New use of Baotouweng decoction in synergistic antibacterial effect of antibiotics

Abstract

The present application relates to the new use of Baotouweng decoction in the synergistic antibacterial effect of ceftriaxone, tetracycline and amikacin antibiotics. In the preparation of the composition or compound preparation, the dosage ratio of Baotouweng decoction and ceftriaxone, tetracycline and amikacin is between 6400:1-12800:1. The FIC index of Baotouweng decoction combined with ceftriaxone, amikacin or tetracycline is 0.1875-0.375, which is less than 0.5, that is, Baotouweng decoction combined with ceftriaxone, amikacin or tetracycline has synergistic antibacterial effect. The present application can effectively reduce the dosage of ceftriaxone, amikacin or tetracycline, and the MIC value of Baotouweng decoction combined with ceftriaxone, amikacin or tetracycline is reduced by 2-16 times compared with that of single use. Baotouweng decoction can be used as an antibacterial drug resistance reversal agent to improve the antibacterial therapeutic effect under the condition of reducing the dosage of antibacterial drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a new use of Pulsatillae Decoction as an antibacterial synergist. BACKGROUND

[0002] Klebsiella pneumoniae (K. pneumoniae) is a gram-negative bacterium widely distributed in nature, and is a zoonotic pathogenic bacterium in the intestinal and respiratory tracts of humans and animals. The bacterium has a fairly wide host range, and can infect K. pneumoniae from humans to various domestic animals, poultry, wild animals and aquatic animals, causing intestinal and respiratory tract infections in humans and various animals and endangering the health of humans and livestock. At present, the control of intestinal and respiratory bacterial infections mainly relies on the use of antibacterial drugs. However, in recent years, with the widespread use of antibacterial drugs, more and more drug-resistant strains have emerged, and the drug resistance spectrum has become wider and wider, so that the cure rate of many antibacterial drugs for bacterial infections has decreased significantly, and the drug resistance of K. pneumoniae has become increasingly serious. At present, K. pneumoniae has become an important pathogenic bacterium commonly seen in clinical infections in the fields of medicine and veterinary medicine, especially the emergence of super broad-spectrum multi-drug resistant strains, which seriously endangers the health of humans and livestock. How to prevent human and animal source K. pneumoniae infection and effectively treat drug-resistant strain infection has become a top priority. Most of urinary tract infections are caused by K. pneumoniae infection, and K. pneumoniae is an important pathogenic bacterium in septicemia with a high mortality rate.

[0003] Pulsatillae Decoction (PD) is a traditional Chinese medicine compound preparation with the effects of clearing heat and detoxifying. It is mainly used for treating dysentery with pus and blood, more redness and less whiteness, abdominal pain, tenesmus, anal burning, thirst and desire to drink water, red tongue with yellow fur, and rapid and thready pulse.

[0004] In recent years, with the increase in the use of antibacterial drugs, and the occurrence of overuse and misuse, the drug resistance rate of clinically isolated bacteria has increased year by year, resulting in a relatively decreased therapeutic effect of antibacterial drugs. Strains resistant to multiple antibacterial drugs have emerged, and there is a serious trend of cross-resistance and multi-drug resistance. Therefore, it is of great clinical application significance to reduce the occurrence of drug resistance by using synergists. Traditional Chinese medicine, as a traditional Chinese medicine, has the characteristics of high efficiency, low toxicity, low residue and low drug resistance. In the case of not increasing the dosage of antibacterial drugs or even reducing the dosage, the use of traditional Chinese medicine compound preparations to effectively treat bacterial infections, especially infections caused by multiple drug-resistant bacteria, has important clinical application value. So far, there has been no report on the use of Pulsatillae Decoction as an antibacterial synergist in antibacterial action. SUMMARY

[0005] The present application aims to provide a new use of Baishouweng decoction in synergistic antibacterial effect of antibacterial agents. In the preparation of compositions or compound preparations, Baishouweng decoction is used for synergistic antibacterial effect of antibacterial agents, and the dosage ratio of Baishouweng decoction to antibacterial agents is 6400:1 or 12800:1, or between 6400:1 and 12800:1.

[0006] The present application aims to provide a new use of Baishouweng decoction in synergistic antibacterial effect of antibacterial agents. In the preparation of compositions or compound preparations, Baishouweng decoction is used for synergistic antibacterial effect of antibacterial agents, and the dosage ratio of Baishouweng decoction to antibacterial agents is 6400:1 or 12800:1, or between 6400:1 and 12800:1.

[0007] The present application aims to provide a new use of Baishouweng decoction in synergistic antibacterial effect of antibacterial agents. In the preparation of compositions or compound preparations, Baishouweng decoction is used for synergistic antibacterial effect of antibacterial agents, and the dosage ratio of Baishouweng decoction to antibacterial agents is 6400:1 or 12800:1, or between 6400:1 and 12800:1.

[0008] The present application aims to provide a new use of Baishouweng decoction in synergistic antibacterial effect of antibacterial agents. In the preparation of compositions or compound preparations, Baishouweng decoction is used for synergistic antibacterial effect of antibacterial agents, and the dosage ratio of Baishouweng decoction to antibacterial agents is 6400:1 or 12800:1, or between 6400:1 and 12800:1.

[0009] The present application aims to provide a new use of Baishouweng decoction in synergistic antibacterial effect of antibacterial agents. In the preparation of compositions or compound preparations, Baishouweng decoction is used for synergistic antibacterial effect of antibacterial agents, and the dosage ratio of Baishouweng decoction to antibacterial agents is 6400:1 or 12800:1, or between 6400:1 and 12800:1.

[0010] Another object of the present application is to provide a new use of Baishouweng decoction in enhancing the effect of amikacin against Klebsiella pneumoniae. In the preparation of a composition or a compound preparation, Baishouweng decoction is used to enhance the antibacterial effect of amikacin. When 1 / 2 MIC (102400 μg / mL) of Baishouweng decoction is added, the MIC of amikacin against Klebsiella pneumoniae is 8-16 μg / mL. The ratio of the amount of Baishouweng decoction to amikacin is 6400:1 or 12800:1, or the ratio of the amount of Baishouweng decoction to amikacin is between 6400:1 and 12800:1.

[0011] Another object of the present application is to provide a use of Baishouweng decoction in the preparation of an antibacterial effect enhancer of ceftriaxone, tetracycline, and / or amikacin against Klebsiella pneumoniae. In the preparation of a composition or a compound preparation, Baishouweng decoction is used to enhance the antibacterial effect of ceftriaxone, tetracycline, and amikacin. The ratio of the amount of Baishouweng decoction to ceftriaxone, tetracycline, and amikacin is 6400:1 or 12800:1, or the ratio of the amount of Baishouweng decoction to ceftriaxone, tetracycline, and amikacin is between 6400:1 and 12800:1.

[0012] Another object of the present application is to provide a use of Baishouweng decoction in the preparation of an antibacterial effect enhancer of ceftriaxone against Klebsiella pneumoniae. In the preparation of a composition or a compound preparation, Baishouweng decoction is used to enhance the antibacterial effect of ceftriaxone. The ratio of the amount of Baishouweng decoction to ceftriaxone is 6400:1 or 12800:1, or the ratio of the amount of Baishouweng decoction to ceftriaxone is between 6400:1 and 12800:1.

[0013] Another object of the present application is to provide a use of Baishouweng decoction in the preparation of an antibacterial effect enhancer of tetracycline against Klebsiella pneumoniae. In the preparation of a composition or a compound preparation, the ratio of the amount of Baishouweng decoction to tetracycline is 6400:1 or 12800:1, or the ratio of the amount of Baishouweng decoction to tetracycline is between 6400:1 and 12800:1.

[0014] Another object of the present application is to provide a use of Baishouweng decoction in the preparation of an antibacterial effect enhancer of amikacin against Klebsiella pneumoniae. In the preparation of a composition or a compound preparation, the ratio of the amount of Baishouweng decoction to amikacin is 6400:1 or 12800:1, or the ratio of the amount of Baishouweng decoction to amikacin is between 6400:1 and 12800:1.

[0015] Another object of the present application is to provide an antibacterial test method for the use of Baishouweng decoction in enhancing the effect of ceftriaxone, tetracycline, and / or amikacin against Klebsiella pneumoniae, which comprises the following steps:

[0016] S1: pretreating Baishouweng decoction;

[0017] S2: mixing the Baishuangan decoction with the antibacterial agent in a ratio to obtain a composition;

[0018] S3: using the composition of step S2 for MIC test, FICI test, or target test.

[0019] The Baishuangan decoction of step S1 is pretreated, including combining two medicinal liquids, reducing pressure concentration, centrifuging for 30 min; taking the supernatant, continuing concentration and sieving, centrifuging again, collecting the medicinal liquid containing 2.048 g of crude drug per milliliter, and then diluting according to actual needs for MIC test or other tests.

[0020] The ratio of the Baishuangan decoction to the antibacterial agent in step S2 is 6400:1-12800:1. Preferably, the ratio of the Baishuangan decoction to the antibacterial agent is 6400:1 or 12800:1.

[0021] The antibacterial agent in step S2 includes ceftriaxone, tetracycline, and amikacin.

[0022] The antibacterial agent in step S2 includes ceftriaxone, tetracycline, and amikacin, or any combination thereof.

[0023] The dosage of the Baishuangan decoction in step S2 is 102400 μg / mL or more than 102400 μg / mL.

[0024] Another object of the present application is to provide an antibacterial composition or Baishuangan decoction antibacterial compound preparation, which comprises the Baishuangan decoction and ceftriaxone, tetracycline, or amikacin.

[0025] The antibacterial composition or Baishuangan decoction antibacterial compound preparation comprises the Baishuangan decoction and ceftriaxone, and the dosage ratio of the Baishuangan decoction to ceftriaxone is 6400:1-12800:1. The antibacterial composition or Baishuangan decoction antibacterial compound preparation comprises the Baishuangan decoction and tetracycline, and the dosage ratio of the Baishuangan decoction to tetracycline is 6400:1-12800:1. The antibacterial composition or Baishuangan decoction antibacterial compound preparation comprises the Baishuangan decoction and amikacin, and the dosage ratio of the Baishuangan decoction to amikacin is 6400:1-12800:1.

[0026] Another object of the present application is to provide a Baishuangan decoction antibacterial compound preparation, which comprises the Baishuangan decoction and ceftriaxone, and the dosage ratio of the Baishuangan decoction to ceftriaxone is 6400:1 or 12800:1, or any ratio between 6400:1 and 12800:1.

[0027] Another object of the present application is to provide a Baishuantong antibacterial compound preparation, which comprises Baishuantong and tetracycline, and the dosage ratio of Baishuantong to tetracycline is 6400:1 or 12800:1, or any ratio between 6400:1 and 12800:1.

[0028] Another object of the present application is to provide a Baishuantong antibacterial compound preparation, which comprises Baishuantong and amikacin, and the dosage ratio of Baishuantong to amikacin is 6400:1 or 12800:1, or any ratio between 6400:1 and 12800:1.

[0029] The term "synergy" as used herein refers to the effect of the two drugs combined being greater than the sum of their individual effects.

[0030] The term "additive effect" as used herein refers to the effect of the two drugs combined being equal to the sum of their individual effects.

[0031] The term "antagonism" as used herein refers to the effect of the two drugs combined being less than the sum of their individual effects.

[0032] The present application has the following advantages: the present application provides a new use of Baishuantong in enhancing the antibacterial effect of antibiotics. (1) Baishuantong combined with ceftriaxone, amikacin, and / or tetracycline shows significant synergistic effect, with FIC index of 0.1875-0.375 (all less than 0.5), i.e. the combination of the two shows obvious antibacterial synergy; (2) the present application can effectively reduce the dosage of ceftriaxone, amikacin, and tetracycline, and the MIC value of Baishuantong combined with ceftriaxone is 4-8 times lower than that of Baishuantong alone. The MIC value of Baishuantong combined with amikacin is 4-16 times lower than that of Baishuantong alone. The MIC value of Baishuantong combined with tetracycline is 2-8 times lower than that of Baishuantong alone; (3) the FIC index of Baishuantong combined with ceftriaxone, amikacin, or tetracycline is 0.1875-0.375, all less than 0.5, i.e. the combination of Baishuantong with ceftriaxone, amikacin, or tetracycline has significant synergistic antibacterial effect. When ceftriaxone, amikacin, tetracycline is combined with Baishuantong, the antibacterial activity of the antibacterial drugs can be significantly enhanced. Thus, the dosage of the antibacterial drugs can be reduced in clinic, and the antibacterial treatment effect can be significantly improved by synergy, which has important clinical application value; (4) the antibacterial drugs of the present application, including ceftriaxone, amikacin, tetracycline, combined with Baishuantong, open up a new method and use for reversing the drug resistance of Klebsiella pneumoniae, and Baishuantong can be used as an antibacterial drug resistance reversing agent, which can improve the antibacterial treatment effect while reducing the dosage of the antibacterial drugs, and further reduce the side effects of the drugs. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 Growth curve changes of Baishuangan decoction on strain Kp20 DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. The present application will be further described below in conjunction with the embodiments, but the protection scope of the present application is not limited to the following embodiments.

[0035] Example 1

[0036] Materials and methods

[0037] 1. Preparation of test drugs

[0038] Ceftriaxone (CRO, purity: 99%), amikacin (AMK, purity: 99%), tetracycline (TET, purity: 98.8%) were purchased from Shanghai Yuan Ye Biological Technology Co., Ltd.

[0039] Preparation of Baishuangan decoction: 90g of 4 kinds of medicinal materials (Bai-tou-weng: Huang-bai: Huang-lian: Qin-pi = 15: 12: 6: 12) were taken according to the prescription proportion of Baishuangan decoction, and 6 times the amount of water was added for soaking, then 10 times the amount of water was added for heating and boiling, and after boiling, the fire was changed to a low fire for 1h of decoction; filtration, the filter residue was added with 8 times the amount of water for decoction for 1h; filtration, the supernatant was taken, and the two times of medicinal liquid were combined, reduced pressure concentration, 3000r / min centrifugation for 30min; take the supernatant, continue to concentrate and sieve, centrifuge again, collect the medicinal liquid containing 2.048g of crude drug per milliliter (concentration of 2.048g / mL, i.e. 2048000μg / mL), and then flow through the air sterilization for 20min, ready for use. Ceftriaxone, amikacin, and tetracycline were respectively prepared into a concentration of 1280μg / mL with deionized water. The test drugs were stored at -20℃. Before the test, the drug storage solution was taken out and melted at room temperature, and fully mixed, and then the pharmacodynamic test was carried out respectively.

[0040] 2. Preparation of mixed drugs

[0041] Preparation of Bai Tou Weng Decoction, according to the proportion of Bai Tou Weng Decoction (Bai Tou Weng: Huang Bai: Huang Lian: Qin Pi = 15: 12: 6: 12) 90g of 4 kinds of medicinal materials were taken, 6 times the amount of water was added for soaking, then 10 times the amount of water was added for heating and boiling, after boiling, the fire was changed to low heat for 1h; filtration, the filter residue was added with 8 times the amount of water for boiling for 1h; filtration, the supernatant was taken, the two times of medicinal liquid were combined, reduced pressure concentration, centrifugation at 3000r / min for 30min; the supernatant was taken, continued to be concentrated and sieved, centrifuged again, collected the medicinal liquid containing 2.048g of crude drug per milliliter (concentration of 2.048g / mL, i.e. 2048000μg / mL), and then flowed through the air for sterilization for 20min, ready for use. Ceftriaxone, amikacin, and tetracycline were respectively prepared into a concentration of 1280μg / mL with deionized water. The test drug was stored at -20℃. Before the test, the drug storage solution was taken out and melted at room temperature, fully mixed, and then the pharmacodynamic test was carried out.

[0042] 2. Strains

[0043] Klebsiella pneumoniae strains isolated clinically: Klebsiella pneumoniae Kp20, Kp24, Kp32 were provided by the Inspection Laboratory of Henan University of Chinese Medicine. Klebsiella pneumoniae was activated on LB medium and cultured at 37℃ for 12-16h, and stored at 4℃ for standby.

[0044] 3. Culture solution

[0045] LB broth medium: produced by Beijing Land Bridge Technology Co., Ltd., 1000ml was prepared according to the instructions, 121℃ high pressure sterilization for 15min, 4℃ storage for standby. LB agar medium: produced by Beijing Land Bridge Technology Co., Ltd., 1000ml was prepared according to the instructions, heated and boiled to dissolve and sterilize, cooled to about 60 degrees, poured into sterilized flat dishes, and stored at 4℃ for standby. MHB broth medium: produced by Beijing Land Bridge Technology Co., Ltd., 1000ml was prepared according to the instructions, 121℃ high pressure sterilization for 15min, 4℃ storage for standby.

[0046] 4. Instruments

[0047] SX-500 high pressure steam sterilizer (Japan Tomy Company); AB204-N type electronic analytical balance (Mettler-Toledo Instruments Shanghai Co., Ltd.); SHA-C type digital water bath constant temperature oscillator (Changzhou Boyuan Experimental Analysis Instrument Factory); DH-600 electric heating constant temperature incubator (Beijing Keyi Yongxing Instrument Co., Ltd.); DZF-6050 type vacuum drying oven (Shanghai Jinghong Experimental Equipment Co., Ltd.); CJ-2F type medical purification workbench SW-CJ-JC super clean workbench (Suzhou Fengshi Experimental Animal Equipment Co., Ltd.).

[0048] 5. Preparation of bacterial solution

[0049] A single colony of K. pneumoniae was picked from LB agar medium stored at 4°C and inoculated into 5 mL of LB broth and incubated at 37°C with 200 rpm shaking to activate the bacteria and make them in exponential growth phase. The bacteria were diluted to 1.0 x 10 5 CFU / mL in MHB broth.

[0050] 6. Preparation of drug sensitivity plate

[0051] The drug sensitivity test was performed using the bacteria solution in Step 5. A sterile 8 x 8 96-well V-shaped reaction plate was used, and 14 μL of freshly prepared 2048000 μg / mL whitehead soup and 126 μL of bacteria solution were added to the first column of the first well, and 14 μL and 126 μL of bacteria solution were added to the second to fourth wells, respectively, and 14 μL of 1280 μg / mL ceftriaxone, amikacin, and tetracycline were added to the fifth to seventh wells, respectively, and 126 μL of bacteria solution containing 1 / 2 MIC whitehead soup (102400 μg / mL) was added, and 140 μL of MHB broth was added to the eighth well as a blank control. 70 μL of bacteria solution was added to each well in the second to twelfth columns, and the mixed bacteria and drug solution in the first column was serially diluted by a factor of two and added to the corresponding second column wells using an 8-way pipettor, and sequentially diluted by a factor of two to the twelfth column, and the 70 μL of mixed solution taken from the twelfth column was discarded. The concentration of whitehead soup used alone was 204800, 102400, 51200, 25600, 12800, 6400, 3200, 1600, 800, 400, 200, 100 μg / mL, respectively. The concentration of each antibacterial drug was 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 μg / mL, respectively. Thereafter, the 96-well drug sensitivity plate was incubated at 37°C for 12-16 h.

[0052] 7. Determination of MIC value

[0053] K. pneumoniae strains Kp20, Kp24, and Kp32 were tested using whitehead soup and antibacterial drugs, respectively. After the 96-well V-shaped reaction plate containing the bacteria was incubated at 37°C for 16-20 h, the MIC (minimum inhibitory concentration) results were read. The drug MIC value (minimum inhibitory concentration) was the drug concentration of the first clear transparent well before the turbid well in the 96-well plate. The above test was performed in triplicate, and when the MIC value could be accurately repeated, it was considered acceptable; when the MIC value differed by more than one concentration, a new test was required until the requirements were met. The results are shown in Tables 1, 2, and 3.

[0054] Table 1 MIC of whitehead soup combined with ceftriaxone for the test strains

[0055]

[0056]

[0057] Note: 1 / 2 MIC Baishuangan Decoction: Baishuangan Decoction added amount is 102400 μg / mL

[0058] As shown in Table 1, the MIC values of Baishuangan Decoction on strains Kp20, Kp24, Kp32 are all 204800 μg / mL, the MIC values of Ceftriaxone on strains Kp20, Kp24, Kp32 are 128, 64, 64 μg / mL respectively, when adding 1 / 2 MIC (102400 μg / mL) Baishuangan Decoction, the MIC of Ceftriaxone on Kp20, Kp24, Kp32 are 16, 8, 16 μg / mL respectively, compared with the MIC values when used alone, the MIC of Kp20, Kp24, Kp32 are decreased by 8, 8, 4 times respectively.

[0059] The results show that when Ceftriaxone is combined with Baishuangan Decoction, it can significantly enhance the antibacterial activity on Klebsiella pneumoniae Kp20, Kp24, Kp32.

[0060] Table 2 MIC of Baishuangan Decoction combined with Amikacin on test strains

[0061]

[0062] Note: 1 / 2 MIC Baishuangan Decoction: Baishuangan Decoction added amount is 102400 μg / mL

[0063] As shown in Table 2, the MIC values of Baishuangan Decoction on strains Kp20, Kp24, Kp32 are all 204800 μg / mL, the MIC values of Amikacin on strains Kp20, Kp24, Kp32 are 128, 64, 64 μg / mL respectively, when adding 1 / 2 MIC (102400 μg / mL) Baishuangan Decoction, the MIC of Amikacin on Kp20, Kp24, Kp32 are 8, 8, 16 μg / mL respectively, compared with the MIC values when used alone, the MIC of Kp20, Kp24, Kp32 are decreased by 16, 8, 4 times respectively. As shown in the results of Table 2, when Amikacin is combined with Baishuangan Decoction, it can significantly enhance the antibacterial activity of tetracycline on Klebsiella pneumoniae.

[0064] Table 3 MIC of Baishuangan Decoction combined with Tetracycline on test strains

[0065]

[0066] Note: 1 / 2 MIC Baishuangan Decoction: Baishuangan Decoction added amount is 102400 μg / mL

[0067] As shown in Table 3, the MIC values of Baishueng Decoction on strains Kp20, Kp24 and Kp32 were all 204800 μg / mL, and the MIC values of tetracycline on strains Kp20, Kp24 and Kp32 were 64, 32 and 64 μg / mL respectively. When 1 / 2 MIC (102400 μg / mL) of Baishueng Decoction was added, the MIC of tetracycline on Kp20, Kp24 and Kp32 were 8, 16 and 8 μg / mL respectively. Compared with the MIC values when tetracycline was used alone, the MIC of Kp20, Kp24 and Kp32 were decreased by 8, 2 and 8 times respectively. The results showed that when tetracycline was combined with Baishueng Decoction, the antibacterial activity of tetracycline on Klebsiella pneumoniae could be significantly enhanced, and the drug resistance of the drug-resistant bacteria to the antibacterial drug could be effectively reversed.

[0068] Example 2

[0069] Materials and methods

[0070] 1. Reagents:

[0071] The same as Example 1.

[0072] 2. Strains

[0073] The same as Example 1.

[0074] 3. Culture solution

[0075] The same as Example 1.

[0076] 4. Instruments

[0077] The same as Example 1.

[0078] 5. Preparation of bacterial solution

[0079] The same as Example 1.

[0080] 6. Preparation of drug sensitivity plate

[0081] On the basis of micro-broth dilution method, we obtained the MIC values of single drug of Pulsatilla decoction (PD), Ceftriaxone (CRO), Amikacin (AMK) and Tetracycline (TET), and selected the concentrations of Pulsatilla decoction (PD) and Ceftriaxone (CRO), Amikacin (AMK), Tetracycline (TET) at 8 times to 1 / 16 times MIC to place in the drug sensitivity chessboard. Specifically, 100 μL of MHB was distributed into 8x8 96-well plates, and then Pulsatilla decoction (PD) and Ceftriaxone (CRO), Amikacin (AMK), Tetracycline (TET) were diluted by 2 times along the horizontal and vertical coordinates, respectively, while the MIC of Pulsatilla decoction (PD), Ceftriaxone (CRO), Amikacin (AMK) and Tetracycline (TET) and negative and positive controls were made. The 96-well plates were placed in a constant temperature incubator at 37°C for 12-16 hours, and then incubated overnight. The results were observed and recorded. The test was repeated three times.

[0082] 7. Determination of FIC index

[0083] The fractional inhibitory concentration index (FICI) was used as the standard for the determination of the chessboard drug sensitivity test. The FICI of two compounds A and B was defined by the following formula:

[0084] FICI = FICA + FICB = MICAB / MICA + MICBA / MICB.

[0085] MICA is the MIC of compound A alone, MICAB is the MIC of compounds A and B, MICB is the MIC of compound B alone, and MICBA is the MIC of compounds B and A. FIC < 0.5 is synergistic, 0.5 ≤ FIC ≤ 1 is additive, 1 < FIC ≤ 2 is irrelevant, and FIC > 2 is antagonistic. The results are shown in Tables 4, 5 and 6.

[0086] Table 4 Combined antibacterial effect of Pulsatilla decoction and Ceftriaxone

[0087]

[0088] As shown in Table 4, the FIC index was 0.1875-0.375, all < 0.5, i.e. the combination of the two showed synergistic effect, and the results showed that the combination of Pulsatilla decoction and Ceftriaxone showed synergistic antibacterial effect on Ceftriaxone, and significantly enhanced the effect of Ceftriaxone on Klebsiella pneumoniae.

[0089] Table 5 Combined antibacterial effect of Pulsatilla decoction and Amikacin

[0090]

[0091] As shown in Table 5, the FIC index is 0.25-0.375, all <0.5, i.e. after combination, the two drugs present synergistic effect. The results show that the combination of Baishuangan Decoction and amikacin presents synergistic antibacterial effect on amikacin, and significantly enhances the effect of amikacin on K. pneumoniae.

[0092] Table 6 Synergistic antibacterial effect of combination of Baishuangan Decoction and tetracycline

[0093]

[0094] As shown in Table 6, the FIC index is 0.1875-0.3125, all <0.5, i.e. after combination, the two drugs present synergistic effect. The results show that the combination of Baishuangan Decoction and tetracycline presents synergistic antibacterial effect on tetracycline, and significantly enhances the antibacterial effect of tetracycline.

[0095] As described above, the combination of Baishuangan Decoction and ceftriaxone, amikacin, and tetracycline all presents significant synergistic effect, the FIC index is 0.1875-0.375, all <0.5, i.e. after combination, the two drugs present significant synergistic antibacterial effect. The results show that Baishuangan Decoction can significantly enhance the antibacterial activity of ceftriaxone, amikacin, and tetracycline on K. pneumoniae Kp20, Kp24, and Kp32. In the case of reduced dosage of ceftriaxone, amikacin, and tetracycline, the combination with Baishuangan Decoction can present synergistic effect, and improve the antibacterial effect of ceftriaxone, amikacin, and tetracycline.

[0096] Example 3

[0097] Materials and methods

[0098] 1. Reagents:

[0099] The same as Example 1.

[0100] 2. Strains

[0101] The same as Example 1.

[0102] 3. Culture solution

[0103] The same as Example 1.

[0104] 4. Instruments

[0105] The same as Example 1.

[0106] 5. Preparation of bacterial solution

[0107] The same as Example 1.

[0108] 6. Growth curve test

[0109] Klebsiella pneumoniae Kp20 was selected for growth curve test. 2.5 g LB broth medium was weighed by electronic balance and dissolved in a 250 mL conical flask containing 100 mL distilled water, which was autoclaved at 120℃ for 20 min, then taken out and dried, and placed in a cool place for standby. Three autoclaved conical flasks containing 100 mL LB broth were prepared, and the white head soup group was labeled 0 h, 1.5 h, 4 h, 6 h, 10 h, 15 h, 20 h, and 25 h, respectively. The final concentration of white head soup in the conical flask was 1 / 2 MIC, and the positive group was 1 / 2 MIC of the final concentration of the antibacterial drug, and then the bacterial liquid of logarithmic growth phase was inoculated and shaken to mix, diluted with LB to a total number of 1.0 x 10 5 CFU / mL, and 5 mL of bacterial liquid in the conical flask was transferred to each test tube, respectively, and cultured in a constant temperature water bath at 37℃ and 100 r / min. The corresponding test tubes were taken out at 0 h, 1.5 h, 4 h, 6 h, 10 h, 15 h, 20 h, and 25 h, respectively, and placed in a 4℃ refrigerator. After all the test tubes were taken out, the OD value was detected at 600 nm in a multifunctional enzyme marker.

[0110] 7. Growth curve drawing

[0111] The growth curve change of white head soup on strain Kp20 is shown in Figure 1 As shown in Figure 1 The results showed that white head soup showed an upward trend on Klebsiella pneumoniae Kp20 within 6 h, and tended to stable growth after 15 h. The results showed that white head soup alone had almost no inhibitory effect on the growth of Klebsiella pneumoniae.

Claims

1. The use of Baishouweng Decoction in the preparation of an amikacin antibacterial effect potentiator for Klebsiella pneumoniae KP20, characterized in that, The amount of amikacin is 8 μg / mL, corresponding to the amount of Baitouweng decoction is 102400 μg / mL.

2. Use according to claim 1, characterized in that, The amount of amikacin is 128 μg / mL when amikacin is used alone, and the amount of Baitouweng decoction is 102400 μg / mL and the amount of amikacin is 8 μg / mL when Baitouweng decoction and amikacin are used simultaneously.

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