Use of a combination of vortioxetine and gemcitabine in the manufacture of a medicament for the treatment of pancreatic cancer

Abstract

The application discloses application of a vortioxetine and gemcitabine composition in preparation of a drug for resisting pancreatic cancer, and the composition is composed of vortioxetine and gemcitabine. The composition of the application can reduce the expression amount of MAOB protein, promote the apoptosis of pancreatic cancer cells, and synergistically increase the sensitivity of pancreatic cancer cell lines in vitro and pancreatic cancer mice in vivo to gemcitabine, so that the chemotherapy effect of pancreatic cancer cells which are resistant to gemcitabine is significantly improved after treatment by the composition. The composition of the application provides a new chemotherapy drug for pancreatic cancer cells which are resistant to gemcitabine treatment.

Description

Technical Field

[0001] This application relates to the field of tumor treatment technology, and in particular to the use of a combination of vortioxetine and gemcitabine in the preparation of an anti-pancreatic cancer drug. Background Technology

[0002] Pancreatic cancer is highly malignant and has a poor prognosis, ranking fourth in global cancer-related mortality, with a five-year survival rate as low as 12%. Surgical resection remains the best treatment option for long-term survival. However, due to the insidious nature of pancreatic cancer symptoms, most patients are already at an advanced stage at initial diagnosis. For locally advanced and metastatic pancreatic cancer patients in good condition, combination chemotherapy is recommended as first-line chemotherapy, with gemcitabine being a commonly used base monotherapy in two-drug combinations. Two-drug combinations include gemcitabine / albumin-bound paclitaxel (GN), gemcitabine / cisplatin (GP), gemcitabine / capecitabine (GX), and gemcitabine / tegafur (GS). Even gemcitabine, a first-line drug for pancreatic cancer treatment, has an objective response rate of less than 20%, severely impacting the prognosis of pancreatic cancer patients. Given the challenges of poor efficacy, drug resistance, and high recurrence rates in clinical pancreatic cancer treatment, there is an urgent need to find more effective treatment options. Summary of the Invention

[0003] In view of this, this application provides an application of a combination of vortioxetine and gemcitabine in the preparation of an anti-pancreatic cancer drug to solve the problems existing in the related art.

[0004] According to a first aspect of the embodiments of this application, the use of a component targeting MAOB in the preparation of an anti-pancreatic cancer drug is provided, said component reducing the expression level of MAOB protein.

[0005] Furthermore, the components include vortioxetine and gemcitabine, with a molar ratio of 100:1 to 80:1.

[0006] Furthermore, the components also include pharmaceutically acceptable excipients.

[0007] According to a second aspect of the present application, a composition for an anti-pancreatic cancer drug is provided, the composition comprising vortioxetine and gemcitabine, wherein the molar concentration ratio of vortioxetine to gemcitabine is 100:1 to 80:1.

[0008] According to a third aspect of the embodiments of this application, an anti-pancreatic cancer drug is provided, which is made from the composition described in the second aspect and pharmaceutically acceptable excipients.

[0009] Furthermore, the dosage form of the drug is a solid dosage form or a liquid dosage form.

[0010] According to a fourth aspect of the present application, the use of a composition of vortioxetine and gemcitabine in the preparation of an anti-pancreatic cancer drug is provided, wherein the pancreatic cancer is ductal adenocarcinoma, and the composition consists of vortioxetine and gemcitabine, wherein the molar concentration ratio of vortioxetine to gemcitabine is 100:1 to 80:1.

[0011] Furthermore, the drug is made from the composition and pharmaceutically acceptable excipients.

[0012] Furthermore, the dosage form of the drug is a solid dosage form or a liquid dosage form.

[0013] The technical solutions provided by the embodiments of this application may include the following beneficial effects:

[0014] As demonstrated by the above embodiments, the composition of the present invention promotes tumor cell apoptosis by reducing the expression level of MAOB and synergistically increases the sensitivity of in vitro pancreatic cancer cell lines and in vivo pancreatic cancer mice to gemcitabine, thereby significantly improving the efficacy of chemotherapy in highly gemcitabine-resistant pancreatic cancer after treatment with the composition. The composition of the present invention provides a novel chemotherapeutic agent for the treatment of gemcitabine-resistant pancreatic cancer.

[0015] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit this application. Attached Figure Description

[0016] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this application and, together with the description, serve to explain the principles of this application.

[0017] Figure 1 This is a diagram of RNA-seq sequencing and drug target screening in an embodiment of the present invention. 1-1 is a volcano diagram of RNA-seq sequencing results after BxPC-3 cells were given vortioxetine alone for 24 hours; 1-2 is a Venn diagram of all potential drug targets obtained by DEGs, Swiss targetprediction and GeneCards, which enriched 5 genes: MAOB, TACR1, HTR1B, SCN5A and TERT.

[0018] Figure 2 2-1 shows the mRNA and protein expression levels of MAOB in two pancreatic cancer cell lines, BxPC-3 and AsPC-1, after treatment with vortioxetine in this embodiment of the invention; 2-1 shows the in vitro RT-qPCR verification experiment when BxPC-3 and AsPC-1 cells were treated with 8 μM and 10 μM of vortioxetine, respectively; 2-2 shows the in vitro MAOB protein expression in BxPC-3 and AsPC-1 cells when treated with 8 μM and 10 μM of vortioxetine, respectively.

[0019] Figure 3 This is the drug synergy index of vortioxetine and gemcitabine in two pancreatic cancer cell lines, BxPC-3 and AsPC-1, in the embodiments of the present invention.

[0020] Figure 4 This invention illustrates the long-term effect of vortioxetine in enhancing gemcitabine's synergistic inhibition of cell proliferation in this embodiment. 4-1 shows the crystal violet staining and photographic results of BxPC-3 and AsPC-1 cells after 14 days of treatment with vortioxetine alone or in combination with gemcitabine. 4-2 shows the results of clonal counting and statistical analysis of BxPC-3 and AsPC-1 cells.

[0021] Figure 5 Figure 5-1 shows the synergistic effect of vortioxetine combined with gemcitabine in promoting apoptosis in pancreatic cancer cells in an embodiment of the present invention. Flow cytometry plots of apoptosis rates in BxPC-3 and AsPC-1 cells after vortioxetine treatment alone or in combination with gemcitabine correspond to statistical analyses of Capan-1 cells, BxPC-3 cells, and AsPC-1 cells after 48 hours of drug treatment, respectively. Statistical analysis of apoptosis rates is shown in Figure 5-2. Flow cytometry analysis of apoptosis-related protein expression in BxPC-3 and AsPC-1 cells after vortioxetine treatment alone or in combination with gemcitabine is also shown in Figure 5-3.

[0022] Figure 6 This is a diagram illustrating the effect of vortioxetine on enhancing gemcitabine's anti-pancreatic cancer cell activity through MAOB sensitization in an embodiment of the present invention. 6-1 shows the significant increase in MAOB protein expression when the two drugs are used in combination; 6-2 shows the mRNA and protein expression of MAOB after siRNA interference with pancreatic cancer cells; and 6-3 shows the therapeutic effect of silencing MAOB and administering gemcitabine at different concentration gradients to enhance gemcitabine's anti-pancreatic cancer activity.

[0023] Figure 7 This is a schematic diagram illustrating the effect of vortioxetine and gemcitabine alone or in combination on tumor growth in a nude mouse model of pancreatic cancer, as described in this embodiment of the invention. 7-1 is a schematic diagram of the relative tumor volume increase; 7-2 is a photograph of the pancreatic cancer mouse model and the separated tumor; 7-3 is a schematic diagram of tumor weight; and 7-4 is a schematic diagram of weight changes in the pancreatic cancer mouse model. Detailed Implementation

[0024] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings. When the following description relates to the drawings, unless otherwise indicated, the same numbers in different drawings denote the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with this application. Rather, they are merely examples of apparatuses and methods consistent with some aspects of this application as detailed in the appended claims.

[0025] The terminology used in this application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The singular forms “a,” “the,” and “the” used in this application and the appended claims are also intended to include the plural forms unless the context clearly indicates otherwise. It should also be understood that the term “and / or” as used herein refers to and includes any or all possible combinations of one or more of the associated listed items.

[0026] Monoamine oxidase (MAOB) is a mitochondrial outer membrane-bound enzyme that breaks down various monoamine neurotransmitters and produces hydrogen peroxide (H2O2). It primarily regulates human mood, behavior, and cognition by modulating the levels of monoamine neurotransmitters such as serotonin and dopamine. MAOB is currently a star molecule in clinical use for treating neurological disorders such as depression, but its mechanism of action in tumors is poorly understood.

[0027] This application demonstrates that vortioxetine can sensitize gemcitabine via MAOB in pancreatic cancer cells, a phenomenon not previously reported. Therefore, by fully utilizing the synergistic effect of combination drugs to address treatment resistance in pancreatic cancer cells, treatment outcomes can be optimized.

[0028] Gemcitabine is a novel cytosine nucleoside derivative with the chemical formula C9H. 11 F₂N₃O₄. Vortioxetine is a chemical substance. Its chemical formula is C₂N₃O₄. 18 H 22 N2S.

[0029] The following examples illustrate this in detail.

[0030] Example 1

[0031] This invention hypothesizes that genes such as MAOB may be key factors in enhancing the anti-pancreatic cancer effect of vortioxetine.

[0032] Reagents and materials:

[0033] Pancreatic cancer cell lines BxPC-3 and AsPC-1 were purchased from Wuhan Pronosei Biotechnology Co., Ltd. Both cell lines were cultured in RPMI-1640 medium (Hangzhou Keyi Biotechnology Co., Ltd., catalog number C0003) containing 10% fetal bovine serum. Culture conditions were 37℃ and 5% CO2. MAOB antibody was purchased from Proteintech (catalog number 12602-1-AP), total RNA extraction reagent Trizol was purchased from Biosharp (BS258A), HiScript IIQ RT SuperMix kit was purchased from Novizan (catalog number R222-01), 2×SYBR qPCR Master Mix kit was purchased from Novizan (catalog number Q311-02), and vortioxetine was purchased from Aladdin (catalog number T2395).

[0034] Experimental Methods: We performed RNA-seq sequencing on the control group and the vortioxetine monotherapy group. Using log2FoldChange<-2, we screened for DEGs with significant changes. Then, we imported the vortioxetine structure into the Swisstarget prediction database to screen for potential drug targets. Using GeneCards, we input the disease name "pancreatic cancer" to obtain potential drug targets for this disease. Finally, we visualized all DEGs obtained from mRNA sequencing, all potential drug targets from the Swisstargetprediction database, and all potential drug targets corresponding to pancreatic cancer in GeneCards using a Venn diagram. Ultimately, we selected genes with significant changes as research subjects.

[0035] To verify these results, we again performed RT-qPCR and Western blot on BxPC-3 and AsPC-1 cells to validate the mRNA and protein expression levels of significantly altered genes. BxPC-3 and AsPC-1 cells were treated with DMSO and vortioxetine (8 μM and 10 μM), respectively, and total RNA and protein were extracted from the cell samples. The total RNA mRNA level was detected by RT-qPCR after extraction. The extracted proteins were relatively quantified using a multi-mode microplate reader. After adjusting the cell protein concentration to be uniform, protein abundance was detected by Western blot.

[0036] Experimental results:

[0037] like Figure 1As shown, the five genes MAOB (P = 5.45E-06), TACR1 (P = 0.0083), HTR1B (P = 6.87E-09), SCN5A (P = 0.0002), and TERT (P = 5.47E-06) were significantly downregulated after vortioxetine treatment. Further analysis revealed that, based on sequencing results, MAOB was the most significantly downregulated gene among the five.

[0038] To verify this result, we again performed RT-qPCR on BxPC-3 and AsPC-1 cells to verify the mRNA expression levels of the five genes. The results showed that only the mRNA expression of the MAOB gene was significantly downregulated after treatment with different concentrations of vortioxetine in both cell lines. Figure 2 (See section 2-1). The experimental results are consistent with the RNA-seq sequencing results. Next, we used Western blotting (WB) to detect the expression levels of MAOB protein after treatment with 8 μM and 10 μM vortioxetine. Consistent with the mRNA level, vortioxetine significantly downregulated the protein expression level of MAOB. Figure 2 (See section 2-2). Therefore, we speculate that MAOB may be a key factor in enhancing the antitumor effect of gemcitabine with vortioxetine.

[0039] Example 2

[0040] This invention has found that vortioxetine can synergistically enhance the toxicity of gemcitabine to pancreatic cancer cell lines.

[0041] Reagents and materials:

[0042] Gemcitabine was purchased from Taoshu (item number T0251), and the other reagents and materials were the same as in Example 1.

[0043] Experimental methods:

[0044] Preparation of vortioxetine, gemcitabine stock solutions and working solutions:

[0045] Using DMSO as solvent, vortioxetine was prepared into a 40 mM stock solution and gemcitabine into a 10 mM stock solution, and stored at -20°C.

[0046] Two groups of BxPC-3 cells were prepared with different molar ratios of vortioxetine and gemcitabine: 100:1 and 80:1. In the 100:1 molar ratio group, vortioxetine concentrations were set at 0, 2, 4, 6, 8, 10, and 20 μM, and gemcitabine concentrations were set at 0, 20, 40, 60, 80, 100, and 200 nM. In the 80:1 molar ratio group, vortioxetine concentrations were set at 0, 2, 4, 6, 8, 10, and 20 μM, and gemcitabine concentrations were set at 0, 25, 50, 75, 100, 125, and 250 nM.

[0047] Similarly, two groups of vortioxetine and gemcitabine concentrations were set in AsPC-1 cells, with molar ratios of 100:1 and 80:1, respectively. In the 100:1 molar ratio group, the vortioxetine concentrations were set at: 0, 2, 4, 6, 8, 10, and 20 μM, and the gemcitabine concentrations were set at: 0, 20, 40, 60, 80, 100, and 200 nM. In the 80:1 molar ratio group, the vortioxetine concentrations were set at: 0, 2, 4, 6, 8, 10, and 20 μM, and the gemcitabine concentrations were set at: 0, 25, 50, 75, 100, 125, and 250 nM.

[0048] Cell culture and drug administration:

[0049] BxPC-3 (2000 cells / well) and AsPC-1 (2000 cells / well) cells were seeded at a specific density in 96-well plates and cultured overnight. Appropriate concentrations of vortioxetine and gemcitabine were added, with the final drug concentrations in both cell lines determined according to experimental procedures.

[0050] Cell viability assay:

[0051] After drug treatment was terminated, 100 μL of pre-cooled 15% TCA was added to each well and fixed at 4°C for 4 hours. Then, the cells were washed three times with distilled water and dried. 50 μL of SRB solution (4 mg / mL, 0.8 g SRB dissolved in 200 mL of 1% acetic acid) was added, and staining was performed for 30 minutes. The SRB was discarded, and the cells were washed with 1% acetic acid and dried in a 37°C oven. Depending on the color intensity, 80-100 μL of 10 mM Tris-HCl (0.6057 g dissolved) was added, and the cells were gently shaken to completely dissolve the crystals. The absorbance (OD value) of each well was measured at 570 nm using a multi-mode microplate reader. The mean and standard deviation of the three replicates were calculated, and the cell viability was determined: Cell viability % = (OD value of drug group / OD value of control group) * 100%.

[0052] Quantitative analysis of the dose-effect of vortioxetine and gemcitabine in combination.

[0053] First, the inhibitory effect (inhibitory%) at each concentration point was calculated. Plotting the concentration of vortioxetine or gemcitabine alone on the x-axis and the corresponding inhibitory effect at each concentration on the y-axis using Prism 9.5, the dose-response curve and its parameters were obtained. Then, the combination index (CI) of vortioxetine and gemcitabine was calculated using Calcusyn software. A CI > 1.2 indicated an antagonistic effect, a CI < 0.8 indicated a synergistic effect, and a CI between 0.8 and 1.2 was considered an additive effect.

[0054] Experimental results:

[0055] This experiment focuses on the molar ratio of 100:1 in BxPC-3 cells and 80:1 in AsPC-1 cells, calculating the combined drug index (CI). The combined drug index (CI) of vortioxetine and gemcitabine on BxPC-1 and AsPC-1 cells is as follows: Figure 3 As shown, vortioxetine can increase the sensitivity of pancreatic cancer to gemcitabine, with CI values ​​all less than 0.8.

[0056] Example 3

[0057] This invention reveals that the combination drugs vortioxetine and gemcitabine can synergistically inhibit the long-term proliferation of pancreatic cancer cells.

[0058] Reagents and materials:

[0059] Crystal violet was purchased from Baizan (catalog number 548-62-9), and the other reagents and materials were the same as in Example 1.

[0060] Experimental methods:

[0061] BxPC-1 and AsPC-1 cells were removed from the cell culture incubator, digested, and single-cell suspensions were prepared. After counting with a hemocytometer, the cells were seeded into 6-well cell culture plates. BxPC-3 cells were seeded at 1 × 10⁶ cells per well. 3 AsPC-1 cells / mL, 2 × 10⁶ cells per well. 3 The experiment was divided into four groups: control group, vortioxetine group, gemcitabine group, and vortioxetine + gemcitabine group. BxPC-3 cells were treated with 4 μM vortioxetine and 40 nM gemcitabine (molar ratio 100:1, consistent with the molar ratio in Example 2). AsPC-1 cells were treated with 2.5 μM vortioxetine and 31.25 nM gemcitabine (molar ratio 80:1, consistent with the molar ratio in Example 2). After 72 hours of seeding, the cells were administered the corresponding drugs according to the groupings. Fresh complete culture medium and supplementary drugs were added every three days. After approximately 14 days of culture, the culture medium was discarded, the cells were washed twice with PBS, and 700 μL of 1% crystal violet staining solution was added. The cells were stained at 4°C in the dark for approximately 30 minutes. Unbound dye was gently washed away with tap water, the cells were dried in an oven, photographed, and the clones were counted.

[0062] Experimental results:

[0063] After treating cells with the drug for 14 days and staining with crystal violet, the clone count was analyzed. Figure 4 Counting is performed in section 4-1. Figure 4In BxPC-3 cells (4-2), the percentage of clones using vortioxetine and gemcitabine alone were 84.90±0.39% and 83.33±0.78%, respectively. After combination therapy, the percentage decreased to 23.70±0.59%, and the combination therapy group (P<0.0001) showed a significant difference compared to both vortioxetine (P<0.001) and gemcitabine (P<0.001) alone. Similar results were observed in AsPC-1 cells. The percentage of clones using vortioxetine and gemcitabine alone were 79.00±0.59% and 83.20±0.59%, respectively. After combination therapy, the percentage decreased to 43.04±1.38%, and the combination therapy group (P<0.0001) showed a significant difference compared to both vortioxetine (P<0.001) and gemcitabine (P<0.01) alone. The results of the clonogenic assay further demonstrated that vortioxetine and gemcitabine, when used together, have a synergistic inhibitory effect on the proliferation of pancreatic cancer cells.

[0064] Example 4

[0065] This invention discovers that the combination drugs vortioxetine and gemcitabine can promote apoptosis in pancreatic cancer.

[0066] Reagents and materials:

[0067] The apoptosis kit was purchased from Linko Biotech (catalog number AP-101), the cleaved-PARP antibody was purchased from Cell Signaling Technology (catalog number D64E10), and the cleaved-Caspase-3 antibody was purchased from Cell Signaling Technology (catalog number 9664S, 5A1E). The remaining reagents and materials were the same as in Example 1.

[0068] Experimental methods:

[0069] Apoptosis rate detection:

[0070] After cell collection and washing, staining was performed. Residual PBS at the bottom of the EP tubes was aspirated, and 100 μL of 1×Binding Buffer was added to each tube. The cells were gently resuspended by pipetting, and dye was added under dark conditions. No dye was added to the unstained group. For the single-stain group, 5 μL of Annexin V-FITC or 10 μL of PI was added. For the double-stain group, both 5 μL of Annexin V-FITC and 10 μL of PI were added. After incubation at room temperature in the dark for 15 min, 1×Binding Buffer was added and mixed well. The cell suspension was then transferred to 5 mL flow cytometry tubes under dark conditions, filtered through a 40 μm cell sieve beforehand, and finally analyzed by flow cytometry.

[0071] Apoptosis protein detection:

[0072] BXPC-3 and CAPAN-1 cells were treated with vortioxetine and / or gemcitabine. A solvent control group, a vortioxetine group, a gemcitabine group, and a vortioxetine and gemcitabine combination group were set up. The effect of vortioxetine alone or in combination on apoptosis proteins was detected by Western blot to evaluate whether the combination drugs vortioxetine and gemcitabine can induce apoptosis and thus inhibit the growth of pancreatic cancer cells.

[0073] Experimental results:

[0074] To investigate the synergistic mechanism by which vortioxetine and gemcitabine kill tumor cells in combination, we used Annexin V-FITC and PI double staining combined with flow cytometry to detect cell apoptosis after drug administration. Figure 5 (5-1) BxPC-3 cells were treated with vortioxetine (8 μM) and / or gemcitabine (200 nM), and AsPC-1 cells were treated with vortioxetine (11 μM) and / or gemcitabine (800 nM). In BxPC-3 cells, the apoptosis rate was 11.33 ± 2.51% in the control group, 17.17 ± 10.37% in the vortioxetine monotherapy group, and 16.35 ± 0.38% in the gemcitabine monotherapy group. The apoptosis rate increased to 50.85 ± 4.51% in the vortioxetine and gemcitabine combination group. Figure 5 The results showed that the combined use group (P<0.0001) was significantly different from the vortioxetine (P<0.001) monotherapy group and the gemcitabine (P<0.001) monotherapy group. Figure 5 (5-2). Similarly, in AsPC-1 cells, the apoptosis rate was 12.49±2.25% in the control group, 18.73±3.13% in the vortioxetine monotherapy group, and 20.02±0.40% in the gemcitabine monotherapy group, while the apoptosis rate increased to 44.72±2.89% in the vortioxetine and gemcitabine combination group. Figure 5 The results showed that the combined use group (P<0.0001) was significantly different from the vortioxetine (P<0.001) monotherapy group and the gemcitabine (P<0.01) monotherapy group. Figure 5 (5-2 in the middle).

[0075] On the other hand, chemotherapy and other cancer treatment strategies typically inhibit tumor cell proliferation and promote apoptosis. Caspase-3 plays a crucial role in apoptosis, and Cleaved-Caspase-3 is the activated form of Caspase-3. PARP cleavage is an important marker of the apoptosis signaling pathway. We found that the expression levels of the apoptosis proteins Cleaved-PARP and Cleaved-Caspase-3 were significantly increased when vortioxetine was used in combination with gemcitabine. Figure 5 (5-3). The above results suggest that the combined use of vortioxetine and gemcitabine can induce the expression of apoptosis-related proteins in pancreatic cancer cells, thereby exerting a synergistic tumor-killing effect by promoting apoptosis in the combined group of cells.

[0076] Example 5

[0077] This invention discovers that vortioxetine enhances the anti-pancreatic cancer effect of gemcitabine through MAOB.

[0078] Reagents and materials:

[0079] siRNA was purchased from Gemma Gene, and jetPRIME transfection reagent was purchased from Polyplus Tranfection SA (catalog number 101000046). The other reagents and materials were the same as in Example 1.

[0080] Experimental methods:

[0081] The siRNA was stored as a lyophilized powder at -20°C. Before use, the siRNA was centrifuged at 12,000 rpm for 1 min at 4°C. The siRNA was dissolved and mixed with the volume of DEPC water indicated in the instructions to obtain a 20 μM stock solution. The solution was aliquoted into 10 μL tubes and stored at -80°C.

[0082] BxPC-3 cells and AsPC-1 cells were digested with trypsin and then administered at 8 × 10⁻⁶ doses. 4 cells / well and 1.5 × 10 5 Cells were seeded in 6-well plates. When the cells reached 30%-40% confluence, siRNA gene silencing transfection was performed. First, 4 μL of siRNA (20 μM) was added to 200 μL of Jet PRIME buffer and gently mixed with an RNAase-free pipette tip. The mixture was incubated at room temperature for 5 min. Then, 4 μL of Jet PRIME rejent was added to the mixture, and after pipetting, it was incubated at room temperature for 15 min. After 15 min, the mixture was transferred to the corresponding 6-well plates and incubated for 48 h before proceeding with subsequent experiments. The remaining experimental procedures, such as RT-qPCR and Western blot, were the same as in Example 1.

[0083] Experimental results:

[0084] We found that compared to vortioxetine alone, the combination of vortioxetine and gemcitabine was required to significantly downregulate MAOB protein expression. Figure 6 (6-1 in the text). Furthermore, after silencing the MAOB gene in both cell lines ( Figure 6 In a study (6-2), BxPC-3 cells were treated with different concentrations of gemcitabine (0, 20, 40, 60, 80, 100 nM), and AsPC-1 cells were treated with different concentrations of gemcitabine (0, 200, 400, 600, 800, 1000 nM). The results showed that, compared with the Control siRNA group, downregulation of MAOB significantly enhanced the anti-pancreatic cancer effect of gemcitabine. Figure 6 (6-3 in the example). Combining the results of RNA-seq in Example 1 and the combined synergistic sensitizing drug effect in Example 2, it is suggested that vortioxetine may inhibit the sensitizing effect of gemcitabine on MAOB in pancreatic cancer.

[0085] Example 6

[0086] This invention found that the combined use of vortioxetine and gemcitabine can increase the toxicity of gemcitabine in a mouse model of xenograft tumors.

[0087] Reagents and materials:

[0088] Female Balb / C nude mice were purchased from Shanghai Slack Laboratory Animal Co., Ltd. and housed at the Laboratory Animal Center of Zhejiang University City College. Other reagents and materials were the same as in Example 1.

[0089] Experimental methods:

[0090] A mouse model of pancreatic cell carcinoma xenograft was established using 4-6 week old Balb / C nude mice. The mice were divided into four groups: solvent control group, vortioxetine group, gemcitabine group, and vortioxetine and gemcitabine combination group, with 5 mice in each group. 5 × 10⁻⁶ mice were used as experimental subjects. 7 A number of BxPC-3 cells were inoculated into the axillae of nude mice. After about half a month, when the average tumor volume reached 100 mm, 3 Treatment of the mouse model began with intraperitoneal injection of gemcitabine 20 mg / kg every three days and oral gavage of vortioxetine 15 mg / kg daily for a total of 23 days. Starting from day 0, the weight of the nude mice was measured daily using a balance, and the tumor volume was measured using calipers. The formula for calculating tumor volume (TV) is: TV = L × W × W / 2 (where L is the long axis of the tumor and W is the short axis). Relative tumor volume (RTV) is: RTV = V t / V0(V tVt represents the tumor volume on day t, and V0 represents the tumor volume on day 0. Nude mice were euthanized humanely on day 23 after drug administration, and the tumors were isolated and stored in 10% neutral formalin for later use.

[0091] Experimental results:

[0092] like Figure 7 As shown in Figures 7-1 to 7-3, in the pancreatic cancer (BXPC-3) mouse model, vortioxetine and gemcitabine alone did not have significant tumor-suppressive effects, while the combination of vortioxetine and gemcitabine showed more significant tumor-suppressive effects. The relative tumor volume and tumor weight in the combination group were lower than those in the single-drug group. Therefore, the combination of vortioxetine and gemcitabine can increase the antitumor activity of gemcitabine in the pancreatic cancer mouse model. Furthermore, there was no significant difference in mouse body weight between the single-drug and combination-drug groups. Figure 7 (7-4) suggests that the combination drugs vortioxetine and gemcitabine have low toxicity and good tolerability in mouse models.

[0093] It should be noted that pancreatic ductal adenocarcinoma cell lines include BxPC-3 and AsPC-1, which are typical representatives of pancreatic ductal adenocarcinoma cell lines. This invention uses BxPC3 and AsPC-1 as examples to illustrate the inhibitory effect of the vortioxetine and gemcitabine combination on pancreatic ductal adenocarcinoma cell lines. Based on these examples, those skilled in the art can undoubtedly determine that the vortioxetine and gemcitabine combination of this invention is also effective against other ductal adenocarcinoma cell lines, such as Capan-1 and PANC-1.

[0094] Other embodiments of this application will readily occur to those skilled in the art upon consideration of the specification and practice of the disclosure herein. This application is intended to cover any variations, uses, or adaptations of this application that follow the general principles of this application and include common knowledge or customary techniques in the art not disclosed herein. The specification and embodiments are to be considered exemplary only, and the true scope and spirit of this application are indicated by the claims.

[0095] It should be understood that this application is not limited to the precise structure described above and shown in the accompanying drawings, and various modifications and changes can be made without departing from its scope. The scope of this application is limited only by the appended claims.

Claims

1. The application of a component targeting MAOB in the preparation of an anti-pancreatic cancer drug, wherein the component reduces the expression level of MAOB protein; wherein the component is vortioxetine and gemcitabine, and the molar ratio is 100:1 to 80:

1.

2. The application according to claim 1, characterized in that, The components also include pharmaceutically acceptable excipients.

3. A composition for preparing an anti-pancreatic cancer drug, characterized in that, The composition comprises vortioxetine and gemcitabine, wherein the molar ratio of vortioxetine to gemcitabine is 100:1 to 80:

1.

4. An anti-pancreatic cancer drug, characterized in that, It is made from the composition of claim 3 and pharmaceutically acceptable excipients.

5. The anti-pancreatic cancer drug according to claim 4, characterized in that, The drug is in the form of a solid dosage form or a liquid dosage form.

6. The use of the combination of vortioxetine and gemcitabine in the preparation of an anti-pancreatic cancer drug, characterized in that, The pancreatic cancer is pancreatic ductal adenocarcinoma, and the composition consists of vortioxetine and gemcitabine, wherein the molar ratio of vortioxetine to gemcitabine is 100:1 to 80:

1.

7. The application according to claim 6, characterized in that, The drug is made from a composition and pharmaceutically acceptable excipients.

8. The application according to claim 6, characterized in that, The drug is in the form of a solid dosage form or a liquid dosage form.

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