A compound preparation and its application in preparing anti-tumor drugs

Abstract

The present invention provides a compound preparation and its application in the preparation of anti-tumor drugs. The compound preparation of the present invention comprises a selenium compound and cisplatin; the selenium compound is one or more of selenocysteine, selenocystine, and sodium selenite acid. The combined use of selenocysteine, selenocysteine, sodium selenite acid and cisplatin can enhance the anti-tumor effect of cisplatin, effectively inhibit tumor cell proliferation and reduce the dosage of cisplatin, while increasing the tumor killing effect, it can also reduce the toxic side effects of cisplatin, and can be used as an active ingredient of anti-tumor drugs for the prevention and treatment of cancer.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a compound preparation and application thereof in the preparation of anti-tumor drugs. Background Art

[0002] In recent years, colon cancer has ranked second among gastrointestinal malignancies due to its high morbidity, mortality and disability rates. Surgery combined with chemotherapy is considered to be an ideal way to treat advanced colon cancer. Cisplatin has become one of the most commonly used chemotherapy drugs for colon cancer due to its broad spectrum, significant efficacy and low price. However, as the duration of cisplatin use increases, cancer cells develop drug resistance, which reduces the therapeutic effect of cisplatin.

[0003] Selenium is one of the essential trace elements for the human body. The bioavailability and biological activity of selenium are closely related to its chemical form. Common forms of selenium include inorganic selenium, such as sodium selenite (Na2SeO3) and sodium selenate (Na2SeO4) and organic selenium, such as selenocysteine ​​(SeCys), selenocystine (SeCys2), methylselenocysteine ​​(MeSeCys) and selenomethionine (SeMet). Studies have shown that selenium has a certain inhibitory effect on many cancers, including lung cancer, bladder cancer, colon cancer, gastric cancer and prostate cancer. At present, the relevant mechanism research work mainly focuses on inorganic selenium (Na2SeO3) alone, and there are few studies on the combination of selenium and chemotherapeutic drugs for cancer. Summary of the invention

[0004] The purpose of the present invention is to provide a compound preparation and its application in the preparation of anti-tumor drugs.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a compound preparation, comprising a selenium compound and cisplatin; the selenium compound is one or more of selenocysteine, selenocystine and sodium selenite.

[0007] Preferably, the weight ratio of the selenium compound to cisplatin is 0.01 to 8:100.

[0008] The invention also provides application of the compound preparation in preparing anti-tumor drugs.

[0009] The present invention also provides application of the compound preparation in preparing a medicine for inhibiting tumor cell proliferation.

[0010] The present invention also provides application of the compound preparation in preparing a medicine for promoting apoptosis of tumor cells.

[0011] Preferably, the concentration of cisplatin used is 50-150 mg / L.

[0012] Preferably, the concentration of the selenium compound is 10 to 8000 μg / L.

[0013] Preferably, the tumor includes one or more of cancer, liver cancer, colon cancer, gastric cancer, esophageal cancer, breast cancer, cervical cancer, and nasopharyngeal cancer.

[0014] By adopting the above technical scheme, the present invention has the following beneficial effects: in the present invention, selenocysteine, selenocystine or sodium selenite is used in combination with cisplatin to significantly reduce the survival rate of human colon cancer cells. Selenocysteine, selenocystine and sodium selenite have the potential to be used in synergistic killing of cancer cells with cisplatin, can be used to prepare anti-tumor drugs, and provide new ideas for the prevention and treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a graph showing the effect of different concentrations of cisplatin on the survival rate of human colon cancer cells;

[0016] Figure 2 The figure shows the effects of different concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocystine on the survival rate of human colon cancer cells;

[0017] Figure 3 The figure shows the effects of different concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocystine combined with cisplatin on the survival rate of human colon cancer cells;

[0018] Figure 4 This graph shows the effects of different concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocystine combined with cisplatin on the cell cycle of human colon cancer cells. DETAILED DESCRIPTION

[0019] The invention provides a compound preparation, comprising a selenium compound and cisplatin; the selenium compound is one or more of selenocysteine, selenocystine and sodium selenite.

[0020] In the present invention, the weight ratio of the selenium compound to cisplatin is 0.01 to 8:100, more preferably 0.1 to 6:100, and even more preferably 2 to 4:100.

[0021] The invention also provides application of the compound preparation in preparing anti-tumor drugs.

[0022] The present invention also provides application of the compound preparation in preparing a medicine for inhibiting tumor cell proliferation.

[0023] The present invention also provides application of the compound preparation in preparing a medicine for promoting apoptosis of tumor cells.

[0024] In the present invention, the tumor includes one or more of cancer, liver cancer, colon cancer, gastric cancer, esophageal cancer, breast cancer, cervical cancer, and nasopharyngeal cancer.

[0025] When inhibiting tumor cell proliferation, the concentration of cisplatin in the culture medium is 50-150 mg / L, more preferably 80-120 mg / L, and more preferably 100 mg / L. The concentrations of selenocysteine, selenocysteine, and sodium selenite are 10-8000 μg / L, more preferably 100-7000 μg / L, more preferably 4000 μg / L, and can also be 1000, 2000, 3000, 5000, or 6000 μg / L.

[0026] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0027] The human colon cancer cells in the examples of the present invention were purchased from Wuhan Pronocell Life Science Co., Ltd.

[0028] Example 1

[0029] (1) Cell culture: Human colon cancer cells were cultured in T25 culture flasks, and the culture flasks were placed in a 37°C incubator containing 5% CO2 for culture and subculture. The culture medium was a DMEM high-glucose medium containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody. When the confluence of human colon cells reached more than 80%, the culture medium in the culture flask was discarded, the cells were washed with PBS, and then quickly digested with 1mL of 0.25% trypsin. After digestion was completed, 2mL of cell culture medium was added to terminate the digestion. The cell suspension was centrifuged at 1000rpm for 3min, the supernatant was discarded, 2mL of cell culture medium was added, and the cells were mixed. The number of living cells was counted using a cell counter and prepared into 1×10 5 Add 100 μL of cell suspension per well to a 96-well plate and continue culturing for 24 hours. After 24 hours, take out the 96-well plate from the incubator and observe whether the cells have attached to the wall.

[0030] (2) Drug treatment: After the cells adhere to the wall, the culture medium in the wells was aspirated and cisplatin solution was added to the wells. The final concentrations of cisplatin solution were 0.5, 50, 100, 200, 300, 400 and 500 mg / L, respectively. The control group added the same volume of culture medium. Six parallel wells were set up for each concentration and cultured for 24 hours. The culture medium in each well was aspirated, and CCK-8 and culture medium were mixed in a volume ratio of 1:9 and added to a 96-well plate. 100 μL was added to each well and incubated at 37°C for 40 minutes. The 96-well plate was set in the microplate reader to oscillate for 15 seconds first, and then the OD value of each well in the plate was measured at a wavelength of 450 nm. The blank group used culture medium instead of cell samples, and the other steps were the same. After the determination was completed, the cell survival rate was calculated according to the following method.

[0031] Cell survival rate (%) = (OD value of experimental group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%.

[0032] The calculation results are as follows Figure 1 As shown. Figure 1 It can be seen that after colon cancer cells were incubated with cisplatin, the survival rate of colon cancer cells decreased, and within the range of 0.5 to 200 mg / L, the higher the cisplatin concentration, the stronger the inhibitory effect on cancer cells.

[0033] Example 2

[0034] The cell culture stage is the same as that in Example 1. After the cells adhere to the wall and the culture medium in the well is sucked out, sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocystine are added to the wells for treatment, and the final concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocysteine ​​are 10, 100, 500, 1000, 2000, 4000, 6000, and 8000 μg / L, respectively. The control group is added with the same volume of culture medium, and 6 replicates are set for each concentration, and the treatment time is 24 hours. After the culture is completed, the operation of Example 1 is repeated and the OD value is measured, and the cell survival rate is calculated. The results are as follows: Figure 2 shown.

[0035] Figure 2 It showed that there was no significant change in the survival rate of colon cancer cells after treatment with different concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocystine.

[0036] Example 3

[0037] The cell culture stage is the same as that in Example 1. After the cells adhere to the wall and the culture medium in the wells is aspirated, sodium selenite and cisplatin, selenomethionine and cisplatin, seleno-methyl-selenocysteine ​​and cisplatin, selenocysteine ​​and cisplatin, selenocysteine ​​and cisplatin, selenocystine and cisplatin are added to the wells for treatment, respectively. The final concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocysteine ​​are 10, 100, 500, 1000, 2000, 4000, 6000, and 8000 μg / L, respectively, and the final concentration of cisplatin is 100 mg / L; cisplatin is added to the control group to a final concentration of 100 mg / L. After 24 hours of treatment, the operation of Example 1 is repeated and the OD value is measured, and the cell survival rate is calculated. The results are shown as follows: Figure 3 shown. Figure 3 The results showed that compared with the control group, the survival rate of colon cancer cells treated with sodium selenite and cisplatin, selenocysteine ​​and cisplatin, and selenocysteine ​​and cisplatin was significantly reduced, while the survival rate of cells treated with selenomethionine and cisplatin, and seleno-methyl-selenocysteine ​​and cisplatin did not change significantly.

[0038] Example 4

[0039] The cell culture stage was the same as that in Example 1. After the cells adhered to the wall and the culture medium in the wells was aspirated, sodium selenite and cisplatin, selenomethionine and cisplatin, seleno-methyl-selenocysteine ​​and cisplatin, selenocysteine ​​and cisplatin, selenocysteine ​​and cisplatin were added to the wells for treatment, respectively, and the final concentrations of sodium selenite, selenomethionine, seleno-methyl-selenocysteine, selenocysteine, and selenocysteine ​​were 2000 μg / L, respectively, and the final concentration of cisplatin was 100 mg / L; cisplatin was added to the control group to a final concentration of 100 mg / L. After 24 hours of treatment, the adherent cells were digested with trypsin according to the conventional method and the cells were collected; washed once with PBS and the supernatant was removed; 0.3 mL of PBS was added to resuspend the cells; the single cell suspension was transferred to 1.2 mL of -20°C anhydrous ethanol, vortexed and shaken vigorously, and placed in a -20°C refrigerator for 1 hour; centrifuged at 300g for 5 minutes, the supernatant was removed, and 1 mL of PBS was added to resuspend the cells, and placed at room temperature for 15 minutes; centrifuged at 300g for 5 minutes, the supernatant was removed, 100 μL of RNaseA was added and the cells were fully suspended, and the cells were bathed at 37°C for 30 minutes; 400 μL of PI solution was added and mixed thoroughly, and incubated at 2-8°C in the dark for 30 minutes; the cell cycle of human colon cancer cells was analyzed by flow cytometry, and the analysis results are shown as follows: Figure 4 shown. Figure 4The results showed that after treating colon cancer cells with sodium selenite and cisplatin, selenocysteine ​​and cisplatin, and selenocysteine ​​and cisplatin, the proportion of colon cancer cells in the G1 phase decreased, and the proportion of colon cancer cells in the S+G2 phase increased, indicating that the cell cycle of colon cancer was blocked in the S phase (DNA synthesis phase) and cell proliferation was inhibited. This indicates that treating colon cancer cells with sodium selenite and cisplatin, selenocysteine ​​and cisplatin, and selenocysteine ​​and cisplatin can inhibit the proliferation of colon cancer cells.

[0040] As can be seen from the above examples, the present invention provides a compound preparation comprising a selenium compound and cisplatin. The compound preparation can effectively inhibit tumor cell proliferation and reduce the dosage of cisplatin.

[0041] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a compound preparation in the preparation of an anti-tumor drug, characterized in that: The compound preparation contains a selenium compound and cisplatin; the selenium compound is selenocysteine; The weight ratio of the selenium compound to cisplatin is 4 to 8:100; The tumor is colon cancer.

2. The use according to claim 1, wherein the compound preparation inhibits tumor cell proliferation and / or promotes tumor cell apoptosis.

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