CN113209272B

CN113209272B - Application of bleomycin and dacarbazine combined medicine in preparation of medicine for treating bile duct cancer

Info

Publication number: CN113209272B
Application number: CN202010577752.XA
Authority: CN
Inventors: 李斌
Original assignee: Alpha Cat Hangzhou Artificial Intelligence Biotechnology Co ltd
Current assignee: Alpha Cat Hangzhou Artificial Intelligence Biotechnology Co ltd
Priority date: 2020-06-23
Filing date: 2020-06-23
Publication date: 2022-02-22
Anticipated expiration: 2040-06-23
Also published as: CN113209272A

Abstract

The invention belongs to the field of medicines, and particularly relates to application of bleomycin and dacarbazine combined medicine in preparation of a medicine for treating bile duct cancer. The invention firstly provides the application of the bleomycin and the dacarbazine combined medicine in preparing the medicine for treating the bile duct cancer, the bleomycin and the dacarbazine 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 two can also save the cost by combining, reduce the economic burden of patients, provide a new way for preventing and treating bile duct cancer, and have wide application prospect in the field of medicine and pharmacology.

Description

Application of bleomycin and dacarbazine combined medicine in preparation of medicine for treating bile duct cancer

The technical field is as follows:

the invention belongs to the field of medicines, and particularly relates to application of bleomycin and dacarbazine combined medicine in preparation of a medicine for treating bile duct cancer.

Background art:

cholangiocarcinoma is a malignant tumor that originates in the epithelial cells of the bile duct, and is second only to liver cancer, a common malignant tumor of the second largest hepatobiliary system. The overall mortality rate for hepatobiliary malignancies is at position 5 in all malignancies. China is a region with high bile duct cancer incidence, and the incidence rate is increasing in recent years. Bile duct cancer is strong in invasiveness, only 30% -40% of patients can be effectively treated by surgical excision during diagnosis, most of patients are in late stage during diagnosis, an effective treatment means is lacked, and prognosis is extremely poor.

Bleomycin (Bleomycin) is a relatively common clinical chemotherapeutic drug. Glycopeptide antibiotic from streptomyces verticillata, a broad-spectrum antitumor drug, and can effectively resist various cancers such as lymphoma, testicular tumor, esophageal cancer and squamous cell carcinoma, lymphoma, testicular tumor, malignant pleural effusion and the like. The bleomycin serving as a clinical antitumor drug has the greatest advantages that the bleomycin has no obvious bone marrow inhibition effect while resisting tumors, and has no inhibition effect on the immune function of an organism. However, like most chemotherapeutic drugs, bleomycin also has significant side effects: prolonged use can lead to pneumonitis-like symptoms and pulmonary fibrosis symptoms, and thus prolonged use of bleomycin in the treatment of tumors is also very dangerous.

Dacarbazine is a novel anti-tumor drug, has broad-spectrum anti-tumor activity and is a first-line drug for treating tumors. The dacarbazine is a triazabenzene derivative, the action mechanism of the dacarbazine is that methyl positive ions are released in vivo to play the role of alkylation, and the dacarbazine is also used as an analogue of a purine nucleotide precursor to inhibit the synthesis of purine nucleotide, and has better anti-tumor metastasis activity.

Reports on the effect of bleomycin in combination with dacarbazine have not yet been made. The invention researches the effect of the combined application of bleomycin and dacarbazine in the treatment of bile duct cancer in a mouse pdx model.

The invention content is as follows:

object of the Invention

The effect of bleomycin and dacarbazine in preparing the medicament for treating the bile duct cancer is provided, so that the anti-tumor curative effect is obviously enhanced, and a basis is provided for new application of old medicaments.

Technical scheme

The invention provides application of bleomycin and dacarbazine combined medicine in preparation of a medicine for treating bile duct cancer.

Therefore, the application of the combination of bleomycin and dacarbazine in preparing the cholangiocarcinoma resisting medicine and the cholangiocarcinoma resisting medicine containing both bleomycin and dacarbazine are all within the protection scope of the invention.

Preferably, the dosage ratio of the bleomycin to the dacarbazine is 2.25 mg/kg: 30 mg/kg.

Advantageous effects

The invention discloses the discovery of the combined medication of bleomycin and dacarbazine in the aspect of inhibiting bile duct cancer tumors for the first time, can realize the synergistic effect, realize the obvious improvement of the anti-tumor effect, and reduce the dosage of the bleomycin and the dacarbazine through the combined medication, thereby reducing the toxic and side effects, and having obvious significance for the application of the combined medication of the bleomycin and the dacarbazine in the aspect of inhibiting bile duct cancer.

Description of the drawings:

FIG. 1 is a graph showing the volume increase of transplanted tumors in experimental mice.

FIG. 2 is the results of HE staining of tumor histopathology following administration of different drugs.

The specific implementation mode is as follows:

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.

Example 1 mouse pdx model establishment

First, experimental material

Matrigel Matrix Matrigel (Corning, USA), RPMI 1640 medium (Biological Industries, Israel), bile duct carcinoma human tumor tissue.

Second, tumor tissue inoculation

1. Thawing the matrix gel, uniformly mixing with 1640 culture medium according to a ratio of 1:1, diluting, and placing on ice for later use.

2. Human tumors grown subcutaneously in nude mice to a sufficient volume were dissected off and placed in petri dishes on ice and poured with appropriate amount of pre-cooled saline.

3. The necrotic parts, blood vessels, the envelope on the surface of the tumor body and the like on the tumor tissue are carefully removed.

4. Cutting the tumor tissue into 1 mm³And mixing the small blocks with the diluted matrigel uniformly for later use.

5. The small tumor mixed with the matrigel is implanted under the skin of the right flank of the nude mouse by a trocar.

6. After 2-4 weeks, the tumor mass stably grows under the skin of the nude mice, the nude mice with proper tumor body size are picked, randomly grouped, and the drug administration is started according to the requirement.

Example 2 mouse graft volume Change

First, experiment grouping

1. Blank group: no drug was administered.

2. Bleomycin group: bleomycin 2.25mg/kg was administered by intraperitoneal injection once every two days.

3. Dacarbazine group: dacarbazine was administered at 30 mg/kg intravenously once every two days.

4. A combination of drugs: the administration of bleomycin by intraperitoneal injection is 2.25mg/kg once every two days; the dacarbazine was administered intravenously at 30 mg/kg once every two days.

Second, result analysis

Tumor size was measured before the start of dosing, and every 4 days thereafter until the end of the experiment. The mice were sacrificed the next day after the dosing was completed by vertebration, and the subcutaneous tumor tissue was dissected and photographed.

TABLE 1 volume Change of transplanted tumors in groups of mice

Group of	Day 0 (mm)³）	Day 4 (mm)³）	Day 8 (mm)³）	Day 12 (mm)³）
					Blank group	67.73	101.23	192.22	248.66
Bleomycin group	60.57	67.81	104.58	154.48
					Dacarbazine group	69.84	75.53	155.34	220.32
Combination drug group	92.15	72.01	96.59	137.81

According to the results shown in fig. 1, compared with the other three groups, the combined group of bleomycin and dacarbazine has the advantages that the volume of the transplanted tumor is obviously smaller, and the volume increase rate is obviously smaller than that of other experimental groups.

Example 3 tumor pathology HE staining

One, HE staining

Fixing the dissected subcutaneous tumor tissue with a fixing solution, and preparing a paraffin section. Dewaxing for 3 minutes according to xylene I, II and III respectively, dewaxing for 2 minutes according to absolute ethyl alcohol I, II, 1 minute according to 95% ethyl alcohol, 90% ethyl alcohol and 80% ethyl alcohol respectively, staining with hematoxylin for 10 minutes, differentiating for 15 seconds according to 1% hydrochloric acid alcohol after water washing, soaking in water for 10 minutes, staining with eosin for 3 minutes, 10 seconds according to 80% ethyl alcohol and 90% ethyl alcohol respectively, 1-2 minutes according to 95% ethyl alcohol, 3 minutes according to absolute ethyl alcohol I, II and III respectively, 3 minutes according to xylene I, II and III respectively, finally sealing with neutral gum, and taking a picture under a microscope.

Second, result analysis

According to the staining results shown in FIG. 2, the blank group, bleomycin group and dacarbazine group had significantly deeper cell nuclear staining, large nuclear-to-cytoplasmic ratio, irregular shape and dense arrangement of tumor cells; the combined medicine group has shallow nuclear staining, loose tumor cell arrangement and more necrotic areas. Therefore, the effect of the combined application of bleomycin and dacarbazine on inhibiting the cancer cells of the bile duct is obvious.

Claims

1. The application of the combined medication of bleomycin and dacarbazine in the preparation of the medicine for treating bile duct cancer is that the dosage proportion of the bleomycin to the dacarbazine is 2.25 mg/kg: 30 mg/kg.