CN107773753B - A kind of medicine containing American cockroach and bleomycin and its application - Google Patents

Abstract

The invention provides a medicine containing Periplaneta americana and bleomycin, and the active components of the medicine include Periplaneta americana and bleomycin. Wherein, the American Periplaneta is the Periplaneta americana extract or the American Periplaneta medicinal powder. The composition of the medicine can be 20,000-50,000 parts of Periplaneta americana extract and 5-25 parts of bleomycin in parts by weight. The components of the medicine can also be 1,000-10,000 parts by weight of Periplaneta americana medicinal powder and 5-25 parts by weight of bleomycin. The present invention also provides the application of the above-mentioned pharmaceutical composition in the preparation of a drug for the treatment of cancer, the drug for the treatment of cancer has obvious curative effect on cancer, and can interfere with the side effects caused by bleomycin in the treatment of cancer, especially The effect of intervening in the formation of pulmonary fibrosis is particularly obvious.

Description

A kind of medicine containing American cockroach and bleomycin and its application

technical field

The invention belongs to the field of medicine, and particularly relates to a medicine containing Periplaneta americana and bleomycin and its application.

Background technique

Cancer, also known as malignant tumor, is a disease caused by a malfunction in the mechanisms that control cell growth and proliferation. In addition to growing out of control, cancer cells can also locally invade surrounding normal tissues and even metastasize to other parts of the body through the body's circulatory system or lymphatic system. Cancer is a general term for a large class of malignant tumors. Cancer cells are characterized by unrestricted and endless proliferation, which consumes a large amount of nutrients in the patient's body. Cancer cells release a variety of toxins, causing a series of symptoms in the human body. Cancer cells can also be transferred to all parts of the body to grow and reproduce. It can lead to weight loss, weakness, anemia, loss of appetite, fever, and severe organ dysfunction.

Bleomycin (BLMs) is a family of glycopeptide antitumor antibiotics composed of a variety of structurally similar active ingredients. In 1966, Japanese scientists Umezawa et al. isolated the natural active ingredient of bleomycin for the first time from Streptomyces verticillus. Bleomycin is widely used in clinical treatment of various cancers such as lung cancer, cervical cancer, vaginal cancer, esophageal cancer, head and neck cancer, and skin squamous cancer, but it has side effects on the kidneys and lungs. Excessive doses can lead to pulmonary fibrosis. . In the process of cancer treatment, the dosage of anticancer drugs is often increased to achieve the purpose of treatment, so that the symptoms of pulmonary fibrosis in patients after the use of bleomycin often appear clinically, and the most common symptom is breathing. Difficulty, after the patient has the side effect of pulmonary fibrosis, it will aggravate the patient's discomfort.

The American cockroach (Periplaneta american) belongs to the class of insects, the family Periplaneta, commonly known as cockroaches. The American scythe was first seen in the "Shen Nong's Materia Medica" as a medicine, and it was used to treat blood-phlegm syndrome, cold and heat, broken accumulation, laryngopharyngeal paralysis, and inner cold without seeds. In the current study, it was found that the ethanol extract of Periplaneta americana had in vitro effects on chronic myelogenous leukemia (K562), myelocytic leukemia (HL60), mouse leukemia (P388D1) cell lines, nasopharyngeal carcinoma (CNE), lung cancer (A549), Oral epithelial cancer (KB) cell line, ovarian cancer (HO8910), cervical cancer (Hela) and prostate cancer (PC3) cell lines, esophageal cancer (Ecal09), gastric adenocarcinoma (BGC823), colon cancer (LS174T) cell lines have growth inhibition.

At present, there is no related report on the combination of American cockroach and bleomycin.

SUMMARY OF THE INVENTION

In view of the above-mentioned technical defects, the present invention provides a medicine containing Periplaneta americana and bleomycin, the medicine has a significant therapeutic effect on cancer, and the medicine is effective in treating pulmonary fibrosis caused by cancer with bleomycin. It has obvious intervention effect and reduces the side effects caused by the use of bleomycin.

The active ingredients of the medicine of the present invention include American cockroach and bleomycin.

Wherein, the American Periplaneta is the Periplaneta americana extract or the American Periplaneta medicinal powder.

Further, the active ingredients of the medicine, in parts by weight, include: 20,000-50,000 parts of Periplaneta americana extract, 5-25 parts of bleomycin; preferably, 20,000 parts of Periplaneta americana extract, bleomycin 10 servings.

The preparation method of described Periplaneta americana extract is:

Weigh Periplaneta americana, pulverize, add 10 to 12 times the weight of the original medicinal material with 70 to 90% ethanol, reflux and extract 3 times at 70 to 95 ° C for 1 to 3 hours, filter, and concentrate the filtrate under reduced pressure to a The specific gravity measured at 60-65°C is 1.10-1.20, add purified water 10-12 times the weight of the concentrate to the concentrate, stir at 70-80°C for 20 minutes, refrigerate and let stand, the upper layer of grease is absorbed by filter paper, and the lower layer is removed. The liquid is filtered, and the filtrate is concentrated under reduced pressure, dried and pulverized to obtain the Periplaneta americana extract.

Further, the active ingredients of the medicine, in parts by weight, include: including: including: 1,000-10,000 parts of Periplaneta americana powder, 5-25 parts of bleomycin; preferably, 3,000 parts of Periplaneta americana powder, bleomycin 10 copies of mycin.

The preparation method of described Periplaneta americana medicinal powder is:

a. Place the adult Periplaneta americana in a vacuum dryer, and dry at 20 to 55°C to a water content of 2 to 5% to obtain the dried adult Periplaneta americana;

b. The dried Periplaneta americana adults obtained in the step a are placed in an ultra-low temperature pulverizer whose temperature is set to -200°C～-50°C and pulverized to 60～300 meshes to obtain Periplaneta americana medicinal powder.

Another object of the present invention is to provide an application of a combination drug of Periplaneta americana and bleomycin in preparing a drug for treating cancer.

Wherein, the cancer includes lung cancer, colon cancer, liver cancer, ovarian cancer, Hodgkin lymphoma, esophageal cancer, hemangioma and other malignant tumors.

Wherein, the drug for treating cancer is a combination drug of Periplaneta americana and bleomycin; the combination drug can also be supplemented with commonly used pharmacy auxiliary materials or auxiliary components.

Wherein, the described Periplaneta americana is Periplaneta americana extract or Periplaneta americana medicinal powder or new rehabilitation liquid.

Wherein, in the medicine for treating cancer, the dosage of Periplaneta americana powder is 1000-10000mg, preferably 3000mg; the dosage of bleomycin is 5-25mg, preferably 10mg.

Wherein, in the medicine for treating cancer, the dosage of Periplaneta americana extract per dosage is 20000-50000 mg, preferably 20000 mg; the dosage of bleomycin per dosage is 5-25 mg, preferably 10 mg.

In the medicine of the present invention, the Periplaneta americana extract or the American Periplaneta medicinal powder and bleomycin are used as active ingredients, and experimental verification shows that the medicine has obvious therapeutic effect on cancer. It is also obtained through experimental verification that the medicament of the present invention can significantly interfere with the formation of pulmonary fibrosis caused by bleomycin, thereby reducing the side effects caused by bleomycin in the treatment of cancer.

The present invention will be described in further detail below by means of specific embodiments, but it is not intended to limit the present invention. According to the above-mentioned content of the present invention, according to common technical knowledge and customary means in the field, without departing from the above-mentioned basic technical idea of the present invention, under the premise, Various other modifications, substitutions or changes may also be made.

Detailed ways

Example 1 Preparation of Periplaneta americana extract

Weigh Periplaneta americana, pulverize, add 80% ethanol 12 times the weight of the original medicinal material, and extract by refluxing for 3 times at 80 ° C, each extraction time is 1 hour, filter, and concentrate the filtrate under reduced pressure to a temperature of 60 ~ 65 ° C. The specific gravity is 1.10 to 1.20, and purified water 12 times the weight of the concentrated solution is added to the concentrated solution, stirred at 80 ° C for 20 minutes, refrigerated and left to stand, the upper layer of grease is absorbed with filter paper, the lower layer of liquid is filtered, and the filtrate is concentrated under reduced pressure, Dry and pulverize to obtain Periplaneta americana extract.

The Periplaneta americana extract can be prepared into liquid preparations, tablets, capsules and the like according to conventional preparation techniques as required.

Example 2 Preparation of Periplaneta americana medicinal powder

Place the adult Periplaneta americana in a vacuum desiccator, and dry it at 45°C to a moisture content of 2% to obtain a dry adult Periplaneta americana; place the dried Periplaneta americana adults in an ultra-low temperature set at -100°C for crushing Pulverize to 200 mesh in a machine to obtain Periplaneta americana medicinal powder.

Example 3

The medicine is a combination medicine of 10 mg of bleomycin and 20 g of the Periplaneta americana extract described in Example 1.

Example 4

The medicine is a combination medicine of 10 mg of bleomycin and 3000 mg of Periplaneta americana medicinal powder described in Example 2.

Example 5

The medicine is a combination medicine of 5 mg of bleomycin and 20 g of the Periplaneta americana extract described in Example 1.

Example 6

The drug is a combination drug of 5 mg of bleomycin and 1000 mg of Periplaneta americana powder described in Example 2.

Example 7

The drug is a combination drug of 25 mg of bleomycin and 50 g of the Periplaneta americana extract described in Example 1.

Example 8

The medicine is a combination medicine of 25 mg of bleomycin and 10 g of Periplaneta americana medicinal powder described in Example 2.

The combination drug of Periplaneta americana and bleomycin described in any one of the above Embodiments 3 to 8 can be added with commonly used pharmacy auxiliary materials or auxiliary components, and made into capsules according to the technical means commonly used in the art , tablets, sustained-release preparations, liquid preparations and other oral preparations.

The application of the medicament of the present invention in the preparation of a medicament for treating cancer will be further described below through experimental examples.

EXPERIMENTAL EXAMPLE Observational study on the tumor inhibition rate and pulmonary fibrosis of the drug of the present invention on colon adenocarcinoma mice

The Periplaneta americana extract described in this experiment is the Periplaneta americana extract described in Example 1, and the described Periplaneta americana medicinal powder is the Periplaneta americana medicinal powder described in Example 2.

1. Materials

1.1 Experimental animals

60 healthy clean-grade BALB/C mice, 6-8 weeks old, weighing 17-25g, were purchased from Chengdu Dashuo Laboratory Animal Co., Ltd.

1.2 Cell lines

Mouse colon adenocarcinoma cell CT26 was a gift from the State Key Laboratory of Biotherapy, Sichuan University.

1.3 Main reagents

Bleomycin hydrochloride for injection (Nippon Chemical Co., Ltd.); superoxide dismutase (SOD), malondialdehyde (MDA), hydroxyproline (HYP) kit (Nanjing Jiancheng Bioengineering Institute).

2. Method

2.1 Cell Culture

The mouse colon adenocarcinoma CT26 cells were routinely cultured, and the cells in the logarithmic growth phase were collected and counted to prepare a 1×106/ml cell suspension for later use.

2.2 Establishment and grouping of mouse colon cancer CT26 cell model

BALB/C mice were taken, and 0.1 ml of cell suspension was subcutaneously inoculated into the groin of each left hind limb, and the status of the mice and tumor growth were observed every day.

2.3 Grouping and administration

The BALB/C mice inoculated with CT26 tumor cells were randomly divided into 6 groups with 10 mice in each group, namely the control group, the bleomycin group, the American cockroach extract group, the American cockroach powder group, and the bleomycin group. +Periplaneta americana extract group, bleomycin + American Periplaneta medicinal powder group. When the volume of CT26 tumor in mice reached visible to the naked eye, administration was started. The administration doses of mice in each group were shown in Table 1, and the administration was administered once a day for 14 days.

Table 1. Dosing mode and dosage of each group

Note: The combination drug group in Table 1 of this experiment uses the combination drugs described in Example 3 and Example 4, respectively, and the dose in Table 1 is the equivalent dose ratio based on the body surface area conversion between humans and animals . 2.4 Tumor volume, tumor weight and tumor inhibition rate

The length and width of the mouse tumor were measured every day, and the weight of the mouse was weighed. The length was the longest part of the mouse tumor, and the width was the shortest part of the mouse tumor. The tumor volume was roughly calculated according to the following formula:

On the 5th day after the drug was stopped, in vivo sampling was carried out, the weight of the mice was weighed, and then the tumor mass was peeled off and weighed, and the tumor inhibition rate was calculated according to the tumor weight, which was calculated according to the following formula:

Tumor inhibition rate=(average tumor weight of control group-average tumor weight of medication group)/average tumor weight of control group×100%. 2.5 Lung coefficient

All mice were sacrificed 5 days after the administration was completed, the body weight was weighed before sacrifice, the femoral artery was bled to death, the trachea was separated and the main bronchus was cut 1.5 cm below the larynx, the whole lung was stripped and taken out, rinsed with ice saline, filtered dry with filter paper, and weighed. Calculate the lung coefficient.

2.6 Pathological examination of lung tissue

The middle lobe of the left lung was taken and fixed in 4% paraformaldehyde solution, routinely embedded in paraffin, sectioned, and the pathological changes of lung tissue and the degree of pulmonary fibrosis (PF) were observed by HE and Massion staining. and grading of pulmonary fibrosis.

2.7 Detection of HPY, SOD, and MDA content in lung tissue

Take the middle lobe of the right lung, rinse it in normal saline, dry the surface water with filter paper, accurately weigh 30-50 mg, and store it in a refrigerator at -80 °C. The activity of SOD and the levels of MDA and HPY in each sample were determined by tissue alkaline hydrolysis and spectrophotometry.

3. Statistical methods

表示。若不符合正态分布，则用非参数检验，若P<0.05，差异有统计学意义。All experimental data were tested for normality using statistical software SPSS17.0. If each group was normally distributed, one-way ANOVA was used to compare differences between groups, and LSD method was used to compare differences between two groups. , the experimental results of each group were used as the mean ± standard deviation

express. If it does not conform to the normal distribution, a nonparametric test is used. If P<0.05, the difference is statistically significant.

4. Results

4.1 Growth of transplanted tumor

n＝10)Table 2 The effects of Periplaneta americana extract, Periplaneta americana medicinal powder and their combined bleomycin on tumor volume and tumor weight of CT26 tumor-bearing mice (

n=10)

Note: Compared with control group (*P<0.05, **P<0.01); compared with bleomycin (▲P<0.05, ▲▲P<0.01).

As can be seen in Table 2: the inhibition rate of the mouse tumor in the combined drug group of 1.5mg/kg bleomycin and 3000mg/kg American cockroach extract reached 80.29%, which was significantly higher than the single use of bleomycin. The inhibition rate of leomycin or Periplaneta americana extract on the tumor of mice; according to the dosage of 1.5mg/kg bleomycin and 450mg/kg Periplaneta americana powder, the inhibition rate of mice tumor reached 82.90 %, significantly higher than the inhibition rate of single use of bleomycin or Periplaneta americana powder on tumor in mice.

4.2 Comparison of lung coefficients of mice in each group

Table 3 Changes in lung coefficient of mice in each group (mg·g-1)

Note: Compared with the control group, (*P<0.05, **P<0.01); compared with the bleomycin group (▲P<0.05, ▲▲P<0.01).

Obtained from Table 3: Compared with the bleomycin group, the lung coefficients of the mice in the combination group of adding the American cockroach extract or the American cockroach powder were significantly lower than those of the mice in the bleomycin group alone. Lung coefficient, the results of this experiment show that the addition of the American cockroach can significantly inhibit the lung injury caused by bleomycin alone.

4.3 Pathological score of lung tissue of mice in each group

Table 4 Grade scores of alveolitis and pulmonary fibrosis in mice in each group

Note: Compared with the control group, (*P<0.05, **P<0.01); compared with bleomycin (▲P<0.05, ▲▲P<0.01).

It can be obtained from the results in Table 4: after the combined administration of bleomycin and the American cockroach of the present invention, the alveolitis and pulmonary fibrosis of mice are obviously improved, indicating that bleomycin and the present invention The combination of the American cockroach can significantly interfere with the side effects of bleomycin, such as alveolitis and pulmonary fibrosis.

4.4 Comparison of SOD activity, MDA and HYP levels in lung tissue of mice in each group

n＝10)Table 5 Comparison of SOD activity, MDA and HYP levels in lung tissue of mice in each group (

n=10)

Note: Compared with the control group, (*P<0.05, **P<0.01); compared with bleomycin (▲P<0.05, ▲▲P<0.01).

It can be obtained from the results in Table 5: compared with the bleomycin group, after bleomycin is used in combination with the Periplaneta americana extract or the Periplaneta americana medicinal powder, SOD (superoxide dismutase) activity can be increased, Reducing the content of MDA (malondialdehyde) and reducing the content of HYP (hydroxyproline) were statistically significant, and the P values were all less than 0.01. This shows that the combination of Periplaneta americana and bleomycin according to the present invention can obviously interfere with this side effect of pulmonary fibrosis caused by bleomycin.

To sum up, the experimental results showed that the inhibition rate of bleomycin and Periplaneta americana in combination was significantly higher than that of bleomycin or Periplaneta americana alone. At the same time, after adding Periplaneta americana, it can significantly interfere with the lung injury and pulmonary fibrosis caused by bleomycin in the treatment of cancer. This shows that the combination of bleomycin and Periplaneta americana has obvious clinical significance in the preparation of cancer drugs, and at the same time solves the problem of related side effects (for example: pulmonary fibrosis) caused by the use of bleomycin.

Claims (3)

1. The combined medicine for treating the colon cancer is characterized in that the active ingredients of the medicine comprise periplaneta americana and bleomycin; wherein the periplaneta americana is periplaneta americana extract or periplaneta americana medicinal powder;

the active ingredients of the medicine are 2000:1 parts by weight of periplaneta americana extract or 300:1 parts by weight of periplaneta americana powder;

the preparation method of the periplaneta americana extract comprises the following steps: weighing periplaneta americana, crushing, adding 70-90% ethanol which is 10-12 times of the weight of the raw medicinal materials, performing reflux extraction for 3 times at 70-95 ℃, performing filtration for 1-3 hours, concentrating the filtrate under reduced pressure until the specific gravity is 1.10-1.20 measured at 60-65 ℃, adding purified water which is 10-12 times of the weight of the concentrated solution into the concentrated solution, stirring for 20 minutes at 70-80 ℃, refrigerating and standing, absorbing the upper layer oil by using filter paper, filtering the lower layer solution, concentrating the filtrate under reduced pressure, drying and crushing to obtain the periplaneta americana extract;

the preparation method of the periplaneta americana medicinal powder comprises the following steps:

a. placing the Periplaneta americana adults in a vacuum dryer, and drying at 20-55 ℃ until the water content is 2-5% to obtain dried Periplaneta americana adults;

b. and (b) placing the dried Periplaneta americana adults obtained in the step a into an ultralow temperature pulverizer set to be-200 ℃ to-50 ℃ to pulverize to 60-300 meshes, and obtaining the Periplaneta americana powder.

2. The use of a combination of periplaneta americana and bleomycin according to claim 1 for the preparation of a medicament for the treatment of colon cancer.

3. The combination according to claim 1, wherein the combination is further added with pharmaceutically acceptable adjuvants or auxiliary components.