CN107773753B - Medicine containing periplaneta americana and bleomycin and application thereof - Google Patents
Medicine containing periplaneta americana and bleomycin and application thereof Download PDFInfo
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- CN107773753B CN107773753B CN201611106195.3A CN201611106195A CN107773753B CN 107773753 B CN107773753 B CN 107773753B CN 201611106195 A CN201611106195 A CN 201611106195A CN 107773753 B CN107773753 B CN 107773753B
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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Abstract
The invention provides a medicine containing periplaneta americana and bleomycin, wherein the active ingredients of the medicine comprise the periplaneta americana and the bleomycin. Wherein the periplaneta americana is periplaneta americana extract or periplaneta americana medicinal powder. The medicine comprises 20000-50000 parts of periplaneta americana extract and 5-25 parts of bleomycin in parts by weight. The medicine can also comprise 1000-10000 parts of periplaneta americana medicinal powder and 5-25 parts of bleomycin in parts by weight. The invention also provides application of the pharmaceutical composition in preparing a medicament for treating cancer, wherein the medicament for treating cancer has obvious curative effect on cancer, can intervene side effect caused by bleomycin treatment on cancer, and particularly has obvious effect on intervening pulmonary fibrosis.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a medicine containing periplaneta americana and bleomycin and application thereof.
Background
Cancer, also known as malignant tumor, is a disease caused by the malfunction of the mechanism controlling cell growth and proliferation. In addition to uncontrolled growth, cancer cells can also locally invade surrounding normal tissues and even migrate to other parts of the body via the systemic circulation or lymphatic system. Cancer is a general term for a large group of malignant tumors. Cancer cells are characterized by unlimited and endless proliferation, so that nutrients in a patient body are greatly consumed, and the cancer cells release various toxins, so that a series of symptoms are produced by the human body; cancer cells can also be transferred to all parts of the body to grow and reproduce, which causes emaciation, weakness, anemia, inappetence, fever, serious organ function impairment and the like.
Bleomycin (BLMs) is a family of glycopeptide antitumor antibiotics consisting of a variety of structurally similar active ingredients. In 1966, Japanese scientist Umezawa et al isolated natural active ingredients of bleomycin from Streptomyces verticillium (Streptomyces verticillius) for the 1 st time. Bleomycin is widely used for clinically treating various cancers such as lung cancer, cervical cancer, vaginal cancer, esophageal cancer, head and neck cancer, skin squamous carcinoma and the like, but has side effects on kidney and lung, and pulmonary fibrosis can be caused by overdose of bleomycin. In the course of cancer treatment, the dosage of anticancer drugs is often increased to achieve the purpose of treatment, so that pulmonary fibrosis symptoms of patients often appear after bleomycin is used in clinic, the most common symptoms are dyspnea, and the patient can aggravate the difficulty of the patients after the side effect of the pulmonary fibrosis appears.
Periplaneta americana (Periplaneta americana) belongs to the class Insecta, the family Blattaceae, and is commonly referred to as a cockroach. American sickle is the first herb in Shen nong Ben Cao Jing as a medicine for treating blood-phlegm syndrome, chills and fever, accumulation, throat and throat impediment, and internal coldness and sterility. In the current research, the ethanol extract of the periplaneta americana has growth inhibition effect on chronic myelogenous leukemia (K562), myelogenous leukemia (HL60), mouse leukemia (P388D1) cell strains, nasopharyngeal Carcinoma (CNE), lung cancer (A549), oral epithelial cancer (KB) cell strains, ovarian cancer (HO8910), cervical cancer (Hela) and prostate cancer (PC3) cell strains, esophageal cancer (Ecal09), gastric adenocarcinoma (BGC823) and colon cancer (LS174T) cell strains in vitro.
At present, no report is found about the drug combination of periplaneta americana and bleomycin.
Disclosure of Invention
Aiming at the technical defects, the invention provides a drug containing periplaneta americana and bleomycin, the drug has obvious treatment effect on cancer, the drug has obvious intervention effect on pulmonary fibrosis caused by cancer treatment by bleomycin, and side effect generated when bleomycin is used is reduced.
The active ingredients of the medicine comprise periplaneta americana and bleomycin.
Wherein the periplaneta americana is periplaneta americana extract or periplaneta americana medicinal powder.
Further, the active ingredients of the medicine comprise, by weight: 20000-50000 parts of periplaneta americana extract and 5-25 parts of bleomycin; preferably 20000 parts of periplaneta americana extract and 10 parts of bleomycin.
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 at 70-95 ℃ for 3 times for 1-3 hours, filtering, 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.
Further, the active ingredients of the medicine comprise, by weight: the method comprises the following steps: the method comprises the following steps: 1000-10000 parts of periplaneta americana medicinal powder and 5-25 parts of bleomycin; preferably 3000 parts of periplaneta americana powder and 10 parts of bleomycin.
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.
The invention also aims to provide application of the periplaneta americana and bleomycin in preparing a medicament for treating cancer.
Wherein the cancer comprises malignant tumors such as lung cancer, colon cancer, liver cancer, ovarian cancer, Hodgkin lymphoma, esophageal cancer, hemangioma and the like.
Wherein the medicament for treating the cancer is a periplaneta americana and bleomycin combined medicament; the combined medicine can be added with auxiliary materials or auxiliary components which are commonly used in pharmacy.
Wherein the periplaneta americana is periplaneta americana extract, periplaneta americana medicinal powder or rehabilitation new liquid.
The amount of the American cockroach medicinal powder used for each time in the medicine for treating the cancer is 1000-10000 mg, preferably 3000 mg; the dosage of the bleomycin for each dosage is 5-25 mg, preferably 10 mg.
Wherein in the medicine for treating cancer, the amount of the periplaneta americana extract used for each dosage is 20000-50000 mg, preferably 20000 mg; the dosage of the bleomycin for each dosage is 5-25 mg, preferably 10 mg.
The medicine provided by the invention takes the periplaneta americana extract or the periplaneta americana powder and bleomycin as active ingredients, and experiments prove that the medicine has an obvious effect of treating cancers. Experiments prove that the medicine can obviously intervene in the formation of pulmonary fibrosis caused by bleomycin, so that the side effect caused by the bleomycin in the treatment of cancer is reduced.
The present invention is described in further detail with reference to the following embodiments, but the present invention is not limited thereto, and various other modifications, substitutions and alterations can be made without departing from the basic technical idea of the present invention based on the above-mentioned contents of the present invention and common technical knowledge and conventional means in the art.
Detailed Description
Example 1 preparation of Periplaneta americana extract
Weighing periplaneta americana, crushing, adding 80% ethanol which is 12 times of the weight of the raw medicinal materials, performing reflux extraction at 80 ℃ for 3 times, wherein the extraction time is 1 hour each time, filtering, concentrating the filtrate under reduced pressure until the specific gravity is 1.10-1.20 at 60-65 ℃, adding purified water which is 12 times of the weight of the concentrated solution into the concentrated solution, stirring for 20 minutes at 80 ℃, refrigerating and standing, absorbing the upper layer of grease by using filter paper, filtering the lower layer of liquid, concentrating the filtrate under reduced pressure, drying and crushing to obtain the periplaneta americana extract.
The periplaneta americana extract can be prepared into liquid preparations, tablets, capsules and the like according to the conventional preparation process.
Example 2 preparation of Periplaneta americana powder
Placing the Periplaneta americana adults in a vacuum dryer, and drying at 45 ℃ until the water content is 2% to obtain dried Periplaneta americana adults; placing the dried Periplaneta americana imago into an ultralow temperature pulverizer with the temperature set to be-100 ℃ to pulverize the Periplaneta americana imago to 200 meshes, and obtaining the Periplaneta americana medicinal powder.
Example 3
The drug is a drug which is combined by 10mg of bleomycin and 20g of the periplaneta americana extract described in example 1.
Example 4
The drug is a drug which is combined by 10mg of bleomycin and 3000mg of the periplaneta americana powder in the embodiment 2.
Example 5
The drug is a drug which is combined by 5mg of bleomycin and 20g of the periplaneta americana extract described in example 1.
Example 6
The drug is a drug which is combined by 5mg of bleomycin and 1000mg of the periplaneta americana powder described in example 2.
Example 7
The medicine is a combination medicine of 25mg of bleomycin and 50g of the periplaneta americana extract described in example 1.
Example 8
The medicine is a combined medicine of 25mg of bleomycin and 10g of periplaneta americana powder described in example 2.
The pharmaceutical composition for combination of periplaneta americana and bleomycin according to any one of embodiments 3 to 8 may be added with pharmaceutically acceptable adjuvants or auxiliary components, and made into oral preparations such as capsules, tablets, sustained release preparations, and liquid preparations by conventional techniques in the art.
The application of the drug of the present invention in preparing a drug for treating cancer will be further described by the following experimental examples.
Experimental example the drug of the present invention has an observation and study on the tumor inhibition rate and pulmonary fibrosis of mice with colonic adenocarcinoma
The periplaneta americana extract in the experiment is the periplaneta americana extract in the embodiment 1, and the periplaneta americana medicinal powder is the periplaneta americana medicinal powder in the embodiment 2.
1. Material
1.1 Experimental animals
Healthy and clean BALB/C mice are 60 mice, 6-8 weeks old and 17-25g in body mass, and are purchased from great laboratory animals Co.
1.2 cell lines
Mouse colon adenocarcinoma cells CT26, a premium laboratory gift of the national biotherapy of Sichuan university.
1.3 Primary reagents
Bleomycin hydrochloride for injection (japan chemicals corporation); superoxide dismutase (SOD), Malondialdehyde (MDA) and Hydroxyproline (HYP) kit (Nanjing institute of bioengineering).
2. Method of producing a composite material
2.1 cell culture
The mouse colon adenocarcinoma CT26 cells are cultured conventionally, and cells in a logarithmic growth phase are collected and counted to prepare a cell suspension of 1 × 106/ml for standby.
2.2 establishment and grouping of mouse colon cancer CT26 cell model
BALB/C mice were inoculated with 0.1ml of cell suspension subcutaneously in the inguinal region of each left hind limb, and the state of the mice and the growth of tumors were observed daily.
2.3 grouping and administration
The BALB/C mice inoculated with CT26 tumor cells were randomly divided into 6 groups, each group containing 10 mice, namely a control group, a bleomycin group, a periplaneta americana extract group, a periplaneta americana powder group, a bleomycin + periplaneta americana extract group and a bleomycin + periplaneta americana powder group. When the tumor volume of mouse CT26 reached visual level, dosing was initiated and the dose administered to each group of mice was as shown in Table 1 once a day for 14 days.
TABLE 1 administration mode and dosage for each group
Note: the combination according to example 3 and example 4 is used in this experimental table 1, and the doses given in table 1 are equivalent dose ratios, converted from the surface area of the human and animal intermediates. 2.4 tumor volume, tumor weight and tumor inhibition rate
Measuring the length and width of the mouse tumor every day, weighing the weight of the mouse, taking the longest part of the mouse tumor as the length, taking the shortest part of the mouse tumor as the width, and roughly calculating the tumor volume according to the following formula:
taking materials in vivo at 5d after stopping administration, weighing the weight of the mice, stripping and weighing tumor masses, calculating the tumor inhibition rate according to the tumor weight, and calculating according to the following formula:
the tumor inhibition ratio was (average tumor weight in control group-average tumor weight in drug-administered group)/average tumor weight in control group × 100%. 2.5 pulmonary factor
All mice were sacrificed 5d after dosing, the weights were weighed before sacrifice, femoral artery was sacrificed by exsanguination, trachea was separated and the main bronchus was cut off 1.5cm below the larynx, the whole lung was taken out by peeling, washed with ice physiological saline, weighed after filter paper was drained, and the lung coefficient was calculated.
2.6 pathological examination of Lung tissue
Fixing the left lung middle leaf in 4% paraformaldehyde solution, embedding in conventional paraffin, slicing, observing lung organization pathological change and Pulmonary Fibrosis (PF) degree by HE and Massion staining, and determining the grade of alveolitis and pulmonary fibrosis by Szapiel and other methods.
2.7 detection of HPY, SOD and MDA content in Lung tissue
Collecting the middle leaf of the right lung, rinsing in normal saline, drying with filter paper, precisely weighing 30-50mg, storing in a refrigerator at-80 deg.C, preparing 10% lung tissue homogenate with normal saline when detecting, and measuring SOD activity and MDA, HPY level in each sample by tissue alkali hydrolysis method and spectrophotometry.
3. Statistical method
All experimental data were tested for normality using statistical software SPSS17.0, if each group is presentAnd (4) performing one-way ANOVA to perform one-way variance analysis to compare differences between groups, and using an LSD method to compare differences between two groups, wherein the experimental results of each group adopt mean values plus or minus standard deviation
And (4) showing. If not, then non-parametric tests are performed, if P is not normal distribution<0.05, the difference was statistically significant.
4. Results
4.1 graft tumor growth
TABLE 2 influence of Periplaneta americana extract, Periplaneta americana powder and bleomycin-associated drug on tumor volume and tumor weight of CT26 tumor-bearing mice (
n＝10)
Note: compared to control group (. P <0.05,. P < 0.01); compared with bleomycin (P <0.05, P < 0.01).
As can be seen in table 2: according to the combined drug group of the bleomycin with the drug concentration of 1.5mg/kg and the periplaneta americana extract with the drug concentration of 3000mg/kg, the inhibition rate of the tumor of the mouse reaches 80.29 percent, which is obviously higher than the inhibition rate of the tumor of the mouse by using the bleomycin or the periplaneta americana extract alone; the inhibition rate of the drug combination group of 1.5mg/kg of bleomycin and 450mg/kg of periplaneta americana powder on the tumor of the mouse reaches 82.90%, which is obviously higher than the inhibition rate of the drug combination group of bleomycin or the periplaneta americana powder on the tumor of the mouse.
4.2 comparison of pulmonary coefficients in groups of mice
TABLE 3 pulmonary factor variation (mg. g-1) for each group of mice
Note: (xvp <0.05,. P <0.01) compared to control; compared with bleomycin group (P <0.05, P < 0.01).
From table 3, we obtain: compared with the bleomycin group, the lung coefficient of mice of the drug combination group added with the periplaneta americana extract or the periplaneta americana powder is obviously lower than that of mice using the bleomycin group alone.
4.3 pathological Scoring of Lung tissue in groups of mice
TABLE 4 groups of mice alveolar inflammation and pulmonary fibrosis grade scores
Note: (xvp <0.05,. P <0.01) compared to control; compared with bleomycin (P <0.05, P < 0.01).
From the results in table 4, it can be obtained: after the bleomycin and the periplaneta americana are jointly used, the pulmonary alveolitis and pulmonary fibrosis of a mouse are obviously improved, and the result shows that the bleomycin and the periplaneta americana can obviously interfere with the side effects of the bleomycin, such as the pulmonary alveolitis and the pulmonary fibrosis, and the like.
4.4 comparison of SOD Activity, MDA and HYP levels in Lung tissues of mice in groups
TABLE 5 comparison of SOD Activity, MDA and HYP levels in Lung tissue of mice in various groups: (
n＝10)
Note: (xvp <0.05,. P <0.01) compared to control; compared with bleomycin (P <0.05, P < 0.01).
From the results in table 5, it can be obtained: compared with bleomycin, after bleomycin is combined with the periplaneta americana extract or the periplaneta americana powder, the activity of SOD (superoxide dismutase) can be improved, the content of MDA (malondialdehyde) is reduced, the content of HYP (hydroxyproline) is reduced, the statistical significance is achieved, and the P value is less than 0.01. Therefore, the periplaneta americana and bleomycin combined drug can obviously interfere the side effect of the pulmonary fibrosis caused by bleomycin after being used.
The experimental results show that the inhibition rate of the bleomycin and the periplaneta americana on the tumor is obviously higher than that of the bleomycin or the periplaneta americana which are singly used. Meanwhile, after the periplaneta americana is added, lung injury and pulmonary fibrosis caused by bleomycin in cancer treatment can be obviously interfered. Therefore, the combined drug of bleomycin and periplaneta americana has obvious clinical significance in preparing the drug for treating cancer, and simultaneously solves the problem of related side effects (such as 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.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109172611B (en) * | 2018-11-09 | 2020-06-16 | 大理大学 | Extraction and purification method of active site for treating pulmonary fibrosis in periplaneta americana |
| CN110664745A (en) * | 2019-11-15 | 2020-01-10 | 重庆希尔安药业有限公司 | Solution preparation and preparation method thereof |
| CN116874553A (en) * | 2023-07-20 | 2023-10-13 | 大理大学 | Preparation method and application of cockroach glycoprotein |
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