Use of duloxetine and chemotherapeutic drug in preparation of pharmaceutical composition for treating or preventing cancer

Abstract

Provided is use of duloxetine and a chemotherapeutic drug in the preparation of a pharmaceutical composition for treating or preventing a cancer, comprising the administration of an effective amount of duloxetine and the chemotherapeutic drug to an individual in need thereof. This administration provides better efficacy than administering duloxetine or the chemotherapeutic drug alone.

Description

Use of duloxetine and chemotherapeutic agents for the preparation of a pharmaceutical composition for treating or preventing cancer TECHNICAL FIELD

[0001] The present disclosure relates to a use of preparing a pharmaceutical composition for treating or preventing cancer, in particular, to a use of duloxetine and chemotherapeutic agents for the preparation of a pharmaceutical composition for treating or preventing cancer. BACKGROUND

[0002] Cancer has long been the leading cause of death worldwide, and the number of people suffering from cancer is rising year by year, so treating cancer is an important issue. Cancer treatment can be divided into surgical treatment, radiation therapy, chemotherapy, and targeted therapy. Cancer cells are characterized by uncontrolled cell proliferation and the potential to invade or metastasize to distant tissues. Clinical manifestations of cancer include weight loss, muscle atrophy, decreased mobility, fatigue, anorexia, satiety, lethargy, pallor, anemia, emaciation, electrolyte imbalance, decreased protein and lipid synthesis, and blood sugar instability. When a patient loses more than 5% of their body weight in 6 months and has the above symptoms, it is called "cachexia", and in severe cases it is commonly known as "skin and bones".

[0003] Despite the progress in cancer treatment over the past few decades, such as surgery, radiotherapy, chemotherapy, and immunotherapy, these treatment methods still have significant side effects. For example, some patients cannot undergo surgical resection of tumors due to their own relationship, or surgery may not completely remove tumor tissue. Furthermore, most existing anticancer agents cannot effectively distinguish between cancer cells and normal cells, so these chemotherapy-related side effects or systemic toxicity caused by individual chemotherapy limit the therapeutic effect of chemotherapy. In addition, as with the aforementioned chemotherapy, radiotherapy can also damage normal tissues, thus limiting its effectiveness. In immunotherapy, cancer cells have developed mechanisms to evade immune responses, so sometimes the effectiveness of immunotherapy for cancer patients is still not significant.

[0004] Therefore, there is an unmet need in the art for developing therapies for treating and inhibiting cancer, and there is an urgent need in the art to develop a method for treating cancer to solve the above problems in the art and meet the clinical needs. SUMMARY

[0005] Use of duloxetine and chemotherapeutic agents for the preparation of a pharmaceutical composition for treating or preventing cancer.

[0006] A method for treating or preventing cancer, comprising administering to a subject in need thereof an effective amount of duloxetine and a chemotherapeutic agent.

[0007] A pharmaceutical composition for treating or preventing cancer, comprising an effective amount of duloxetine and a chemotherapeutic agent.

[0008] BRIEF DESCRIPTION OF DRAWINGS

[0009] The present disclosure will be more readily understood by reference to the following description, taken in conjunction with the accompanying drawings.

[0010] Figure 1 shows the effect of duloxetine and docetaxel on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC50 of docetaxel (0.004 μΜ); G2: ½ IC50 of docetaxel (0.002 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of docetaxel (0.002 μΜ); G5: duloxetine (5 μΜ) + IC50 of docetaxel (0.004 μΜ). 50 (0.004 μΜ);G2: ½ IC50 of docetaxel (0.002 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of docetaxel (0.002 μΜ); G5: duloxetine (5 μΜ) + IC50 of docetaxel (0.004 μΜ). 50 (0.002 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of docetaxel (0.002 μΜ); G5: duloxetine (5 μΜ) + IC50 of docetaxel (0.004 μΜ). 50 (0.002 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of docetaxel (0.002 μΜ); G5: duloxetine (5 μΜ) + IC50 of docetaxel (0.004 μΜ). 50 (0.004 μΜ);G2: ½ IC50 of docetaxel (0.002 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of docetaxel (0.002 μΜ); G5: duloxetine (5 μΜ) + IC50 of docetaxel (0.004 μΜ).

[0011] Figure 2 shows the effect of duloxetine and cisplatin on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC50 of cisplatin (3 μΜ); G2: ½ IC50 of cisplatin (1.5 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cisplatin (1.5 μΜ); G5: duloxetine (5 μΜ) + IC50 of cisplatin (3 μΜ). 50 (3 μΜ);G2: ½ IC50 of cisplatin (1.5 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cisplatin (1.5 μΜ); G5: duloxetine (5 μΜ) + IC50 of cisplatin (3 μΜ). 50 (1.5 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cisplatin (1.5 μΜ); G5: duloxetine (5 μΜ) + IC50 of cisplatin (3 μΜ). 50 (1.5 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cisplatin (1.5 μΜ); G5: duloxetine (5 μΜ) + IC50 of cisplatin (3 μΜ).

[0012] Figure 3 shows the effect of duloxetine and cyclophosphamide on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC50 of cyclophosphamide (40 μΜ); G2: ½ IC50 of cyclophosphamide (20 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cyclophosphamide (20 μΜ); G5: duloxetine (5 μΜ) + IC50 of cyclophosphamide (40 μΜ). 50 (40 μΜ);G2: ½ IC50 of cyclophosphamide (20 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cyclophosphamide (20 μΜ); G5: duloxetine (5 μΜ) + IC50 of cyclophosphamide (40 μΜ). 50 (20 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cyclophosphamide (20 μΜ); G5: duloxetine (5 μΜ) + IC50 of cyclophosphamide (40 μΜ). 50 (20 μΜ);G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cyclophosphamide (20 μΜ); G5: duloxetine (5 μΜ) + IC50 of cyclophosphamide (40 μΜ). 50 (40 μΜ);G2: ½ IC50 of cyclophosphamide (20 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of cyclophosphamide (20 μΜ); G5: duloxetine (5 μΜ) + IC50 of cyclophosphamide (40 μΜ).

[0013] Figure 4 shows the effect of duloxetine and 5-fluorouracil on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC50 of 5-fluorouracil (400 μΜ); G2: ½ IC50 of 5-fluorouracil (200 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of 5-fluorouracil (200 μΜ); G5: duloxetine (5 μΜ) + IC50 of 5-fluorouracil (400 μΜ). 50 (400 μΜ);G2: ½ IC50 of 5-fluorouracil (200 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC50 of 5-fluorouracil (200 μΜ); G5: duloxetine (5 μΜ) + IC50 of 5-fluorouracil (400 μΜ). 50G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + 5-fluorouracil IC 50 G5: Duloxetine (5 μΜ) + 5-fluorouracil IC 50 (400 μΜ).

[0014] Figure 5 shows the effect of duloxetine and doxorubicin on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC of doxorubicin 50 (0.5 μΜ); G2: ½ IC of doxorubicin 50 (0.25 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of doxorubicin 50 (0.25 μΜ); G5: Duloxetine (5 μΜ) + IC of doxorubicin 50 (0.5 μΜ).

[0015] Figure 6 shows the effect of duloxetine and temozolomide on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC of temozolomide 50 (400 μΜ); G2: ½ IC of temozolomide 50 (200 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of temozolomide 50 (400 μΜ); G5: Duloxetine (5 μΜ) + IC of temozolomide 50 (200 μΜ).

[0016] Figure 7 shows the effect of duloxetine and osimertinib (AZD-9291) on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC of osimertinib 50 (5 μΜ); G2: ½ IC of osimertinib 50 (2.5 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of osimertinib 50 (2.5 μΜ); G5: Duloxetine (5 μΜ) + IC of osimertinib 50 (5 μΜ).

[0017] Figure 8 shows the effect of duloxetine and etoposide on triple negative breast cancer (MDA-MB-231 cell line). G1 : IC of etoposide 50 (100 μΜ); G2: ½ IC of etoposide 50G3: Duloxetine (5 μM); G4: Duloxetine (5 μM) + 1 / 2 IC of Etoposide 50 G5: Duloxetine (5 μM) + IC of Etoposide 50 (100 μM).

[0018] Figure 9 shows the effect of Duloxetine and Docetaxel on non-small cell lung cancer (A549 cell line). G1 : IC of Docetaxel 50 (0.004 μM); G2: 1 / 2 IC of Docetaxel 50 (0.002 μM); G3: Duloxetine (5 μM); G4: Duloxetine (5 μM) + 1 / 2 IC of Docetaxel 50 (0.002 μM); G5: Duloxetine (5 μM) + IC of Docetaxel 50 (0.004 μM).

[0019] Figure 10 shows the effect of Duloxetine and 5-Fluorouracil on non-small cell lung cancer (A549 cell line). G1 : IC of 5-Fluorouracil 50 (400 μM); G2: 1 / 2 IC of 5-Fluorouracil 50 (200 μM); G3: Duloxetine (5 μM); G4: Duloxetine (5 μM) + 1 / 2 IC of 5-Fluorouracil 50 (200 μM); G5: Duloxetine (5 μM) + IC of 5-Fluorouracil 50 (400 μM).

[0020] Figure 11 shows the effect of Duloxetine and Doxorubicin on non-small cell lung cancer (A549 cell line). G1 : IC of Doxorubicin 50 (0.5 μM); G2: 1 / 2 IC of Doxorubicin 50 (0.25 μM); G3: Duloxetine (5 μM); G4: Duloxetine (5 μM) + 1 / 2 IC of Doxorubicin 50 (0.25 μM); G5: Duloxetine (5 μM) + IC of Doxorubicin 50 (0.5 μM).

[0021] Figure 12 shows the effect of Duloxetine and Temozolomide on non-small cell lung cancer (A549 cell line). G1 : IC of Temozolomide 50 (400 μM); G2: 1 / 2 IC of Temozolomide 50 (200 μM); G3: Duloxetine (5 μM); G4: Duloxetine (5 μM) + 1 / 2 IC of Temozolomide 50 (400 μM); G5: Duloxetine (5 μM) + IC of Temozolomide50 (200 μΜ).

[0022] Figure 13 shows the effect of Duloxetine and Osimertinib (AZD-9291) on non-small cell lung cancer (A549 cell line). G1 : IC of Osimertinib 50 (5 μΜ); G2: ½ IC of Osimertinib 50 (2.5 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of Osimertinib 50 (2.5 μΜ); G5: Duloxetine (5 μΜ) + IC of Osimertinib 50 (5 μΜ).

[0023] Figure 14 shows the effect of Duloxetine and Sorafenib on non-small cell lung cancer (A549 cell line). G1 : IC of Sorafenib 50 (20 μΜ); G2: ½ IC of Sorafenib 50 (10 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of Sorafenib 50 (10 μΜ); G5: Duloxetine (5 μΜ) + IC of Sorafenib 50 (20 μΜ).

[0024] Figure 15 shows the effect of Duloxetine and Etoposide on non-small cell lung cancer (A549 cell line). G1 : IC of Etoposide 50 (100 μΜ); G2: ½ IC of Etoposide 50 (50 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of Etoposide 50 (50 μΜ); G5: Duloxetine (5 μΜ) + IC of Etoposide 50 (100 μΜ).

[0025] Figure 16 shows the effect of Duloxetine and Cisplatin on pancreatic cancer (Mia-PaCa2 cell line). G1 : IC of Cisplatin 50 (3 μΜ); G2: ½ IC of Cisplatin 50 (1.5 μΜ); G3: Duloxetine (5 μΜ); G4: Duloxetine (5 μΜ) + ½ IC of Cisplatin 50 (3 μΜ); G5: Duloxetine (5 μΜ) + IC of Cisplatin 50 (1.5 μΜ).

[0026] Figure 17 shows the effect of Duloxetine and Doxorubicin on pancreatic cancer (Mia-PaCa2 cell line). G1 : IC of Doxorubicin50 (0.5 μΜ); G2: ½ IC of doxorubicin 50 (0.25 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of doxorubicin 50 (0.25 μΜ); G5: duloxetine (5 μΜ) + IC of doxorubicin 50 (0.5 μΜ).

[0027] Figure 18 shows the effect of duloxetine and osimertinib (AZD-9291) on pancreatic cancer (Mia-PaCa2 cell line). G1 : IC of osimertinib 50 (5 μΜ); G2: ½ IC of osimertinib 50 (2.5 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of osimertinib 50 (2.5 μΜ); G5: duloxetine (5 μΜ) + IC of osimertinib 50 (5 μΜ).

[0028] Figure 19 shows the effect of duloxetine and etoposide on pancreatic cancer (Mia-PaCa2 cell line). G1 : IC of etoposide 50 (100 μΜ); G2: ½ IC of etoposide 50 (50 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of etoposide 50 (50 μΜ); G5: duloxetine (5 μΜ) + IC of etoposide 50 (100 μΜ).

[0029] Figure 20 shows the effect of duloxetine and docetaxel on liver cancer (PLC / PRF / 5 cell line). G1 : IC of docetaxel 50 (0.004 μΜ); G2: ½ IC of docetaxel 50 (0.002 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of docetaxel 50 (0.002 μΜ); G5: duloxetine (5 μΜ) + IC of docetaxel 50 (0.004 μΜ).

[0030] Figure 21 shows the effect of duloxetine and cisplatin on liver cancer (PLC / PRF / 5 cell line). G1 : IC of cisplatin 50 (3 μΜ); G2: ½ IC of cisplatin 50 (1.5 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of cisplatin50 (3 μM); G5: duloxetine (5 μM) + IC of cisplatin 50 (1.5 μM).

[0031] Figure 22 shows the effect of duloxetine and 5-fluorouracil on liver cancer (PLC / PRF / 5 cell line). G1 : IC of 5-fluorouracil 50 (400 μM); G2: ½ IC of 5-fluorouracil 50 (200 μM); G3: duloxetine (5 μM); G4: duloxetine (5 μM) + ½ IC of 5-fluorouracil 50 (200 μM); G5: duloxetine (5 μM) + IC of 5-fluorouracil 50 (400 μM).

[0032] Figure 23 shows the effect of duloxetine and doxorubicin on liver cancer (PLC / PRF / 5 cell line). G1 : IC of doxorubicin 50 (0.5 μM); G2: ½ IC of doxorubicin 50 (0.25 μM); G3: duloxetine (5 μM); G4: duloxetine (5 μM) + ½ IC of doxorubicin 50 (0.25 μM); G5: duloxetine (5 μM) + IC of doxorubicin 50 (0.5 μM).

[0033] Figure 24 shows the effect of duloxetine and sorafenib on liver cancer (PLC / PRF / 5 cell line). G1 : IC of sorafenib 50 (20 μM); G2; ½ IC of sorafenib 50 (10 μM); G3: duloxetine (5 μM); G4: duloxetine (5 μM) + ½ IC of sorafenib 50 (10 μM); G5: duloxetine (5 μM) + IC of sorafenib 50 (20 μM).

[0034] Figure 25 shows the effect of duloxetine and etoposide on liver cancer (PLC / PRF / 5 cell line). G1 : IC of etoposide 50 (100 μM); G2: ½ IC of etoposide 50 (50 μM); G3: duloxetine (5 μM); G4: duloxetine (5 μM) + ½ IC of etoposide 50 (50 μM); G5: duloxetine (5 μM) + IC of etoposide 50 (100 μM).

[0035] Figure 26 shows the effect of duloxetine and docetaxel on glioblastoma (LN-229 cell line). G1 : IC of docetaxel 50 (0.004 μΜ); G2: ½ IC of docetaxel 50 (0.002 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of docetaxel 50 (0.002 μΜ); G5: duloxetine (5 μΜ) + IC of docetaxel 50 (0.004 μΜ).

[0036] Figure 27 shows the effect of duloxetine and cyclophosphamide on glioblastoma (LN-229 cell line). G1 : IC of cyclophosphamide 50 (40 μΜ); G2: ½ IC of cyclophosphamide 50 (20 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of cyclophosphamide 50 (20 μΜ); G5: duloxetine (5 μΜ) + IC of cyclophosphamide 50 (40 μΜ).

[0037] Figure 28 shows the effect of duloxetine and 5-fluorouracil on glioblastoma (LN-229 cell line). G1 : IC of 5-fluorouracil 50 (400 μΜ); G2: ½ IC of 5-fluorouracil 50 (200 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of 5-fluorouracil 50 (200 μΜ); G5: duloxetine (5 μΜ) + IC of 5-fluorouracil 50 (400 μΜ).

[0038] Figure 29 shows the effect of duloxetine and temozolomide on glioblastoma (LN-229 cell line). G1 : IC of temozolomide 50 (400 μΜ); G2: ½ IC of temozolomide 50 (200 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + ½ IC of temozolomide 50 (400 μΜ); G5: duloxetine (5 μΜ) + IC of temozolomide 50 (200 μΜ).

[0039] Figure 30 shows the effect of duloxetine and osimertinib (AZD-9291 ) on glioblastoma (LN-229 cell line). G1 : IC of osimertinib 50(5 μΜ); G2: 1 / 2 IC of osimertinib 50 (2.5 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + 1 / 2 IC of osimertinib 50 (2.5 μΜ); G5: duloxetine (5 μΜ) + IC of osimertinib 50 (5 μΜ).

[0040] Figure 31 shows the effect of duloxetine and etoposide on liver cancer (PLC / PRF / 5 cell line). G1 : IC of etoposide 50 (100 μΜ); G2: 1 / 2 IC of etoposide 50 (50 μΜ); G3: duloxetine (5 μΜ); G4: duloxetine (5 μΜ) + 1 / 2 IC of etoposide 50 (50 μΜ); G5: duloxetine (5 μΜ) + IC of etoposide 50 (100 μΜ).

[0041] Figure 32 shows the tumor shrinkage rate of individuals in the duloxetine + neoadjuvant chemotherapy (NACT) group after 24 weeks of treatment. R001, R002, R003, R004, R005, R006 and R008 represent the tumor shrinkage rate of 7 subjects, respectively, and 22.65% represents the average tumor shrinkage rate. R002, R004 and R008 are complete response (CR), so they are in the same line.

[0042] Figure 33 shows the tumor shrinkage rate of the NACT group and the duloxetine + NACT group. ***: P < 0.001.

[0043] Figure 34 shows the waterfall plot of the tumor shrinkage rate of the NACT group and the duloxetine + NACT group.

[0044] Figure 35 shows the changes in the tumor computed tomography images of the complete response (CR) subjects in the duloxetine + NACT group before treatment (left half) / after treatment (right half), and the changes in the tumor computed tomography images of the partial response (PR) subjects in the duloxetine + NACT group before treatment (left half) / after treatment (right half).

[0045] Figure 36 shows the objective response rates (ORR) of the duloxetine + NACT group. CR: complete response; PR: partial response.

[0046] Figure 37 shows the forkhead box protein P3 (Foxp3) expression of peripheral blood mononuclear cells (upper panel) and regression analysis of Foxp3-V1 and tumor shrinkage rate (lower panel) in duloxetine + NACT group before and after treatment. V1: visit 1, V8: visit 8; V16: visit 16; V18: visit 18. R001, R002, R003, R004, R005, and R006 represent the Foxp3 expression of 6 subjects, respectively.

[0047] Figure 38 shows the indoleamine 2,3-dioxygenase 1 (IDO-1) expression of peripheral blood mononuclear cells (upper panel) and regression analysis of IDO-1-V1 and tumor shrinkage rate (lower panel) in duloxetine + NACT group before and after treatment. V1: visit 1, V8: visit 8; V16: visit 16; V18: visit 18. R001, R002, R003, R004, R005, and R006 represent the IDO-1 expression of 6 subjects, respectively.

[0048] Figure 39 shows the Immunoglobulin kappa constant (IGKC) expression of peripheral blood mononuclear cells (upper panel) and regression analysis of IGKC-V18 and tumor shrinkage rate (lower panel) in duloxetine + NACT group before and after treatment. V1: visit 1, V8: visit 8; V16: visit 16; V18: visit 18. R001, R002, R003, R004, R005, and R006 represent the IGKC expression of 6 subjects, respectively. DETAILED DESCRIPTION

[0049] The following examples are provided to illustrate the present disclosure in detail. Those skilled in the art can easily understand the advantages and effects of the present disclosure after reading the present specification, and can also implement or apply them in other different embodiments. Therefore, the following examples for performing the present disclosure can be modified and / or changed without departing from the scope of different aspects and applications thereof, and any element or method within the scope of the present disclosure can be combined with any other element or method described in any embodiment of the present disclosure.

[0050] The articles "a," "an," and "the" each refer to one or more (i.e., at least one) of the grammatical object of the article. The terms "or" and "and / or" are used herein interchangeably and are each to be interpreted as an inclusive-inclusive rather than an exclusive-inclusive. The term "includes" is used herein to mean, and is used interchangeably with, the phrase "includes but is not limited to." As used herein, the term "about" is used to allow for variations normally associated with typical measurement of equivalents in the particular field. For example, "about" can encompass within its meaning ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% of the stated value. When "about" precedes a series of numbers or a range, it is understood that "about" can modify each number in the series or range. For example, numerical values are intended to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% from the stated value. As used herein, numerical ranges include any number that falls within the range, including the maximum and minimum values of the range, and can be combined to form a sub-range. For example, it is understood that a numerical range "20 to 30%" includes any sub-range between the minimum of 20% and the maximum of 30%, such as a sub-range from 20% to 25%, from 25% to 30%, and from 22.5% to 27.5%. Such variations can occur, for example, due to experimental error, typical errors in measuring or handling a compound, composition, concentrate, or formulation, differences in the source, manufacture, or purity of starting materials or ingredients used in the present disclosure, or similar considerations.

[0051] As described herein, "individual" is used to refer to any vertebrate, including, but not limited to, a human or a mammal, such as a deer, a mule, an elk, a black-tailed deer. In some preferred embodiments, the individual is a mammal, such as a human or a non-human mammal, for example, a domesticated mammal, such as a dog, a cat, a horse, a rat, a mouse, and the like, or a livestock mammal, such as a cow, a sheep, a pig, a deer, and the like.

[0052] As described herein, the terms "comprise", "include", "have", "contain", "involving", and any other variant thereof are intended to cover non-exclusive inclusions. For example, when a target is described as "comprising" a limiting factor, unless otherwise specified, other components, elements, members, structures, regions, portions, devices, systems, steps, or connection relationships, and the like can be additionally included, and other limiting factors should not be excluded.

[0053] As described herein, the term "effective amount" refers to an amount of an active agent or a pharmaceutical composition sufficient to produce a prophylactic or therapeutic effect in an individual in need thereof. In some embodiments, an effective amount of a composition causes prevention or reduction of appearance and / or symptoms associated with an undesirable condition. The effective amount can be varied by one of ordinary skill in the art depending on the use of the excipient, the route of administration, the possibility of co-usage with other therapeutic treatments, or the condition to be treated, but the present disclosure is not limited thereto.

[0054] As described herein, the term "administering" refers to the introduction of an active ingredient into an individual by a method or route such that at least a portion of the active ingredient is positioned at a desired site to produce a desired effect. For example, the active ingredients of the present disclosure can be administered to an individual by means of injection or topical application, but the present disclosure is not so limited. Administration of the compositions of the present disclosure can be performed in the systemic or local environment of the individual. For example, the site of topical administration can be any site in the body where tissue development is desired or beneficial, such as: a joint, a surgical site, a segmental bone gap or site of non-union, a wound, an ulcer, or an inflammatory rash.

[0055] As described herein, the term "preventing" refers to prophylactic or preventative measures against a disease, symptom, or condition, such as, but not limited to, the application or administration of one or more active agents to an individual who is not yet diagnosed with the disease, symptom, or condition, but who can be susceptible to or otherwise at risk of developing the disease, symptom, or condition.

[0056] As described herein, the term "treating" refers to obtaining a desired pharmacological or physiologic effect, such as, but not limited to, inhibiting the growth of cancer cells or reducing the size of a lesion tumor. The effect can be prophylactic in terms of completely or partially preventing a condition, symptom, disease, or disorder, and / or can be therapeutic in terms of a partial or complete cure for a condition and / or adverse effect attributable to the condition or disease.

[0057] As described herein, the term "pharmaceutically acceptable carrier or excipient" refers to a pharmaceutically-acceptable material, composition, or carrier, such as a liquid or solid filler, diluent, solvent, or encapsulation material. In some embodiments, each component is "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of a cosmetic or pharmaceutical formulation, and suitable for use in contact with the tissue or organ of an individual (e.g., a human or animal) without excessive toxicity, irritation, allergic response, immunogenicity, or other problem or complication, commensurate with a reasonable benefit / risk ratio. See Remington: The Science and Practice of Pharmacy, 22ndEd.; Allen, Ed.; Philadelphia, PA, 2012; Handbook of Pharmaceutical Excipients, 7thEd.; Rowe et al., Eds.; Pharmaceutical Press and American Pharmaceutical Association: 2012; Handbook of Pharmaceutical Additives, 3rdEd.; Ash and Ash, Eds.; Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2ndEd.; Gibson, Ed.; CRC Press LLC: Boca Raton, FL, 2009.

[0058] As described herein, the term "IC 50"IC50" refers to the half-inhibitory concentration (or half-inhibitory rate). It is a very important data in the standard curve of indirect competitive ELISA (icELISA), which is an S-shaped curve. In icELISA, the optical density (OD value) of the control group without adding drugs is defined as B0, and the OD value of the experimental group with drugs is B, and B / B0% is called the binding rate. The concentration of the drug corresponding to 50% of the binding rate is called IC 50 . Generally, the smaller the value of IC 50 , the stronger the inhibitory effect of the drug.

[0059] As used herein, the term "tumor" includes all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.

[0060] As described herein, the term "cancer" includes diseases of skin tissue, organs, blood and blood vessels, such as, but not limited to, bladder cancer, bone cancer, blood cancer, brain cancer, breast cancer, cervical cancer, chest cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, mouth cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, stomach cancer, testicular cancer, throat cancer, and uterine cancer. Specific cancers include, but are not limited to, advanced malignancy, amyloidosis, neuroblastoma, meningioma, hemangiopericytoma, multiple brain metastases, glioblastoma multiforme, glioblastoma, brain stem glioma, poor prognosis malignant brain tumor, malignant glioma, recurrent malignant glioma, pleomorphic xanthoastrocytoma, pleomorphic oligodendroglioma, neuroendocrine tumor, rectal adenocarcinoma, Dukes C & D colorectal cancer, unresectable colorectal cancer, metastatic hepatocellular carcinoma, Kaposi's sarcoma, karyotypically acute myeloblasts leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, cutaneous B-cell lymphoma, diffuse large B-cell lymphoma, low grade follicular lymphoma, malignant melanoma, malignant mesothelioma, malignant pleural mesothelioma syndrome, peritoneal cancer, papillary serous carcinoma, gynecological sarcoma, soft tissue sarcoma, scleroderma, cutaneous vasculitis, Langerhans cell histiocytosis, leiomyosarcoma, fibrodysplasia ossificans progressiva, hormone refractory prostate cancer, high risk resected soft tissue sarcoma, unresectable hepatocellular carcinoma, Waldenstrom's macroglobulinemia, and indolent myeloma, inert myeloma, fallopian tube cancer, androgen independent prostate cancer, androgen dependent stage IV non-metastatic prostate cancer, hormone-insensitive prostate cancer, chemotherapy-insensitive prostate cancer, urachal cancer, papillary thyroid cancer, follicular thyroid cancer, medullary thyroid cancer, and leiomyoma.

[0061] As described herein, expression of Ki-67 protein is closely associated with breast cancer proliferation in the development of breast cancer, and expression of Ki-67 is a known prognostic and outcome indicator. Tumors can be classified according to Ki-67 index, and tumors with high Ki-67 index have a large number of proliferating cells, and thus can grow faster. Ki-67 can be detected by immunohistochemistry (IHC) method at present.

[0062] As described herein, forkhead box protein P3 (Foxp3) is a member of the forkhead transcription factor family. Expression of Foxp3 is associated with poor prognosis. Foxp3 has been widely recognized as a gene associated with breast cancer and prostate cancer in in vitro and in vivo studies.

[0063] As described herein, indoleamine 2,3-dioxygenase 1 (IDO-1) is a tryptophan catabolism enzyme, and IDO-1 is considered not only an immunomodulator during pregnancy, but also as an immunomodulator in autoimmune diseases, chronic inflammation, and tumor immunity. In addition, IDO-1 has been considered a novel cancer immunotherapy target in recent years.

[0064] As described herein, immunoglobulin kappa constant (IGKC) can currently be used as a single and powerful immunomarker for predicting metastasis-free survival and chemotherapy response.

[0065] In at least one embodiment of the present disclosure, an effective amount of duloxetine and a chemotherapeutic drug can be administered to an individual in need thereof.

[0066] In at least one embodiment of the present disclosure, a chemotherapeutic drug can be administered to an individual first, and then duloxetine can be administered.

[0067] In at least one embodiment of the present disclosure, the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma.

[0068] In some embodiments, the breast cancer can be triple-negative breast cancer.

[0069] In at least one embodiment of the present disclosure, the chemotherapeutic drug can be at least one selected from the group consisting of an alkylating antineoplastic agent, an antimitotic agent, a DNA intercalating agent, a topoisomerase inhibitor, a DNA cleaving agent, an antimetabolites agent, and a tyrosine kinase inhibitor.

[0070] In at least one embodiment of the present disclosure, the chemotherapeutic drug can comprise an anti-mitotic agent, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma. In some embodiments, the anti-mitotic agent can be at least one selected from the group consisting of Docetaxel, Vinblastine, Vincristine, and Hupehenine. In some embodiments of the present disclosure, the chemotherapeutic drug can be Docetaxel, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma.

[0071] In at least one embodiment of the present disclosure, the chemotherapeutic drug can comprise an alkylating antineoplastic agent, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma. In some embodiments, the alkylating antineoplastic agent can be at least one selected from the group consisting of Cisplatin, Cyclophosphamide, Temozolomide, Nitrogen mustards, Chlormethine, Uramustine, Melphalan, Chlorambucil, Ifosfamide, Bendamustine, Nitrosoureas, Carmustine, Lomustine, Streptozocin, Alkyl sulfonates, Busulfan, Platinum, Carboplatin, Dicycloplatin, Eptaplatin, Lobaplatin, Miriplatin, Nedaplatin, Oxaliplatin, Picoplatin, Satraplatin, Triplatin tetranitrate, Triazenes, Dacarbazine, Mitozolomide, Procarbazine, and Altretamine. In some embodiments, the alkylating antineoplastic agent can be Cisplatin, and the cancer can be at least one selected from the group consisting of breast cancer, pancreatic cancer, and liver cancer. In some embodiments, the chemotherapeutic drug can be Cyclophosphamide, and the cancer can be breast cancer and / or glioblastoma. In some embodiments, the chemotherapeutic drug can be Temozolomide, and the cancer can be breast cancer and / or non-small cell lung cancer.

[0072] In at least one embodiment of the present disclosure, the chemotherapeutic drug can comprise an antimetabolite agent, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma. In some embodiments, the antimetabolite agent is at least one selected from the group consisting of 5-Fluorouracil, Cytarabine, 6-Mercaptopurine, and Methotrexate. In some embodiments, the antimetabolite agent can be 5-Fluorouracil, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma.

[0073] In at least one embodiment of the present disclosure, the chemotherapeutic drug can comprise a DNA intercalator or a DNA cleaving agent, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, and liver cancer. In some embodiments, the DNA intercalator can be at least one selected from the group consisting of Doxorubicin, Actinomycin D, and Daunorubicin, and the DNA cleaving agent can be at least one selected from the group consisting of Doxorubicin, Bleomycin, and Daunorubicin. In some embodiments, the DNA intercalator or the DNA cleaving agent can be Doxorubicin, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, and liver cancer.

[0074] In at least one embodiment of the disclosure, the chemotherapeutic drug can comprise a tyrosine kinase inhibitor, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma. In some embodiments, the tyrosine kinase inhibitor can be at least one selected from the group consisting of Osimertinib, Sorafenib, Lenvatinib, Imatinib, Sunitinib, Erlotinib, Gefitinib, Dasatinib, and Lapatinib. In some embodiments, the cancer is a cancer with low or absent expression of argininosuccinate synthase 1 (ASS1), such as, but not limited to, a liver cancer with low or absent expression of ASS1 or a breast cancer with low or absent expression of ASS1. In some embodiments, the tyrosine kinase inhibitor can be Osimertinib, and the cancer can be at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, and glioblastoma. In some embodiments, the tyrosine kinase inhibitor can be Sorafenib, and the cancer can be non-small cell lung cancer and / or liver cancer. In some embodiments, the tyrosine kinase inhibitor can be Lenvatinib, and the cancer can be liver cancer.

[0075] In at least one embodiment of the disclosure, the chemotherapeutic drug can comprise a topoisomerase inhibitor, but not etoposide. In some embodiments, the topoisomerase inhibitor can be at least one selected from the group consisting of Camptothecin, Topotecan, Irinotecan, and Podophyllotoxin. In some embodiments, the topoisomerase inhibitor comprises a type 1 topoisomerase inhibitor, but not a type 2 topoisomerase inhibitor. In some embodiments, the topoisomerase inhibitor does not comprise etoposide.

[0076] In at least one embodiment of the disclosure, the pharmaceutical composition can comprise a pharmaceutically acceptable carrier or excipient.

[0077] In at least one embodiment of the disclosure, the pharmaceutical composition can be formulated into a dosage form selected from the group consisting of a tablet, a capsule, an injection, a lozenge, a powder, a granule, and any combination thereof.

[0078] In at least one embodiment of the disclosure, the chemotherapeutic drug can be at least one selected from the group consisting of Docetaxel, Cisplatin, Cyclophosphamide, 5-Fluorouracil, Doxorubicin, Temozolomide, Osimertinib, Sorafenib, Lenvatinib, and any combination thereof.

[0079] In at least one embodiment of the disclosure, the pharmaceutical composition of the disclosure can reduce Ki-67.

[0080] In at least one embodiment of the disclosure, the pharmaceutical composition of the disclosure can reduce tumor by at least 40% (for example, but not limited to, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%).

[0081] In at least one embodiment of the disclosure, the pharmaceutical composition of the disclosure shows that 1) Docetaxel combined with Duloxetine produces better efficacy on triple-negative breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma than Docetaxel alone; 2) Cisplatin combined with Duloxetine produces better efficacy on triple-negative breast cancer, pancreatic cancer, and liver cancer than Cisplatin alone; 3) Cyclophosphamide combined with Duloxetine produces better efficacy on triple-negative breast cancer and glioblastoma than Cyclophosphamide alone; 4) 5-Fluorouracil combined with Duloxetine produces better efficacy on triple-negative breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma than 5-Fluorouracil alone; 5) Doxorubicin combined with Duloxetine produces better efficacy on triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer, and liver cancer than Doxorubicin alone; 6) Temozolomide combined with Duloxetine produces better efficacy on triple-negative breast cancer and non-small cell lung cancer than Temozolomide alone; 7) Osimertinib combined with Duloxetine produces better efficacy on triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer, and glioblastoma than Osimertinib alone; and 8) Sorafenib combined with Duloxetine produces better efficacy on non-small cell lung cancer and liver cancer than Sorafenib alone; 9) Lenvatinib combined with Duloxetine produces better efficacy on liver cancer than Lenvatinib alone; and 10) Etoposide combined with Duloxetine does not produce better efficacy on treating triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma than Etoposide alone.

[0082] In at least one embodiment of the present disclosure, the results show that duloxetine + NACT combination can effectively improve tumor shrinkage rate, increase the number of complete remission, and reduce Ki-67. In some embodiments of the present disclosure, the relative tumor reduction of the duloxetine + NACT group is 22.65%, the tumor shrinkage rate of the duloxetine + NACT group is higher than that of the NACT group, the tumor shrinkage rate of the duloxetine + NACT group is higher than that of the NACT group, the proportion of individuals with CR (42.9%) in the duloxetine + NACT group is higher than that (0%) in the NACT group, individuals with less Foxp3 concentration before treatment (Foxp3-V1) are more likely to achieve better tumor shrinkage rate, individuals with less IDO-1 concentration before treatment (IDO-1-V1) are more likely to achieve better tumor shrinkage rate, and individuals with less IGKC concentration after treatment (IGKC-V18) are more likely to achieve better tumor shrinkage rate. In some embodiments, individuals with XIRP2 gene mutations can exhibit better efficacy, and the mutations can be, but are not limited to, single nucleotide variants (SNVs). In some embodiments, individuals with complete remission (CR) all exhibit XIRP2 gene SNVs, but individuals with partial remission (PR) do not exhibit XIRP2 gene SNVs.

[0083] In some specific embodiments of the present disclosure, XIRP2 gene mutations (such as, but not limited to, SNVs), Foxp3 expression, and / or IGKC expression of individuals can be detected before the individuals are administered the combination therapy of the present disclosure, and the combination therapy of the present disclosure is administered to individuals who exhibit XIRP2 variations, reduced Foxp3 expression, and / or reduced IGKC expression for the purpose of precision medicine. In other specific embodiments of the present disclosure, XIRP2 gene mutations (such as, but not limited to, SNVs), Foxp3 expression, and / or IGKC expression of individuals can be detected after the individuals are administered the combination therapy of the present disclosure, and if the individuals exhibit XIRP2 variations, reduced Foxp3 expression, and / or reduced IGKC expression, it is determined that they have better prognostic efficacy.

[0084] EMBODIMENT

[0085] The exemplary embodiments of the present disclosure are further described below, which should not be interpreted as limiting the scope of the present disclosure.

[0086] In the present disclosure, the concentrations of the example drugs in the embodiments are listed in Table 1, and the information of the cancer cell models is listed in Table 2.

[0087] Table 1 Concentrations of drugs in embodiments of the present disclosure

[0088] Table 2 Cancer cell models in the present disclosure

[0089] Example 1: Establishment of cell lines and cancer cell survival assay method

[0090] Please refer to Table 2, the cell lines of different cancer types were subcultured, and after calculating the cell number, 1x10 4Cell number, 24 hours later, drug was added (as shown in Table 1), 72 hours later, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was added, 90 minutes later, optical density value at 570 nm wavelength was detected. Data was calculated as cell inhibition rate (as 100% - cell survival rate). If G5 value is higher than G1, or G4 value is higher than G2, then the combination result is better than using chemotherapy drugs alone. Results are shown in Figures 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 and 31, refer to Tables 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 (experimental conditions are also described below the table), 13 and 14 below, and the results of Example 1 are shown in Table 14. The results show that: 1) compared with docetaxel alone, docetaxel combined with duloxetine has better efficacy on triple-negative breast cancer, non-small cell lung cancer, liver cancer and glioblastoma; 2) compared with cisplatin alone, cisplatin combined with duloxetine has better efficacy on triple-negative breast cancer, pancreatic cancer and liver cancer; 3) compared with cyclophosphamide alone, cyclophosphamide combined with duloxetine has better efficacy on triple-negative breast cancer and glioblastoma; 4) compared with 5-fluorouracil alone, 5-fluorouracil combined with duloxetine has better efficacy on triple-negative breast cancer, non-small cell lung cancer, liver cancer and glioblastoma; 5) compared with doxorubicin alone, doxorubicin combined with duloxetine has better efficacy on triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer and liver cancer; 6) compared with temozolomide alone, temozolomide combined with duloxetine has better efficacy on triple-negative breast cancer and non-small cell lung cancer; 7) compared with osimertinib alone, osimertinib combined with duloxetine has better efficacy on triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer and glioblastoma; and 8) compared with sorafenib alone, sorafenib combined with duloxetine has better efficacy on non-small cell lung cancer and liver cancer; 9) compared with lenvatinib alone, lenvatinib combined with duloxetine has better efficacy on liver cancer; and 10) compared with etoposide alone, etoposide combined with duloxetine does not have better efficacy on triple-negative breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer and glioblastoma.

[0091] Table 3 Results of cancer cell inhibition rate of docetaxel and duloxetine combination in the present disclosure

[0092] Y: better than the chemotherapeutic drug alone

[0093] N: not better than the chemotherapeutic drug alone

[0094] Table 4 Results of cancer cell inhibition rate of cisplatin and duloxetine combination in the present disclosure

[0095] Y: better than the chemotherapeutic drug alone

[0096] N: not better than the chemotherapeutic drug alone

[0097] Table 5 Results of cancer cell inhibition rate of cyclophosphamide (IC50) and duloxetine combination in the present disclosure

[0098] Y: better than the chemotherapeutic drug alone

[0099] N: not better than the chemotherapeutic drug alone

[0100] Table 6 Results of cancer cell inhibition rate of 5-fluorouracil (1 / 2 IC50) and duloxetine combination in the present disclosure

[0101] Y: better than the chemotherapeutic drug alone

[0102] N: not better than the chemotherapeutic drug alone

[0103] Table 7 Results of cancer cell inhibition rate of doxorubicin and duloxetine combination in the present disclosure

[0104] Y: better than the chemotherapeutic drug alone

[0105] N: not better than the chemotherapeutic drug alone

[0106] Table 8 Results of cancer cell inhibition rate of temozolomide and duloxetine combination in the present disclosure

[0107] Y: better than the chemotherapeutic drug alone

[0108] N: not better than the chemotherapeutic drug alone

[0109] Table 9 Results of cancer cell inhibition rate of osimertinib (IC50) and duloxetine combination in the present disclosure 50

[0110] Y: better than the chemotherapeutic drug alone

[0111] N: not better than the chemotherapeutic drug alone ​

[0112] Table 10 Cancer cell inhibition rate results of the combination of osimertinib (1 / 2 IC 50 ) and duloxetine in the present disclosure

[0113] Y: superior to the use of the chemotherapeutic drug alone

[0114] N: not superior to the use of the chemotherapeutic drug alone

[0115] Table 11 Cancer cell inhibition rate results of the combination of sorafenib (1 / 2 IC 50 ) and duloxetine in the present disclosure

[0116] Y: superior to the use of the chemotherapeutic drug alone

[0117] N: not superior to the use of the chemotherapeutic drug alone

[0118] Table 12 Cancer cell inhibition rate of the combination of lenvatinib and duloxetine in the present disclosure

[0119] Y: superior to the use of the chemotherapeutic drug alone

[0120] N: not superior to the use of the chemotherapeutic drug alone

[0121] The experimental conditions of Table 12 above are as follows:

[0122] The PLC / PRF / 5 cells or SK-HEP-1 cells were subcultured, and after counting the cell number, 1 x 10 4 The cell number was calculated, and after 24 hours, the drugs were added: 10 μM lenvatinib and / or 10 μM duloxetine were used for PLC / PRF / 5 cells, and 6.25 μM lenvatinib and / or 10 μM duloxetine (experimental group), 10 μM chloroquine and / or 10 μM duloxetine (control group) were used for SK-HEP-1 cells. After 72 hours, MTT was added, and after 90 minutes, the optical density value at 570 nm was detected. The data were calculated as the cell inhibition rate (100% - cell survival rate).

[0123] As can be seen from Table 12 above, the combination of lenvatinib and duloxetine (experimental group) had a cancer cell inhibition rate as high as 76%, which was much higher than the cancer cell inhibition rate of 33% of the combination of lenvatinib and chloroquine (control group). In addition, except for the liver cancer cell line SK-HEP-1, the cell inhibition rate of the combination of lenvatinib and duloxetine for another liver cancer cell line PLC / PRF / 5 was also superior to that of each of lenvatinib and duloxetine used alone (cell inhibition rate of lenvatinib: 18%; cell inhibition rate of duloxetine: 19%; cell inhibition rate of lenvatinib + duloxetine: 41%).

[0124] Argininosuccinate synthetase 1 (ASS1) is an enzyme that catalyzes the synthesis of argininosuccinate from aspartate and citrulline. ASS1 is highly expressed in liver, but lowly expressed or absent in some cancer cells, such as but not limited to hepatoma cell line SK-HEP-1, which makes these cells have exogenous arginine auxotrophy. Generally, hepatoma cells with low or absent ASS1 expression have higher invasiveness, proliferation, and poor prognosis. Compared with hepatoma cell line SK-HEP-1, hepatoma cell line PLC / PRF / 5 has normal or high ASS1 expression. As shown in the above examples, the combination of lenvatinib and duloxetine has a synergistic inhibitory effect on hepatoma cell line SK-HEP-1 with low or absent ASS1 expression and hepatoma cell line PLC / PRF / 5 with normal or high ASS1 expression. However, compared with hepatoma cell line PLC / PRF / 5 with normal or high ASS1 expression (cell inhibition rate: 41%), the combination of lenvatinib and duloxetine has a better inhibitory effect on hepatoma cell line SK-HEP-1 with low or absent ASS1 expression (cell inhibition rate: 76%). Therefore, the combination therapy of the present disclosure provides a new treatment for cancers with low or absent ASS1 expression. Although the above example is based on hepatoma cell line SK-HEP-1, the present disclosure is not limited to hepatoma, for example, triple-negative breast cancer cell line MDA-MB-231 also belongs to breast cancer cells with low or absent ASS1 expression, and the combination therapy of the present disclosure also has a good cell inhibition rate on MDA-MB-231.

[0125] Table 13 Cancer cell inhibition rate results of the combination of etoposide and duloxetine in the present disclosure

[0126] Y: better than using a chemotherapy drug alone

[0127] N: not better than using a chemotherapy drug alone

[0128] Table 14 General table of cancer cell inhibition rate analysis in the present disclosure

[0129] Y: better than using a chemotherapy drug alone

[0130] N: not better than using a chemotherapy drug alone

[0131] Example 2 Clinical trial

[0132] Inclusion criteria: Female subjects with newly diagnosed stage II breast cancer with triple negative tumors > 2 cm in size who were > 20 years of age and had not undergone a mastectomy or received systemic chemotherapy. NACT: First, 4 cycles of docetaxel were administered, followed by 4 cycles of cyclophosphamide and doxorubicin, and finally surgical resection of the lesion. Duloxetine + NACT: After clinical inclusion, duloxetine was administered starting one week before the administration of the chemotherapy drugs, and after the end of the chemotherapy, the drug was reduced for two weeks, and finally surgical resection of the lesion.

[0133] Methods for detecting Foxp3 expression and IDO-1 expression

[0134] The pre-treatment process is described as follows. First, collect 4 tubes of whole blood samples with ethylenediaminetetraacetic acid (EDTA) anticoagulant. Perform pre-treatment on the same day at room temperature. Centrifuge (340g, 5min) two tubes of whole blood from the subject, remove the supernatant (plasma), and place it in a 2mL microcentrifuge tube. Label the sample number, take a photo, and check for hemolysis. Store at -80°C. After the 4 tubes of samples from the same subject are collected, perform ELISA testing. The third tube of whole blood is not centrifuged and is stored at 4°C for later use. The fourth tube is pre-treated for flow cytometry experiments. Take IX RBC (red blood cell) lysis buffer and dilute it 10 times with deionized and distilled water (dd water). Mix the whole blood gently and evenly, take 500μl, and add 9.5mL of IX RBC lysis buffer. Mix well and avoid light for 15min. After the reaction is complete, centrifuge (340g, 5min) to remove the supernatant. Add 200μl of staining buffer to disperse the cells on the wall of the tube, and transfer them to a 1.75mL microcentrifuge tube. Centrifuge (340g, 5min) to remove the supernatant. Add 0.5mL of fixation buffer and avoid light for 20min. After fixation is complete, centrifuge (340g, 5min) to remove the supernatant. Add 0.5mL of staining buffer and store at 4°C. The antibodies used in this disclosure are listed in Table 15. The chemicals used in this disclosure are listed in Table 16.

[0135] Staining and machine

[0136] Centrifuge (340g, 5min) the pre-treated sample to remove the supernatant, add 300μl of staining buffer to resuspend it again, mix it evenly, and then divide it equally into three tubes, each with a volume of 100μl.

[0137] Foxp3 staining was performed in the 2nd tube (permeabilization). 100 μl of sample was centrifuged (340g, 5 min) and the supernatant was removed. 200 μl of intracellular staining permeabilization buffer was added, mixed well for permeabilization, and left to stand for 10 min. Centrifugation (340g, 5 min) was performed, and the supernatant was removed. After resuspension with 100 μl of staining buffer, the corresponding staining volume of antibody Foxp3 was taken, mixed well, and left to stand for 20 min in the dark. 400 μl of staining buffer was added to make the volume 500 μl when loaded onto the machine. Flow cytometry analysis was performed (Novocyte 3000, Agilent), and 10,000 white blood cells were selected for analysis.

[0138] IDO-1 staining was performed in the 3rd tube (permeabilization). 100 μl of sample was centrifuged (340g, 5 min) and the supernatant was removed. 200 μl of intracellular staining permeabilization buffer was added, mixed well for permeabilization, and left to stand for 10 min. Centrifugation (340g, 5 min) was performed, and the supernatant was removed. After resuspension with 100 μl of staining buffer, the corresponding staining volume of antibody IDO-1 was taken, mixed well, and left to stand for 20 min in the dark. 400 μl of staining buffer was added to make the volume 500 μl when loaded onto the machine. Flow cytometry analysis was performed (Novocyte 3000, Agilent), and 10,000 white blood cells were selected for analysis.

[0139] Table 15 Antibodies used in the present disclosure

[0140] Table 16 Chemicals used in the present disclosure

[0141] Method for detecting IGKC expression

[0142] The method is described in detail as follows. Whole blood samples were collected in EDTA-containing anticoagulant purple head tubes, a total of 4 tubes, and pretreated at room temperature on the same day. Two tubes of whole blood from the patient were centrifuged (340g, 5 min), and the plasma was removed and placed in a 2 mL microcentrifuge tube. The sample number was labeled and photographed, and hemolysis was observed. The target content was analyzed by ELISA kit, and the optical density value at 450 nm wavelength was detected by spectrophotometer (M200 PRO, TECAN). The experiment was completed in a 96-well plate, and the standard and sample in the sequence dilution were planned in the plate in duplicate. According to the known concentration of the standard after sequence dilution, a function equation (calculated by software, usually approximated to a binary first-order equation graph) can be obtained with the corresponding optical density value. The OD value of the sample measured was brought into the formula, and the concentration value of the sample was obtained.

[0143] Exclusion criteria, the following people are not suitable for participating in the embodiments of the disclosure:

[0144] History of chemotherapy for any malignancy; history of radiation therapy for breast cancer; second primary malignancy not in remission five years ago, except for non-melanoma skin cancer or cervical carcinoma in situ; poor general condition (not suitable for receiving dose-intensive, dose-intensified anthracycline (taxane) targeted drug combination chemotherapy); history of suicide or self-harm behavior in the past five years.

[0145] The following drugs are not suitable for participating in the embodiments of the disclosure: Monoamine oxidase inhibitors (MAO-I), use of drugs within 14 days before registration. Simultaneous use with phenothiazines (including thioridazine), propafenone, flecainide, triptans, MAO-I, selective serotonin reuptake inhibitors (SSRI), serotonin-norepinephrine reuptake inhibitors (SNRI) or tricyclic antidepressants.

[0146] Individuals with the following cardiovascular diseases or individuals with autoimmune diseases or using immunosuppressive agents (except steroids) are not suitable for participating in the embodiments of the disclosure: symptomatic congestive heart failure, myocardial infarction, severe or unstable angina pectoris (within 6 months before the screening day), high risk of arrhythmia, uncontrolled hypertension.

[0147] The following individuals are not suitable for participating in the embodiments of the disclosure: Patients with the following diseases: liver dysfunction, chronic liver disease, severe kidney disease; symptomatic chronic lung disease, symptomatic restrictive lung disease, interstitial pneumonia or other lung function abnormalities that may affect patient safety. Uncontrolled angle-closure glaucoma or clinically significant coagulopathy; patients with uncontrolled bacterial or viral infections, or active or recent (within 6 months) fungal infections at the time of joining the trial; patients with active central nervous system disease at the time of joining the trial; currently have primary mental illness (schizophrenia, psychosis) or have a history of suicidal thoughts, bipolar disorder or seizure disorders.

[0148] Serologically positive for human immunodeficiency virus (HIV) infection or HIV positive, hepatitis B virus (HBV) or hepatitis C virus (HCV) acute phase; patients known to be hypersensitive to any of the study treatment drugs, including excipients of the study drug. History of alcohol abuse in the past year, i.e., more than 14 standard drinks per week for men or more than 7 standard drinks per week for women (one standard drink is defined as 360 ml of beer, 150 ml of wine, or 45 ml of 40% distilled liquor, such as rum, whiskey, brandy, etc.).

[0149] Patients who are unable to use effective contraception from screening until 6 months after the end of the study and after withdrawal from the study. Pregnant or lactating women, or women planning to become pregnant or to breastfeed. Any condition that, in the opinion of the investigator, would affect the study treatment assessment, patient safety, or interpretation of the study results.

[0150] Individuals who have participated in another clinical trial and received a trial product within 28 days prior to enrollment in the current example, or received a pre-study cellular therapy / transplant, unless otherwise approved by the principal investigator, are not eligible to participate in the present example.

[0151] Referring to FIG. 32, the relative tumor reduction in the duloxetine + NACT group was 22.65%. Referring to FIG. 33, the tumor shrinkage rate in the duloxetine + NACT group was higher than that in the NACT group. Referring to FIG. 34, the tumor shrinkage waterfall plot showed that the tumor shrinkage ratio in the duloxetine + NACT group was higher than that in the NACT group. Referring to FIGS. 35 and 36, the results showed that the proportion of individuals with CR (42.9%) in the duloxetine + NACT group was higher than that in the NACT group (0%). Referring to FIG. 37, the results showed that the Foxp3 expression in peripheral blood mononuclear cells before treatment (upper panel) in the duloxetine + NACT group had a trend of negative correlation with the tumor shrinkage rate (R2=0.728, P=0.031, lower panel), indicating that individuals with less Foxp3 concentration before treatment (Foxp3-V1) were more likely to achieve better tumor shrinkage. Referring to FIG. 38, the IDO-1 expression in peripheral blood mononuclear cells before treatment (upper panel) in the duloxetine + NACT group had a trend of negative correlation with the tumor shrinkage rate (R2=0.579, P=0.079, lower panel), indicating that individuals with less IDO-1 concentration before treatment (IDO-1-V1) were more likely to achieve better tumor shrinkage. Referring to FIG. 39, the higher IGKC expression in peripheral blood mononuclear cells after treatment (upper panel) in the duloxetine + NACT group had a trend of negative correlation with the tumor shrinkage rate, indicating that individuals with less IGKC concentration after treatment (IGKC-V18) were more likely to achieve better tumor shrinkage (R2=0.796, P=0.017, lower panel).

[0152] As shown in Table 17 below, in the present embodiment, the NACT group (N=6) and the duloxetine+NACT group (N=7) were similar in terms of the basic information of the individuals before treatment.

[0153] Table 17 Basic information of the NACT group (N=6) and the duloxetine+NACT group (N=7) before treatment

[0154] BMI: body mass index

[0155] ECOG score: Eastern Cooperative Oncology Group score, which is an index for evaluating the activity and physical condition of cancer patients and is commonly used in clinical trials and treatment decisions for cancer. In cancer clinical trials, ECOG scores of 0 (indicating that the subject is completely normal and has no difference from healthy people, and can perform all daily activities) or 1 (indicating that the subject is slightly limited and can engage in light physical activities such as general household chores or office work, but cannot engage in heavy physical activities) are often listed as inclusion criteria to ensure that the subjects have certain physical conditions. TNM stage number: an international standard for staging malignant tumors / cancer, which consists of three main parameters: T (describing the size and local invasion of the primary tumor), N (describing the presence or absence and extent of regional lymph node metastasis), and M (describing distant metastasis). Each parameter is further subdivided by number according to its severity, for example: T0 to T4, N0 to N3, M0 to M1. TNM IIa: T0 to T2; N0 to N1; M0. TNM IIb: T2 to T3; N0 to N1; M0. TNM IIIc: any of T, N3, and M0.

[0156] SD: standard deviation

[0157] As shown in Table 18 below, in the present embodiment, the medication adherence of the duloxetine+NACT group was about 80 to 90%.

[0158] Table 18 Medication adherence of the duloxetine+NACT group (N=7)

[0159] SD: standard deviation

[0160] As shown in Table 19 below, in the present embodiment, after treatment, the duloxetine+NACT group showed significant decreases in tumor size, blood pressure, and Ki-67 (%). According to Table 20 below, the average tumor shrinkage rate of the duloxetine+NACT group was higher than that of the NACT group.

[0161] Table 19 Change in body weight, blood pressure, and tumor size after treatment in NACT group vs. duloxetine + NACT group

[0162] *Statistical analysis was performed for only 5 individuals due to lack of ki-67 data for 1 individual in NACT group.

[0163] Table 20 Mean tumor shrinkage rate in NACT group vs. duloxetine + NACT group

[0164] NACT: neoadjuvant chemotherapy

[0165] Referring to FIG. 35 and Table 21, it can be seen that the NACT group had only partial remission. In the duloxetine + NACT group, 3 individuals had complete remission and 4 individuals had partial remission. It can be seen that the duloxetine + NACT group had better results than the NACT group.

[0166] Table 21 Objective response rate and disease control rate

[0167] The Response Evaluation Criteria in Solid Tumors (RECIST) is an international standard for evaluating the response of solid tumors to cancer treatment and is widely used for drug efficacy evaluation in clinical trials and as one of the treatment response indicators. RECIST mainly relies on changes in tumor lesion diameter measured by imaging to objectively classify the treatment response of patients, which is divided into: (1) complete remission (Complete Response, CR): complete disappearance of lesions; (2) partial remission (Partial Response, PR): total lesion diameter is reduced by at least 30%; (3) stable disease (Stable Disease, SD): tumor changes do not meet the PR or PD criteria; (4) progressive disease (Progressive Disease, PD): total lesion diameter increases by at least 20% or new lesions appear.

[0168] Table 21 above only includes individuals who can be evaluated for RECIST response. NACT: neoadjuvant chemotherapy group. Objective response rate (ORR) data from computed tomography (CT) image results.

[0169] In addition, in the duloxetine + NACT group, it was found that 3 CR individuals had single nucleotide variations (SNVs) in the XIRP2 gene, while 4 PR individuals did not have SNVs, indicating that the XIRP2 gene can be used as a key indicator for cancer prediction, treatment, and prognosis evaluation in the embodiments of the present disclosure.

[0170] Referring to Tables 22, 23, and 24, the hematological report, biochemical report, and urine analysis of the duloxetine + NACT group showed that duloxetine + NACT intervention had no adverse effects on the individuals.

[0171] Table 22 Hematological report of the duloxetine + NACT group

[0172] RBC: red blood cells. WBC: white blood cells. DC: differential count. Neu: neutrophils. Eos: eosinophils. Baso: basophils. Mono: monocytes. RDW: red blood cell distribution width. MCV: mean corpuscular volume. MCHC: mean corpuscular hemoglobin concentration. ANC: absolute neutrophil count. SD: standard deviation.

[0173] Table 23 Biochemical report of the duloxetine + NACT group

[0174] BUN: blood urea nitrogen. TG: triglyceride. SD: standard deviation.

[0175] AST: aspartate aminotransferase. SGOT: serum glutamic-oxaloacetic transaminase. γ-GT: γ-glutamyltransferase. LDH: lactate dehydrogenase.

[0176] Table 24 Urine analysis of the duloxetine + NACT group

[0177] SD: standard deviation.

[0178] The results of this embodiment show that duloxetine + NACT combination can effectively improve tumor shrinkage rate, increase the number of complete remission, and reduce Ki-67.

[0179] The above only describes some embodiments of the present disclosure, and those skilled in the art will easily understand that various modifications and changes can be made to each embodiment without departing from the teachings of the present disclosure. Therefore, any equivalent changes and modifications made in accordance with the claims attached to the present disclosure shall be within the scope of the present disclosure.

Claims

1. Use of duloxetine and a chemotherapeutic agent for the manufacture of a pharmaceutical composition for treating or preventing cancer.

2. The use of claim 1, comprising administering to a subject in need thereof an effective amount of the duloxetine and the chemotherapeutic agent.

3. The use of claim 2, wherein, The chemotherapeutic agent is administered to the subject prior to the duloxetine.

4. The use of claim 1, wherein, The cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma.

5. The use of claim 4, wherein, The breast cancer is triple-negative breast cancer.

6. The use of claim 1, wherein, The chemotherapeutic agent is at least one selected from the group consisting of an alkylating antineoplastic agent, an antimitotic agent, a DNA intercalating agent, a topoisomerase inhibitor, a DNA cleaving agent, an antimetabolite, and a tyrosine kinase inhibitor.

7. The use of claim 6, wherein, The chemotherapeutic agent comprises the antimitotic agent, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma.

8. The use of claim 7, wherein, The antimitotic agent is at least one selected from the group consisting of docetaxel, vinblastine, vincristine, and hainwoodine.

9. The use of claim 6, wherein, The chemotherapeutic agent comprises the alkylating antineoplastic agent, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma.

10. The use of claim 9, wherein, The alkylating antineoplastic agent is at least one selected from the group consisting of cisplatin, cyclophosphamide, temozolomide, mechlorethamine, mechlorethamine, uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, nitrosoureas, carmustine, cyclophosphamide, streptozotocin, alkyl sulfonate, busulfan, platinum, carboplatin, dicycloplatin, iproplatin, lobaplatin, nedaplatin, oxaliplatin, picoplatin, satraplatin, triplatin, triazenes, dacarbazine, mitozolomide, procarbazine, and hexamethylmelamine.

11. The use of claim 6, wherein, The chemotherapeutic agent comprises the antimetabolite, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, liver cancer, and glioblastoma.

12. The use of claim 11, wherein, The antimetabolite is at least one selected from the group consisting of 5-fluorouracil, cytarabine, 6-mercaptopurine, and methotrexate.

13. The use of claim 6, wherein, The chemotherapeutic agent comprises the DNA intercalating agent or the DNA cleaving agent, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, and liver cancer.

14. The use of claim 13, wherein, The DNA intercalating agent is at least one selected from the group consisting of doxorubicin, dactinomycin, and daunorubicin, and the DNA cleaving agent is at least one selected from the group consisting of doxorubicin, bleomycin, and daunorubicin.

15. The use of claim 6, wherein, The chemotherapeutic agent comprises the tyrosine kinase inhibitor, and the cancer is at least one selected from the group consisting of breast cancer, non-small cell lung cancer, pancreatic cancer, liver cancer, and glioblastoma.

16. The use of claim 15, wherein, The tyrosine kinase inhibitor is at least one selected from the group consisting of osimertinib, sorafenib, lenvatinib, imatinib, sunitinib, erlotinib, gefitinib, dasatinib, and lapatinib.

17. The use of claim 16, wherein, The cancer is a cancer with low or absent expression of argininosuccinate synthetase 1.

18. The use of claim 6, wherein, The chemotherapeutic agent comprises the topoisomerase inhibitor, but does not comprise etoposide.

19. The use of claim 18, wherein, The topoisomerase inhibitor is at least one selected from the group consisting of camptothecin, topotecan, irinotecan, and podophyllotoxin.

20. The use of claim 2, wherein, The individual has a mutation in the XIRP2 gene, reduced expression of forkhead box protein P3, and / or reduced expression of immunoglobulin kappa constant region.

21. The use of claim 20, further comprising detecting a mutation in the XIRP2 gene, expression of forkhead box protein P3, and / or expression of immunoglobulin kappa constant region in the individual prior to administering the effective amount of the duloxetine and the chemotherapeutic agent.

Images