阅读说明：本技术 一种青霉素增效减量联合抑菌剂及其制备方法 (Synergistic and decrement combined bacteriostatic agent for penicillin and preparation method thereof ) 是由 尚军 张洁 陈海娟 张国燕 拉本 于 2020-12-31 设计创作，主要内容包括：本发明提供一种青霉素增效减量联合抑菌剂及其制备方法,包括青霉素、熊果酸、齐墩果酸和芹菜素中的两种或多种药物的组合。其中,青霉素：熊果酸：齐墩果酸：芹菜素＝10μg/mL:80μg/mL:100μg/mL:20μg/mL；青霉素：芹菜素：齐墩果酸＝5μg/mL:10μg/mL:20μg/mL；青霉素：熊果酸：芹菜素＝5μg/mL:50μg/mL:5μg/mL；青霉素：熊果酸：齐墩果酸＝5μg/mL:100μg/mL:20μg/mL；青霉素：齐墩果酸＝5μg/mL:40μg/mL；青霉素：熊果酸＝5μg/mL:100μg/mL；青霉素：芹菜素＝5μg/mL:10μg/mL等组合能够显著抑制对金黄色葡萄球菌或大肠杆菌。本发明所述药物联合方式均能降低青霉素用量,提高抑菌效果；其中几种药物组合方式能够显著的降低青霉素用量,大大减少青霉素的用量。(The invention provides a synergistic and decrement combined bacteriostatic agent of penicillin and a preparation method thereof, comprising the combination of two or more drugs of penicillin, ursolic acid, oleanolic acid and apigenin. Wherein, penicillin: ursolic acid: oleanolic acid: apigenin is 10 mu g/mL, 80 mu g/mL, 100 mu g/mL, 20 mu g/mL; penicillin: apigenin: oleanolic acid 5 mug/mL, 10 mug/mL, 20 mug/mL; penicillin: ursolic acid: 5 mu g/mL of apigenin, 50 mu g/mL of apigenin, and 5 mu g/mL of apigenin; penicillin: ursolic acid: oleanolic acid 5 mug/mL 100 mug/mL 20 mug/mL; penicillin: oleanolic acid is 5 mug/mL and 40 mug/mL; penicillin: 5 mug/mL of ursolic acid, 100 mug/mL; penicillin: the combination of 5 mu g/mL apigenin and 10 mu g/mL can obviously inhibit staphylococcus aureus or escherichia coli. The drug combination mode of the invention can reduce the dosage of penicillin and improve the bacteriostatic effect; the combination mode of several medicines can obviously reduce the dosage of the penicillin, and greatly reduce the dosage of the penicillin.)

1. A penicillin combined bacteriostatic agent, which is characterized by comprising the following components in part by weight: penicillin: ursolic acid: oleanolic acid: apigenin 10 μ g/mL, 80 μ g/mL, 100 μ g/mL, 20 μ g/mL.

2. The combined bacteriostatic agent for penicillin is characterized in that the combined bacteriostatic agent adopts any one of the following medicinal combination modes: (1) penicillin: apigenin: oleanolic acid 5 mug/mL, 10 mug/mL, 20 mug/mL; (2) penicillin: ursolic acid: 5 mu g/mL of apigenin, 50 mu g/mL of apigenin, and 5 mu g/mL of apigenin; (3) penicillin: ursolic acid: oleanolic acid 5. mu.g/mL, 100. mu.g/mL, 20. mu.g/mL.

3. The combined bacteriostatic agent for penicillin is characterized in that the combined bacteriostatic agent adopts any one of the following medicinal combination modes: (1) penicillin: oleanolic acid is 5 mug/mL and 40 mug/mL; (2) penicillin: 5 mug/mL of ursolic acid, 100 mug/mL; (3) penicillin: apigenin is 5 mu g/mL and 10 mu g/mL.

4. Penicillin combination bacteriostatic agent according to any one of the claims 1-3, characterized in that the bacteriostatic agent is an inhibitory effect against Staphylococcus aureus or Escherichia coli.

5. A combined bacteriostatic agent for enhancing and reducing penicillin and a preparation method thereof comprise the following steps: (ii) a (1) Selecting materials and instruments; (2) preparing an antibacterial agent and a culture medium; (3) preparing 1% TTC staining solution; (4) preparing a bacterial liquid; (5) determining the MIC of a single medicine; (6) measuring the antibacterial effect of the combination of the penicillin and one of the medicines; (7) measuring the antibacterial effect of the combination of the penicillin and the two medicaments; (8) the antibacterial effect of the combination of the penicillin and the three medicines is measured; (9) experimental results and analysis.

Technical Field

The invention relates to an antibiotic combined medicine, in particular to a penicillin synergistic decrement combined bacteriostatic agent and a preparation method thereof.

Background

Pneumonia belongs to one of common clinical diseases, can bring serious harm to human health, and has certain influence on the quality of life of people. The disease can be caused by microorganisms such as viruses, chlamydia, fungi, bacteria, etc., such as bacterial and viral infection, stimulation of physicochemical factors, allergy or drug factors. In recent years, western medicines are mainly used for treating the bacterial pneumonia which is the biggest factor threatening the health of children and old people, and a standard accepted treatment therapy is an antibacterial medicine. With the continuous variation of bacterial viruses, the produced antibiotics and vaccines play a certain role, but the fatality rate and the morbidity rate of the bacterial pneumonia are still in positive correlation. At present, the aging society of China is continuously developed, the frequency and the degree of pneumonia of the old people are increased, diagnosis is delayed due to the fact that organ systems are mistakenly aged, and the morbidity and mortality are increased. Therefore, the research on the combined use of antibiotics and other medicines is particularly important for reducing the side effect of the medicines.

Antibiotics are commonly used drugs for the treatment of infectious diseases, however, in recent years, antibiotic abuse has received increasing attention. Abuse of antibacterial drugs easily causes the generation of drug-resistant strains, the virulence of pathogenic bacteria is easily enhanced under certain conditions to destroy the normal flora of a host, and the improper use of the antibacterial drugs easily causes the problems of serious adverse reactions, double infection and the like. In the prior art, the report of preparing compound medicines by combining oleanolic acid, ursolic acid and apigenin with penicillin is not found.

Disclosure of Invention

In view of the defects of the prior art, the invention provides a synergistic and decrement combined bacteriostatic agent for penicillin and a preparation method thereof. In order to achieve the purpose, the invention adopts the following technical scheme:

1. a penicillin combination bacteriostatic agent comprising the combination of: penicillin, ursolic acid, oleanolic acid, and apigenin.

2. A penicillin combination bacteriostatic agent comprising: when penicillin: apigenin: oleanolic acid 5 μ g/mL 10 μ g/mL 20 μ g/mL, penicillin: ursolic acid: apigenin 5 μ g/mL 50 μ g/mL 5 μ g/mL, penicillin: ursolic acid: when the oleanolic acid is 5 mu g/mL, 100 mu g/mL and 20 mu g/mL, the antibacterial effect on staphylococcus aureus is best.

3. A penicillin combination bacteriostatic agent comprising the combination of: when penicillin: oleanolic acid 5 μ g/mL 40 μ g/mL, penicillin: 5 mug/mL ursolic acid, 100 mug/mL penicillin: when the apigenin is 5 mu g/mL and 10 mu g/mL, the bacteriostatic effect on staphylococcus aureus is the best.

4. A combined bacteriostatic agent for enhancing and reducing penicillin and a preparation method thereof comprise the following steps: (ii) a (1) Selecting materials and instruments; (2) preparing an antibacterial agent and a culture medium; (ii) a (3) Preparing 1% TTC staining solution; (4) preparing a bacterial liquid; (5) determining the MIC of a single medicine; (6) measuring the antibacterial effect of the combination of the penicillin and one of the medicines; (7) measuring the antibacterial effect of the combination of the penicillin and the two medicaments; (8) the antibacterial effect of the combination of the penicillin and the three medicines is measured; (9) experimental results and analysis.

Has the advantages that: the experiment preliminarily analyzes the effect of the combined medicine on treating the bacterial pneumonia by researching the action mechanism of the combined medicine of the active ingredients for treating the bacterial pneumonia and the antibiotics. The results show that the expected result of inhibiting the growth of bacteria can be realized by using the penicillin and the oleanolic acid, the ursolic acid and the apigenin in combination according to different proportions. Because the tested bacteria utilized in the experimental design have two types of gram-positive bacteria and gram-negative bacteria, the primary judgment shows that the drug effect exerted by the combined drug is consistent with that of broad-spectrum bacteria, and the effect of broad-spectrum bacteria on treating diseases can be achieved. The Chinese herbal medicine components are complex, the Chinese herbal medicine components have little toxic and side effects, the antibacterial effect is strong, the antibacterial mechanism has multiple ways, the antibacterial result is influenced by different internal and external environments, and the drug resistance of bacteria is not easy to induce. The antibiotic has rich resources in China, can reduce the dosage when being used together with the antibiotic, and improves the curative effect of the antibiotic. The active ingredients with better clinical bacteriostatic effect are screened under the in vitro drug condition and reasonably combined with the antibacterial drugs, so that the dosage of the Chinese herbal medicines and the antibacterial drugs can be greatly reduced, the clinical curative effect is improved, and the generation of drug-resistant strains and drug residues caused by abuse of antibiotics can be avoided or reduced.

The drug combination mode of the invention can reduce the dosage of penicillin and improve the bacteriostatic effect. Wherein, penicillin: ursolic acid: oleanolic acid: apigenin is 10 mu g/mL, 80 mu g/mL, 100 mu g/mL, 20 mu g/mL; penicillin: apigenin: oleanolic acid 5 mug/mL, 10 mug/mL, 20 mug/mL; penicillin: ursolic acid: 5 mu g/mL of apigenin, 50 mu g/mL of apigenin, and 5 mu g/mL of apigenin; penicillin: ursolic acid: oleanolic acid 5 mug/mL 100 mug/mL 20 mug/mL; penicillin: oleanolic acid is 5 mug/mL and 40 mug/mL; penicillin: 5 mug/mL of ursolic acid, 100 mug/mL; penicillin: the combination of 5 mu g/mL apigenin and 10 mu g/mL can obviously inhibit staphylococcus aureus or escherichia coli. The combination mode of the medicines can obviously reduce the dosage of the penicillin, and greatly reduce the dosage of the penicillin.

Drawings

FIG. 1 is a graph comparing single and combination administration for Staphylococcus aureus;

FIG. 2 is a graph comparing the individual and combined administration for E.coli;

FIG. 3 is a graph comparing ursolic acid and oleanolic acid.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention are described in detail below with reference to the accompanying drawings. Examples of these preferred embodiments are illustrated in the accompanying drawings. The embodiments of the invention shown in the drawings and described in accordance with the drawings are exemplary only, and the invention is not limited to these embodiments. It should be noted that, in order to avoid obscuring the technical solution of the present invention with unnecessary details, only the structures and/or processing steps closely related to the solution according to the present invention are shown in the drawings, and other details that are not relevant are omitted.

Example 1

The embodiment provides a synergistic and decrement combined bacteriostatic agent for penicillin and a preparation method thereof, and the preparation method comprises the following steps:

1. materials and instruments

Compound 1, ursolic acid control (vicker biotechnology limited, sichuan); compound 2, oleanolic acid control (vicker biotechnology limited, sichuan province); compound 3, apigenin control (vikkci biotechnology limited, sichuan); antibiotics, penicillins; strains, staphylococcus aureus, escherichia coli; culture medium, nutrient agar, MH broth; dimethyl sulfoxide solution, Shi chemical Co., Ltd, Tianjin; normal saline, Sichuan Kelun pharmaceutical Co., Ltd; triphenyltetrazolium chloride, Teijin chemical reagents research institute, Inc.; disodium hydrogen phosphate, remote chemical agents ltd, Tianjin; potassium dihydrogen phosphate, remote chemical agents of Tianjin, Inc.; electronic analytical balance, shanghai yueping scientific instruments ltd; clean bench, Tensted instruments, Inc., Tianjin; a constant temperature water bath, repulped instruments teaching limited, Changzhou; vertical pressure steam sterilizer, LS-35HD Jiangyin Binjiang corporation; ultrapure water machine, model: WP-UP-Y; high-temperature sterilization box, model: DZF-6050; refrigerator, hail 539L; microplate fast oscillators, linbel technologies ltd; a constant temperature water-proof incubator, HPX-250 Xiang Instrument centrifuge, Inc.; non-removable 96-well plates, Greiner, germany; oxford cup, Beijing Jitai Yuan science and technology Co., Ltd; vernier calipers are available in large-scale tool factories.

2. Preparation of antibacterial agent and culture medium

Weighing 6mg of the required medicine by using an electronic analytical balance, preparing into 25% DMSO (dimethylsulfoxide) medicine stock solutions with initial concentrations of 1200 mu g/mL respectively, fully shaking up and dissolving, and storing at 4 ℃ for later use as test medicine solutions. Weighing 21.0g of the product, heating and dissolving in 1000ml of distilled water, and autoclaving at 121 ℃ for 30 minutes for later use. The nutrient agar is prepared into 500ml nutrient agar solution with distilled water according to a certain proportion, the constant temperature is 121 ℃, and the sterilization time is 30 min.

Preparation of 3.1% TTC staining solution

Dissolving 1.973g of disodium hydrogen phosphate and 0.436g of potassium dihydrogen phosphate in 1L of deionized water, fully mixing and dissolving, and adjusting the pH value to 6.5-7.5 to obtain the required phosphoric acid buffer solution. According to the required TTC working solution concentration, 1g of TTC powder is added into 100ml of mixed phosphate buffer solution, and then 1% of TTC staining solution is obtained. It was filled in a brown bottle and stored at 4 ℃ for further use.

4. Preparation of bacterial liquid

The experiment selects gram-positive bacteria (staphylococcus aureus) and gram-negative bacteria (escherichia coli) as tested strains respectively, and whether the antibacterial effects on different strains are the same when the extract and the penicillin are jointly used is observed and analyzed, so that whether the antibacterial characteristics of the broad-spectrum antibiotics are achieved is preliminarily judged. After different tested bacteria (staphylococcus aureus and escherichia coli) which are frozen and stored in a refrigerator at 4 ℃ and are in a dormant period are put in a constant-temperature incubator at 37 ℃ for resuscitation for 24 hours, a proper amount of bacterial colonies are picked in physiological saline by an inoculating loop in a super-clean workbench, and bacterial liquid which is diluted into 1 x 105cfu/ml of strain in a logarithmic growth phase is prepared for later use.

5. Determination of MIC for Single drug

Sterile 96-well plates (sterilized on a clean bench) were taken, and 90. mu.L of MH broth was added to each well of the 96-well plates. The prepared same liquid medicine is added into three rows B/C/D (equivalent to repeated control) with different concentrations of 10 mu L respectively. Staphylococcus aureus is selected as a test strain, and 90 mu L of diluted bacterial liquid is added into each hole. Three rows of E/F/G (equivalent to repeated control) are added with different concentrations of another liquid medicine for the same treatment. A negative control (adding only the blank broth without adding the bacteria solution) was performed in column 2 and a positive control (adding only the bacteria solution without adding the liquid solution) was performed in column 3 on the same plate. After the operation was completed, 10. mu.L of triphenyltetrazolium chloride (TTC)10 mg/mL (1%) was added dropwise to each well. All the loaded 96-well plates were placed on a microplate rapid shaker for 15 minutes. After finishing the culture, placing the mixture in an incubator at the constant temperature of 37 ℃ for culturing for 18-24h, taking out and observing results, wherein bacterial growth holes are red, and the minimum concentration of the broth clear sterile growth holes is the MIC of the medicine.

6. Determination of antibacterial effect of combination of penicillin and one of medicines

The dilution of the combination was determined by checkerboard dilution based on the MIC concentrations of drug A (penicillin) and drug B (either extract), and 6 concentrations of each were selected. After incubation for 18-24h at 37 ℃, the bacterial growth cells are observed to be red. And screening out the concentration ratio without bacterial growth, placing the mixture in a solid culture medium containing the same bacteria, incubating for 24 hours at 37 ℃, measuring the size of an inhibition zone, and comparing the size with that of the penicillin alone to determine the concentration of the penicillin at the optimal combination effect.

And (4) analyzing results: the minimum inhibitory concentrations MIC A drug order and MIC B drug order of the two drugs which are applied independently are recorded, and the combination of the MIC A drug and the MIC B drug when the two drugs are combined at the optimal combined effect is selected to calculate the FIC index. By using the method, penicillin is respectively subjected to experiments with oleanolic acid, ursolic acid and apigenin, and the optimal combined dilution is respectively selected. FIC ═ MIC/A single time of A drug combination + MIC/A single time of A drug combination

TABLE 1 FIC index analysis

Table 1 FIC index analysis

7. Determination of antibacterial effect of combination of penicillin and two drugs

Taking a sterile 96-well plate (sterilized by a clean bench), and adding 90 mu L of broth culture medium, 90 mu L of bacterial liquid, 10 mu L of prepared liquid medicine and 10 mu L of TTC staining solution into the corresponding wells according to the operation method. Culturing at 37 deg.C for 18-24h, taking out and observing. The concentration ratio without bacterial growth was selected and each drug combination was repeated three times for a total of three drug combinations. Placing the penicillin in a solid culture medium containing the same bacteria, incubating for 24h at 37 ℃, and measuring the size of an inhibition zone so as to determine the combined inhibition effect of the penicillin and the two drugs. And (4) analyzing results: the combined MIC of the three drugs is greater than that of any single drug, namely the antagonist effect, the same effect as the MIC of one of the single drugs is irrelevant, and the effect is less than that of any single drug, namely the synergistic effect.

8. Antibacterial effect determination of joint combination of penicillin and three drugs

Respectively using MIC concentrations of penicillin, oleanolic acid, ursolic acid and apigenin as optimal concentrations to prepare liquid medicine, placing the liquid medicine in a solid culture medium containing the same bacteria, observing and culturing the bacteria at 37 ℃ for 24 hours, and measuring the size of a bacteriostatic circle to further determine the bacteriostatic effect.

9. Results and analysis of the experiments

(1) MIC determination of different drugs against Staphylococcus aureus

Each drug was formulated at 1200. mu.g/mL, 1000. mu.g/mL, 800. mu.g/mL, 600. mu.g/mL, 400. mu.g/mL, 200. mu.g/mL, 100. mu.g/mL, 90. mu.g/mL, 80. mu.g/mL, 70. mu.g/mL, 60. mu.g/mL, 50. mu.g/mL, 40. mu.g/mL, 30. mu.g/mL, 20. mu.g/mL, 15. mu.g/mL, 10. mu.g/mL, 5. mu.g/mL, respectively, for 17 different concentrations. The MIC value of each drug was finally determined by the above experimental procedure. The results are shown in Table 2. As can be seen from the table, the MIC concentrations for Staphylococcus aureus were different for different drugs.

TABLE 2 MIC assay results for different drugs against Staphylococcus aureus

Table 2 MIC determination results of different drugs against Staphylococcus aureus

(2) Antibacterial effect of combination of penicillin and one of medicines

According to the above results, the dilution of the combination of the two drugs was determined by the checkerboard dilution method, and the maximum concentration of each antibacterial drug was 2 times the MIC. The drug concentrations are shown in tables 3, 4 and 5.

TABLE 3 dilution of penicillin in combination with oleanolic acid

Table 3 Dilution of penicillin and oleanolic acid combination

TABLE 4 dilution of the combination of penicillin and ursolic acid

Table 4 Dilution of penicillin and ursolic acid combination

TABLE 5 dilution of the combination of penicillin and apigenin

Table 5 Dilution of penicillin and apigenin combination

By carrying out experiments on different concentration ratios in the table and observing the size of the inhibition zone, the optimal concentration ratio of the combination of the penicillin and one of the medicines is obtained. When penicillin: oleanolic acid 5 μ g/mL 40 μ g/mL, penicillin: 5 mug/mL ursolic acid, 100 mug/mL penicillin: when the apigenin is 5 mu g/mL and 10 mu g/mL, the bacteriostatic effect on staphylococcus aureus is the best. According to the concentration ratio, the escherichia coli is further subjected to bacteriostasis experiment operation, and the difference of the bacteriostasis effects of the penicillin and one of the medicines on different strains is compared, and the results are shown in table 6.

TABLE 6 comparison of the antibacterial effects of the combination of the two drugs at the optimum concentrations on different strains

Table 6 Comparison of the antibacterial effect of the combination of two drugs at the optimum concentration on different strain

Note:^**represents P<0.01, significantly different from the blank group;^Arepresents P<0.01, compared with the single medicine, the difference is extremely remarkable.

As can be seen from the table, the two drugs have very significant differences in the antibacterial effect on different strains. Compared with the whole body, the bacteriostatic effect of the escherichia coli is more obvious by different drug concentrations. When the penicillin is used together with the oleanolic acid and the apigenin, the additive effect is shown on staphylococcus aureus and escherichia coli, namely the combined drug effect of the two antibacterial drugs is consistent with the sum of the antibacterial effects of the two drugs with the same concentration. When the penicillin and the ursolic acid are used in combination, the combined drug has irrelevant effects on staphylococcus aureus and escherichia coli, namely, the drug activity of the combined drug is the same as that of the single drug, which indicates that the combined drug of the penicillin and the ursolic acid is possibly influenced by other factors, and the phenomenon needs further deep research.

(3) Antibacterial effect of combination of penicillin and two drugs

The penicillin and two of the medicines are used together in three combination modes, namely: penicillin, apigenin, oleanolic acid; penicillin, ursolic acid, apigenin, penicillin, ursolic acid and oleanolic acid. Based on the minimum dosage of 5 mug/mL of penicillin, the antibacterial test is carried out according to the combination ratio of different dosages of other medicines. Finally, it is obtained when penicillin: apigenin: oleanolic acid 5 μ g/mL 10 μ g/mL 20 μ g/mL, penicillin: ursolic acid: apigenin 5 μ g/mL 50 μ g/mL 5 μ g/mL, penicillin: ursolic acid: when the oleanolic acid is 5 mu g/mL, 100 mu g/mL and 20 mu g/mL, the antibacterial effect on staphylococcus aureus is best. According to the concentration ratio, the escherichia coli is further subjected to bacteriostasis experiment operation, and the difference of the bacteriostasis effects of the penicillin and the two drugs on different strains when the two drugs are used together is compared, and the results are shown in table 7.

TABLE 7 comparison of the antibacterial effects of the combination of the three drugs at the optimum concentrations on different strains

Table 7 Comparison of the antibacterial effects of the three drugs in combination with the optimal concentration on different strains

Note:^**represents P<0.01, significantly different from the blank group;^Arepresents P<0.01, compared with the single medicine, the difference is extremely remarkable.

As can be seen from the table, the differences of the bacteriostatic effects of the three drugs in combination on different strains are very obvious. Compared with the whole, when the penicillin is used together with the apigenin and the oleanolic acid and the penicillin, the ursolic acid and the apigenin are used together, the antibacterial effect on escherichia coli is more obvious than that on staphylococcus aureus; when the penicillin, the ursolic acid and the apigenin are used together, the bacteriostatic effect on staphylococcus aureus is slightly better. From the combination index, no matter which combination mode is used, the obvious synergistic effect is shown, namely the MIC concentration of the three medicines in the combined use is smaller than that of each medicine in the independent use.

(4) Antibacterial effect of penicillin and three medicines

The results obtained after antibacterial experiments were performed on staphylococcus aureus and escherichia coli, respectively, by preparing a liquid medicine with MIC concentrations of penicillin, oleanolic acid, ursolic acid, and apigenin as optimum concentrations, are shown in table 8.

TABLE 8 comparison of the inhibitory effects of the combination of the MIC concentrations of the four drugs on different strains

Table 8 Comparison of antibacterial effects of four drugs in combination with MIC concentration on different strains

Note:^**represents P<0.01, significantly different from the blank group;^Arepresents P<0.01, compared with the single medicine, the difference is extremely remarkable.

As can be seen from the table, the differences of the bacteriostatic effects of the combination of the four medicaments on different strains are very obvious. But the bacteriostatic effect of different drug concentrations is more obvious than that of Escherichia coli.

(5) Comparison of independent medication and combined medication of four medicines

Tables 9 and 10 record the bacteriostatic effects of different concentrations of penicillin, ursolic acid, oleanolic acid, and apigenin, respectively, on two different strains when administered alone. As can be seen from the table, the higher the concentration of each drug when taken alone, the more obvious the bacteriostatic effect. Compared with a blank group, the drug combination shows extremely obvious difference, and the concentration of the screened single drug has better bacteriostatic effect.

TABLE 94 bacteriostatic effect of different concentrations of the drugs on Staphylococcus aureus when used alone

Table 9 The antibacterial effects of 4drugs at different concentrations on Staphylococcus aureus when used alone

Note: denotes P <0.01, the difference is very significant.

Bacteriostatic effect of different concentrations of the table 104 drugs on escherichia coli when used alone

Table 10 The antibacterial effect of 4kinds of drugs in different concentrations on E.coli when used alone

Note: denotes P <0.01, the difference is very significant.

The size of the zone of inhibition for the individual and combined administration was compared for different strains to obtain the bar graphs shown in fig. 1 and 2. As can be seen from the figure, the bacteriostatic effect after the combined medication is more obvious than that of the four medicaments which are taken separately on the premise of selecting the minimum concentration of 5 mug/mL of penicillin. The antibacterial effect of the combined drug of penicillin and two drugs is better than that of the combined drug of one of the two drugs aiming at gram-positive bacteria (staphylococcus aureus). However, the combined use of penicillin, ursolic acid and oleanolic acid for gram-negative bacteria (Escherichia coli) has an antibacterial effect obviously inferior to that of the combined use of penicillin, ursolic acid and oleanolic acid. By reviewing the relevant literature, and further analysis in conjunction with drug-bacterial targets in network pharmacology, the following conclusions were drawn:

ursolic acid and oleanolic acid are triterpenes, and belong to isomers (as shown in figure 3). The experimental results of Wu Fang, etc. show that the curative effect of the same drug combination with the same action mechanism is not enhanced, but toxicity is increased or the same target of bacteria is competed to generate antagonism. From table 11, it can be found that the drug targets for treating diseases of the two drugs are basically consistent, and for treating escherichia coli, ursolic acid and oleanolic acid compete for 4 targets of MAPK8, MAPK3, ACE and IL6 simultaneously, so that the combined drug of penicillin, ursolic acid and oleanolic acid shows antagonism, and the antibacterial effect is poorer than that of the combined drug of penicillin, ursolic acid and oleanolic acid.

TABLE 11 Ursolic acid and Oleanolic acid drug-bacterial target analysis

Table 11 Analysis of ursolic acid and oleanolic acid drugs-bacterial targets

The foregoing is directed to embodiments of the present application and it is noted that numerous modifications and adaptations may be made by those skilled in the art without departing from the principles of the present application and are intended to be within the scope of the present application.