Application of paclitaxel in enhancing antibacterial effect of tigecycline

Abstract

The invention relates to a novel application of paclitaxel in enhancing the antibacterial effect of tigecycline antibiotics. When the composition or the compound preparation is prepared, paclitaxel is used for enhancing the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is (16: 1)-(256: 1). The paclitaxel and the tigecycline are combined to show a remarkable synergistic effect, and the FICI is 0.1875-0.3125 (both less than 0.5), namely the paclitaxel and the tigecycline are combined to show the remarkable synergistic effect. The use amount of tigecycline can be effectively reduced by using the combined paclitaxel, and the use amount of tigecycline independently used is 1-16 times higher than that of the combined use of paclitaxel and tigecycline.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a new use of paclitaxel as a tigecycline synergist. Background Art

[0002] Tigecycline (Tigecycline TGC) belongs to a new generation of antibiotics, which is a glycylcycline antibiotic. It was approved for marketing in the United States in 2005 and in my country in early 2012. The main mechanism of action of tigecycline is similar to that of other antibiotics. It is through reversible binding to the helical region (H34) on the 30S subunit of the bacterial ribosome to prevent the entry of tRNA and inhibit bacterial protein translation (i.e., the extension of the peptide chain), which ultimately blocks bacterial protein synthesis and restricts bacterial growth. However, due to the addition of an N-alkyl-glycylamino side chain to the C9 position of the D ring of tigecycline, its ability to bind to the ribosome is enhanced, and its antibacterial spectrum is expanded. Its ability to inhibit protein synthesis is 3 to 20 times that of other drugs. Tigecycline can effectively overcome the main mechanisms of antibiotic resistance, such as the specific efflux pump Tet (A) and the ribosome protection protein Tet (M).

[0003] The anticancer drug Paclitaxel (PTX) has a molecular formula of C47H51NO14, a molecular weight of 853.91 g / mol, a melting point of 213°C, and is a white crystalline powder. It is a tricyclic diterpene structure and a taxane chemotherapy product belonging to the taxane class. It exists naturally in the bark and needles of Taxus brevifolia and has high efficiency, low toxicity, and a broad spectrum of effects. The structural formula of Paclitaxel is shown below:

[0004]

[0005] In recent years, with the increase in the use of tigecycline and the occurrence of abuse of other antibiotics, the therapeutic effect of tigecycline has decreased relatively, and tigecycline resistance has emerged. According to the "EUCAST EU Drug Susceptibility Test Standard", when the MIC value of tigecycline against Enterobacteriaceae is 4μg / mL, the bacteria show resistance. At present, tigecycline resistance is showing a significant upward trend. Therefore, it is of great clinical significance to reduce the occurrence of its resistance peak through synergists. So far, there has been no research on using paclitaxel as a synergist for tigecycline. Summary of the invention

[0006] The present invention aims to provide a new use of paclitaxel in enhancing the antibacterial effect of an antibacterial agent. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of an antibacterial agent, and the amount of paclitaxel is 128 μg / mL, and the amount of tigecycline is 4 μg / mL or 8 μg / mL. Preferably, when the amount of paclitaxel is 128 μg / mL, the amount of tigecycline is 4-8 μg / mL.

[0007] A new use of paclitaxel in enhancing the antibacterial effect of an antibacterial agent. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 16:1 or 32:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 16:1 and 32:1.

[0008] A new use of paclitaxel in enhancing the antibacterial effect of an antibacterial agent, when preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 64:1, or 128:1, or 256:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 64:1-256:1.

[0009] A new use of paclitaxel in enhancing the antibacterial effect of an antibacterial agent. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline. When the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1 μg / mL, or 2 μg / mL, or 4 μg / mL. Preferably, when the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1-4 μg / mL.

[0010] A new use of paclitaxel in enhancing the antibacterial effect of antibacterial agents. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is between 16:1 and 256:1.

[0011] Another object of the present invention is to provide a new use of paclitaxel in enhancing the antibacterial effect of tigecycline, wherein the antibacterial includes anti-Escherichia coli and Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of the antibacterial agent, and the dosage of paclitaxel is 128 μg / mL, and the dosage of tigecycline is 4 μg / mL or 8 μg / mL. Preferably, when the dosage of paclitaxel is 128 μg / mL, the dosage of tigecycline is 4-8 μg / mL. A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Escherichia coli and Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is 16:1 or 32:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is between 16:1-32:1.

[0012] A novel use of paclitaxel in enhancing the antibacterial effect of tigecycline against Escherichia coli and Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline. When the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1 μg / mL, or 2 μg / mL, or 4 μg / mL. Preferably, when the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1-4 μg / mL.

[0013] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Escherichia coli and Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is 16:1, or 32:1, or 64:1, or 128:1, or 256:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline against Escherichia coli and Klebsiella pneumoniae, and the dosage ratio of paclitaxel to tigecycline is between 64:1-256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-128:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 128:1-256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 16:1-32:1.

[0014] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Escherichia coli and Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is between 16:1 and 256:1.

[0015] Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage of tigecycline is 4-8 μg / mL and the dosage of paclitaxel is 128 μg / mL. Preferably, the dosage of tigecycline is 1-4 μg / mL and the dosage of paclitaxel is 256 μg / mL.

[0016] Another object of the present invention is to provide a new use of paclitaxel in enhancing the anti-Escherichia coli effect of tigecycline. In the preparation of a composition or a compound preparation, paclitaxel is used to enhance the anti-Escherichia coli effect of tigecycline, and the amount of paclitaxel is 128 μg / mL, and the amount of tigecycline is 4 μg / mL or 8 μg / mL. Preferably, when the amount of paclitaxel is 128 μg / mL, the amount of tigecycline is 4-8 μg / mL.

[0017] The present invention provides a new use of paclitaxel in enhancing the anti-Escherichia coli effect of tigecycline. The anti-Escherichia coli bacterial strains include strains ZZ9DT16R, ZC9CTR2, and JXCE34. Paclitaxel is used to enhance the anti-Escherichia coli effect of tigecycline against Escherichia coli strains ZZ9DT16R, ZC9CTR2, and JXCE34. The dosage of paclitaxel is 128 μg / mL, and the dosage of tigecycline is 4 μg / mL or 8 μg / mL. Preferably, when the dosage of paclitaxel is 128 μg / mL, the dosage of tigecycline is 4-8 μg / mL. Preferably, the dosage ratio of paclitaxel to tigecycline is 16:1 or 32:1; preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the dosage ratio of paclitaxel to tigecycline is between 16:1-32:1.

[0018] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 16:1 or 32:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 16:1 and 32:1.

[0019] A new use of paclitaxel in enhancing the anti-Escherichia coli effect of tigecycline. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline. When the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1 μg / mL, or 2 μg / mL, or 4 μg / mL. Preferably, when the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 1-4 μg / mL.

[0020] A new use of paclitaxel in enhancing the anti-Escherichia coli effect of tigecycline. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 64:1, 128:1, or 256:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 64:1-256:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 64:1-128:1. Preferably, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is between 128:1-256:1.

[0021] Another object of the present invention is to provide a new use of paclitaxel in enhancing the effect of tigecycline against Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the effect of tigecycline against Klebsiella pneumoniae, and the amount of paclitaxel is 512 μg / mL, and the amount of tigecycline is 4 μg / mL. Preferably, when the amount of paclitaxel is 128 μg / mL, the amount of tigecycline is 4 μg / mL.

[0022] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 32:1 or 64:1. Preferably, when paclitaxel is used to enhance the antibacterial effect of tigecycline, the ratio of paclitaxel to tigecycline is between 32:1 and 64:1.

[0023] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline. When the amount of paclitaxel is 256 μg / mL, the amount of tigecycline is 4 μg / mL. Preferably, when the amount of paclitaxel is 128 μg / mL, the amount of tigecycline is 4 μg / mL.

[0024] A new use of paclitaxel in enhancing the antibacterial effect of tigecycline against Klebsiella pneumoniae. When preparing a composition or a compound preparation, paclitaxel is used to enhance the antibacterial effect of tigecycline, and the ratio of paclitaxel to tigecycline is 64:1 or 32:1. Preferably, when paclitaxel is used to enhance the antibacterial effect of tigecycline, the ratio of paclitaxel to tigecycline is between 32:1 and 64:1.

[0025] Another object of the present invention is to provide a use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on bacteria, wherein the ratio of paclitaxel to tigecycline is 16:1 or 32:1, or 64:1. Preferably, the ratio of paclitaxel to tigecycline is between 16:1-64:1; preferably, the ratio of paclitaxel to tigecycline is between 32:1-64:1.

[0026] The invention discloses an application of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on bacteria, wherein the dosage ratio of paclitaxel to tigecycline is 32:1, 64:1, 128:1 or 256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 32:1 and 256:1.

[0027] Another object of the present invention is to provide a use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli and Klebsiella pneumoniae, wherein the ratio of paclitaxel to tigecycline is 16:1 or 32:1, or 64:1. Preferably, the ratio of paclitaxel to tigecycline is between 16:1-64:1; preferably, the ratio of paclitaxel to tigecycline is between 32:1-64:1.

[0028] The use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli and Klebsiella pneumoniae, wherein the Escherichia coli strains are ZZ9DT16R, ZC9CTR2, JXCE34, and the Klebsiella pneumoniae strain is HC139, wherein the dosage ratio of paclitaxel to tigecycline is 32:1, or 64:1, or 128:1, or 256:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-256:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-128:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 128:1-256:1. The use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli and Klebsiella pneumoniae, wherein the Escherichia coli strains are ZZ9DT16R, ZC9CTR2, and JXCE34, and the Klebsiella pneumoniae strain is HC139, wherein the dosage ratio of paclitaxel to tigecycline is 16:1, 32:1, or 64:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 16:1-64:1, or the dosage ratio of paclitaxel to tigecycline is between 16:1-32:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 32:1-64:1.

[0029] The invention discloses an application of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli and Klebsiella pneumoniae, wherein the dosage ratio of paclitaxel to tigecycline is 16:1, 32:1, 64:1, 128:1 or 256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 16:1 and 256:1; preferably, the dosage ratio of paclitaxel to tigecycline is between 32:1 and 256:1.

[0030] Another object of the present invention is to provide a use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli, wherein the ratio of paclitaxel to tigecycline is 16:1 or 32:1. Preferably, the ratio of paclitaxel to tigecycline is between 16:1-32:1. The use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli, the Escherichia coli strains include strains ZZ9DT16R, ZC9CTR2, JXCE34, wherein the ratio of paclitaxel to tigecycline is 64:1 or 128:1, or 256:1. Preferably, the ratio of paclitaxel to tigecycline is between 64:1-256:1. Preferably, the ratio of paclitaxel to tigecycline is between 64:1-128:1. Preferably, the ratio of paclitaxel to tigecycline is between 128:1-256:1.

[0031] Another object of the present invention is to provide a use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Klebsiella pneumoniae, wherein the ratio of paclitaxel to tigecycline is 32:1 or 64:1; preferably, the ratio of paclitaxel to tigecycline is between 32:1-64:1. The use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Klebsiella pneumoniae, the Klebsiella pneumoniae strain includes strain HC139, wherein the ratio of paclitaxel to tigecycline is 32:1 or 64:1; preferably, the ratio of paclitaxel to tigecycline is between 32:1-64:1.

[0032] Another object of the present invention is to provide a paclitaxel antibacterial composition, the antibacterial composition comprising paclitaxel and tigecycline, the paclitaxel antibacterial composition is used to fight against Escherichia coli or Klebsiella pneumoniae, wherein the ratio of paclitaxel to tigecycline is 16:1, 32:1, or 256:1. Preferably, the ratio of paclitaxel to tigecycline in the antibacterial composition is between 16:1-256:1; preferably, the ratio of paclitaxel to tigecycline in the antibacterial composition is between 32:1-256:1.

[0033] Another object of the present invention is to provide a paclitaxel antibacterial composition, the antibacterial composition comprises paclitaxel and tigecycline, the paclitaxel antibacterial composition is used to fight Escherichia coli or Klebsiella pneumoniae, wherein the dosage ratio of paclitaxel to tigecycline is 32:1, 64:1, or 128:1 or 256:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-256:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 64:1-128:1. Preferably, the dosage ratio of paclitaxel to tigecycline is between 128:1-256:1.

[0034] Another object of the present invention is to provide a paclitaxel antibacterial compound preparation, the compound preparation comprises paclitaxel and tigecycline, the paclitaxel antibacterial compound preparation is used to fight against Escherichia coli or Klebsiella pneumoniae, wherein the dosage ratio of paclitaxel to tigecycline is 16:1, 32:1, or 64:1, or 128:1 or 256:1. Preferably, the dosage ratio of paclitaxel to tigecycline is any ratio between 16:1-256:1.

[0035] Another object of the present invention is to provide a paclitaxel antibacterial compound preparation, the compound preparation comprises paclitaxel and tigecycline, the paclitaxel antibacterial compound preparation is used to fight against Escherichia coli or Klebsiella pneumoniae, and the dosage ratio of paclitaxel to tigecycline is 16:1 or 32:1. Preferably, the dosage ratio of paclitaxel to tigecycline is any ratio between 16:1-32:1.

[0036] Another object of the present invention is to provide an antibacterial test method or a target test method for the use of paclitaxel in enhancing the anti-Escherichia coli activity of tigecycline, the method comprising the following steps:

[0037] S1: dissolving paclitaxel and tigecycline in a solvent;

[0038] S2: Paclitaxel and tigecycline are mixed in proportion to obtain a composition;

[0039] S3: using the composition of step S2 for MIC test, FICI test, time-kill curve test, or target test.

[0040] The solvent described in step S1 includes dimethyl sulfoxide or deionized water.

[0041] In step S2, the ratio of paclitaxel to tigecycline is 16:1. Preferably, the ratio of paclitaxel to tigecycline is 16:1.

[0042] When the dosage of paclitaxel in step S3 is 128 μg / mL or 256 μg / mL, the dosage of tigecycline is 1-8 μg / mL. Preferably, when the dosage of paclitaxel is 256 μg / mL, the dosage of tigecycline is 1 μg / mL, 2 μg / mL, or 4 μg / mL. Preferably, when the dosage of paclitaxel is 256 μg / mL, the dosage of tigecycline is 1-4 μg / mL.

[0043] The term "synergistic effect" as used herein means that the effect of two drugs combined is greater than the algebraic sum of the effects of the individual drugs.

[0044] As used herein, the term "additive effect" means that the effect of two drugs taken together is equal to the algebraic sum of their individual effects.

[0045] As used herein, the term "antagonism" means that the effect of two drugs taken together is less than the sum of their individual effects.

[0046] The term "anti-E. coli effect" herein means inhibiting the proliferation or growth of E. coli, or killing E. coli.

[0047] The term "anti-Klebsiella pneumoniae effect" herein refers to inhibiting the proliferation or growth of Klebsiella pneumoniae, or killing Klebsiella pneumoniae.

[0048] The present invention has the following beneficial effects: 1. When the dosage of paclitaxel is 128 μg / mL or 256 μg / mL, the dosage of tigecycline is 1-8 μg / mL, which can significantly enhance the antibacterial activity of tigecycline against the large intestine. 2. The combination of paclitaxel and tigecycline both show significant synergistic effects, with FICI of 0.1875-0.3125 (all less than 0.5), that is, the combination of the two shows significant synergistic effects. 3. The dosage of tigecycline can be effectively reduced by using paclitaxel in combination. The dosage of tigecycline used alone is 1 to 16 times higher than that of paclitaxel and tigecycline in combination, and the drug resistance of bacteria to tigecycline is synergistically improved. 4. The combination of paclitaxel and tigecycline can play a synergistic role in reversing drug resistance, with FICI of 0.1875-0.3125, and the combined use of drugs kills all bacteria after 12 hours. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 This is the time-killing curve of paclitaxel and tigecycline against strain ZZ9DT16R. DETAILED DESCRIPTION

[0050] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0051] Example 1

[0052] Materials and methods

[0053] 1. Drug testing

[0054] Paclitaxel (PTX): Shanghai MacLean Biochemical Technology Co., Ltd., purity 98%

[0055] Tigecycline (TGC): Shanghai Yuanye Biotechnology Co., Ltd., purity 98%

[0056] Paclitaxel was prepared with dimethyl sulfoxide to a concentration of 5120 μg / mL. Tigecycline was prepared with deionized water to a concentration of 1280 μg / mL. The tested drugs were stored at -20°C. Before the test, the drug storage solution was taken out and thawed at room temperature, mixed thoroughly, and the pharmacodynamic tests were performed separately.

[0057] 2. Strains

[0058] The clinically isolated Escherichia coli strains were Escherichia coli ZZ9DT16R, ZC9CTR2, and JX21CE34, and the clinically isolated Klebsiella pneumoniae and Klebsiella pneumoniae HC139 were provided by the Pharmacology Laboratory of Henan Agricultural University. Escherichia coli and Klebsiella pneumoniae were activated by streaking on MacConkey medium, cultured at 37°C for 16 to 18 hours, and stored at 4°C for later use.

[0059] 3. Culture medium

[0060] LB broth medium: produced by Beijing Aoboxing Biotechnology Co., Ltd., prepare 1000 ml according to the instructions of this product, sterilize at 121℃ for 15 minutes, and store at 4℃ for later use. SS agar medium: produced by Beijing Aoboxing Biotechnology Co., Ltd., prepare 1000 ml according to the instructions of this product, heat and boil to fully dissolve and sterilize, cool to 60 degrees, pour into sterilized plates and store at 4℃ for later use. MHB broth medium: produced by Beijing Aoboxing Biotechnology Co., Ltd., prepare 1000 ml according to the instructions of this product, sterilize at 121℃ for 15 minutes, and store at 4℃ for later use.

[0061] 4. Instruments

[0062] High-pressure steam autoclave (Shanghai Shen'an Medical Instrument Factory); AB204-N electronic analytical balance (Mettler-Tolly Instrument Shanghai Co., Ltd.); HZQ-R air bath oscillator (Beijing Liuyi Instrument Factory); DH-600 electric constant temperature incubator (Beijing Kewei Yongxing Instrument Co., Ltd.); DHG-9140A vacuum drying oven (Shanghai Yiheng Experimental Instrument Co., Ltd.); SW-CJ-JC clean bench (Suzhou Antai Air Technology Co., Ltd.).

[0063] 5. Preparation of Bacterial Liquid

[0064] Pick a single colony of Escherichia coli or Klebsiella pneumoniae from the MacConkey medium stored at 4°C, inoculate it into 5 mL of LB broth, and culture it at 37°C with shaking at 180 rpm to keep the bacteria in the late exponential growth phase. Take the bacterial solution into MHB broth and dilute the original bacterial solution to 10 5 CFU / mL. Add corresponding concentrations of paclitaxel to the bacterial suspension for later use. The final concentrations of paclitaxel in the bacterial suspension are: 128μg / mL and 256μg / mL.

[0065] 6. Preparation of Drug Sensitivity Plate

[0066] Use the bacterial suspension in step 5 to conduct drug sensitivity test. Take a sterile 96-well reaction plate, add 126 μL of freshly prepared bacterial suspension containing paclitaxel and 14 μL of 1280 μg / mL tigecycline to well 1 in the first row; add 70 μL of freshly prepared bacterial suspension containing paclitaxel to wells 2-12; add 70 μL of freshly prepared bacterial suspension containing paclitaxel to wells 1-7 in the second row, and add 70 μL of MHB broth to well 8 as a blank control. Wells 1-12 in the first row and wells 1-6 in the second row were serially diluted to make the final concentrations of tigecycline in wells 1-12 in the first row and wells 1-6 in the second row 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125, 0.015625, 0.0078125, 0.00390625 μg / mL, respectively; well 7 in the second row was a drug-free MHB bacterial suspension as a negative control; well 8 was a drug-free bacterial suspension as a positive control. The drug sensitivity plate was cultured at 37°C for 16-20 hours.

[0067] 7. Determination of MIC value

[0068] After the 96-well plate containing bacteria was cultured at 37°C for 16-20 hours, the MIC (minimum inhibitory concentration) results were read. The drug MIC value (minimum inhibitory concentration) is the drug concentration in the clear and transparent well before the turbid well in the 96-well plate. The above experiments were all performed in parallel 3 times, and only when the MIC value could be accurately repeated was it accepted; when the MIC value differed by more than one concentration, a new test was required until it met the requirements. The results are shown in Table 1.

[0069] Table 1 MIC of paclitaxel combined with tigecycline against the tested strains

[0070]

[0071] Note: 1 / 2MIC paclitaxel: the amount of paclitaxel added is 256μg / mL; 1 / 4MIC paclitaxel: the amount of paclitaxel added is 128μg / mL;

[0072] The results showed that the MIC values ​​of tigecycline against strains ZZ9DT16R, ZC9CTR2, JXCE34 and HC139 were 16, 4, 32 and 16 μg / mL, respectively; the MIC value of paclitaxel against strains ZZ9DT16R, ZC9CTR2, JXCE34 and HC139 was 512 μg / mL. When 1 / 2MIC (256 μg / mL) and 1 / 4MIC (128 μg / mL) paclitaxel were added, the MIC of tigecycline against ZZ9DT16R was 4 and 8 μg / mL, respectively; the MIC for ZC9CTR2 was 1, and 4 μg / mL, respectively; the MIC for JXCE34 was 2, and 4 μg / mL, respectively; and the MIC for HC139 was 4, and 4 μg / mL, respectively.

[0073] Compared with the MIC value of tigecycline used alone, when 256μg / mL and 128μg / mL of paclitaxel were added, the MIC of tigecycline against strain ZZ9DT16R decreased by 4 and 2 times, respectively; the MIC of tigecycline against strain ZC9CTR2 decreased by 4 and 1 times, respectively; the MIC of tigecycline against strain JXCE34 decreased by 16 and 8 times, respectively; the MIC of tigecycline against strain HC139 decreased by 4 times.

[0074] The results showed that when paclitaxel was used to enhance the antibacterial effect of tigecycline, the dosage ratio of paclitaxel to tigecycline was 16:1, 32:1, 64:1, 128:1 or 256:1, which reduced the dosage of tigecycline by 1 to 16 times compared with that of tigecycline alone.

[0075] The results showed that paclitaxel at concentrations of 256 μg / mL and 128 μg / mL could significantly enhance the antibacterial activity of tigecycline against Escherichia coli ZZ9DT16R, ZC9CTR2, JXCE34, and Klebsiella pneumoniae HC139.

[0076] Example 2

[0077] Materials and methods

[0078] 1. Drug testing:

[0079] Same as Example 1.

[0080] 2. Strains

[0081] Same as Example 1.

[0082] 3. Culture medium

[0083] Same as Example 1.

[0084] 4. Instruments

[0085] Same as Example 1.

[0086] 5. Preparation of Bacterial Liquid

[0087] Same as Example 1.

[0088] 6. Preparation of Drug Sensitivity Plate

[0089] Based on the microbroth dilution method, we obtained the MIC values ​​of PTX and TGC alone, and selected PTX and TGC for multiple dilution. Specifically, 100 μL of MHB was distributed into 8×8 96-well plates, and then TGC and PTX were diluted 2 times along the horizontal and vertical axes, respectively, and PTX and TGC single-drug MICs as well as negative and positive controls were made at the same time. (Place the 96-well plate in a constant temperature incubator at 37°C for 16 to 18 hours, and incubate it statically, and observe and record the results. The experiment was repeated three times.

[0090] The fractional inhibitory concentration index (FICI) was used as the criterion for the chessboard susceptibility test. The FICI of two compounds A and B was defined by the following formula:

[0091] FICI=FIC A +FIC B =MIC AB / MIC A +MIC BA / MIC B .

[0092] MIC A is the MIC of compound A alone, MIC AB is the MIC of compound A in combination with MIC B is the MIC of compound B, MIC BA is the MIC when compound B is combined. FICI < 0.5 indicates synergistic effect; 0.5 ≤ FICI ≤ 1 indicates additive effect; 1 < FICI ≤ 2 indicates irrelevant effect; FICI > 2 indicates antagonistic effect. The results are shown in Table 2.

[0093] Table 2 Antibacterial effect of paclitaxel and tigecycline combined

[0094]

[0095] The results showed that paclitaxel alone had a weak antibacterial effect on the tested strains, and when used in combination with TGC, it showed a synergistic effect on TGC, with FICI ranging from 0.1875 to 0.3125, all <0.5, that is, the combination of the two showed a synergistic effect.

[0096] Example 3

[0097] Materials and methods

[0098] 1. Drug testing:

[0099] Same as Example 1.

[0100] 2. Strains

[0101] Same as Example 1.

[0102] 3. Culture medium

[0103] Same as Example 1.

[0104] 4. Instruments

[0105] Same as Example 1.

[0106] 5. Preparation of Bacterial Liquid

[0107] Same as Example 1.

[0108] 6. Time-killing curve

[0109] Escherichia coli ZZ9DT16R was selected for time-kill curve test. A single colony was inoculated into fresh LB broth, placed in a shaker at 37°C, 180r / min, and cultured overnight; then the bacterial solution was transferred to 10mL fresh LB broth at a ratio of 1:100 and divided into 5 test groups:

[0110] Group A: Control;

[0111] Group B: 1 / 2MIC TGC;

[0112] Group C: 1 / 2MIC PTX;

[0113] Group D: 1 / 2MIC PTX+1 / 2MIC TGC;

[0114] Group E: 1 / 4MIC PTX+1 / 2MIC TGC.

[0115] After grouping, place the cells at 37°C and shake culture at 180 r / min. Take 100 μL of the bacterial suspension at 0 h, 2 h, 4 h, 8 h, 12 h, and 24 h, respectively. Dilute the 100 μL bacterial suspension 10-fold with PBS buffer, spread 100 μL of the dilution on the LB agar plate, and culture at 37°C for 16 to 18 h. Finally, count the colonies on the agar plate, with time as the horizontal axis, and log 10 The time-killing curve was drawn with CFU / mL as the ordinate, and the experiment was repeated three times.

[0116] Result determination: At 24 hours, the number of colonies in the combination group decreased by ≥2 log compared with the single drug. 10 The results are defined as synergistic. Figure 1 shown.

[0117] The experimental results showed that when the single drug 1 / 2MIC PTX was used, the strain grew slowly in the first 12 hours, indicating that PTX had a weak antibacterial effect on the strain, and the number of strains decreased at 24 hours, indicating a weak bactericidal effect. 1 / 2MIC TGC initially showed a certain bactericidal effect on the strain, but the number of strains increased rapidly between 4 hours and 24 hours; 1 / 2MIC TGC + 1 / 4MIC PTX showed a significant bactericidal effect from 2 to 24 hours, and the number of colonies decreased by more than 2log 10 CFU / mL. The bactericidal effect of 1 / 2MIC TGC+1 / 2MIC PTX was more significant, and the bacteria could be completely killed in 12 hours. The time-kill curve results were basically consistent with the combined drug sensitivity results, that is, the antibacterial effect of PTX alone was weak, and the combination of 1 / 2MIC PTX or 1 / 4MIC PTX and TGC could significantly improve the antibacterial activity of TGC against drug-resistant bacteria, showing a good synergistic reversal of drug resistance.

[0118] The above experiments showed that when the concentration of paclitaxel was 128μg / mL and 256μg / mL, it could significantly enhance the antibacterial activity of tigecycline against Escherichia coli ZZ9DT16R, ZC9CTR2, JX21CE34 and Klebsiella pneumoniae HC139. The combination of paclitaxel and tigecycline showed significant synergistic effect, with FICI of 0.1875-0.3125, both <0.5, that is, the combination of the two showed significant synergistic effect. The results of the time-kill curve showed that the combination showed significant bactericidal effect, among which 1 / 2MICPTX+1 / 2MIC TGC killed all the bacteria in 12h, which was basically consistent with the combined drug sensitivity results, that is, the antibacterial effect of PTX alone was weak, and the combination of 1 / 2MIC PTX or 1 / 4MIC PTX and TGC could significantly improve the antibacterial activity of TGC against drug-resistant bacteria, showing a good synergistic reversal of drug resistance.

[0119] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. The use of paclitaxel in the preparation of a synergist for the antibacterial effect of tigecycline on Escherichia coli and Klebsiella pneumoniae, characterized in that: The dosage ratio of paclitaxel to tigecycline is between 32:1-256:

1.

2. The use according to claim 1, characterized in that: The dosage ratio of paclitaxel to tigecycline was 16:1 or 32:

1.

3. The use according to claim 1, characterized in that: The dosage ratio of paclitaxel to tigecycline is between 16:1 and 32:

1.

4. The use according to claim 1, characterized in that: The dosage ratio of paclitaxel to tigecycline was 64:1, 128:1 or 256:

1.

5. The use according to claim 1, characterized in that: The dosage ratio of paclitaxel to tigecycline is between 64:1-256:1, or the dosage ratio of paclitaxel to tigecycline is between 64:1-128:

1.

6. A paclitaxel antibacterial composition, comprising paclitaxel and tigecycline, wherein the ratio of paclitaxel to tigecycline is 64:1-256:

1.

7. The antibacterial composition according to claim 6, characterized in that The dosage ratio of paclitaxel to tigecycline is 64:1 or 256:

1.

8. The antibacterial composition according to claim 6, characterized in that The dosage ratio of paclitaxel to tigecycline is between 64:1 and 128:

1.

9. The antibacterial composition according to claim 6, characterized in that The dosage ratio of paclitaxel to tigecycline is between 128:1 and 256:

1.

10. The paclitaxel antibacterial composition according to claim 6, characterized in that: The dosage ratio of paclitaxel to tigecycline is 32:1, 64:1, or 128:1 or 256:1.

Images