CN111821303A - Application of vortioxetine and its salts in the preparation of antitumor drugs - Google Patents

Abstract

The invention discloses application of vortioxetine and salts thereof in preparation of antitumor drugs. In particular to application of vortioxetine and hydrobromide thereof in preparing a medicament for treating esophageal cancer, wherein the chemical name of vortioxetine hydrobromide is as follows: 1- [2- (2, 4-Dimethylphenylthio) phenyl]Piperazine hydrobromide, CAS number: 960203-27-4, formula: c₁₈H₂₃BrN₂S, molecular weight: 379.358. experiments prove that the vortioxetine hydrobromide can play a role in inhibiting the growth and transformation of esophageal squamous cell carcinoma cells when being used for the esophageal squamous cell carcinoma cells (KYSE 150 cells and KYSE450 cells), and the appropriate concentration of the vortioxetine hydrobromide capable of inhibiting the proliferation and transformation of the esophageal squamous cell carcinoma cells is 0.5-4 mu M.

Description

Application of vortioxetine and its salts in the preparation of antitumor drugs

technical field

The invention belongs to the field of medicine, in particular to the application of vortioxetine hydrogen and its salts in the preparation of antitumor drugs.

Background technique

Treatment of esophageal cancer includes surgery, radiotherapy and chemotherapy, with surgery being the first choice. Preoperative systemic chemotherapy or concurrent chemoradiotherapy can significantly reduce tumor stage, tumor activity, reduce tumor volume, and facilitate surgical resection. It can not only improve the resection rate, but also eliminate potential micrometastases in other parts, which can effectively improve the local control rate. Combination of radiotherapy and chemotherapy can not only improve the sensitivity of tumor tissue to radiotherapy, but also improve the efficacy of chemotherapy drugs. For people with advanced cancer whose tumor has spread beyond the esophagus, chemotherapy alone may be used to help relieve the signs and symptoms caused by the tumor.

Drug reuse, also known as new use of old drugs, refers to the use of existing drugs to treat some new diseases and to find new indications for existing drugs. It has the characteristics of high efficiency and low cost, and can also greatly reduce the cost of treatment. Typical new uses of traditional medicines are aspirin and metformin. Aspirin was originally used as an antipyretic, analgesic, and antirheumatic. However, with the deepening of research, it has also been found that it can reduce blood viscosity, can be used to prevent and treat myocardial infarction, and has a role in colorectal cancer, liver cancer, tumors, infectious diseases, nervous system diseases and many other aspects. Metformin is the most commonly used drug for the treatment of type 2 diabetes. In recent years, metformin has been found to have potential effects such as anti-tumor, anti-aging, cardiovascular protection, neuroprotection or selective treatment of polycystic ovary syndrome. The research and development cost of new anti-cancer drugs is high. It usually takes 10-15 years to develop a new drug. However, the global tumor burden is getting heavier and heavier. It is increasingly difficult to find new and affordable anti-cancer drugs through the new use of old drugs. more important.

Antidepressant drugs have a long clinical application time and good safety, and can be used for drug reuse. In addition to antidepressant effects, antidepressant drugs have been reported to have effective anticancer effects on a variety of malignant tumors. For example, fluoxetine can significantly reduce the incidence of pituitary adenomas in rats and fibroadenomas in female rats, and has antioxidant effects, inhibiting skin carcinogenesis in mice; citalopram has anti-tumor effects on colon tumors in mice and rats Effects; Clomipramine can enhance the cytotoxicity of antitumor drugs to drug-resistant tumors; Clogiline can inhibit the growth of prostate cancer in mice through its anti-proliferative effect. Vortioxetine hydrobromide is a new clinical antidepressant. Its main pharmacological action is to antagonize 5-HT ₃ , 5-HT ₇ , 5-HT _1D receptors, partially stimulate 5-HT _1B receptors, and stimulate 5-HT 1B receptors. As a 5-HT _1A receptor agent and inhibition of 5-HT transporter, vortioxetine hydrobromide has a positive effect in the treatment of acute depression, and can effectively prevent the recurrence of depression in maintenance treatment. But there are no studies between vortioxetine hydrobromide and tumors. The incidence of depression in cancer patients is high. If vortioxetine hydrobromide has antidepressant and anticancer effects at the same time, it will have good clinical application value.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide an application of vortioxetine and its salts in the preparation of antitumor drugs, in particular to the application of vortioxetine hydrobromide in the preparation of drugs for treating esophageal cancer.

Based on the above purpose, the present invention adopts the following technical scheme: a FDA-approved medicinal chemical named 1-[2-(2,4-dimethylphenylthio)phenyl]piperazine hydrobromide (Chinese name: wo Tioxetine hydrobromide, English name: VortioxetineHydrobromide, CAS number: 960203-27-4, molecular formula: C ₁₈ H ₂₃ BrN ₂ S , molecular weight: 379.358, trade name: Xindayue, vortioxetine hydrobromide )

The tumors are all mammalian tumors.

The antitumor drugs include vortioxetine, pharmaceutically acceptable salts, esters of vortioxetine or their combinations or combinations with other compounds and drugs.

"Anti-tumor drugs" can be used to prevent tumor occurrence, treat tumors, and prevent tumor recurrence.

Further, vortioxetine hydrobromide can inhibit the proliferation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450 at a concentration of 0.5-4 μM.

When vortioxetine hydrobromide is used in esophageal cancer human tumor xenograft model, when it is below 12mg/kg/day, it has less toxic and side effects on animals, and it is between 1.5mg/kg/day and 12mg/kg/day. Can significantly inhibit tumor growth.

Wherein, for vortioxetine, pharmaceutically acceptable salts, esters or combinations thereof, the appropriate daily dose range for humans is 0.1-2 mg/kg/day, and animals are the corresponding converted doses.

In summary, when used to treat tumor patients, vortioxetine hydrobromide or a pharmaceutically acceptable salt thereof is present at a concentration of from about 0.5% to about 90% by weight of the total composition, ie, in an amount sufficient to provide the desired unit dose quantity exists.

Vortioxetine hydrobromide has been used clinically as an antidepressant drug, which does not require clinical safety assessment and has a good application prospect.

The present invention discovers for the first time the inhibitory effect of vortioxetine hydrobromide on tumor cells, especially the inhibitory effect on esophageal squamous cell carcinoma cells. The results showed that vortioxetine hydrobromide could inhibit the proliferation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450. Thus, it is shown that vortioxetine hydrobromide has a significant inhibitory effect on esophageal squamous cell carcinoma cells. Therefore, vortioxetine hydrobromide provides a reference for the preparation of antitumor drugs, and the prevention, treatment and recurrence of tumors.

Description of drawings

Fig. 1 is the chemical structure of vortioxetine hydrobromide;

Figure 2 shows the cytotoxic effect of vortioxetine hydrobromide on esophageal squamous cell carcinoma cells KYSE150 and KYSE450 in the concentration range of 1-50 μM;

Figure 3 shows the inhibitory effect of vortioxetine hydrobromide on the proliferation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450; * p < 0.05, ** p < 0.01, *** p <0.001;

Figure 4 shows that vortioxetine hydrobromide inhibits the formation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450 clones. The figure shows the statistical results of the number of clones in the dosing and control groups; * p < 0.05, ** p < 0.01, ** * p <0.001;

Figure 5 shows the changes in tumor tissue volume and weight in mice treated with vortioxetine hydrobromide. Among them, vortioxetine hydrobromide can significantly inhibit the growth of tumors in SCID esophageal squamous cell carcinoma PDX model mice at a normal oral dose in adults (converted to an oral dose of 1.5 mg/kg/d in mice), and No effect on mouse body weight; * p < 0.05, ** p < 0.01, *** p < 0.001.

Detailed ways

The technical solutions of the present invention will be further described in detail below with reference to the embodiments, but the protection scope of the present invention is not limited thereto.

Materials and Methods

1. Materials

1.1 Cells and Reagents

Esophageal squamous cell carcinoma cells KYSE150 and KYSE450 were obtained from the Department of Pathophysiology, School of Basic Medicine, Zhengzhou University, and vortioxetine hydrobromide was purchased from Beijing Bailingwei Technology Co., Ltd. with a purity of 98.5%.

1.2 Instruments and equipment: high-content cell imaging analysis system (GE Healthcare), micropipette (Eppendorf, Germany), ultra-clean workbench (Suzhou Purification Co., Ltd.), cell incubator (ThermoScientific Company, USA), 4 ℃ refrigerator ( Henan Haier Electric Co., Ltd.), analytical balance (Ohaus Instrument Co., Ltd.).

2 methods

2.1 Cytotoxicity test

Esophageal squamous cell carcinoma cells were seeded in a 96-well plate with about 8,000 cells per well for KYSE150 and 12,000 cells per well for KYSE450, and placed in a 37°C, 5% CO ₂ incubator for 12-16 h. Dissolve the drug powder with DMSO, prepare vortioxetine hydrobromide with different concentrations (0mM, 3.125mM, 6.25mM, 12.5mM, 25mM, 50mM, 75mM, 100mM), and add 0.5μL of each Vortioxetine hydrobromide in 500 μL of cell culture medium (5 replicate wells for each concentration, 100 μL of culture medium was added to each well, KYSE150 cells: 10% FBS/RPMI-1640; KYSE450 cells: 10% FBS/DMEM), vortioxetine hydrobromide The final concentrations of salt in the medium were 0 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 75 μM, 100 μM, and cells were treated for 24 h and 48 h. After taking out, add 100 μL of 4% paraformaldehyde solution to each well of the 96-well plate, fix the cells for 30 min at room temperature, and then add DAPI staining solution (DAPI:PBS according to the ratio of 1:1000) at 37°C, 5% CO ₂ . incubator for 30 min. The number of cells was counted by a high-content cell imaging analysis system, and the statistical results were analyzed. The results are shown in Figure 2.

2.2 Cell proliferation experiment

Esophageal squamous cell carcinoma cells were seeded in a 96-well plate with about 3000 cells per well for KYSE150 and about 5000 cells per well for KYSE450, and placed in a 37°C, 5% CO ₂ incubator for 12-16 h. Dissolve the drug powder with DMSO and prepare it into different concentrations (0mM, 0.5mM, 1mM, 2mM, 4mM) of vortioxetine hydrobromide, and add 0.5μL of each to 500μL of cell culture medium (each concentration Set 5 duplicate wells, add 100 μL of medium to each well, KYSE150 cells: 10% FBS/RPMI-1640; KYSE450 cells: 10% FBS/DMEM), the final concentration of vortioxetine hydrobromide in the medium is Cells were treated with 0 μM, 0.5 μM, 1 μM, 2 μM, and 4 μM for 24, 48, 72, and 96 h. After taking out, add 100 μL of 4% paraformaldehyde solution to each well of the 96-well plate, fix the cells for 30 min at room temperature, and then add DAPI staining solution (DAPI:PBS according to the ratio of 1:1000) at 37°C, 5% CO ₂ . incubator for 30 min. The number of cells was counted by a high-content cell imaging analysis system, and the statistical results were analyzed. The results are shown in Figure 3.

2.3 Cell anchorage-independent growth experiments

The cells (8000 cells per well) were seeded on a 6-well plate, the lower layer of gel was agarose gel with the corresponding concentration of vortioxetine hydrobromide, and the upper layer of gel was agarose with the corresponding concentration of vortioxetine hydrobromide added. Salt and esophageal squamous cell carcinoma cells KYSE150, KYSE450. The solidified agar was then placed in a 37°C, 5% _CO2 incubator for 7-14 days. The high content cell imaging analysis system was used to count the number of cell clones, and the statistical results were analyzed. The results are shown in Figure 4.

2.4 Construction of esophageal squamous cell carcinoma humanized xenograft model (PDX immunodeficiency mouse model)

Female SCID mice aged 6-8 weeks were selected and implanted into the back of esophageal squamous cell carcinoma EG20 tumor tissue after anesthesia (patient, male, 46 years old, 2042083, T2NOMOII, moderately differentiated squamous cell carcinoma, obtained from Henan Cancer Hospital). When the growth state of the mice was normal, the tumor-bearing mice were randomly divided into 3 groups, and the first group was administered with normal saline as the control group. Vortioxetine hydrobromide clinical drug vortioxetine hydrobromide tablets (trade name: Xindayue) were ground into powder and dissolved in normal saline to form the required concentration as the drug group. The second group was the vortioxetine hydrobromide low-dose group (1.5 mg/kg/d), and the third group was the vortioxetine hydrobromide high-dose group (12 mg/kg/d). The groups were given oral administration of vortioxetine hydrobromide every day, weighed twice a week and measured the tumor volume once a week, tumor volume=(long diameter × short diameter × short diameter)/2. When the tumor tissue of the mice in the control group reached 1000 mm ³ , the mice were sacrificed and the tumor tissue was taken out. The data obtained from the experiment were plotted and analyzed by SPSS to calculate whether there was a difference and whether it was statistically significant. The results are shown in Figure 5.

Experimental results

1) Vortioxetine hydrobromide has a cytotoxic effect on esophageal squamous cell carcinoma cells. The results in Figure 2 show that vortioxetine hydrobromide has a cytotoxic effect on esophageal squamous cell carcinoma cells KYSE150 at a concentration range of 1 μM-50 μM. , the cytotoxic effect of KYSE450; for the cell survival rate at different time points when the control group was 100%, it can be seen from Fig. The effect of cancer cell lines KYSE 150 and KYSE 450 was very small, so 4 μM in cells was chosen as the highest dose in subsequent experiments in esophageal cancer cell lines, and at 0-4 μM, vortioxetine hydrobromide had no effect on KYSE 150, KYSE 450 Cell inhibition was time and dose dependent.

2) Tumor cell proliferation curve of vortioxetine hydrobromide at different time points in esophageal squamous cell carcinoma cells (see Figure 3). The results in Figure 3 show that vortioxetine hydrobromide can inhibit the proliferation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450 cells in the concentration range of 0.5-4 μM after culturing for 72 hours, * p < 0.05, ** p < 0.01, *** p < 0.001.

3) Figure 4 shows that vortioxetine hydrobromide inhibits the colony formation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450. Among them, with the increase of drug concentration, the number of clones decreased significantly, and the clones became significantly smaller. Figure 4 shows the statistical results of the number of clones in the dosing and control groups, as well as the clone micrographs of the corresponding groups. Specifically, vortioxetine hydrobromide at a concentration of 0.5 μM to 4 μM can significantly inhibit the colony formation of KYSE450 cells. The colony formation of KYSE150 cells was significantly inhibited at the concentration of 2~4μM. * p < 0.05, ** p < 0.01, *** p < 0.001.

4) Figure 5 shows the changes in the volume and weight of tumor tissue in mice treated with vortioxetine hydrobromide. After the toxicity test, it was found that after oral administration of vortioxetine hydrobromide, the body weight of mice in both high-dose and low-dose groups did not change significantly, indicating that vortioxetine hydrobromide had no obvious toxic effect on mice. . Among them, vortioxetine hydrobromide can significantly inhibit the SCID esophageal squamous cell carcinoma PDX model at the normal oral dose for adults (1.5 mg/kg/d and 12 mg/kg/d in mice). Tumor growth in mice, tumor tissue volume and weight were significantly reduced, thus indicating that vortioxetine hydrobromide can inhibit the proliferation of esophageal squamous cell carcinoma in vivo. * p < 0.05, ** p < 0.01, *** p < 0.001.

To sum up, it can be seen that the present invention has found that vortioxetine hydrobromide has the cytotoxic effect on esophageal squamous cell carcinoma cells KYSE150 and KYSE450, and can inhibit the proliferation of esophageal squamous cell carcinoma cells KYSE150 and KYSE450, and inhibit the esophageal squamous cell carcinoma cells. The formation of KYSE150, KYSE450 clones, and can inhibit the proliferation of esophageal squamous cell carcinoma in vivo.

The above examples are only used to illustrate the preferred embodiments of the present invention, but the present invention is not limited to the above-mentioned embodiments, and any modifications made within the spirit and principles of the present invention within the scope of knowledge possessed by those of ordinary skill in the art , equivalent substitutions and improvements, etc., shall be regarded as the protection scope of this application.

Claims (6)

1. The application of the vortioxetine and the salts thereof in preparing the antitumor drug is characterized in that the antitumor drug is a drug for treating esophageal cancer.

2. The use of claim 1, wherein vortioxetine and its salts are used in the preparation of a medicament for inhibiting esophageal squamous cell proliferation.

3. The use of claim 2, wherein vortioxetine and its salts are capable of inhibiting the proliferation of esophageal squamous cell carcinoma cells and the number and size of colony formations at a concentration of 0.5 μ Μ to 4 μ Μ.

4. The use as claimed in claim 3 wherein the esophageal squamous cancer cells are KYSE150 cells and/or KYSE450 cells.

5. The use of claim 1, wherein vortioxetine and its salts are used for the preparation of a medicament for inhibiting tumor growth in a humanized transplanted tumor model of esophageal cancer.

6. The use of claim 5, wherein vortioxetine and its salts inhibit the growth of esophageal cancer humanized graft tumor model tumors at a concentration of 1.5 mg/kg/day to 12 mg/kg/day.

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