Application of combined medicine composition in preparation of medicine for resisting transforming follicular lymphoma

Abstract

The invention relates to application of a combined pharmaceutical composition in preparation of a medicine for resisting transforming follicular lymphoma. Active components in the combined pharmaceutical composition comprise a first component and a second component, the first component is selected from any one or a combination of at least two of sitagliflozin, pharmaceutically acceptable salt, isomer, solvate and metabolite of sitagliflozin; and the second component is selected from any one or a combination of at least two of chidamide, pharmaceutically acceptable salts, isomers, solvates and metabolites of the chidamide. Research finds that the combined use of the two active components not only can reduce the dosage of each drug and improve the medication safety, but also has the effect of improving or treating convertible follicular lymphoma more remarkably than that of a single drug active component, and plays a role in synergistic promotion. The invention provides an effective drug combination strategy for improvement or treatment of convertible follicular lymphoma, and has very significant significance.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and relates to a new combined drug intervention strategy for transformed follicular lymphoma, and particularly to the application of a combined drug composition in the preparation of a drug for treating transformed follicular lymphoma. Background Art

[0002] Follicular Lymphoma (FL) is a common indolent lymphoma, and there is currently no cure. Particularly critically, from the time of diagnosis, some patients will evolve into histologically more aggressive lymphomas and will ultimately transform into aggressive lymphomas, and this type is called transformed follicular lymphoma (t-FL). The prognosis of t-FL is poor, and its death risk is 5 to 6 times that of non-transformed FL, which is the main factor for the death of current FL patients and is also the difficulty and focus in the treatment of FL.

[0003] The current treatment regimens recommended by the guidelines for t-FL mainly include immunochemotherapy, hematopoietic stem cell transplantation, cell therapy, and clinical trials. Previous studies have shown that autologous hematopoietic stem cell transplantation (auto-HCT) can optimize the prognosis of t-FL. However, the latest clinical studies suggest that auto-HCT mainly has an improvement effect on patients who have not used rituximab, and the vast majority of current t-FL patients have used rituximab combined with chemotherapy in previous treatments. The CAR-T therapy has a significant effect on t-FL, but the price of CAR-T is high, making it unaffordable for the vast majority of patients. In particular, it should be noted that the median age of onset of FL patients is 60 years old, and the age of patients will be even older after transformation. Most t-FL patients cannot tolerate high-dose chemotherapy and auto-HCT. Therefore, there is an urgent need in clinical practice to explore a safe and effective regimen to improve the efficacy of t-FL and its prognosis.

[0004] Chidamide, as the only new histone deacetylase (HDAC) inhibitor approved for marketing in China at present, can specifically target subtypes 1, 2, and 3 of class I HDACs and subtype 10 of class IIb HDACs, promote the increase in the acetylation levels of histone H3 and H4, and ultimately activate gene transcription, thereby causing changes in the gene expression of multiple signal transduction pathways (i.e., epigenetic changes), further inhibiting the proliferation of tumor cells and inducing their apoptosis.

[0005] Chiglitazar, as a novel peroxisome proliferator-activated receptor (PPAR) pan-agonist, can regulate glucose, lipid, and energy metabolism, coordinate environmental signals and metabolic pathways, and maintain glucose homeostasis. Clinically, it is mainly used for the treatment of type 2 diabetes.

[0006] It is not clear whether the combined use of these two drugs can synergistically inhibit the progression of t-FL. Therefore, developing a new drug combination therapy that can effectively kill t-FL has important clinical value for the treatment of t-FL. Summary of the Invention

[0007] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a new combined drug intervention strategy for transformed follicular lymphoma, specifically to provide the use of a combined drug composition in the preparation of a drug for treating transformed follicular lymphoma.

[0008] To achieve the purpose of this invention, the following technical solutions are adopted:

[0009] In the first aspect, the present invention provides the use of a combined drug composition in the preparation of a drug for treating transformed follicular lymphoma. The active components in the combined drug composition include a first component and a second component. The first component is selected from any one or at least two combinations of ciglitazone, its pharmaceutically acceptable salts, isomers, solvates, and metabolites. The second component is selected from any one or at least two combinations of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0010] The present invention has developed a completely new drug use strategy for improving or treating transformed follicular lymphoma. Specifically, ciglitazone (or its pharmaceutically acceptable salts, isomers, solvates, metabolites) is combined with chidamide (or its pharmaceutically acceptable salts, isomers, solvates, metabolites) for intervening in the progression of transformed follicular lymphoma.

[0011] The present invention's research has found that the combined use of these two active components can not only reduce the dosage of each drug and improve drug safety, but also has a more significant effect on improving or treating transformed follicular lymphoma than a single drug active component, showing a synergistic promotion effect. The present invention first proves through transformed follicular lymphoma cell lines that it can significantly inhibit the proliferation of transformed follicular lymphoma cells and induce their apoptosis. Through the CDX mouse model, it is proved that the combined drug composition can inhibit the tumorigenesis process in mice and improve the survival rate of mice. Through transcriptome sequencing analysis, it shows that the combined drug composition can induce mitophagy and affect signal pathways such as glycolysis / gluconeogenesis. The present invention provides an effective drug combination strategy for the improvement or treatment of transformed follicular lymphoma, which has very significant significance.

[0012] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylate include alkali metal salts, alkaline earth metal salts, ammonium salts or quaternary ammonium salts.

[0013] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylate include sodium salts or potassium salts.

[0014] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylate is sodium ciglitazone.

[0015] Preferably, the combined pharmaceutical composition further contains pharmaceutically acceptable excipients.

[0016] Preferably, the combined pharmaceutical composition of the present invention can be administered alone or in combination with excipients to form a suitable dosage form for administration. The pharmaceutically acceptable excipients include any one or a combination of at least two of carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, bacteriostatic agents or buffers.

[0017] Preferably, the combined pharmaceutical composition is a single compound preparation or a combination of two separate preparations. When the combined pharmaceutical composition is a combination of two separate preparations, the two separate preparations are administered simultaneously or sequentially.

[0018] The combined pharmaceutical composition can be in the form of a single compound preparation or a combination of two separate preparations; when it is a combination of two separate preparations, the administration method can be simultaneous administration, cross-administration or sequential administration.

[0019] Preferably, the preparation is any pharmaceutically acceptable dosage form, such as tablets, powders, suspensions, granules, capsules, solutions, enemas, emulsions, etc.

[0020] In a second aspect, the present invention provides an application of a combined pharmaceutical composition in the preparation of a transforming follicular lymphoma cell proliferation inhibitor. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone carboxylate, its pharmaceutically acceptable salts, isomers, solvates, metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, metabolites.

[0021] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylate include alkali metal salts, alkaline earth metal salts, ammonium salts or quaternary ammonium salts.

[0022] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylate include sodium salts or potassium salts.

[0023] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylic acid is sodium ciglitazone.

[0024] In a third aspect, the present invention provides the use of a combined pharmaceutical composition for the non-therapeutic preparation of an inhibitor of the proliferation of transformed follicular lymphoma cells. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone carboxylic acid, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0025] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include alkali metal salts, alkaline earth metal salts, ammonium salts, or quaternary ammonium salts.

[0026] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include sodium salts or potassium salts.

[0027] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylic acid is sodium ciglitazone.

[0028] According to the research results of the present invention, the combined pharmaceutical composition has a significant effect of inhibiting the proliferation of transformed follicular lymphoma cells. Therefore, this result indicates that the combined pharmaceutical composition can be used as a reagent for in vitro experiments in the scientific research field, rather than for the treatment of diseases, such as studying the growth and other metabolic mechanisms or behaviors of transformed follicular lymphoma cells, screening drugs for the treatment of follicular lymphoma, etc.

[0029] Preferably, the transformed follicular lymphoma cells include SC-1 cells and / or DHL4 cells.

[0030] In a fourth aspect, the present invention provides the use of a combined pharmaceutical composition for the preparation of a promoter of apoptosis of transformed follicular lymphoma cells. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone carboxylic acid, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0031] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include alkali metal salts, alkaline earth metal salts, ammonium salts, or quaternary ammonium salts.

[0032] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include sodium salts or potassium salts.

[0033] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylic acid is sodium ciglitazone.

[0034] In a fifth aspect, the present invention provides the use of a combined pharmaceutical composition for non-therapeutic purposes in the preparation of an agent for promoting apoptosis of transformed follicular lymphoma cells. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone carboxylic acid, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0035] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include alkali metal salts, alkaline earth metal salts, ammonium salts, or quaternary ammonium salts.

[0036] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include sodium salts or potassium salts.

[0037] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylic acid is sodium ciglitazone.

[0038] According to the research results of the present invention, the combined pharmaceutical composition has a significant effect of promoting apoptosis of transformed follicular lymphoma cells. Therefore, this result indicates that the combined pharmaceutical composition can be used as a reagent for in vitro experiments in the scientific research field, rather than for the treatment of diseases, such as studying the apoptosis and other metabolic mechanisms or behaviors of transformed follicular lymphoma cells, screening drugs for the treatment of follicular lymphoma, etc.

[0039] Preferably, the transformed follicular lymphoma cells include SC-1 cells and / or DHL4 cells.

[0040] In a sixth aspect, the present invention provides the use of a combined pharmaceutical composition in the preparation of a preparation for inducing mitophagy of transformed follicular lymphoma cells. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone carboxylic acid, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0041] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include alkali metal salts, alkaline earth metal salts, ammonium salts, or quaternary ammonium salts.

[0042] Preferably, the pharmaceutically acceptable salts of ciglitazone carboxylic acid include sodium salts or potassium salts.

[0043] Preferably, the pharmaceutically acceptable salt of ciglitazone carboxylic acid is sodium ciglitazone.

[0044] In a seventh aspect, the present invention provides the use of a combined pharmaceutical composition for non-therapeutic purposes in the preparation of a preparation for inducing mitophagy in transformed follicular lymphoma cells. The active components in the combined pharmaceutical composition include a first component and a second component; the first component is selected from any one or a combination of at least two of ciglitazone, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; the second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

[0045] Preferably, the pharmaceutically acceptable salts of ciglitazone include alkali metal salts, alkaline earth metal salts, ammonium salts or quaternary ammonium salts.

[0046] Preferably, the pharmaceutically acceptable salts of ciglitazone include sodium salts or potassium salts.

[0047] Preferably, the pharmaceutically acceptable salt of ciglitazone is sodium ciglitazone.

[0048] According to the research results of the present invention, the combined pharmaceutical composition has a significant effect of inducing mitophagy in transformed follicular lymphoma cells. Therefore, the results indicate that the combined pharmaceutical composition can be used as a reagent for in vitro experiments in the scientific research field, rather than for the treatment of diseases, such as studying the mitophagy behavior or other metabolic mechanisms of transformed follicular lymphoma cells.

[0049] Compared with the prior art, the present invention has the following beneficial effects:

[0050] The present invention has developed a brand-new medication strategy for improving or treating transformed follicular lymphoma. Specifically, ciglitazone (or its pharmaceutically acceptable salts, isomers, solvates, metabolites) is combined with chidamide (or its pharmaceutically acceptable salts, isomers, solvates, metabolites) for use in intervening in the progression of transformed follicular lymphoma.

[0051] The present invention has found through research that the combination of the above two active components can not only reduce the dosage of each drug and improve the drug safety, but also has a more significant effect of improving or treating transformed follicular lymphoma than a single drug active component, achieving a synergistic promotion effect. The present invention first proves through a transformed follicular lymphoma cell line that it can significantly inhibit the proliferation of transformed follicular lymphoma cells and induce apoptosis; through a CDX mouse model, it is proved that the combined pharmaceutical composition can inhibit the tumorigenesis process of mice and improve the survival rate of mice; through transcriptome sequencing analysis, it shows that the combined pharmaceutical composition can induce mitophagy and affect signal pathways such as glycolysis / gluconeogenesis. The present invention provides an effective drug combination strategy for the improvement or treatment of transformed follicular lymphoma, which has very significant significance. Description of the Drawings

[0052] Figure 1A It is a statistical result diagram of the cell proliferation inhibition rate after treating SC-1 cells with ciglitazone sodium combined with chidamide;

[0053] Figure 1B It is a statistical result diagram of the cell proliferation inhibition rate after treating DHL4 cells with ciglitazone sodium combined with chidamide;

[0054] Figure 2A It is a flow cytometry detection result diagram of the apoptosis level after treating SC-1 cells with ciglitazone sodium combined with chidamide;

[0055] Figure 2B It is a statistical result diagram of the apoptosis rate after treating SC-1 cells with ciglitazone sodium combined with chidamide;

[0056] Figure 3A It is a flow cytometry detection result diagram of the apoptosis level after treating DHL4 cells with ciglitazone sodium combined with chidamide;

[0057] Figure 3B It is a statistical result diagram of the apoptosis rate after treating DHL4 cells with ciglitazone sodium combined with chidamide;

[0058] Figure 4A It is a comparison result diagram of the tumor weights of each group of mice after treating the CDX mouse model with ciglitazone sodium combined with chidamide;

[0059] Figure 4B It is a comparison result diagram of the tumor volumes of each group of mice after treating the CDX mouse model with ciglitazone sodium combined with chidamide;

[0060] Figure 4C It is an anatomical appearance diagram of the tumors of each group of mice after treating the CDX mouse model with ciglitazone sodium combined with chidamide;

[0061] Figure 5 It is a result diagram of transcriptome sequencing analysis after treating SC-1 cells with ciglitazone sodium combined with chidamide;

[0062] Figure 6 It is a Western blot result diagram of the expression levels of mitophagy-related proteins after treating SC-1 cells and DHL4 cells with ciglitazone sodium combined with chidamide. Detailed Embodiments

[0063] The technical solution of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations of the present invention.

[0064] The drugs Chiglitazar involved in the following examples were provided by Shenzhen Chipscreen Biosciences Co., Ltd.; the drug CS055 was provided by Shenzhen Chipscreen Biosciences Co., Ltd.; the transformed follicular lymphoma cell lines (including SC-1 cells and DHL4 cells) were provided by the Institute of Hematology, School of Medicine, Xiamen University.

[0065] Example 1

[0066] Inhibitory effect of the combined drug composition on the proliferation of t-FL cell lines

[0067] The operation method was as follows: Take t-FL cell lines (including SC-1 and DHL4 cell lines) in the logarithmic growth phase and inoculate them into 96-well plates. Set up the Chiglitazar single-drug group, CS055 single-drug group, Chiglitazar and CS055 combination group (Combo), and control group; 4 The single-drug concentrations of Chiglitazar in the experimental group cells were 2 μM, 4 μM, 8 μM, 16 μM, and 32 μM respectively. The single-drug concentrations of CS055 in the experimental group cells were 0.2 μM, 0.4 μM, 0.8 μM, 1.6 μM, and 3.2 μM respectively. The combined concentrations of Chiglitazar and CS055 were 2 μM + 0.2 μM, 4 μM + 0.4 μM, 8 μM + 0.8 μM, 16 μM + 1.6 μM, and 32 μM + 3.2 μM (the former is Chiglitazar and the latter is CS055). Among them, the control group cells were treated with the same volume of DMSO; After gently oscillating and mixing the corresponding volume of the drug or DMSO with the cells in the above 96-well cell culture plate, culture them in a cell culture incubator (Thermo) for 24 h, and then detect the cell proliferation level using a CCK8 kit (MCE, Shanghai).

[0068] The results of the cell proliferation inhibition rate of each group are as

[0069] shown in Figure 1A (SC-1) and Figure 1B (DHL4);

[0070] From Figure 1A and Figure 1B the results shown, compared with higher concentrations of Chiglitazar single drug or higher concentrations of CS055 single drug, when Chiglitazar and CS055 are used in combination, it can significantly reduce the usage amounts of the two drugs on the basis of improving the inhibition level, that is, it can ensure both low drug toxicity and side effects and excellent anti-transformed follicular lymphoma effects.

[0071] Example 2

[0072] Apoptosis-inducing effect of the combined drug composition on the t-FL cell line (SC-1 cell line)

[0073] The operation method is as follows: Take 1×10 5 t-FL cell line (SC-1 cell line) in the logarithmic growth phase was inoculated into a 24-well plate, and a single-drug group of Chiglitazar, a single-drug group of CS055, a combined group of Chiglitazar and CS055 (Combo), and a control group were set up;

[0074] The single-drug concentrations of Chiglitazar in the experimental group cells were 8 μM, 16 μM, and 32 μM respectively, and the single-drug concentrations of CS055 in the experimental group cells were 0.8 μM, 1.6 μM, and 3.2 μM respectively. The combined concentrations of Chiglitazar and CS055 were 8 μM + 0.8 μM, 16 μM + 1.6 μM, and 32 μM + 3.2 μM respectively (the former is Chiglitazar and the latter is CS055). The control group cells were treated with the same volume of DMSO. After gently oscillating and mixing the corresponding volume of the drug or DMSO with the cells in the above 24-well cell culture plate, the cells were cultured in a cell culture incubator for 24 h, then centrifuged at 4 °C and 300 g for 5 min to collect the cells, washed once with PBS, and then the apoptosis level of the cells was detected by AnnexinV / PI (Thermofisher, USA) flow cytometry staining method and the apoptosis rate was counted.

[0075] Figure 2A and Figure 2B They are respectively the flow cytometry detection result graph of the apoptosis level of SC-1 cells treated with the combination of Chiglitazar and CS055 for 24 h and the statistical result graph of the apoptosis rate.

[0076] From the results shown in the above figure, it can be seen that compared with a higher concentration of Chiglitazar alone or a higher concentration of CS055 alone, when Chiglitazar and CS055 are used in combination, it can significantly reduce the usage amounts of the two drugs on the basis of improving the promotion of t-FL cell apoptosis, that is, it can ensure both low drug toxicity and side effects and excellent anti-transformed follicular lymphoma effect.

[0077] Example 3

[0078] Apoptosis-inducing effect of the combined drug composition on the t-FL cell line (DHL4 cell line)

[0079] The operation method is as follows: Take 1×10 5 t-FL cell line (DHL4 cell line) in the logarithmic growth phase was inoculated into a 24-well plate, and a single-drug group of Chiglitazar, a single-drug group of CS055, a combined group of Chiglitazar and CS055 (Combo), and a control group were set up;

[0080] The single-drug concentrations of chiglitazar in the experimental group cells were 8 μM, 16 μM, and 32 μM respectively, and the single-drug concentrations of chidamide in the experimental group cells were 0.8 μM, 1.6 μM, and 3.2 μM respectively. The combined concentrations of chiglitazar and chidamide were 8 μM + 0.8 μM, 16 μM + 1.6 μM, and 32 μM + 3.2 μM respectively (the former is chiglitazar and the latter is chidamide). Among them, the control group cells were treated with the same volume of DMSO; after gently oscillating and mixing the corresponding volume of the drug or DMSO with the cells in the above 24-well cell culture plate, they were cultured in a cell incubator for 24 h, then centrifuged at 4 °C and 300 g for 5 min to collect the cells, washed once with PBS, and then the apoptosis level of the cells was detected by AnnexinV / PI (Thermofisher, USA) flow cytometry staining method and the apoptosis rate was statistically analyzed.

[0081] Figure 3A and Figure 3B They are respectively the flow cytometry detection result chart of the apoptosis level of DHL4 cells after being treated with the combination of chiglitazar and chidamide for 24 h and the statistical result chart of the apoptosis rate.

[0082] It can be seen from the results shown in the above figure that compared with the single drug of chiglitazar at a higher concentration or the single drug of chidamide at a higher concentration, when chiglitazar and chidamide are used in combination, it can significantly reduce the usage amounts of the two drugs on the basis of improving the promotion of t-FL cell apoptosis level, that is, it can ensure both low drug toxicity and side effects and excellent anti-transformed follicular lymphoma effect at the same time.

[0083] Example 4

[0084] Effect of the combined drug composition on the tumorigenesis process of t-FL

[0085] The specific operation method is as follows:

[0086] (1) Set up a single-drug group of chiglitazar (Chiglitazar), a single-drug group of chidamide (CS055), a combined group of chiglitazar and chidamide (Combo), and a control group; among them, chiglitazar was formulated into a 100 mM stock solution with dimethyl sulfoxide (DMSO), and chidamide was formulated into a 10 mM stock solution with DMSO, and diluted with DMSO to the corresponding working concentration when in use.

[0087] (2) Construct a CDX mouse model

[0088] After irradiating CB17-SCID mice with 1 Gy of γ-ray, collect 1×10 6 Logarithmic growth phase t-FL cell line SC-1 cells were subcutaneously injected into CB17-SCID mice for subcutaneous xenograft tumors to establish a CDX mouse model.

[0089] (3) Administration of the drugs started 7 days after injection. The dose of ciglitazone sodium was 10 mg / kg / day, and the dose of chidamide was 5 mg / kg / day. The drugs were administered continuously for 14 days. The tumorigenesis progress in mice was detected by measuring the size of subcutaneous tumors. The starting day of drug administration was recorded as day 0, and the mice were sacrificed 14 days later for sample collection. Photos were taken and the samples were weighed to statistically analyze the volume and weight of the tumors. Among them, CB17-SCID mice were purchased from Jiangsu Genscript Biotech Co., Ltd. and were raised in the experimental animal center.

[0090] The statistical results of the tumor weights of mice in each group are shown in Figure 4A, and the statistical results of the tumor volumes are shown in Figure 4B as shown, and the anatomical appearance of the tumors is shown in Figure 4C as shown. The above experimental results show that, compared with the single-drug group, the combination of ciglitazone sodium and chidamide has a better effect in inhibiting the tumorigenesis process of the CDX model.

[0091] Example 5

[0092] Transcriptome sequencing experiment

[0093] The operation method is as follows:

[0094] Take 1×10 6 t-FL cell line (SC-1 cells) in the logarithmic growth phase and inoculate them into 6-cm cell culture dishes. Set a control group and a combination group of ciglitazone sodium and chidamide (Combo), with three replicates in each group;

[0095] The concentration of the combination group of ciglitazone sodium and chidamide is 16 μM + 1.6 μM (the former is ciglitazone sodium and the latter is chidamide). Among them, the cells in the control group were treated with the same volume of DMSO; after gently oscillating and mixing the corresponding volume of the drug or DMSO with the cells in the above cell culture dishes, the cells were cultured in a cell incubator for 12 h, then centrifuged at 4°C and 300 g for 5 min to collect the cells, washed once with PBS, and after discarding the supernatant, they were snap-frozen in liquid nitrogen and stored at -80°C, and sent to Kangce Biotechnology Company for transcriptome sequencing analysis.

[0096] Figure 5 For the analysis result graph, it can be seen from the figure that the t-FL cells treated with the combination of ciglitazone sodium and chidamide were KEGG-enriched to signal pathways such as mitophagy, glycolysis / gluconeogenesis, and one-carbon metabolism. The heatmap results also showed that the proteins related to mitophagy (the expression of VDAC1 and MFN2 was down-regulated, and the genes such as PINK1, PARK2, BNIP3L, SQSTM1, and BECLIN1 were up-regulated).

[0097] Example 6

[0098] Inductive effect of the combined pharmaceutical composition on mitophagy of t-FL cell line

[0099] The operation method is as follows: Take 1×10 6 t-FL cell lines in the logarithmic growth phase (SC-1 cells and DHL4 cells) were inoculated into 6-cm cell culture dishes, and groups were set up including a single-agent group of Chiglitazar, a single-agent group of CS055, a combined group of Chiglitazar and CS055 (Combo), and a control group;

[0100] The single-agent concentration of Chiglitazar in the experimental group cells was 16 μM respectively, the single-agent concentration of CS055 in the experimental group cells was 1.6 μM respectively, and the concentration in the combined group of Chiglitazar and CS055 was 16 μM + 1.6 μM (the former is Chiglitazar and the latter is CS055). Among them, the control group cells were treated with the same volume of DMSO; After gently oscillating and mixing the corresponding volume of the drug or DMSO with the cells in the above cell culture dishes, the cells were cultured in a cell incubator for 12 h, then centrifuged at 4 °C and 300 g for 5 min to collect the cells, washed once with PBS, lysed on ice for 1 h with 200 μL of RIPA lysis buffer (Thermo, USA), and the total protein was extracted for Western blot to detect the expression levels of proteins related to mitophagy.

[0101] Figure 6 The results of Western blot showed that, compared with the single-agent groups, the effect of inducing mitophagy in t-FL cell lines was more significant when Chiglitazar and CS055 were used in combination. Among them, Parkin, Beclin1, Bnip3L, and LC3BⅡ / LC3BⅠ were up-regulated, and the expressions of P62, MFN2, VDAC1, and BCL-XL were down-regulated, suggesting enhanced mitophagy. The combination of the two drugs may induce the death of t-FL cells through mitophagy.

[0102] The applicant declares that the present invention illustrates the technical solution of the present invention through the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the present invention product, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0103] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0104] In addition, it should be noted that, in the above specific embodiments, the various specific technical features described can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the present invention will not separately describe various possible combination manners.

Claims

1. Use of a combined pharmaceutical composition in the preparation of a medicament for treating transformed follicular lymphoma, characterized in that, The active components in the combined pharmaceutical composition include a first component and a second component; The first component is selected from any one or a combination of at least two of sitagliptin carboxylate, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; The second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

2. The application according to claim 1, characterized in that, The pharmaceutically acceptable salts of sitagliptin carboxylate include alkali metal salts, alkaline earth metal salts, ammonium salts, or quaternary ammonium salts; Preferably, the pharmaceutically acceptable salts of sitagliptin carboxylate include sodium salts or potassium salts.

3. The application according to claim 1, characterized in that, The combined pharmaceutical composition further contains pharmaceutically acceptable excipients.

4. The application according to claim 3, wherein The pharmaceutically acceptable excipients include any one or a combination of at least two of carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, bacteriostatic agents, or buffers.

5. The application according to claim 1, characterized in that, The combined pharmaceutical composition is a single compound preparation or a combination of two separate preparations; Preferably, the combined pharmaceutical composition is a combination of two separate preparations, and the two separate preparations are administered simultaneously or sequentially; Preferably, the preparation is any pharmaceutically acceptable dosage form.

6. Use of a combined pharmaceutical composition in the preparation of a transforming follicular lymphoma cell proliferation inhibitor, characterized in that, The active components in the combined pharmaceutical composition include a first component and a second component; The first component is selected from any one or a combination of at least two of sitagliptin carboxylate, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; The second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

7. The application according to claim 6, wherein The transformed follicular lymphoma cells include SC-1 cells and / or DHL4 cells.

8. Use of a combined pharmaceutical composition in the preparation of an apoptosis promoter for transformed follicular lymphoma cells, characterized in that, The active components in the combined pharmaceutical composition include a first component and a second component; The first component is selected from any one or a combination of at least two of sitagliptin carboxylate, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; The second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

9. The application according to claim 8, wherein The transformed follicular lymphoma cells include SC-1 cells and / or DHL4 cells.

10. Use of a combined pharmaceutical composition in the preparation of a preparation for inducing mitophagy in transformed follicular lymphoma cells, characterized in that, The active components in the combined pharmaceutical composition include a first component and a second component; The first component is selected from any one or a combination of at least two of sitagliptin carboxylate, its pharmaceutically acceptable salts, isomers, solvates, and metabolites; The second component is selected from any one or a combination of at least two of chidamide, its pharmaceutically acceptable salts, isomers, solvates, and metabolites.

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