CN112823794A

CN112823794A - Application of metronidazole and taxol in preparation of lung cancer treatment drug

Info

Publication number: CN112823794A
Application number: CN201911140550.2A
Authority: CN
Inventors: 李斌; 国栋
Original assignee: Alpha Cat Hangzhou Artificial Intelligence Biotechnology Co ltd
Current assignee: Alpha Cat Hangzhou Artificial Intelligence Biotechnology Co ltd
Priority date: 2019-11-20
Filing date: 2019-11-20
Publication date: 2021-05-21

Abstract

The invention belongs to the field of medicines, and particularly relates to application of metronidazole and taxol in preparation of a medicine for treating lung cancer. The invention firstly proposes the application of the metronidazole and the taxol in preparing the lung cancer treatment drug, the metronidazole and the taxol have obvious synergistic effect, the curative effect is effectively improved, the curative effect is more obvious compared with that of a single component, and the killing property to tumor cells is improved; effectively reduces the dosage, thereby reducing the toxic and side effects. The combined use of the two can also save the cost, reduce the economic burden of patients, provide a new way for preventing and treating lung cancer, and have wide application prospect in the field of medicine and pharmacology.

Description

Application of metronidazole and taxol in preparation of lung cancer treatment drug

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of metronidazole and taxol in preparation of a medicine for treating lung cancer.

Background

Lung cancer is short for "primary bronchogenic lung cancer", a malignant tumor that originates in respiratory epithelial cells (bronchi, bronchioles and alveoli). According to the pathological changes of tissues, the cancer can be divided into small cell cancer and non-small cell cancer. High incidence rate, high death rate, high growth rate and low cure rate of lung cancer. Surgery is the most effective treatment for patients with early stage lung cancer, but most patients are already advanced at the time of diagnosis. Chemotherapy is the primary treatment for metastatic lung cancer, but with its attendant serious side effects and drug resistance. Even today, the molecular targeted therapy for lung cancer is changing day by day, the overall 5-year survival rate for lung cancer is still less than 20%.

Paclitaxel is a complex tetracyclic diterpenoid compound, which was first extracted from the bark of Taxus brevifolia by Wani et al in 1971, and the structure of the taxus brevifolia was determined and found to have a wide range of anti-malignant tumor effects. At present, paclitaxel is widely used for treating lung cancer, breast cancer, ovarian cancer, partial head and neck tumors and digestive tract malignant tumors in clinic, but still produces some toxic and side effects, such as: alopecia, bone marrow transplantation, facial flushing, muscle and joint soreness, etc.

Metronidazole is an anti-anaerobe and anti-trichomonad drug collected in Chinese pharmacopoeia, has good activity on gram-negative anaerobes including bacteroides fragilis and clostridium, and also has activity on gram-positive anaerobes including clostridium and sensitive strains of fungi. It is mainly used for treating or preventing systemic or topical infection caused by above anaerobic bacteria, such as anaerobic bacteria infection of abdominal cavity, digestive tract, female reproductive system, lower respiratory tract, skin and soft tissue, bone and joint, etc., and also has therapeutic effect on septicemia, endocarditis, meningeal infection and colitis caused by antibiotic.

Reports on the effect of metronidazole in combination with paclitaxel have not yet been made. The invention researches the effect of metronidazole in sensitizing paclitaxel to treat lung cancer in a human lung cancer model and a mouse.

Disclosure of Invention

Object of the Invention

The application of metronidazole sensitization taxol in preparing the medicine for treating lung cancer is provided, the curative effect of the compound is enhanced, and a basis is provided for new application of old medicines.

Technical scheme

The invention provides application of metronidazole and taxol in preparing a medicine for treating lung cancer.

Therefore, the application of the metronidazole and the paclitaxel in the aspect of preparing the anti-lung cancer medicament and the anti-lung cancer medicament containing both the metronidazole and the paclitaxel are within the protection scope of the invention.

Preferably, the lung cancer is human lung cancer cell a 549.

Preferably, the dosage ratio of metronidazole to paclitaxel is 100-200 mug/ml: 140 to 280 mu g/ml.

Particularly preferably, the dosage ratio of the metronidazole and the paclitaxel is 200 mug/ml: 280. mu.g/ml.

Advantageous effects

The invention discloses the discovery of metronidazole on improving the sensitivity of lung cancer cells to paclitaxel for the first time, can realize the synergistic effect, realize the obvious improvement of the anti-tumor effect, and reduce the dosage of paclitaxel by combined medication so as to reduce the toxic and side effect, thereby having obvious significance on the application of metronidazole and paclitaxel combined medication to resisting lung cancer.

Drawings

FIG. 1 is a graph showing the results of the inhibition analysis of lung cancer cells treated with metronidazole alone by the MTT method.

FIG. 2 is a graph showing the results of the MTT method for detecting the inhibition rate of paclitaxel alone or metronidazole and paclitaxel combined after treating lung cancer cell A549.

FIG. 3 is a morphological comparison of transplanted tumors in groups of nude mice after administration of different drugs.

Detailed Description

The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

The kit materials used in the following examples are all commercially available unless otherwise specified.

Example 1 MTT method for detecting sensitivity of Lung cancer cells to Metronidazole

1. Experimental Material

(1) Medicine preparation: and (4) metronidazole. The chemical structural formula of metronidazole is shown as follows:

(2) lung cancer cell: human lung cancer cell a 549.

(3) Commercial MTT kit.

2. Experiment grouping

(1) Control group: the blank control, i.e., lung cancer cells, were not subjected to any drug treatment.

(2) Experimental groups: lung cancer cells were treated with different concentrations of metronidazole.

3. MTT method for detecting sensitivity of tumor cells to metronidazole

(1) The method comprises the steps of inoculating 5000-9000 cells of human lung cancer cells A549 per well to a 96-well plate, and respectively adding different concentrations of metronidazole PBS (phosphate buffer solution) after the cells adhere to the wall. After 24 hours of incubation, 10. mu.l of 5mg/ml MTT was added to each well for another 4 hours, and then 100. mu.l of DMSO was added to each well after discarding the culture solution, and the absorbance of each well was read at 490nm using a microplate reader. Statistical analysis was performed on the data.

(2) The analysis result is shown in figure 1, which indicates that metronidazole has no obvious inhibition effect on lung cancer cells.

Example 2 Effect of paclitaxel alone or combination of Metronidazole and paclitaxel on the inhibition Rate of Lung cancer cells

1. Experimental Material

(1) Medicine preparation: metronidazole and taxol.

(2) Lung cancer cell: human lung cancer cell a 549.

(3) Commercial MTT kit.

2. Experiment grouping

(1) Control group: lung cancer cells were treated with paclitaxel only.

(2) Experimental groups: different concentrations of metronidazole and paclitaxel were used in combination to treat lung cancer cells.

3. MTT method for detecting sensitivity of lung cancer cells to metronidazole and paclitaxel combined drug

(1) The method comprises the steps of inoculating 5000-9000 cells of human lung cancer cells A549 per well to a 96-well plate, and respectively adding PBS solutions with different proportions of metronidazole and paclitaxel after the cells adhere to the wall. After 24 hours of incubation, 10. mu.l of 5mg/ml MTT was added to each well for another 4 hours, and then 100. mu.l of DMSO was added to each well after discarding the culture solution, and the absorbance of each well was read at 490nm using a microplate reader. Statistical analysis was performed on the data.

(2) The analysis result is shown in fig. 2, compared with the single drug, the metronidazole is added on the basis of the use of the paclitaxel, the inhibition effect of the metronidazole on the tumor is weak, the inhibition rate of the combined drug on the tumor is far greater than the sum of the tumor inhibition rates of the paclitaxel and the metronidazole, and the combination of the metronidazole and the paclitaxel obviously improves the treatment effect of the lung cancer and realizes the synergistic treatment effect.

Example 3 mouse experiments

1. Experimental Material

(1) Medicine preparation: metronidazole and taxol.

(2) The weight of the nude mice is 18-20 g/mouse, and the weight of the nude mice is half of that of the nude mice in 4-6 weeks.

2. Experiment grouping

(1) Control group: no treatment was performed.

(2) Metronidazole group: metronidazole 180mg/kg is given, and the stomach is drenched for administration once a day.

(3) A paclitaxel group: paclitaxel 80mg/kg was administered by intragastric administration once a day.

(4) Metronidazole and paclitaxel combination: the dosage and administration method of metronidazole and paclitaxel are the same as above two groups.

3. The treatment with drug administration allows observation of the weight and volume changes of the transplanted tumor in mice.

(1) A549 cells are adherently grown in RPMI-1640 complete medium containing 10% fetal calf serum at 37 ℃ and 5% CO₂And performing conventional culture in a saturated humidity incubator, and changing the culture solution for passage once in 2-3 days. Collecting and concentrating to 2 × 10⁶Injecting 0.2ml of the cell suspension into the shoulder and back subcutaneous parts of the nude mice to form subcutaneous bulges with the diameter of about 3mm, and randomly dividing the nude mice into 4 groups, namely a control group and metronidazole when observing for one week till the length diameter of a tumor body is 8-10 mmAzole group, paclitaxel group, metronidazole and paclitaxel combination group. The treatment was carried out according to the dose of each group, and the survival state of the nude mice was observed.

(2) The nude mice were sacrificed 5 weeks after cell injection, and the transplanted tumor was isolated, and it can be seen from fig. 3 that the transplanted tumor was round or oval, uneven in surface, irregular, red in color and neutral in texture. Compared with the control group, the metronidazole group and the paclitaxel group, the metronidazole and paclitaxel combined group has obviously smaller volume of the transplanted tumor, and the growth speed of the volume of the transplanted tumor of the combined group is obviously lower than that of other groups.

(3) The isolated transplanted tumors were weighed, the tumor weight and tumor inhibition rate of each group are shown in table 1, and the volume of the transplanted tumors was measured, and the tumor volume and tumor inhibition rate of each group are shown in table 2. As can be seen from tables 1 and 2, the metronidazole and paclitaxel combination significantly reduced the weight and volume of the transplanted tumor as compared to the other three groups, and it can be seen from the above that the combination of metronidazole and paclitaxel had a significant effect on inhibiting lung cancer cells.

TABLE 1 weight and tumor inhibition rate of nude mice transplanted tumor of each group

Group of	Weight of transplanted tumor (g)	Tumor inhibition Rate (%)
			Control group	0.831±0.049
Metronidazole group	0.746±0.024	10.2
			Paclitaxel group	0.552±0.016	33.6
Metronidazole and taxol	0.329±0.033	60.4

TABLE 2 transplant tumor volume and tumor inhibition rate of each group of nude mice

Group of	Volume of transplanted tumor (mm)³）	Tumor inhibition Rate (%)
			Control group	1487.330±0.088
Metronidazole group	1301.580±0.025	12.5
			Paclitaxel group	1045.397±0.014	29.7
Metronidazole and taxol	743.579±0.044	50.0

Claims

1. Application of metronidazole and taxol in preparing medicine for treating lung cancer is disclosed.

2. Use according to claim 1, characterized in that: the dosage ratio of metronidazole to paclitaxel is 100-200 mug/ml: 140 to 280 mu g/ml.

3. Use according to claim 1, characterized in that: the dosage ratio of metronidazole to paclitaxel is 200 mug/ml: 280. mu.g/ml.

4. A preparation for treating lung cancer is characterized in that: the composition consists of the composition as claimed in claims 1 to 3 and pharmaceutically acceptable auxiliary materials.

5. The formulation of claim 4, wherein the formulation is for treating lung cancer: the preparation is in the form of injection, tablet, granule or capsule.