similar Myc expression, Myc downstream was further activated, suggesting that there may be key factors that can regulate Myc transcriptional activity.

[0036] Example 2 Example 2: High expression of NAA10 in diffuse large B-cell lymphoma is a key factor regulating Myc activation. The analysis results from Example 1 suggest the possible existence of key factors regulating Myc activation in patients with diffuse large B-cell lymphoma; therefore, further research is needed. Figure 1 Based on the analysis of the B / A+ and D / C groups, differentially expressed genes with different transcriptional activities were identified among patients with similar Myc expression. This resulted in the identification of 592 differentially expressed genes (Foldchange>1.8) that could regulate Myc transcriptional activity. Further, to identify key genes regulating the malignant progression of diffuse large B-cell lymphoma, this result was combined with the upregulated 4114 gene (Foldchange>2) in tumor tissues and normal tissues (diffuse large B-cell lymphoma patients), the highly dependent 895 gene (Cripsr<-1) in diffuse large B-cell lymphoma cell lines, and the 1706 gene, which is specifically highly expressed in diffuse large B-cell lymphoma cancer types. Multivariate analysis was then conducted, ultimately identifying 30 potential genes, such as... Figure 2 As shown in A. Subsequently, the five-year survival rates of the above 30 genes in patients with diffuse large B-cell lymphoma under high and low expression states were analyzed. Figure 2 As shown in Figure B, high expression of NAA10 was found to be closely associated with poor prognosis in patients with diffuse large B-cell lymphoma, suggesting that NAA10 may be a key factor regulating Myc transcriptional activation in diffuse large B-cell lymphoma.

[0037] The above analysis suggests that NAA10 can promote Myc transcriptional activity in patients with diffuse large B-cell lymphoma and is specifically highly expressed in this disease. Further validation using the GES5844 lymphoma dataset revealed poor prognostic statistical results. Figure 3 As shown, this also confirms that high NAA10 expression leads to poor prognosis in patients (P<0.05). Furthermore, patients in this dataset were divided into two groups based on Myc expression: high Myc expression and low Myc expression. These two groups were then ranked according to NAA10 expression, and their clustering heatmaps are shown below. Figure 4 As shown in Figure A, the results indicate that high NAA10 expression further activates Myc transcriptional activity in both patients with high and low Myc expression. Subsequent prognostic analyses of these four patient groups were performed, and their survival curves are shown in Figure A. Figure 4As shown in Figure B, the results indicate that the prognosis was worst when both NAA10 and Myc were highly expressed (P<0.05).

[0038] All of the above results confirm that NAA10 participates in the disease progression of diffuse large B-cell lymphoma by regulating Myc transcriptional activity, and its high expression is closely related to poor prognosis in patients with diffuse large B-cell lymphoma.

[0039] Example 3 Example 3 confirms that NAA10 promotes Myc transcriptional activity in diffuse large B-cell lymphoma. To further clarify the regulatory role of NAA10 on Myc transcriptional activity, a firefly luciferin reporter gene assay was used for investigation. Figure 5 As shown, the results indicated that the activation fold of Myc alone was 2.73 ± 0.13 times that of the control group (P < 0.001), and overexpression of NAA10 further promoted the transcriptional activation level of Myc, with an activation fold of 3.32 ± 0.13 times (P < 0.001).

[0040] Furthermore, RNA-seq experiments were used to comprehensively analyze the gene expression changes induced by shNAA10. OCI-LY10 cells (diffuse large B-cell lymphoma cell line) were infected with viruses packaging shCtrl, shNAA10 #1, and shNAA10 #2 plasmids via lentiviral infection. Cells were collected on day four after viral infection for RNA-seq sequencing analysis. Gene set enrichment analysis (GSEA) was introduced to examine the impact of NAA10 silencing on key signaling pathways. Hallmark pathway enrichment analysis was performed on the gene expression matrices of shCtrl and shNAA10, such as... Figure 6 As shown in AB, nine Hallmark pathways with significant differences (P<0.05) were screened out, and it was found that downstream target genes of Myc were highly enriched in the shCtrl group (NES=1.511 and P=0.0047 and NES=1.596 and P=0.0084), further suggesting that NAA10 is a key factor promoting Myc transcriptional activity.

[0041] Example 4 Example 4: NAA10 promotes malignant progression of diffuse large B-cell lymphoma Based on the transcriptional activation effect of NAA10 on Myc confirmed in Example 3 above, to investigate whether NAA10 plays an important role in diffuse large B-cell lymphoma, a cell model overexpressing NAA10 was constructed in normal BaF3 B cells via lentiviral infection. The effect of NAA10 overexpression was then examined using RT-PCR. Figure 7 As shown in Figure A, the results indicated that NAA10 was significantly overexpressed compared to the control group, with an overexpression fold of 58.91 ± 13.37 (P < 0.01). Based on this, the pro-cancer effect of NAA10 was investigated using counting and clonogenic assays, and the results are shown in Figure A. Figure 7 As shown in Figure BC, overexpression of NAA10 significantly promoted the proliferation (P<0.001) and clonogenic capacity of diffuse large B-cell lymphoma cells compared to the control group. Therefore, it can be considered that NAA10, which is specifically highly expressed in diffuse large B-cell lymphoma, is a key protein promoting the malignant progression of diffuse large B-cell lymphoma.

[0042] The important role of NAA10 in the malignant progression of diffuse large B-cell lymphoma (DBL) suggests that targeted intervention with NAA10 may improve the prognosis of DBL patients. Two specific NAA10-targeting shRNAs were designed to investigate the effect of NAA10 silencing on the malignant progression of DBL. The specific shRNAs were used in the NAA10-high-expressing cell lines OCI-LY10 and SU-DHL-10 to assess the effect of NAA10 silencing on cell proliferation. Silent OCI-LY10 and SU-DHL-10 cells were seeded at the same density in six-well plates and cultured. Cell counts were performed at specified time points (days 1, 3, 5, and 7). The results are shown below. Figure 8 As shown in Figure A, the results indicated that both shNAA10 symptomatic agents significantly inhibited the proliferation of OCI-LY10 and SU-DHL-10 cells (P<0.001). These results confirm that silencing NAA10 can significantly inhibit the proliferation of diffuse large B-cell lymphoma cells.

[0043] Soft agar assay is an important method for evaluating cell clonal capacity, providing harsher survival conditions for cell growth and aiding in the assessment of tumor cell malignancy. This study investigated the effect of silencing NAA10 on the clonogenic capacity of diffuse large B-cell lymphoma cells using a soft agar assay. Lentiviral infection was used to introduce shNAA10-carrying virus into OCI-LY10 and SU-DHL-10 cells to silence NAA10. Cells were seeded at a density of 2,000 cells / well in six-well plates. After cell colony formation, staining was performed using NBT. Figure 8As shown in Figure B, where the right figure represents the clone count results of the left figure, in OCI-LY10, compared to shCtrl, the clone fold of shNAA10 #1 was 0.18±0.08, and the clone fold of shNAA10 #2 was 0.17±0.11; in SU-DHL-10, compared to shCtrl, the clone fold of shNAA10 #1 was 0.07±0.04, and the clone fold of shNAA10 #2 was 0.29±0.06. These results confirm that silencing NAA10 can significantly inhibit the clonogenic ability of diffuse large B-cell lymphoma cells.

[0044] In summary, it is clear that NAA10 is an important factor in controlling the malignant progression of diffuse large B-cell lymphoma.

[0045] Example 5 Example 5: Effects of silencing NAA10 on progression and survival time of diffuse large B-cell lymphoma in mice. Examples 3 and 4 above confirmed the inhibitory effect of silencing NAA10 on diffuse large B-cell lymphoma in vitro. Further, a mouse model of diffuse large B-cell lymphoma was constructed to evaluate the effect of silencing NAA10 on the malignant progression and survival time of diffuse large B-cell lymphoma cells in mice. An OCI-LY10 cell line stably expressing the luciferase tag protein was constructed. The luciferase expressed by the luciferase plasmid can react with the imaging solution in vivo to produce fluorescence, allowing visualization of the proliferation of diffuse large B-cell lymphoma cells in mice. OCI-LY10 cells expressing luciferase protein were infected with lentiviruses shCtrl and shNAA10, and the infected cells were transplanted into NSG mice via tail vein injection to construct a xenograft tumor model. After successful cell inoculation, under anesthesia, each NSG mouse was injected with the imaging solution on days 10, 16, and 20 to assess tumor progression. In vivo bioluminescence imaging results in mice showed that silencing NAA10 significantly inhibited the in vivo proliferation of OCI-LY10 cells; fluorescence intensities detected on days 10, 16, and 20 were quantified and statistically analyzed using Living-image software, such as... Figure 9 As shown in Figure A, the results showed that the fluorescence value of the NAA10 silencing group was significantly lower than that of the control group (P<0.001), confirming that silencing NAA10 can inhibit the malignant progression of diffuse large B-cell lymphoma in vivo.

[0046] Continuous monitoring of mouse body weight changes showed that mice in the shCtrl group, due to the development of diffuse large B-cell lymphoma, experienced weight loss around day 18 after cell inoculation, while the shNAA10 group showed no significant change. Furthermore, continuously recorded mouse survival time showed that mice in the shCtrl group died successively between days 21 and 26 post-inoculation, while mice in the shNAA10 group had a longer overall survival time, dying successively between days 24 and 35 post-inoculation. Figure 9 As shown in B, silencing NAA10 confirmed that it can delay disease progression and prolong survival in mice with diffuse large B-cell lymphoma, suggesting that targeting NAA10 may be a potential therapeutic strategy for diffuse large B-cell lymphoma.

[0047] Example 6 Example 6: Myricetin as a specific NAA10 inhibitor Example 5 above demonstrates that NAA10 plays a crucial role in the malignant progression of diffuse large B-cell lymphoma, suggesting that targeting NAA10 may be a potential therapeutic strategy for diffuse large B-cell lymphoma. However, there are currently no reports on NAA10 inhibitors. Therefore, a NAA10 inhibitor screening model was further constructed to discover compounds that can specifically inhibit NAA10. Specifically, a BaF3 cell model overexpressing NAA10 was constructed by lentiviral infection. Control group cells and NAA10-overexpressing group cells were seeded at the same cell density into 96-well plates. The cells were then treated with drugs from a library of natural small molecule compounds, and changes in cell proliferation were detected. Compounds that could significantly inhibit the proliferation of NAA10-overexpressing cells were screened. The results are as follows: Figure 10 As shown, the results indicate that Myricetin significantly inhibited the proliferation of NAA10-overexpressing cells (inhibition rate 64.94%), with minimal impact on control cells (inhibition rate 35.09%). This screening experiment confirms that Myricetin may have a specific inhibitory effect on NAA10-overexpressing cells while exhibiting good safety, suggesting its potential as a specific inhibitor of NAA10 for the treatment of diffuse large B-cell lymphoma.

[0048] Example 7 Example 7: Myricetin inhibits the autoacetylation of NAA10. Current research indicates that NAA10, as an N-α-acetyltransferase, can regulate its enzymatic activity through autoacetylation. Autoacetylation can alter the conformation of NAA10, thereby affecting its substrate binding ability and catalytic efficiency. This regulatory effect allows NAA10 to flexibly perform its functions in different cellular environments.

[0049] Example 6 above confirmed that Myricetin can inhibit the proliferation of cells overexpressing NAA10. To investigate whether Myricetin exerts its effect by inhibiting the autoacetylation of NAA10, a NAA10 protein purification plasmid was constructed, and the mechanism of action was simulated by purifying the target NAA10 protein. The purified NAA10 protein and Myricetin compound were co-incubated, followed by incubation with AcCoa for 7 hours. The results are as follows... Figure 11 As shown in the figure. Western blotting results showed that the acetylation level of NAA10 decreased significantly with increasing drug concentration after Myricetin treatment, while the total protein level did not change significantly. This confirms that Myricetin can inhibit the autoacetylation of NAA10, and this inhibitory effect may inhibit the progression of diffuse large B-cell lymphoma by regulating the function of NAA10.

[0050] Example 8 Example 8: Myricetin inhibits NAA10-induced transcriptional activation of Myc. Example 6 above confirmed the inhibitory effect of Myricetin on NAA10 overexpressing cells. NAA10 is closely related to Myc transcriptional activity. To further verify the effect of Myricetin on NAA10-activated Myc transcriptional activity, NAA10, Myc, and the downstream E-box sequence of Myc were transfected into HEK-293T cells. Myricetin was administered 12 hours after transfection. After 24 hours of drug treatment, the effect of Myricetin on Myc transcriptional activity was detected using a luciferase reporter gene assay. The results are as follows: Figure 12 As shown in Figures AB, the results of the firefly luciferase reporter gene assay confirmed that Myricetin significantly inhibited the activation of Myc transcriptional activity by NAA10. Specifically, without Myricetin treatment, the activation factor of Myc transcription by NAA10 was 5.32 ± 0.94 (P < 0.05), while after Myricetin treatment, the activation factor of Myc transcriptional activity by NAA10 was significantly reduced, with an activation factor of 1.74 ± 0.52 (P < 0.01).

[0051] Example 9 Example 9: Myricetin may exert its anti-diffuse large B-cell lymphoma effect by inhibiting NAA10 autoacetylation and its activation of Myc transcriptional activity. Based on Examples 6 and 7, to further clarify the effect of Myricetin's inhibitory effect on NAA10 on diffuse large B-cell lymphoma cells, the U2932 cell model of diffuse large B-cell lymphoma was used to investigate the effect of different concentrations of Myricetin on cell proliferation. U2932 cells from the lentivirus-infected control group and the NAA10-overexpressing group were seeded in 96-well plates and treated with different concentration gradients of Myricetin (0, 0.625, 1.25, 2.5, 5, and 10 μM). The results are as follows: Figure 13 As shown in Figure A, the results indicate that the survival rate of U2932 cells overexpressing NAA10 decreased with increasing Myricetin treatment concentration, while the survival rate of control cells was less affected. Simultaneously, the inhibitory effect of Myricetin on cell proliferation was investigated using the OCI-LY10 lymphoma cell line, which highly expresses NAA10. The results are shown in Figure A. Figure 13 As shown in Figure B, the results indicate that Myricetin can dose-dependently inhibit the proliferation of OCI-LY10 cells expressing NAA10.

[0052] Example 10 Example 10 A pharmaceutical composition The NAA10 inhibitors and their pharmaceutically acceptable salts, esters, prodrugs, or isomers provided in this embodiment can be combined with one or more pharmaceutically acceptable carriers, excipients, or diluents to prepare pharmaceutical compositions. Pharmaceutically acceptable carriers, excipients, and diluents are known in the art and include, but are not limited to, water, salt solutions, alcohols, polyethylene glycol, glycerol, gelatin, carbohydrates, phosphates, surfactants, lipids, buffers, suspending agents, binders, fillers, etc. The pharmaceutical compositions of this invention can be prepared into various dosage forms as needed, such as injections, tablets, capsules, granules, oral solutions, etc., to suit different routes of administration, such as intravenous injection, oral administration, subcutaneous injection, etc. Common methods for preparing pharmaceutical compositions are well known to those skilled in the art. The dosage of the active ingredient in the pharmaceutical composition can be adjusted according to factors such as the patient's age, weight, disease severity, and route of administration; the specific dosage is determined based on the clinical situation.

[0053] For those skilled in the art, other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations, but obvious variations or modifications derived therefrom are still within the scope of protection of the claims of this invention.

Claims

1. The use of an acetyltransferase NAA10 inhibitor in the preparation of a drug for the treatment of lymphoma.

2. The application according to claim 1, characterized in that, The lymphoma is non-Hodgkin's lymphoma.

3. The application according to claim 2, characterized in that, The lymphoma is a diffuse large B-cell lymphoma.

4. The application according to claim 1, characterized in that, The NAA10 inhibitor can reduce intracellular Myc transcriptional activity.

5. The application according to any one of claims 1 to 4, characterized in that, The NAA10 inhibitor is selected from natural small molecule compounds.

6. The application according to claim 5, characterized in that, The natural small molecule compound is a flavonoid.

7. The application according to claim 6, characterized in that, The flavonoid compound is myricetin.

8. The application according to claim 7, characterized in that, The myricetin exists in its pharmaceutically acceptable salt, ester, prodrug, or isomer forms.

9. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a therapeutically effective amount of an acetyltransferase NAA10 inhibitor according to any one of claims 1 to 8, and one or more pharmaceutically acceptable carriers, excipients, or diluents.

10. The pharmaceutical composition according to claim 9, characterized in that, The NAA10 inhibitor is myricetin or its pharmaceutically acceptable salt, ester, prodrug, or isomer.

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