CN108126201B - Application of Enterobacterial Antibiotics in Preparation of Drugs for Prevention and Treatment of Complications of Chemotherapy - Google Patents

Abstract

The invention discloses the application of intestinal bacteria antibiotics in the preparation of drugs for preventing and treating complications of chemotherapy. Because no intestinal bacteria enter the blood circulation to grow and proliferate, thereby reducing the release of a large number of toxins and inflammatory factors, antagonizing non-specific inflammatory responses, inhibiting and blocking the damage of pathogenic factors to circulating tissue cells, eliminating local inflammatory responses, Thereby reducing inflammation as well as the side effects of chemotherapy drugs, which has an unexpected preventive effect on slowing damage. The present invention inhibits the release of endotoxin and inflammatory factors in the damaged intestinal tissue by microbial bacteria by administering a preparation containing antibiotics, thereby activating systemic tissue damage; at the same time, the antibiotics used in the present invention are small-molecule drugs, which are easy to obtain and low in price. Stable in nature, easy to store and transport.

Description

Application of Enterobacterial Antibiotics in Preparation of Drugs for Prevention and Treatment of Complications of Chemotherapy

technical field

The invention relates to the application of an intestinal bacteria antibiotic in preparing a drug for preventing and treating chemotherapy complications, in particular to the application of an intestinal gram-negative bacteria antibiotic in preparing a drug for preventing and treating systemic inflammatory response syndrome caused by chemotherapy drugs , belonging to the field of biomedical technology.

Background technique

Malignant tumors are one of the common diseases that seriously threaten human health. At present, the research on tumor has made great progress. With the continuous introduction of anti-tumor drugs and the application of various combined chemotherapy, the life cycle of patients with malignant tumors has been extended. However, various drugs and therapies have limitations, and it is difficult to To achieve the desired effect, and prone to drug resistance and side effects. Among them, myelosuppression and gastrointestinal reactions are the most common. This is mainly because the digestive tract mucosa is one of the early damaged tissues. Therefore, maintaining the function and structural integrity of the intestinal mucosa has also become a research hotspot for preventing and treating side effects of chemotherapy.

An important and common complication of antitumor therapy, almost all chemotherapeutic drugs have the potential to cause emesis, and chemotherapeutic drugs can easily cause stomatitis, oral ulcers, and esophagitis. The most common drugs that cause mucositis are methotrexate, antibiotics and fluorouracil. Treatment is mainly symptomatic. For example, local pain relief can be used for oral ulcer pain. Chemotherapy drugs also often cause diarrhea. For obvious diarrhea, intestinal mucosal protective agents such as Yimengtai and Smecta can be given; if necessary, temporary fasting and antibiotics are used to prevent and treat intestinal infection, so as to facilitate the return of intestinal mucosa to normal.

Most intestinal bacteria are gram-negative bacteria that produce endotoxins and cause disease by endotoxins. Common gram-negative bacteria include Shigella, Typhi, Escherichia coli, Proteus, Pseudomonas aeruginosa, Bacillus pertussis, Vibrio cholera, and Meningococcus. The main antibiotics against Gram-negative bacteria include aminoglycosides, rifamycins, and polymyxins, etc., and their mechanisms of action are also different. For example, aminoglycosides can bind to bacterial ribosomes, thereby interfering with bacterial proteins. Synthetic process, with strong antibacterial activity against gram-negative bacilli.

At present, the commonly used antibiotics for enteric Gram-negative bacteria include streptomycin, gentamicin, kanamycin, amikacin, tobramycin, etc. Systemic inflammatory response syndrome (SIRS) is a systemic inflammatory response mainly caused by bacterial infection by releasing endotoxin in the blood circulation and activating inflammatory factors.

SUMMARY OF THE INVENTION

In view of the problems existing in the above-mentioned prior art, the present invention provides an application of an intestinal bacteria antibiotic in the preparation of a drug for preventing and treating complications of chemotherapy, which can effectively prevent and treat systemic inflammatory response syndrome caused by chemotherapy.

In order to achieve the above purpose, the application of the intestinal bacteria antibiotics used in the present invention in the preparation of medicines for preventing and treating chemotherapy complications.

As an improvement, the intestinal bacteria antibiotic is specifically an intestinal gram-negative bacteria antibiotic.

As an improvement, the antibiotic for enteric gram-negative bacteria adopts any one or a mixture of gentamicin, levofloxacin, ampicillin and amikamycin.

As an improvement, the chemotherapy complication is specifically systemic inflammatory response syndrome caused by chemotherapy drugs.

As an improvement, the chemotherapeutic drug is any one or a mixture of cisplatin, pentafluorouracil, camptothecin, paclitaxel, Herceptin, erlotinib, IL-12, CART, and PD-1.

As an improvement, the medicine for preventing and treating chemotherapy complications is prepared by using intestinal bacteria antibiotics as active ingredients and adding a water-soluble organic carrier.

As an improvement, the water-soluble organic carrier adopts any one of dextrin, corn oil or dimethyl sulfoxide.

As an improvement, the medicament is an oral formulation or an injectable formulation.

As an improvement, the administration dose of the intestinal bacteria antibiotic is 0.05g/kg-10g/kg, which is administered daily.

As a further improvement, the administration dose of the intestinal bacteria antibiotic is 0.5g/kg-10g/kg, which is administered daily.

The principle of the invention is as follows: the intestinal gram-negative bacteria antibiotics can remove the main gram-negative bacteria in the intestinal tract, and when the chemotherapeutic drugs cause intestinal mucosal damage, because no intestinal bacteria enter the blood circulation to grow and proliferate , thereby reducing the release of a large number of toxins and inflammatory factors, antagonizing non-specific inflammatory responses, inhibiting and blocking the damage of pathogenic factors to cells in circulating tissues, eliminating local inflammatory responses, thereby reducing inflammatory responses and the side effects of chemotherapy drugs. Slowing damage has unexpected preventive effects.

Compared with the prior art, the present invention inhibits microbial bacteria from releasing endotoxin and inflammatory factors in damaged intestinal tissue by administering a preparation containing antibiotics, thereby activating systemic tissue damage; at the same time, the antibiotics used in the present invention are small-molecule drugs , easy to obtain, low price, stable in nature, convenient for storage and transportation, and has very important clinical application value in the prevention and treatment of tissue damage caused by chemotherapy drugs.

Description of drawings

Fig. 1 is the content of TNF-α in the serum of chemotherapeutic animals in the embodiment of the present invention;

FIG. 2 is a schematic diagram of intestinal mucosal injury in chemotherapy animals in the embodiment of the present invention.

Detailed ways

In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below through the accompanying drawings and embodiments. However, it should be understood that the specific embodiments described herein are only used to explain the present invention, and not to limit the scope of the present invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field of the present invention, and the terms used herein in the description of the present invention are only for describing specific implementations The examples are not intended to limit the invention.

The application of the intestinal bacteria antibiotics provided by the invention in the preparation of medicines for preventing and treating complications of chemotherapy.

As an improvement, the intestinal bacteria antibiotic is specifically an intestinal gram-negative bacteria antibiotic.

As a further improvement, the enteric Gram-negative bacteria antibiotic adopts any one or a mixture of gentamicin, levofloxacin, ampicillin and amikamycin. Pretreatments such as gentamicin can clear gut microbes and slow extensive tissue damage caused by the release of endotoxins and inflammatory mediators triggered by chemotherapeutic drugs. Prevention and treatment of systemic inflammatory injury caused by chemotherapy drugs.

As an improvement, the chemotherapy complication is specifically systemic inflammatory response syndrome caused by chemotherapy drugs.

As an improvement, the chemotherapeutic drugs are cisplatin (25mg/kg), pentafluorouracil (10mg/kg), camptothecin (10mg/kg), paclitaxel (20mg/kg), herceptin (15mg/kg), Rotinib (150 mg/kg), IL-12 (100 μl injection, 0.5 μg/ml), CART (100 μl subcutaneously inoculated with 6×10 ⁶ CART cells on the left dorsal side), PD-1 (15 mg/kg). Malignant tumors caused by chemotherapy drugs include liver cancer, ovarian cancer, esophageal cancer, lymphoma, malignant melanoma, pancreatic cancer and breast cancer.

As an improvement, the medicine for preventing and treating chemotherapy complications is prepared by using intestinal bacteria antibiotics as active ingredients and adding a water-soluble organic carrier.

As an improvement, the water-soluble organic carrier adopts any one of dextrin, corn oil or dimethyl sulfoxide.

As an improvement, the medicament is an oral formulation or an injectable formulation. The antibiotic drug of the present invention can be administered orally, intravenously, intraperitoneally or any other way that can deliver an effective dose of the active substance.

As an improvement, the daily dosage of the intestinal bacteria antibiotic is 0.05g/kg-10g/kg. As a further improvement, the daily dosage of the intestinal bacteria antibiotic is 0.5g/kg-10g/kg. A suitable dose is one that will yield the desired final amount. Different doses may also be required to treat different diseases. An effective amount of an antibiotic in the present invention is an amount that can significantly reduce the degree of damage caused by the chemotherapeutic drug. A researcher of ordinary skill will be able to determine the most effective dose and timing of administration of the drugs of the present invention taking into account the mode of administration, drug metabolism, and several other pharmacokinetic parameters such as drug distribution, clearance, and the like.

The present invention is exemplified by observing the antagonizing effect of intestinal bacteria antibiotics on toxic reactions such as tissue function damage caused by large doses of antitumor drugs. The anti-tumor drugs used in the present invention are divided into three categories according to the mechanism of action: (i) cytotoxic drugs: cisplatin, pentafluorouracil, camptothecin, paclitaxel; (ii) molecular targeted drugs: Herceptin, errotti (iii) Immunotherapy drugs: IL-12, CART, PD-1.

The ICR experimental mice used in the present invention are used to simulate the main pathological process of human beings in the study of chemotherapeutic drug toxicity resistance. Animals herein include but are not limited to mice, rats, domesticated animals including but not limited to cats, dogs, and other animals such as but not limited to cows, sheep, pigs, horses, primates such as but not limited to monkeys and people.

The following examples are used to explain the specific details of the invention, but should not be construed as limiting the functional scope of the invention.

Example 1

The application of the intestinal bacteria antibiotic of the present invention in the preparation of a drug for preventing and treating chemotherapy complications, the specific preparation steps are as follows:

1) water-soluble antibiotics are prepared into a stock solution of a certain concentration with PBS, and stored at -20°C, wherein the water-soluble antibiotics adopted in the present embodiment are respectively gentamicin and levofloxacin;

2) Dilute the stock solution into drinking water, and prepare antibiotic drinking water with a concentration of 5 g/L at 1.25 g/kg.

Example 2

Application of oral intestinal bacteria antibiotics in the preparation and prevention of systemic inflammatory response syndrome caused by chemotherapy drugs.

1. Experimental animals

Male ICR mice, weighing 18-22 g, were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. (SPF grade) and raised in the Experimental Animal Center of Nanjing University.

2. Grouping and processing

2.1 Experimental grouping

Mice were randomly divided into normal group, chemotherapy drug group, gentamicin group, levofloxacin group, gentamicin + chemotherapy drug group, levofloxacin + chemotherapy drug group, a total of six groups.

2.2 Experimental processing

a) The gentamicin group, the levofloxacin group, the gentamicin + chemotherapy drug group, and the levofloxacin + chemotherapy drug group were given drinking water containing antibiotics, respectively, and the normal group and the chemotherapy drug group were given the corresponding amount of drinking water;

b) Two weeks later, the chemotherapy drug group, the gentamicin + chemotherapy drug group, and the levofloxacin + chemotherapy drug group were given intraperitoneal injections containing chemotherapy drugs cisplatin (25mg/kg), pentafluorouracil (10mg/kg), camptothecin (10mg/kg), Paclitaxel (20mg/kg), Herceptin (15mg/kg), Erlotinib (150mg/kg), IL-12 (100μl injection, 0.5μg/ml), CART (left dorsal Subcutaneously inoculate 100 μl of CART cells with a total number of 6 × 10 ⁶ ), PD-1 (15 mg/kg) in physiological saline solution, administered once a day for 3 consecutive days;

c) Collect samples on the 2nd day after drug withdrawal, remove the eyeballs after fasting for 12 hours, and collect blood to detect the content of endotoxin and TNF-α in serum; Colon tissue was fixed, embedded, and stained with HE.

The specific treatment is shown in Table 1 below.

Table 1 The treatment process of each experimental group

2.3 The performance effect of gut bacteria antibiotics on systemic inflammatory response syndrome

A) Effects of gentamicin and levofloxacin on the body weight of chemotherapy animals

Table 2 lists the effects of gentamicin and levofloxacin on the body weight of mice in each group treated with chemotherapeutic drugs.

Table 2 The effect of intestinal bacteria antibiotics on the weight change of mice with chemotherapy drugs (g)

Data are presented as means, and significant differences were determined by ANOVA test. *represents P≤0.05.

As can be seen in Table 2, the weight gain trend of the two groups (C, E) pretreated with antibiotics was consistent with that of the normal group (A); the weight of mice in the chemotherapy drug group (B) given chemotherapy drugs decreased significantly; Compared with the chemotherapy drug group (B), the body weight of the mice in the gentamicin + chemotherapy drug group (D) and levofloxacin + chemotherapy drug group (F) increased, and the weight loss of the mice was small, and there were significant differences . It can be seen from the above results that gentamicin and levofloxacin have certain chemoprotective effects on chemotherapy mice.

B) Effects of gentamicin and levofloxacin on the overall survival rate of chemotherapy animals

To further investigate whether gentamicin and levofloxacin can affect the overall survival rate of animals after chemotherapy, all mice in the chemotherapy drug paclitaxel group died as the node. The results are shown in Table 3.

Table 3 The effect of intestinal bacteria antibiotics on the overall survival rate of chemotherapy drugs in mice

Data are presented as mean ± standard deviation, and significant differences were determined by ANOVA test. *represents P≤0.05.

It can be seen from Table 3 that the gentamicin and levofloxacin treatment groups (D, F) can prolong the average survival days of chemotherapy mice, and the survival rate is significantly higher than that of the chemotherapy drug group (B). It is suggested that gut bacteria antibiotics can significantly improve the overall survival rate of chemotherapy animals.

C) Effects of gentamicin and levofloxacin on serum endotoxin in chemotherapy animals

Endotoxin, the main component of the outer membrane of Gram-negative bacteria, has a strong ability to induce systemic inflammatory response syndrome. Eyeball blood was collected from mice in each group on the 2nd and 4th day, and serum endotoxin content was determined by ELISA method. The results are shown in Table 4 below.

Table 4 Influence of intestinal bacteria antibiotics on endotoxin content of chemotherapeutic drugs in mice (EU/ml)

Data are presented as mean ± standard deviation, and significant differences were determined by ANOVA test. *represents P≤0.05.

On the second day of the experiment, the endotoxin levels of the chemotherapy drug group and the antibiotic treatment group were significantly increased, and there was a statistical difference compared with the normal group; on the fourth day of the experiment, the endotoxin level of the intestinal bacteria treatment group was lower than that of the chemotherapy drug group, with a statistical difference. , indicating that gentamicin and levofloxacin can reduce the damage of intestinal mucosa

D) Effects of gentamicin and levofloxacin on serum TNF-α in DOX chemotherapy animals

Eyeball blood was collected from the mice in each group on the 4th day of drug treatment. After separating the serum, the serum endotoxin content was determined by ELISA. As shown in Figure 1, the results showed that the intestinal bacteria treatment group significantly decreased the content of the inflammatory factor TNF-α in the serum of the mice treated with chemotherapy drugs, with a statistical difference, indicating that gentamicin and levofloxacin can reduce the systemic inflammatory response syndrome.

E) Effects of gentamicin and levofloxacin on intestinal mucosa of animals undergoing chemotherapy

The colons of the mice in each group were taken out on the 4th day after drug treatment, and the damage of the colon was detected by H&E pathological observation with the naked eye. The results are shown in Figure 2. Compared with the normal control group, the chemotherapeutic drug group showed a more serious injury state and a large number of inflammatory cells infiltrated, while the mucosal injury state and inflammatory response of the intestinal bacteria treatment group were significantly slowed down, suggesting that gentamicin , Levofloxacin has the function of reducing systemic inflammatory response syndrome and protecting tissue from damage.

The above descriptions are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements or improvements made within the spirit and principles of the present invention shall be included in the protection of the present invention. within the range.

Claims (5)

1. the application of intestinal bacteria antibiotics in the preparation of drugs for preventing and treating chemotherapy complications, it is characterized in that, described intestinal bacteria antibiotics are specifically intestinal gram-negative bacteria antibiotics, and described intestinal bacteria antibiotics are gram-negative bacteria antibiotics. Any one or both of damacin and levofloxacin, the chemotherapy complication is systemic inflammatory response syndrome; intestinal gram-negative bacteria antibiotics clear the flora of gram-negative bacilli in the intestine, when chemotherapy After the drug causes intestinal mucosal damage, the systemic inflammatory response syndrome is eliminated by the intestinal gram-negative bacteria antibiotic, and the administration dose of the intestinal bacteria antibiotic is 0.5g/kg-10g/kg, which is administered daily. 2. application according to claim 1, is characterized in that, described chemotherapeutic drug is cisplatin, pentafluorouracil, camptothecin, paclitaxel, Herceptin, erlotinib, IL-12, CART, PD- 1 any one or a combination of several. 3. application according to claim 1, is characterized in that, described medicine for preventing and treating chemotherapy complications is the medicament prepared by taking intestinal bacteria antibiotics as active ingredients, and adding water-soluble organic carrier. 4. application according to claim 3, is characterized in that, described water-soluble organic carrier adopts any one in dextrin, corn oil or dimethyl sulfoxide. 5. The use according to claim 3 or 4, wherein the medicament is an oral preparation or an injection preparation.

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