Antibacterial peptide composition for synergistically inhibiting pseudomonas aeruginosa and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and discloses an antibacterial peptide composition for synergistically inhibiting pseudomonas aeruginosa and application thereof. The antibacterial peptide composition is composed of an antibiotic and an antibacterial peptide with an amino acid sequence as shown in SEQ ID NO.1. The antibiotic and the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.1 have a synergistic effect, so that the effect of inhibiting proliferation of the multi-drug-resistant pseudomonas aeruginosa is remarkably improved, the effect of inhibiting the forming ability of a pseudomonas aeruginosa biofilm is shown, the development of drug resistance is slowed down, and the antibacterial peptide has the characteristics of high synthesis efficiency and low production cost and is suitable for industrial production. An efficient and safe anti-infection treatment scheme is provided for clinical treatment and overcoming of the problem that traditional antibiotics are increasingly serious in drug resistance, and great clinical application potential is achieved for immunocompromised patients.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and specifically relates to an antimicrobial peptide composition that synergistically inhibits Pseudomonas aeruginosa and its application. Background Technology

[0002] Pseudomonas aeruginosa (P.) aeruginosa *Clostridium difficile* is a Gram-negative opportunistic pathogen that coexists in both the environment and the human body, exhibiting significant genetic diversity and environmental adaptability. In recent years, multidrug-resistant and extensively drug-resistant strains have become increasingly prevalent in clinical isolates, demonstrating high tolerance to multiple antibiotics. This is primarily due to the synergistic effects of multiple factors, including enhanced outer membrane barrier, activated efflux pump system, biofilm maturation, and the production of antibiotic-degrading enzymes. These "clinically variant" strains can cause severe infections of the lungs, wounds, and urinary tract, significantly increasing treatment complexity and posing a major threat to patient survival and healthcare resources.

[0003] Antibiotics have played a decisive role in the treatment of infections, but drug resistance has become a core global challenge. As pathogens continue to evolve, many traditional drugs are gradually losing their effectiveness, while the development of new antibiotics lags far behind the emergence of drug-resistant bacteria, making treatment options increasingly limited. Therefore, finding novel antibacterial strategies that can replace or complement antibiotics is urgently needed.

[0004] Antimicrobial peptides (AMPs) are important small-molecule functional peptides in the body's innate immunity, possessing advantages such as broad-spectrum antibacterial activity, rapid onset of action, and low resistance induction. They exhibit significant activity against drug-resistant pathogens such as Pseudomonas aeruginosa. AMPs achieve bactericidal effects through multiple pathways, including membrane disruption, inhibition of biofilm formation, and interference with key intracellular processes. However, natural antimicrobial peptides still have certain shortcomings in terms of stability and potential cytotoxicity, limiting their direct use as drugs. Novel antimicrobial peptides based on structural modification and computational design are gradually becoming an effective direction for solving these problems.

[0005] Combination therapy is a key strategy for combating drug-resistant bacteria. Under the premise of complementary mechanisms, the combined treatment of antibiotics and antimicrobial peptides can produce a synergistic antibacterial effect, not only improving bactericidal efficiency but also reducing individual dosages, minimizing toxic side effects, and delaying the development of drug resistance. However, combinations that can stably achieve synergistic effects remain limited, especially against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Therefore, the systematic development of antimicrobial peptide compositions with synergistic activity and the validation of their application value in refractory infections are of great significance for overcoming current bottlenecks in anti-infective therapy. Summary of the Invention

[0006] The primary objective of this invention is to address the shortcomings of existing technologies by providing an antimicrobial peptide composition that synergistically inhibits Pseudomonas aeruginosa. This antimicrobial peptide composition combines antibiotics with antimicrobial peptides, significantly enhancing antimicrobial efficacy and slowing the development of drug resistance, thus offering a solution to the crisis of drug-resistant bacteria caused by antibiotic overuse.

[0007] Another object of the present invention is to provide the application of the above-mentioned antimicrobial peptide composition that synergistically inhibits Pseudomonas aeruginosa.

[0008] The objective of this invention is achieved through the following technical solution: An antimicrobial peptide composition for synergistic inhibition of Pseudomonas aeruginosa, comprising an antibiotic and an antimicrobial peptide; preferably, the antibiotic and the antimicrobial peptide are composed in a mass ratio of 2-64:1-4.

[0009] The amino acid sequence of the antimicrobial peptide is shown below: VKTGNRAKVKKVIKGLKKASKLHAGQAKTLKKVLGNGKKKRSKSRN.

[0010] The antibiotic is at least one of ciprofloxacin (CIP), amikacin (AMK), aztreonam (ATM), levofloxacin (LEVO), cefepime (FEB), tobramycin (TOB), meropenem (MEM), and ceftazidime (CAZ); preferably at least one of aztreonam (ATM), levofloxacin (LEVO), cefepime (FEB), meropenem (MEM), and ceftazidime (CAZ).

[0011] The *Pseudomonas aeruginosa* is preferably a multidrug-resistant *Pseudomonas aeruginosa*; more preferably, it is at least one of *Pseudomonas aeruginosa* 19-89, *Pseudomonas aeruginosa* 19-130, *Pseudomonas aeruginosa* 3-2, and *Pseudomonas aeruginosa* 19-129.

[0012] Preferably, when the antibiotic is aztreonam, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-89, Pseudomonas aeruginosa 19-130, or Pseudomonas aeruginosa 3-2; the antibiotic and the antimicrobial peptide are mixed in a mass ratio of 4-64:1-4.

[0013] When the antibiotic is levofloxacin, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-89; the antibiotic and the antimicrobial peptide are mixed in a mass ratio of 8:4.

[0014] When the antibiotic is cefepime, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130 or Pseudomonas aeruginosa 19-129; the antibiotic and the antimicrobial peptide are mixed in a mass ratio of 2:4.

[0015] When the antibiotic is meropenem, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130; the antibiotic and the antimicrobial peptide are mixed in a mass ratio of 4:4.

[0016] When the antibiotic is ceftazidime, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130 or Pseudomonas aeruginosa 3-2; the antibiotic and the antimicrobial peptide are mixed in a mass ratio of 2:2 to 4.

[0017] The above-mentioned antimicrobial peptide composition that synergistically inhibits Pseudomonas aeruginosa is used in the preparation of anti-Pseudomonas aeruginosa formulations.

[0018] The term "anti-Pseudomonas aeruginosa" refers to inhibiting the growth of Pseudomonas aeruginosa and inhibiting the formation of Pseudomonas aeruginosa biofilm.

[0019] Implementing this invention has the following beneficial effects: This invention provides an innovative antimicrobial peptide and antibiotic composition that significantly enhances the inhibition of multidrug-resistant Pseudomonas aeruginosa proliferation through synergistic effects. It exhibits an inhibitory effect on the biofilm formation ability of Pseudomonas aeruginosa, slowing down the development of drug resistance. It features high synthesis efficiency and low production cost, providing a highly efficient and safe anti-infective treatment option for clinical treatment and overcoming the increasingly serious problem of traditional antibiotic resistance. It has great clinical application potential for patients with weakened immune function. Attached Figure Description

[0020] Figure 1 This is a chromatogram of the HPLC report results for chemically synthesized antimicrobial peptides.

[0021] Figure 2 This is a graph showing the MS report results of chemically synthesized antimicrobial peptides.

[0022] Figure 3 This is a graph showing the MIC test results of the tested pathogenic microorganisms against commonly used antibiotics.

[0023] Figure 4 In vitro bactericidal kinetic curves of the corresponding antibiotics, antimicrobial peptide PsCDS-1, and the combination of the two against multidrug-resistant Pseudomonas aeruginosa 19-89.

[0024] Figure 5 The resistance development curves for multidrug-resistant PsCDS-19-89 using corresponding antibiotics, antimicrobial peptide PsCDS-1, and the combination of the two.

[0025] Figure 6 The figure shows the effect of using corresponding antibiotics, antimicrobial peptide PsCDS-1, and the combination of the two on the biofilm formation ability of multidrug-resistant Pseudomonas aeruginosa 19-89. Detailed Implementation

[0026] The present invention will be further described in detail below with reference to embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto. It should be noted that any processes not specifically described in detail below are those that can be implemented or understood by those skilled in the art by referring to existing technology. Reagents or instruments whose manufacturers are not specified are considered to be conventional products that can be purchased commercially.

[0027] Example 1: Conventional Peptide Solid-Phase Synthesis of Antimicrobial Peptide PsCDS-1 The antimicrobial peptide PsCDS-1 of this invention is synthesized using conventional solid-phase peptide synthesis, and its sequence is: VKTGNRAKVKKVIKGLKKASKLHAGQAKTLKKVLGNGKKKRSKSRN.

[0028] Sequence characteristics: The sequence type is an amino acid sequence containing 46 amino acid residues, with a molecular weight of 5012.062 Da and a net charge of 17.509.

[0029] The specific synthesis steps of the antimicrobial peptide PsCDS-1 are as follows: (1) Resin swelling: Weigh 2-chlorotriphenylmethyl chloride resin (2ChlorotritylChloride Resin, SUNRESIN Company, China, catalog number LXSS03-1-1204) with a degree of substitution of 0.1 mmol, put the resin into the reaction tube, add DCM (dichloromethane) solvent, the amount of DCM added is calculated as 15 mL / g resin, and shake for 40 min.

[0030] (2) Add the first amino acid: filter out the DCM solvent by sand core, add 3 times the resin mass of Fmoc-Arg(Pbf)-OH amino acid, then add 1 times the resin mass of 2,4,6-trimethylpyridine, and finally add DCM (15 mL / g, i.e., the amount added is calculated based on 15 mL / g resin) to dissolve, shake for 12 h, and then wash 3 times with DMF (dimethylformamide).

[0031] (3) Deprotection: Add 10 mL of 20% v / v piperidine DMF solution, the amount of piperidine DMF solution added is calculated as 15 mL / g resin, shake for 6 min and then remove the piperidine DMF solution, add another 10 mL of 20% v / v piperidine DMF solution, the amount of piperidine DMF solution added is calculated as 15 mL / g resin, shake for 20 min.

[0032] (4) Detection: Remove the piperidine DMF solution, take a dozen resin grains, wash them three times with ethanol, add one drop each of ninhydrin and phenol solution, heat at 105°C~110°C for 3 minutes, and a deep blue color indicates a positive reaction.

[0033] (5) First cleaning: Clean with DMF 3 times in sequence, with the amount of DMF used each time calculated as 10 mL / g resin.

[0034] (6) Sealing: Add methanol (the amount added is calculated based on 10 mL / g resin) and DIEA (N,N diisopropylethylamine) at 3 times the resin mass, shake for 20 min, and then wash with DMF 3 times.

[0035] (7) Condensation (to produce the second amino acid Lysine in the sequence): Add 3 times the resin mass of Fmoc-Lys(boc)-OH amino acid, 2.7 times the resin mass of HCTU (6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate), and 10 times the resin mass of DIEA (N,N-diisopropylethylamine) to the reaction tube, dissolve in 5 mL of DMF, and react for 1 h.

[0036] (8) Second wash: Wash three times with DMF (the amount added is calculated based on 10 mL / g resin) and repeat the above operation, linking the amino acids in the sequence from left to right.

[0037] (9) After the last amino acid is ligated, the protection is removed and the resin is washed as follows: DMF (10 mL / g resin) twice, DCM (10 mL / g resin) twice, and then dried for 8 min.

[0038] (10) Cutting peptides from resin: The amount of cutting solution is calculated at 10 mL / g resin, and the solution is shaken at a constant temperature for 150 min. (The cutting solution is prepared by volume percentage and the composition is as follows: TFA (trifluoroacetic acid) 95% v / v, water 2.5% v / v, TIS (triisopropylsilane) 2.5% v / v.

[0039] (11) Drying and washing: Dry the lysis buffer as much as possible with nitrogen gas, precipitate it with ether, wash it with ether six times, and then evaporate it at room temperature to obtain crude peptide.

[0040] (12) HPLC-MS analysis and identification of peptides: A. Prepare a solution with a concentration of 1 mg / mL using crude peptide; B. Filter the solution using a 0.45 μm filter membrane; C. Analysis: Analyze 10 μL of the solution using HPLC-MS. The mobile phase is water and acetonitrile, the time is 25 min, isocratic elution is performed, and the HPLC is equilibrated with an isocratic gradient for 5 min before injection. The gradient is 40% v / v water and 60% v / v acetonitrile.

[0041] The purified solution was freeze-dried to obtain a white powdery polypeptide, which was then sealed and stored at -20°C.

[0042] HPLC report such as Figure 1 As shown, the MS test report is as follows: Figure 2As shown in the figure, the target polypeptide with a purity greater than 90% was prepared.

[0043] Example 2: Determination of the minimum inhibitory concentration and minimum bactericidal concentration of the antimicrobial peptide PsCDS-1 against clinical isolates The multidrug-resistant Pseudomonas aeruginosa in this invention are all from the South China University of Technology-Finch Biotechnology Microbial Coating Joint Laboratory.

[0044] The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of antimicrobial peptides were determined according to the methods developed by the Clinical and Laboratory Standards Institute (CLSI), with slight modifications as needed.

[0045] The specific implementation involved diluting the candidate antimicrobial peptide with a bacterial suspension cultured to the logarithmic growth phase in a 96-well microplate using Mueller-Hinton broth to achieve a final bacterial suspension concentration of 5 × 10⁻⁶. 5 CFU / mL, add 90 μL of bacterial suspension to each well, then add 10 μL of PsCDS-1 serially diluted 2-fold and mix thoroughly to achieve a final concentration range of 128 to 0.25 μg / mL. PBS was used as a negative control. Each experiment was repeated three times, and the average value was taken. The mixture was incubated at 37°C for 16–20 hours until visibly turbid bacterial suspension appeared in the negative control wells. The absorbance at 600 nm was measured using a microplate reader. The MIC was defined as the lowest peptide concentration at which no bacterial growth was observed. Subsequently, 10 μL of the bacterial suspension with no obvious bacterial growth was inoculated onto MH agar plates and incubated at 37°C for 16 h. MBC was defined as the lowest peptide concentration at which more than 99.9% of bacteria were killed. The results are shown in Table 1.

[0046] Table 1. MIC and MBC of antimicrobial peptide PsCDS-1 against tested pathogenic microorganisms

[0047] As shown in Table 1, the antimicrobial peptide PsCDS-1 exhibits good antibacterial activity against multidrug-resistant Pseudomonas aeruginosa 19-89, 19-130, 19-1293-2, 19-126, 19-144, 19-163, and 12077, especially showing significant inhibitory effects against 3-2, 19-126, 19-144, 19-163, and 12077.

[0048] Example 3: Determination of Fractional Inhibitory Concentration (FIC) and Fractional Inhibitory Concentration Index (FICI) for Combined Drug Use According to the Clinical and Laboratory Standards Institute (CLSI) guidelines and recommendations for the selection of antimicrobial agents against Pseudomonas aeruginosa, the antibiotics used in this embodiment include ciprofloxacin (CIP), amikacin (AMK), aztreonam (ATM), levofloxacin (LEVO), cefepime (FEB), tobramycin (TOB), meropenem (MEM), and ceftazidime (CAZ). The pathogen resistance phenotype results in this invention are as follows... Figure 3 As shown, the MIC measurement method is the same as in Example 2.

[0049] This embodiment uses a checkerboard method in 96-well plates to study the synergistic effect between PsCDS-1 and antibiotics. Specifically, the single-drug MIC of the antibiotics was determined using MH broth in a system completely consistent with the conditions described below. Subsequently, in the combination experiments, serially diluted PsCDS-1 was arranged vertically in two-fold increments, with the concentrations of the antibiotics and antimicrobial peptides set at 2–0.125 times their respective single-drug MICs. Then, the concentration of the Pseudomonas aeruginosa suspension cultured to the logarithmic growth phase was adjusted to 10 using fresh MH broth. 5 CFU / mL and mixed thoroughly with the antibiotic / antimicrobial peptide mixture. Set up a PBS-mixed bacterial suspension as a blank control.

[0050] After incubating 96-well microplates at 37°C for 16–20 hours, the absorbance at 600 nm was measured using a microplate reader. The concentration corresponding to each well where no bacterial growth was observed was defined as the MIC of the combined drug. Interactions between antimicrobial agents were determined by calculating the partial inhibitory concentration index (FICI), as shown in the following formula:

[0051] MIC A / （A+B） This indicates the MIC (Minimum Intake) of drug A in the combination test. A Indicates the MIC of drug A in independent trials; MIC B / （A+B） This indicates the MIC (Minimum Intake) of drug B in the combination trial. B This indicates the MIC of drug B in an independent trial.

[0052] The evaluation criteria for the combined use of PsCDS-1 and antibiotics are as follows: when FICI ≤ 0.5, the combination is considered synergistic; when 0.5 < FICI ≤ 1, it is considered a partial enhancing effect (additive effect); when 1 < FICI ≤ 4, it is considered no difference or additive effect; when FICI > 4, it is considered an antagonistic effect.

[0053] Table 2 shows the MIC and FICI of PsCDS-1 in combination with antibiotics, both as a single agent and in combination. As shown in Table 2, the combination of the antimicrobial peptide PsCDS-1 with multiple antibiotics can produce synergistic or enhancing bactericidal effects against multidrug-resistant Pseudomonas aeruginosa. Specifically, the combination of PsCDS-1 with aztreonam, levofloxacin, cefepime, and meropenem shows synergistic or enhancing effects against various multidrug-resistant Pseudomonas aeruginosa strains. Most combinations exhibit enhancing effects, which in turn alter the MIC values ​​of resistant strains, reducing them from the resistant range to the sensitive range. Through these combinations, multidrug-resistant bacteria can regain their inhibitory effect against antibiotics that have already developed resistance.

[0054] According to the results in Table 2, the optimal synergistic combination was the antibacterial effect of PsCDS-1 and levofloxacin (LEVO) against 19-89. Based on the CLSI antimicrobial susceptibility testing performance standards, 19-89 showed resistance to all eight antibiotics used. The combination of levofloxacin and PsCDS-1 significantly enhanced the antibacterial effect against 19-89 and exhibited a synergistic effect. Analysis revealed that levofloxacin, a broad-spectrum fluoroquinolone bactericide, primarily inhibits bacterial type II topoisomerase, stabilizing DNA at the intermediate stage of cleavage and rejoining and inducing necrotic DNA damage, thereby blocking DNA replication and transcription. PsCDS-1, on the other hand, primarily acts by disrupting the cell membrane. The complementary mechanisms of their target mechanisms may be one reason for the synergistic effect of their combination.

[0055] Table 2 Results of PsCDS-1 combined with antibiotics

[0056] Example 4: Bactericidal kinetics of combined drug use Using 19-89 as the pathogen, the bactericidal kinetic curves of antibiotics and antimicrobial peptide PsCDS-1 used alone and in combination were measured to reflect the combined effect. In this example, levofloxacin was selected as the test antibiotic.

[0057] The concentration of the activated Pseudomonas aeruginosa suspension cultured to the logarithmic growth phase was adjusted to 10 using fresh MH broth. 5The concentration of CFU / mL was determined, and the baseline (T0) was set from the initial sample count before drug administration. The samples were then mixed with the antimicrobial agents. The antimicrobial agents were PsCDS-1 (64 μg / mL), levofloxacin (16 μg / mL), and PsCDS-1 + levofloxacin. The final concentrations for the combination groups were 1×MIC (PsCDS-1 2 μg / mL, levofloxacin 4 μg / mL) or 2×MIC (PsCDS-1 4 μg / mL, levofloxacin 8 μg / mL). A PBS-treated bacterial suspension served as a blank control. The mixture was incubated at 37°C, and samples were taken at different time points. After appropriate dilution, the samples were spotted onto MH agar plates and incubated at 37°C before colony counting.

[0058] The results are as follows Figure 4 Compared with the control group, the bacterial concentrations of the antimicrobial peptide PsCDS-1 group, levofloxacin group, and combination group were significantly reduced, showing a significant inhibitory effect on pathogenic microorganisms. Moreover, the bacterial concentrations of the PsCDS-1 group and the combination group were close to 0 after 30 min of incubation, indicating that PsCDS-1 and the combination group have rapid bactericidal activity, which may enable them to quickly destroy bacterial cells and reduce the possibility of bacterial adaptation or the development of drug resistance. This may be of great significance in various application scenarios.

[0059] Example 5: Drug resistance development experiment with combination therapy Using the method described by Kim et al. with appropriate modifications, and with 19-89 as the test pathogen, the development of resistance to PsCDS-1 and levofloxacin, alone and in combination, was assessed using a continuous culture method. Specifically, the initial MIC was determined using the same measurement protocol as for the minimum inhibitory concentration (MIC). Then, based on the MIC results, bacterial suspensions from wells at sub-inhibitory concentrations were taken, and the next generation MIC was determined using the same method. This process was repeated up to the 10th generation to obtain the MIC versus passage development curve.

[0060] Figure 5 The development of resistance to levofloxacin, antimicrobial peptide PsCDS-1, and the combination of the two was statistically analyzed. After 10 generations of culture, the minimum inhibitory concentration of levofloxacin against the tested pathogens increased by about 8 times. The MIC values ​​of PsCDS-1 and the combination group remained stable, fluctuating only within the range of 1-3 times. Figure 5 As can be seen, the minimum inhibitory concentration (MIC) of the levofloxacin group increased very rapidly, indicating that drug resistance developed quickly. The MIC of the antimicrobial peptide PsCDS-1 group and the combination group increased less, and remained at a low level within ten generations. This suggests that the combination of antimicrobial peptide PsCDS-1 and levofloxacin can significantly slow down the development of bacterial resistance by reducing the concentration of antibiotics used. This is of great significance for inhibiting the occurrence and development of drug-resistant bacteria.

[0061] Example 6: Drug combination therapy to inhibit biofilm formation The effects of antimicrobial peptides PsCDS-1 and levofloxacin, used alone and in combination, on the biofilm formation ability of 19-89 bacteria were investigated using the crystal violet biofilm staining method. Activated clinical strains of *Pseudomonas aeruginosa* were inoculated into culture media supplemented with PsCDS-1, levofloxacin, and the combined drug, respectively, at final concentrations of 1×MIC, 0.5×MIC, and 0.25×MIC. The cultures were incubated at 37°C until biofilm maturation. The plates were washed five times with PBS solution, air-dried, and then 200 μL of anhydrous methanol was added to each well. The plates were incubated at room temperature for 15 minutes to fix the biofilm. The plates were stained with 1% crystal violet at room temperature for 10 minutes, and excess staining solution was washed away with deionized water. The staining was then decolorized with anhydrous ethanol, and the absorbance was measured at 590 nm. PBS was used as a control group instead of the antimicrobial agents.

[0062] Modified according to the method of Stepanovic et al., based on OD. 590 Classification of Pseudomonas aeruginosa's biofilm-forming ability: Critical OD (ODc) = Mean value of negative control group OD0 + 3 SD; Relative value (SI) = Average OD of biofilm / ODc; When 0 < SI ≤ 2, the biofilm formation ability of the strain is weak; when 2 < SI ≤ 4, the biofilm formation ability is moderate; when SI > 4, it indicates that the biofilm formation ability is strong.

[0063] Biofilm formation as follows Figure 6 As shown, strain 19-89 is a *Pseudomonas aeruginosa* mutant with strong biofilm formation ability, while the control group maintained a biofilm formation ability of SI ≥ 4. Levofloxacin, PsCDS-1, and the combination therapy all showed biofilm inhibition. The PsCDS-1 and LEV+PsCDS-1 groups showed significantly reduced biofilm formation ability compared to the control and levofloxacin groups, with inhibition exceeding 50%, suppressing biofilm formation to an SI < 2 level. Furthermore, the combination therapy group maintained a strong inhibitory effect even at a concentration of 0.25 × MIC. These results indicate that both levofloxacin and the antimicrobial peptide PsCDS-1 can inhibit biofilm formation in *Pseudomonas aeruginosa*, but the combined use of the two has a more significant inhibitory effect, and it can still exert an inhibitory effect at lower concentrations.

[0064] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A synergistic antibacterial peptide composition for inhibiting Pseudomonas aeruginosa, characterized by: The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition comprises an antibiotic and an antibacterial peptide; The amino acid sequence of the antibacterial peptide is shown as SEQ ID NO.

1. The antibiotic is at least one of ciprofloxacin, amikacin, aztreonam, levofloxacin, cefepime, tobramycin, meropenem and ceftazidime.

2. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 1, wherein: The Pseudomonas aeruginosa is multidrug-resistant Pseudomonas aeruginosa. The antibiotic is at least one of aztreonam, levofloxacin, cefepime, meropenem and ceftazidime.

3. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 2, wherein: The Pseudomonas aeruginosa is at least one of Pseudomonas aeruginosa 19-89, Pseudomonas aeruginosa 19-130, Pseudomonas aeruginosa 3-2 and Pseudomonas aeruginosa 19-129.

4. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 3, wherein: When the antibiotic is aztreonam, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-89, Pseudomonas aeruginosa 19-130 or Pseudomonas aeruginosa 3-2.

5. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 3, wherein: When the antibiotic is levofloxacin, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-89.

6. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 3, wherein: When the antibiotic is cefepime, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130 or Pseudomonas aeruginosa 19-129.

7. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 3, wherein: When the antibiotic is meropenem, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130.

8. The synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to claim 3, wherein: When the antibiotic is ceftazidime, the Pseudomonas aeruginosa is Pseudomonas aeruginosa 19-130 or Pseudomonas aeruginosa 3-2.

9. Use of the synergistically inhibiting Pseudomonas aeruginosa antibacterial peptide composition according to any one of claims 1-8 in the preparation of an anti-Pseudomonas aeruginosa preparation.

10. Use according to claim 9, characterized in that: The anti-Pseudomonas aeruginosa refers to inhibiting the growth of Pseudomonas aeruginosa and inhibiting the formation of Pseudomonas aeruginosa biofilm.

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