Application of combined L-lys and N1-MNA in synergistic inhibition of growth of glioma

Abstract

The invention belongs to the technical field of glioma treatment, and discloses an application of combination of L-lys and N1-MNA in synergistic inhibition of glioma growth, L-lys and N1-MNA inhibit glioma growth through a synergistic mechanism of microenvironment regulation and metabolic signaling pathway blocking, L-lys inhibits glioma growth through arginine sweep, N1-MNA inhibits glioma growth through arginine sweep, N1-MNA inhibits glioma growth through arginine sweep, N1-MNA inhibits glioma growth through arginine sweep, and N1-MNA inhibits glioma growth through arginine sweep. N1-MNA and L-lys synergistically inhibit the growth of glioma by inhibiting a Sirt1 / Akt / mTOR pathway, and the invention has the following beneficial effects: 1, the growth of glioma is inhibited by a synergistic mechanism of L-lys and N1-MNA microenvironment regulation and metabolic signal pathway blocking, the growth of glioma is inhibited by L-lys through arginine sweep, the growth of glioma is synergistically inhibited by N1-MNA and L-lys by inhibiting the Sirt1 / Akt / mTOR pathway, and the growth of glioma is inhibited by L-lys through arginine sweep, and the growth of glioma is inhibited by N1-MNA and L-lys by inhibiting Sirt1 / Akt / mTOR pathway; the combination of the two can significantly enhance the anti-tumor effect. 2, the combination of L-lys and N1-MNA can significantly improve the tumor inhibition rate, has no significant weight loss (indicating low toxicity), is significantly superior to the'curative effect-toxicity 'balance of traditional chemotherapy, and avoids the drug resistance problem of targeted drugs.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of glioma treatment, in particular to application of L-lys and N1-MNA in synergistically inhibiting growth of glioma. BACKGROUND

[0002] Glioma is the most common malignant tumor in the central nervous system, and has the characteristics of high invasiveness, high recurrence rate and treatment resistance, and the existing treatment methods are difficult to significantly improve the prognosis of patients.

[0003] Glioma accounts for more than 80% of central nervous system malignant tumors, among which glioblastoma multiforme (GBM) has the highest malignancy, and the median survival of patients is only 12-15 months. The current standard treatment regimen is mainly surgical resection combined with radiotherapy and chemotherapy (temozolomide), but due to the infiltrative growth of tumor cells, the limitation of drug delivery by the blood-brain barrier and the drug resistance mediated by tumor stem cells, the treatment effect is still not ideal.

[0004] In recent years, targeting tumor metabolic abnormalities and regulating tumor microenvironment have become a new direction for glioma treatment, and intestinal microbial metabolites, as key "messenger molecules" connecting intestinal flora and tumor progression, have gradually attracted attention in the field of anti-tumor.

[0005] However, the role and mechanism of intestinal microbial metabolites on glioma are still relatively scarce, especially the synergistic effect between different metabolites has not been reported. SUMMARY

[0006] To solve the above problems, the application provides application of L-lys and N1-MNA in synergistically inhibiting growth of glioma, and the application is realized by the following technical solutions.

[0007] The application of L-lys and N1-MNA in synergistically inhibiting growth of glioma, L-lys and N1-MNA inhibit the growth of glioma through the synergistic mechanism of microenvironment regulation and metabolic signal pathway blockage, L-lys is L-lysine, and N1-MNA is N1-methylnicotinamide.

[0008] As a further scheme of the application, L-lys and N1-MNA synergistically inhibit the growth of glioma through the following mechanisms:

[0009] Microenvironment regulation, L-lys inhibits the growth of glioma through arginine predation;

[0010] Metabolic signal pathway blockage, N1-MNA and L-lys synergistically inhibit the growth of glioma by inhibiting the Sirt1 / Akt / mTOR pathway.

[0011] As a further aspect of the present application, L-lys and N1-MNA jointly inhibit glioma cell proliferation, migration and invasion.

[0012] As a further aspect of the present application, L-lys and N1-MNA jointly inhibit glioma tumor growth.

[0013] As a further aspect of the present application, N1-MNA reduces the ability of glycolysis in glioma cells.

[0014] A drug for inhibiting growth of neuroglioma, the drug comprising an effective dose of L-lys and N1-MNA.

[0015] As a further aspect of the present application, the drug comprises a pharmaceutically acceptable carrier or diluent, and the administration mode of the drug includes but is not limited to oral administration, intravenous injection, intraperitoneal injection or local administration.

[0016] The beneficial effects of the present application are as follows:

[0017] 1. The synergistic mechanism of L-lys and N1-MNA microenvironment regulation and metabolic signal pathway blocking inhibits growth of neuroglioma, L-lys inhibits growth of neuroglioma through arginine deprivation, N1-MNA and L-lys synergistically inhibit growth of neuroglioma by inhibiting the Sirt1 / Akt / mTOR pathway, and the combination of the two can significantly enhance the anti-tumor effect.

[0018] 2. The combination of L-lys and N1-MNA can significantly improve the tumor inhibition rate, and there is no significant weight loss (indicating low toxicity), which is significantly better than the "efficacy-toxicity" balance of traditional chemotherapy, and at the same time avoids the problem of drug resistance of targeted drugs. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the present application, the drawings needed in the following specific embodiment description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0020] Figure 1 : Schematic diagram of the effect of L-lys and N1-MNA on U251 cell proliferation;

[0021] Figure 2 : Schematic diagram of the effect of L-lys and N1-MNA on U87 cells and other cells;

[0022] Figure 3 : Schematic diagram of the effect of L-lys and N1-MNA on growth of glioma tumor in nude mice;

[0023] Figure 4 : Schematic diagram of the effect of L-lys and N1-MNA on glycolysis and matrix metalloproteinase;

[0024] Figure 5 : Schematic diagram of the effect of L-lys and N1-MNA on glioma cell antitumor immunity;

[0025] Figure 6 : Schematic diagram of L-lys and N1-MNA inhibiting Sirt1 / Akt / mTOR pathway. DETAILED DESCRIPTION

[0026] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0027] The effects of L-lys and N1-methylnicotinamide in synergistically inhibiting the growth of glioma are verified in the present application through the following experiments.

[0028] 1. Cell culture

[0029] Human glioma cell lines U251, U87, mouse normal astrocyte C8-D1A, mouse neuron cell HT22, and mouse microglial cell BV-2 cells (Shanghai Cell Bank of Chinese Academy of Sciences) were cultured in DMEM medium containing 10% fetal bovine serum (FBS, Gibco) and 1% penicillin-streptomycin (HyClone) in a 37℃, 5% CO2 incubator.

[0030] 2. CCK8 method for detecting cell proliferation

[0031] U251 and U87 cells were inoculated in a 96-well plate at a concentration of 5×10 3 cells / well, and after 24h of culture, the following were added:

[0032] Different concentrations of L-lys (CAS: 56-87-1, HY-N0469, MCE, 0, 2.5, 5, 10, and 20mM), wherein CAS is a unique numerical identifier, HY-N0469 is a product catalog number assigned by the supplier, and MCE is the name of the supplier. This L-lys is referred to as H9;

[0033] Different concentrations of N1-MNA (CAS: 114-33-0, HY-124124, MCE, 0, 1, 5, 10, and 20μM). This N1-MNA is referred to as G5;

[0034] The two were combined (H9+G5, 1 / 1, 2.5 / 2.5, 5 / 5, 10 / 10 (mM / μM), with 3 replicates per group. After 48 h of culture, 10 μL of CCK8 solution (Beyotime) was added, and incubation continued for 2 h. The absorbance (OD value) at 450 nm was measured using a microplate reader, and cell viability and IC50 were calculated.

[0035] 3. Cell scratch assay to detect migration ability

[0036] U251 cells were seeded in 6-well plates. After confluence, the cells were scratched with a 200 μL pipette tip, washed with PBS, and then added to serum-free medium containing the drug. The cells were photographed at 0, 12, and 24 h, and the scratch healing rate (migration rate) was calculated using ImageJ software.

[0037] 4. Transwell assay for invasiveness

[0038] The upper chamber of the Transwell (Corning) apparatus is coated with Matrigel (BD Biosciences, Cat. No.: 354234, 1:8 dilution, final concentration 200 μg / mL), the lower chamber is added with culture medium containing 20% ​​FBS, and the upper chamber is inoculated with 1×10⁶ cells / mL. 5 U251 cells treated with the drug were cultured for 24 hours, then fixed, stained with crystal violet, and the number of cells that penetrated the membrane was counted.

[0039] 5. Real-time PCR

[0040] Tumor tissues from each group of mice were collected and placed in tubes containing an appropriate amount of Trizol (TIANGEN, DP424, China). Two 3mm enzyme-free magnetic beads were added to each tube. The tumor tissues were homogenized using a tissue homogenizer, and total RNA was extracted. A reverse transcription system (Takara, RR036A, Japan) was prepared, incubated at 37°C for 15 min, then at 85°C for 5 s to inactivate the reverse transcriptase, and the reverse transcription reaction was terminated to obtain the reverse transcription product. The cDNA samples were diluted appropriately and a cDNA reaction system was prepared for testing (TIANLONG, Gentier 96E). 2 -ΔΔCt The method was used to calculate the differences in mRNA transcription levels of the target gene.

[0041] 6. Blood-brain permeability test

[0042] Male SD rats aged 6-8 weeks were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd. and housed in a specific pathogen-free (SPF) environment. After one week of acclimatization, the rats were randomly divided into three groups: normal control group (administered via gavage with phosphate-buffered saline (PBS); N1-MNA group (administered via gavage with N1-methylnicotinamide (N1-MNA) at a dose of 50 mg / kg); and L-lys group (administered via gavage with L-lys at a dose of 100 mg / kg). At 0, 1, 2, and 4 hours after gavage, the concentrations of N1-MNA and L-lys in rat blood and brain tissue were detected using liquid chromatography-mass spectrometry (LC-MS).

[0043] 7. ELISA

[0044] U251 cells were used at a rate of 2 × 10 5 Cells were seeded in 6-well plates. After 48 hours of drug treatment, cell lysates were collected. Following the instructions of the EGFR ELISA kit (R&D Systems), the OD value at 450 nm was measured, and the EGFR protein content was calculated based on the standard curve. Methods for detecting the concentrations of MMP-9, MMP-2, and IL-10 in tumor tissues were also discussed.

[0045] The concentration of matrix metalloproteinase-9 (MMP-9) in tumor tissue was detected by enzyme-linked immunosorbent assay (ELISA), and the procedure was strictly performed in accordance with the instructions of the kit manufacturer (Biotechne).

[0046] The concentration of matrix metalloproteinase-2 (MMP-2) was determined using an ELISA-type assay system (manufacturer: Amersham Biosciences). This assay system comprises a monoclonal antibody capture system and uses MMP-2-specific substrates, with a detection specificity range of 190 pg / mL to 0.5 ng / mL.

[0047] The level of interleukin-10 (IL-10) in tumor tissue was detected using an ELISA kit (manufacturer: R&D Systems). The procedure was performed in accordance with the instructions provided by the kit manufacturer.

[0048] 8. Detection of hexokinase (HK-2) and lactate dehydrogenase (LDH) activities

[0049] Hexokinase (HK-2) activity was detected according to the instrument instructions (Solarbio, catalog number BC0745). The specific steps were as follows: After ultrasonic disruption, the lysate was centrifuged at 8000×g for 10 minutes. In the assay system (final concentration containing specified gradient concentrations of glucose, 6.6 mM adenosine triphosphate (ATP), 0.5 mg / mL nicotinamide adenine dinucleotide phosphate (NADP), and 0.5 U / mL glucose-6-phosphate dehydrogenase (G6PDH), pH 8.5), HK-2 activity was determined by the ability of the whole-cell lysate to phosphorylate glucose within 5 minutes. Glucose-6-phosphate (G6P) production was indirectly measured by reducing nicotinamide adenine dinucleotide phosphate (NADPH) catalyzed by glucose-6-phosphate dehydrogenase (G6PDH)—the absorbance was measured at 340 nm using a spectrophotometer to reflect the amount of G6P produced. Lactate dehydrogenase (LDH) activity was detected using an LDH assay kit (Jiancheng Biotechnology Co., Ltd., catalog number A020-2), strictly following the manufacturer's instructions. Silent information regulator 1 (SIRT1) enzyme activity was detected using a commercial kit from Abcam, Cambridge, UK (catalog number ab156065), according to the manufacturer's instructions. The inhibition rate of SIRT1 enzyme activity by the samples was calculated as the ratio of fluorescence intensity between the sample group and the control group.

[0050] 9. Construction of a subcutaneous xenograft model of glioma

[0051] Male BALB / c nude mice (4-5 weeks old, weighing 15-20g, Hunan Slack Jingda Experimental Animal Co., Ltd.) were housed in an SPF-grade environment with free access to food and water. U251 cells (1×10⁻⁶) were introduced. 7 L-lys (50 mg / kg, gavage) was injected subcutaneously into the right ventral dorsal region of nude mice. When the tumor volume reached 50 mm³, the nude mice were randomly divided into 4 groups (n=6): control group (physiological saline), L-lys group (50 mg / kg, gavage), N1-MNA group (10 mg / kg, intraperitoneal injection), and combination group (L-lys 50 mg / kg + N1-MNA 10 mg / kg). The drugs were administered once every 2 days for 4 weeks.

[0052] 10. Tumor growth monitoring and sample collection

[0053] Measure tumor volume (V = length × width) every 6 days. 2 / 2) and the weight of nude mice were used to plot tumor growth curves and calculate the tumor growth inhibition rate (TGI = [1 - relative tumor volume of experimental group (RTV) / RTV of control group] × 100%, RTV = Vt / V0, where Vt is the volume at the experimental time point and V0 is the initial volume of drug administration). At the end of the experiment, nude mice were sacrificed, and tumor tissue was dissected. Part of the tissue was used for HE staining, and part was used for Western blot detection.

[0054] 11. Western blot detection of signaling pathway proteins

[0055] Tumor tissue or cell lysates were extracted, and the expression of p-Akt, p-mTOR, and Sirt1 proteins was detected. GAPDH was used as an internal control, and the grayscale values ​​were quantitatively analyzed using ImageJ. Anti-Sirt1, p-Akt, and anti-p-mTOR antibodies (Abcam) were also used.

[0056] 12. Flow cytometry

[0057] After trypsin digestion of tumor tissue, anti-CD206 antibody was added and incubated. The number of CD206+ cells was counted by flow cytometry, and CD206+ positive cells were analyzed by ImageJ.

[0058] 13. Arginine content detection

[0059] The arginine content in cancerous tissue was determined by high performance liquid chromatography (HPLC) in cell culture medium and tumor tissue homogenate. The chromatographic column was a C18 column (250 mm × 4.6 mm), the mobile phase was methanol-0.1% formic acid water (5:95), and the detection wavelength was 220 nm.

[0060] 14. Statistical Analysis

[0061] Statistical analysis was performed using GraphPad Prism 9.0 software. Data are expressed as mean ± standard deviation (x ± s). One-way ANOVA was used for comparisons among multiple groups, and LSD-t test was used for pairwise comparisons. P < 0.05 was considered statistically significant.

[0062] Based on the above experiments, the roles of L-lys and N1-methylnicotinamide in synergistically inhibiting glioma growth were analyzed.

[0063] 1. L-lys combined with N1-MNA inhibits the proliferation, migration, and invasion of glioma cells.

[0064] like Figure 1 As shown, Figure 1 middle:

[0065] A. Schematic diagram of the effects of G5, H9, and G5+H9 on U251 cell survival; B. Schematic diagram of the effects of G5, H9, and G5+H9 on EGFR mRNA and protein expression in U251 cells; C. Schematic diagram of the effects of G5, H9, and G5+H9 on U251 cell migration; D. Schematic diagram of the effects of G5, H9, and G5+H9 on U251 cell invasion ability; E. Statistical graph of the effects of G5, H9, and G5+H9 on U251 cell migration; F. Statistical graph of the effects of G5, H9, and G5+H9 on U251 cell invasion.

[0066] Figure 1 In the intervals, at **, p < 0.01; at ***, p < 0.001; at ****, p < 0.0001.

[0067] The CCK8 results showed that the inhibitory effects of L-lys and N1-MNA on U251 cell proliferation were concentration-dependent. Figure 1 A) In U251 cells, the IC50 of N1-MNA 50 IC50 with a concentration of 5.2 μM and L-lys 50 The value is 8.7mM, and the IC is processed together. 50 When the concentration was reduced to 3.5 mmol / L, the cell viability was significantly lower than that of the treatment alone (p < 0.01, Figure 1A).

[0068] RT-PCR and ELISA results showed that treatment with L-lys (5 mM) or N1-MNA (5 μM) alone significantly reduced EGFR mRNA and protein expression in U251 cells, and the EGFR content further decreased after combined treatment with L-lys and N1-MNA (2.5 mM + 2.5 μM) (Figure 1B).

[0069] Cell scratch assays showed no significant difference in migration rates among the groups at 12 h, but at 24 h, the migration rates of L-lys and N1-MNA and the combined treatment group were significantly lower than those of the control group (Figure 1C, E).

[0070] Transwell assays showed that the L-lys, N1-MNA and combined treatment groups had significantly fewer antibodies against the control group (Figures 1D and F), indicating that L-lys or N1-MNA and the combination of both can enhance the inhibition of glioma cell invasion.

[0071] like Figure 2 As shown, Figure 2 middle:

[0072] A. Schematic diagram of the effects of G5, H9, and G5+H9 on the survival rate of U87 cells; B. Schematic diagram of the effects of G5+H9 on the proliferation of CTX TNA2, HT-22, and BV-2 cells; C. Schematic diagram of the structure and bioinformatics analysis of L-lys; D. Schematic diagram of the structure and bioinformatics analysis of N1-MNA; E. Schematic diagram of plasma and intracerebral concentrations at different time points after gavage administration of 100 mg / kg L-Lys; F. Schematic diagram of plasma and intracerebral concentrations at different time points after gavage administration of 50 mg / kg N1-MNA.

[0073] Figure 2 At **, p < 0.01; at ***, p < 0.001.

[0074] Further CCK8 results showed that, in U87 cells, the inhibitory effects of L-lys and N1-MNA on cell proliferation were also concentration-dependent. Figure 2 A), N1-MNA IC 50 IC50 with a concentration of 5.1 μM and L-lys 50 The concentration is 5.4 mM, and the IC is after joint processing. 50 When the concentration dropped to 3.3 mmol / L, cell viability was significantly lower than that of the treatment alone (Figure 2A).

[0075] L-lys, N1-MNA, and their combined treatment had no effect on the proliferation of normal astrocytes C8-D1A and neurons HT-22 within the treatment concentration range, but had an inhibitory effect on microglia BV-2 at combined treatment concentrations of 5 / 5 and 10 / 10 (L-lys / N1-MNA: mM / μM) (Figure 2B).

[0076] Figures 2C and 2D show the structures of L-lys and N1-MNA, respectively. Figure 2 (The image above C) and bioinformatics analysis ( Figure 2 (See the image below for C).

[0077] Figure 2 E showed that when L-Lys was administered by gavage at a dose of 100 mg / kg, the time to peak concentration was about 1 hour, and the time to peak concentration in the brain was about 2 hours. The brain concentration was (7.44±0.89)% of the blood concentration.

[0078] Figure 2 F shows that when 50 mg / kg N1-MNA was administered by gavage, the time to peak concentration was about 1 hour, and the time to peak concentration in the brain was about 2 hours. The brain concentration was (18.33±1.56)% of the blood concentration.

[0079] 2. L-lys combined with N1-MNA inhibits the growth of glioma xenografts.

[0080] like Figure 3 As shown, Figure 3 In the diagram: A, tumor growth curve; B, tumor volume diagram at 24 days for each group; C, TGI diagram of tumor at each time point.

[0081] Among them, M is the control group; M+G5 is the N1-MNA administration group; M+H9 is the L-Lys administration group; and M+G5 / H9 is the combined administration group.

[0082] Figure 3 At **, p < 0.01; at ***, p < 0.001.

[0083] There was no significant difference in body weight among the nude mice in each group during the administration period (p>0.05), indicating that the drug has good safety.

[0084] Tumor growth curves showed that the tumor volume in the L-lys, N1-MNA and combined treatment groups was significantly smaller than that in the control group starting from day 12. At the experimental endpoint (day 24), the tumor volume in the L-lys, N1-MNA and combined treatment groups was significantly smaller than that in the control group, and the tumor volume in the combined group was further reduced compared with the single treatment group (Figures 3A-C).

[0085] The TGI results showed that at 24 days, the TGI of the combined group was significantly higher than that of the L-lys group and the N1-MNA treatment group alone. Figure 3 D).

[0086] 3. N1-MNA reduces the glycolytic capacity of glioma cells.

[0087] like Figure 4 As shown, Figure 4 A. Schematic diagram of HK-2 mRNA and HK-2 activity (expressed as NADH production); B. Schematic diagram of LDH-A mRNA and LDH-A activity; C. Schematic diagram of MMP-2 expression in tissues; D. Schematic diagram of MMP-9 expression in tissues.

[0088] Figure 4 In the intervals, * indicates p < 0.05; ** indicates p < 0.01; *** and **** indicate p < 0.001.

[0089] L-lys had no effect on HK-2 mRNA or HK-2 activity in tumor tissue. N1-MNA treatment alone downregulated HK-2 enzyme activity, and the enzyme activity expression was further reduced in the combined treatment group (Figure 4A).

[0090] L-lys had no effect on LDH-A mRNA or enzyme activity. N1-MNA treatment alone downregulated LDH-A enzyme activity (P<0.05), and the enzyme activity was further reduced in the combined treatment group (Figure 4B). RT-PCR and ELISA results showed that N1-MNA significantly promoted MMP-2 expression (Figure 4C). RT-PCR and ELISA results showed that N1-MNA had no effect on MMP-9 expression, but L-lys significantly inhibited MMP-9 expression, showing a synergistic effect. Figure 4 D).

[0091] 4. L-lys inhibits glioma growth through arginine depletion.

[0092] like Figure 5 As shown, Figure 5 In Chinese: A, Schematic diagram of intracellular arginine concentration; B, Schematic diagram of IL-10 expression detection; C, CD206 + Schematic diagram illustrating the impact of mRNA; D, detection of CD206 + Schematic diagram of a cell; E, CD206 + Schematic diagram of cytological statistical analysis.

[0093] Figure 5 In the intervals, at **, p < 0.01; at ***, p < 0.001; at ****, p < 0.0001.

[0094] HPLC results showed that the arginine content in tumor tissue was significantly reduced after L-lys treatment (Figure 5A).

[0095] RT-PCR and ELISA results showed that L-lys significantly inhibited IL-10 secretion in tumor tissue (Figure 5B).

[0096] RT-PCR and flow cytometry showed that CD206 in the L-lys group + The expression of CD206 was significantly reduced, and the combined treatment group showed a decrease in CD206 expression in tumor tissue. + The number of cells was significantly less than in the treatment-alone group (Figure 5 CE).

[0097] 5. N1-MNA and L-lys synergistically inhibit glioma growth by suppressing the Sirt1 / Akt / mTOR pathway.

[0098] like Figure 6 As shown, Figure 6 A. Schematic diagram of Sirt1 mRNA, protein, and activity; B. Detection results of Sirt1 / p-Akt / p-mTOR; C. Schematic diagram of statistical analysis of Sirt1 / p-Akt / p-mTOR molecules.

[0099] Figure 6 In the intervals, * indicates p < 0.05; ** indicates p < 0.01; *** indicates p < 0.001.

[0100] Tumor tissue samples were collected, and Real-time PCR, Western blot, and enzyme activity assays showed that L-lys treatment alone had no effect on Sirt1 mRNA, protein, or enzyme activity. However, N1-MNA treatment alone significantly downregulated Sirt1 enzyme activity, and combined treatment further reduced Sirt1 enzyme activity (Figures 6A-C).

[0101] Western blot results showed that both L-lys and N1-MNA treatment alone significantly downregulated p-Akt and p-mTOR expression, and combined treatment further reduced p-mTOR protein expression (Figures 6B and 6C). This suggests that N1-MNA and L-lys exert their anti-tumor effects synergistically by inhibiting the Akt / mTOR pathway.

[0102] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to any specific implementation. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention.

Claims

1. The application of combined L-lys and N1-MNA in synergistic inhibition of glioma growth, characterized in that, L-lys and N1-MNA inhibit glioma growth through a synergistic mechanism of microenvironment regulation and metabolic signaling pathway blockade. L-lys is L-lysine, and N1-MNA is N1-methylnicotinamide.

2. The application of the combined use of L-lys and N1-MNA in synergistic inhibition of glioma growth according to claim 1, characterized in that, L-lys and N1-MNA synergistically inhibit glioma growth through the following mechanisms: Microenvironment regulation: L-lys inhibits glioma growth through arginine depletion; By blocking metabolic signaling pathways, N1-MNA and L-lys synergistically inhibit glioma growth by inhibiting the Sirt1 / Akt / mTOR pathway.

3. The application of the combined use of L-lys and N1-MNA according to claim 1 in the synergistic inhibition of glioma growth, characterized in that, L-lys combined with N1-MNA inhibits the proliferation, migration, and invasion of glioma cells.

4. The application of the combined use of L-lys and N1-MNA according to claim 1 in the synergistic inhibition of glioma growth, characterized in that, L-lys combined with N1-MNA inhibits the growth of glioma xenografts.

5. The application of the combined use of L-lys and N1-MNA according to claim 1 in the synergistic inhibition of glioma growth, characterized in that, N1-MNA reduces the glycolytic capacity of glioma cells.

6. A drug for inhibiting the growth of gliomas, characterized in that, The drug contains an effective dose of L-lys and N1-MNA.

7. A drug for inhibiting the growth of gliomas, characterized in that, The drug contains a pharmaceutically acceptable carrier or diluent, and the methods of administration include, but are not limited to, oral, intravenous, intraperitoneal, or local administration.

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