CN116549448A - Application of vitamin B6 in preparation of medicines for improving sensitivity of bacteria to antibiotics - Google Patents

Application of vitamin B6 in preparation of medicines for improving sensitivity of bacteria to antibiotics Download PDF

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Publication number
CN116549448A
CN116549448A CN202310577199.3A CN202310577199A CN116549448A CN 116549448 A CN116549448 A CN 116549448A CN 202310577199 A CN202310577199 A CN 202310577199A CN 116549448 A CN116549448 A CN 116549448A
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vitamin
effective concentration
chloramphenicol
antibiotics
antibiotic
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王超
阎贝贝
孟庆磊
孙明英
王锡荣
彭宣宪
董学飒
安丽
张龙岗
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Jinan Children's Hospital Shandong University Affiliated Children's Hospital
Shandong Freshwater Fisheries Research Institute
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Jinan Children's Hospital Shandong University Affiliated Children's Hospital
Shandong Freshwater Fisheries Research Institute
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4415—Pyridoxine, i.e. Vitamin B6
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/429—Thiazoles condensed with heterocyclic ring systems
    • A61K31/43—Compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula, e.g. penicillins, penems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention discloses an application of vitamin B6 in preparing medicines for improving the sensitivity of bacteria to antibiotics, wherein the bacteria are Edwardsiella tarda; the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin. The invention also discloses application of the vitamin B6 combined antibiotics in preparation of antibacterial drugs. The invention also discloses an antibacterial drug, which comprises the active ingredients of vitamin B6 and antibiotics. According to the invention, the research shows that vitamin B6 can improve the sensitivity of Edwardsiella tarda multi-resistant bacteria to antibiotics, so that the problem of bacterial drug resistance is solved. The combination of vitamin B6 and antibiotics can obviously improve the sterilization effect of the antibiotics, and has better effect and higher safety. The invention has better application prospect.

Description

Application of vitamin B6 in preparation of medicines for improving sensitivity of bacteria to antibiotics
Technical Field
The invention relates to application of vitamin B6 in preparation of a medicament for improving the sensitivity of bacteria to antibiotics, and belongs to the technical field of medicines.
Background
Antibiotic resistance (antibiotic resistance) refers to the tolerance of bacteria to the otherwise sensitive antibiotics, so that the normal dosage of antibiotics cannot exert the due bactericidal effect, and finally the therapeutic effect of the antibiotics is obviously weakened. Bacterial resistance seriously jeopardizes the physical health of human beings and the sustainable development of the breeding industry, but effective prevention and treatment measures for drug-resistant bacteria are lacking at present.
Recent researches prove that the small molecular metabolites can improve the sterilization efficiency of antibiotics, which is an innovative field for researching and developing the sterilization of drug-resistant bacteria. For example, chinese patent application CN 106668011A discloses a new use of vitamin C, namely, an application of vitamin C in preparing bacteriostatic or bactericidal drugs containing β -lactam antibiotics, bacteriostatic or bactericidal drugs capable of improving sensitivity of bacteria to β -lactam antibiotics, and bacteriostatic or bactericidal drugs capable of reducing resistance of bacteria to β -lactam antibiotics, respectively. Chinese patent No. CN 104721176A discloses the use of alpha-ketoglutarate to increase the sensitivity of bacteria to antibiotics. Chinese patent No. CN 107929741A discloses the use of L-aspartic acid to increase the susceptibility of bacteria to antibiotics. Chinese patent No. CN 108042521A discloses the use of L-arginine to increase the susceptibility of bacteria to antibiotics. The micromolecular metabolite and the antibiotics are used cooperatively, so that the originally ineffective antibiotic dose can kill drug-resistant bacteria, and the method has important clinical significance and production value for treating the infection of the drug-resistant bacteria.
Edwardsiella tarda (Edwardsiella tarda, et), also known as Edwardsiella tarda or Edwardsiella tarda, is a gram-negative bacterium, belongs to the Edwardsiella genus (Edwardsiella) of the Enterobacteriaceae family, is one of the earlier found members of the Edwardsiella bacterial family, is a model species of the Edwardsiella genus, and has a model strain ATCC 15947 (DSM 30052) and has a G+C mol% in DNA of 55 to 58 (Tm, bd). The host range of the strain is very wide, and the strain can infect fishes, amphibians, reptiles and mammals, and human beings are also hosts of the strain. The strain is a main pathogenic bacteria for aquaculture, can infect carps, tilapia, eel, paralichthys olivaceus, chinese soft-shelled turtles and the like, and causes great harm to aquaculture.
Vitamin B6 (Vitamin B6), also known as pyridoxine, is a water-soluble Vitamin, which is easily destroyed by light or alkali, is not resistant to high temperature, is easily dissolved in water and ethanol, and is stable in acid liquor. Vitamin B6, a coenzyme for certain enzymes in the organism, is involved in various metabolic reactions, particularly closely related to amino acid metabolism, mainly in the conversion of sulfur-containing amino acids, synthesis of nicotinic acid, glucose metabolism, lipid metabolism, and in the synthesis of various neurotransmitters. The animal cannot synthesize vitamin B6, is mainly obtained through food and intestinal flora, and has wide sources. Clinically, vitamin B6 preparation is used for preventing and treating vomiting of pregnancy and vomiting of radiation sickness. There is no report on vitamin B6 in terms of increasing the susceptibility of bacteria to antibiotics.
Disclosure of Invention
Aiming at the prior art, the invention provides a new application of vitamin B6, namely an application in preparing medicines for improving the sensitivity of bacteria to antibiotics. The invention also provides application of the vitamin B6 combined antibiotics in preparation of antibacterial drugs, and also provides an antibacterial drug.
The invention is realized by the following technical scheme:
use of vitamin B6 for the preparation of a medicament for increasing the sensitivity of bacteria to antibiotics, said bacteria being edwardsiella tarda; the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
The application of vitamin B6 combined with antibiotics in the preparation of antibacterial drugs, wherein the bacteria are Edwardsiella tarda; the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
Further, the effective concentration of vitamin B6 is 1.25-20 mM. Further, when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 100-2500 mg/L, preferably 2000-mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5-mM. When the antibiotic is ampicillin, the effective concentration of ampicillin is 200-400 mg/L, preferably 300 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM. When the antibiotic is kanamycin, the effective concentration of kanamycin is 600-1000 mg/L, preferably 800 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM. When the antibiotic is ciprofloxacin, the effective concentration of ciprofloxacin is 2-5 mg/L, preferably 3 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM.
An antibacterial agent contains vitamin B6 and antibiotics as effective components; the bacteria are Edwardsiella tarda, and the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
Further, the effective concentration of vitamin B6 is 1.25-20 mM. Further, when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 100-2500 mg/L, preferably 2000-mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5-mM. When the antibiotic is ampicillin, the effective concentration of ampicillin is 200-400 mg/L, preferably 300 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM. When the antibiotic is kanamycin, the effective concentration of kanamycin is 600-1000 mg/L, preferably 800 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM. When the antibiotic is ciprofloxacin, the effective concentration of ciprofloxacin is 2-5 mg/L, preferably 3 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM, preferably 2.5 mM.
Further, the antibacterial agent also comprises a pharmaceutically acceptable carrier, such as hydrogel, mesoporous silica coating, metal organic framework, and the like. The dosage form of the antibacterial agent may be any known dosage form such as injection, tablet, capsule, aerosol, etc.
Further, the antibacterial drug consists of vitamin B6, chloramphenicol and water, wherein the concentration of the chloramphenicol is 100-2500 mg/L, preferably 2000-mg/L, and the concentration of the vitamin B6 is 1.25-20 mM, preferably 2.5-mM.
In order to overcome the problem of bacterial drug resistance, edwardsiella tarda is taken as a research object, and research shows that: vitamin B6 can improve the sensitivity of Edwardsiella tarda multi-drug resistant bacteria to antibiotics (chloramphenicol, ampicillin, kanamycin and ciprofloxacin), especially chloramphenicol, and vitamin B6 of 2.5 mM can improve the sterilization efficiency of chloramphenicol by 48.5 times.
The invention discovers that vitamin B6 can improve the sensitivity of Edwardsiella tarda multi-resistant bacteria to antibiotics for the first time, thereby overcoming the problem of bacterial drug resistance. The combination of vitamin B6 and antibiotics can obviously improve the sterilization effect of the antibiotics, and has better effect and higher safety compared with the method of using the antibiotics as antibacterial drugs. The invention has better application prospect.
Drawings
Fig. 1: multiple of resistance of edwardsiella tarda ATCC 15947 to different antibiotics.
Fig. 2: statistical graphs of survival rate of Edwardsiella tarda after addition of vitamin B6 and chloramphenicol.
Fig. 3: absorbance measurement of edwardsiella tarda.
Fig. 4: statistical graphs of survival rate of Edwardsiella tarda after addition of vitamin B6 and chloramphenicol at different concentrations.
Fig. 5: statistical graphs of survival rate of Edwardsiella tarda after addition of chloramphenicol and different concentrations of vitamin B1.
Fig. 6: statistical graphs of survival rate of Edwardsiella tarda after addition of chloramphenicol and different concentrations of vitamin B3.
Fig. 7: figure of survival of Edwardsiella tarda supplemented with ampicillin and vitamin B6 in example 3.
Fig. 8: statistical graphs of survival rate of Edwardsiella tarda supplemented with kanamycin and vitamin B6 in example 3.
Fig. 9: statistical graphs of survival rate of Edwardsiella tarda with ciprofloxacin and vitamin B6 added in example 3.
Detailed Description
The invention is further illustrated below with reference to examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art will appreciate that various changes and modifications can be made to the invention without departing from the spirit and scope thereof.
The instruments, reagents and materials used in the examples below are conventional instruments, reagents and materials known in the art and are commercially available. The experimental methods, detection methods, and the like in the examples described below are conventional experimental methods and detection methods known in the prior art unless otherwise specified.
Example 1 vitamin B6 enhances sensitivity of Edwardsiella tarda to chloramphenicol
(1) Determination of Edwardsiella tarda ATCC 15947 resistance
The minimum inhibitory concentrations of various antibiotics on the Edwardsiella tarda ATCC 1597antibiotic-sensitive strain and the drug-resistant strain are detected respectively, and the results are shown in figure 1, wherein the minimum inhibitory concentrations of chloramphenicol, ampicillin, kanamycin and ciprofloxacin on the Edwardsiella tarda ATCC 15947-sensitive strain are respectively 0.5 mg/L, 5 mg/L, 5 mg/L and 0.002 mg/L, the minimum inhibitory concentrations of the drug-resistant strain are greatly improved relative to the sensitive strain, and the improvement times are respectively 200, 100 and 100 times, so that the Edwardsiella tarda ATCC 15977 is a multi-drug-resistant strain.
(2) Preparation of test specimens
Single colonies were picked from TSA plates, transferred to 1 mL of TSB medium, and shake cultured at 37℃and 200 rpm for 24 h to saturation. Centrifuging (5000 g,5 min) to collect thallus, washing thallus with 0.85% physiological saline, suspending thallus in M9 culture medium, and regulating bacterial liquid OD 600 nm To 0.2, the mixture was dispensed into 1 mL Ep tubes for further use.
Vitamin B6 used in the present invention was purchased from sigma (P5669-5G).
(3) Vitamin B6 enhances the sensitivity of Edwardsiella tarda ATCC 15947 to chloramphenicol
Taking the bacterial liquid prepared in the step (2), and dividing the bacterial liquid into 4 groups (each group is 0.1 mL,3 biological repeats): experimental group, control group 1, control group 2 and control group 3, wherein chloramphenicol and vitamin B6 are added in the experimental group, M9 culture medium is added to 1 mL, the final concentration of chloramphenicol is 2000 mg/L, and the final concentration of vitamin B6 is 2.5 mM; control 1 was not supplemented with chloramphenicol and vitamin B6, M9 medium to 1 mL; control group 2 added vitamin B6 (no chloramphenicol), M9 medium to 1 mL, vitamin B6 to a final concentration of 2.5 mM; control 3 was supplemented with chloramphenicol (vitamin B6 was not added) and M9 medium was added to 1 mL at a final concentration of 2000 mg/L.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
As a result, as shown in FIG. 2, the survival rate of the control group 3 was 25.5%, and the survival rate of the experimental group was 0.661%. It was found that the sensitivity of Edwardsiella tarda to chloramphenicol increased by 38.6-fold after the addition of vitamin B6 (survival rate decreased from 25.5% with chloramphenicol alone to 0.661% after the addition of vitamin B6 and chloramphenicol). The results show that: after vitamin B6 is added, the sensitivity of Edwardsiella tarda to chloramphenicol can be obviously improved.
(4) Vitamin B6 enhances sensitivity of Edwardsiella tarda ATCC 15947 to chloramphenicol with concentration dependence
The bacterial liquid prepared in the step (2) is taken out to be 0.01 part (mL) (4 parts) and added to a TSB culture medium containing 100 mg/L chloramphenicol in a quantity of 0.3 part (mL). Then, vitamin B6 was added at final concentrations of 0, 1, 2 and 4 mM, and the mixture was subjected to stationary culture at 37℃for 7 h, absorbance was measured per hour, and a growth inhibition curve was drawn.
As a result, as shown in FIG. 3, when the chloramphenicol concentration was 100 mg/L and the vitamin B6 addition concentrations were 0, 1, 2, and 4 mM, the absorbance values were measured at 4 hours and were 0.150, 0.128, 0.119, and 0.102, respectively; absorbance values were measured at 5 hours of 0.181, 0.158, 0.144 and 0.117, respectively; absorbance values were measured at 6 hours of 0.205, 0.184, 0.164 and 0.125, respectively; absorbance values were measured at 7 hours of 0.226, 0.201, 0.184 and 0.134, respectively.
The results show that: within a certain range, the sterilization efficiency of chloramphenicol against bacteria is gradually increased with the increase of the concentration of added vitamin B6, and compared with the low concentration (1 mM), the high concentration (4 mM) of vitamin B6 can help chloramphenicol to inhibit the bacterial growth more remarkably.
Taking bacterial liquid 0.1 mL (6 parts) prepared in the step (2), adding chloramphenicol, and adding M9 culture medium to 1 mL, wherein the final concentration of chloramphenicol is 2000 mg/L. Vitamin B6 was then added at final concentrations of 0, 1.25, 2.5, 5, 10, 20 mM, respectively, and incubated in a 37 ℃ incubator for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate was calculated (as above, the survival rate of control group 1 of step 3 was 100%).
As shown in FIG. 4, when the chloramphenicol concentration was 2000 mg/L, the survival rate of vitamin B6 was 35.5%, the chloramphenicol sterilization efficiency was 12.0-fold (survival rate was reduced to 2.97%) by adding vitamin B6 of 1.25 mM, 48.5-fold (survival rate was reduced to 0.732%) by adding vitamin B6 of 2.5 mM, 35.9-fold (survival rate was reduced to 0.990%) by adding vitamin B6 of 5 mM, 7.55-fold (survival rate was reduced to 4.70%) by adding vitamin B6 of 10 mM, and 4.65-fold (survival rate was reduced to 7.64%) by adding vitamin B6 of 20 mM.
The results show that: with the increase of vitamin B6 concentration, the survival rate of bacteria is firstly reduced and then increased, and the sterilization efficiency is highest at the concentration of 2.5 and mM.
Example 2 vitamins B1, B3 do not increase the sensitivity of Edwardsiella tarda to chloramphenicol
To verify whether other B vitamins increase the sensitivity of edwardsiella tarda to chloramphenicol, bacterial samples were prepared according to the procedure of step (2) of example 1.
(1) Prepared bacterial samples were taken and divided into 7 groups (0.1 mL per group): experimental groups 1, 2, 3, 4 and 5, control group 1 and control group 2, wherein chloramphenicol and vitamin B1 are added in experimental groups 1, 2, 3, 4 and 5, M9 culture medium is added to 1 mL, the final concentration of chloramphenicol is 2000 mg/L, and the final concentrations of vitamin B1 are 1.25, 2.5, 5, 10 and 20 mM respectively; control 1 was not supplemented with chloramphenicol and vitamin B1, M9 medium to 1 mL; control group 2 was supplemented with chloramphenicol (vitamin B1 was not added) to a final concentration of 2000 mg/L with M9 medium to 1 mL.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
As a result, as shown in FIG. 5, the survival rate of the bacteria was gradually increased when vitamin B1 was added at different concentrations to a 2000 mg/L chloramphenicol medium, as compared with the case where vitamin B1 was not added.
(2) Prepared bacterial samples were taken and divided into 7 groups (0.1 mL per group): experimental groups 1, 2, 3, 4 and 5, control group 1 and control group 2, wherein chloramphenicol and vitamin B3 are added in experimental groups 1, 2, 3, 4 and 5, M9 culture medium is added to 1 mL, the final concentration of chloramphenicol is 2000 mg/L, and the final concentrations of vitamin B3 are 1.25, 2.5, 5, 10 and 20 mM respectively; control 1 was not supplemented with chloramphenicol and vitamin B3, M9 medium was added to 1 mL; control group 2 was supplemented with chloramphenicol (vitamin B3 was not added) and M9 medium was added to 1 mL at a final concentration of 2000 mg/L.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
As a result, as shown in FIG. 6, the survival rate of the bacteria was gradually increased when vitamin B3 was added at different concentrations to a medium containing 2000 mg/L chloramphenicol, as compared with the case where vitamin B3 was not added.
The above results indicate that neither vitamin B1 nor B3 can increase the sensitivity of Edwardsiella tarda to chloramphenicol, and that chloramphenicol also decreases the sterilizing efficiency of Edwardsiella tarda. This suggests that not all B vitamins can increase the bactericidal efficiency of chloramphenicol.
Example 3 vitamin B6 enhances sensitivity of Edwardsiella tarda to other antibiotics
To verify whether vitamin B6 can increase the sensitivity of edwardsiella tarda to other antibiotics, bacterial samples were prepared according to the procedure of step (2) of example 1.
(1) Prepared bacterial samples were taken and divided into 3 groups (0.1 mL per group): an experimental group, a control group 1 and a control group 2, wherein ampicillin and vitamin B6 are added in the experimental group, M9 culture medium is added to 1 mL, the final concentration of ampicillin is 300 mg/L, and the final concentration of vitamin B6 is 2.5 mM; control 1 was not added ampicillin and vitamin B6, M9 medium to 1 mL; control group 2 was supplemented with ampicillin (without vitamin B6) and M9 medium to 1 mL, with a final ampicillin concentration of 300 mg/L.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
(2) Prepared bacterial samples were taken and divided into 3 groups (0.1 mL per group): experimental group, control group 1, control group 2, wherein the experimental group was added kanamycin and vitamin B6, M9 medium was added to 1 mL, the final concentration of kanamycin was 800 mg/L, and the final concentration of vitamin B6 was 2.5 mM; control 1 was not supplemented with kanamycin and vitamin B6, M9 medium to 1 mL; control group 2 was supplemented with kanamycin (vitamin B6 was not added) and M9 medium was added to 1 mL, the final concentration of kanamycin being 800 mg/L.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
(3) Prepared bacterial samples were taken and divided into 3 groups (0.1 mL per group): the experimental group, the control group 1 and the control group 2, wherein the experimental group is added with ciprofloxacin and vitamin B6, M9 culture medium is added to 1 mL, the final concentration of the ciprofloxacin is 3 mg/L, and the final concentration of the vitamin B6 is 2.5 mM; control 1 was not added ciprofloxacin and vitamin B6, M9 medium was added to 1 mL; control group 2 was supplemented with ciprofloxacin (vitamin B6 was not added) and M9 medium was added to 1 mL, with a final ciprofloxacin concentration of 3 mg/L.
Bacterial solutions of each experimental group and control group were incubated in a constant temperature incubator at 37℃for 5 h. TSA plate colony counts were performed on 0.02. 0.02 mL bacterial liquid, the colony numbers were counted, and the survival rate (based on 100% of the survival rate of control group 1) was calculated.
As shown in FIGS. 7 to 9, after addition of vitamin B6, edwardsiella tarda had a 5.50-fold increase in ampicillin sensitivity (survival rate decreased from 51.6% with ampicillin alone to 9.39% with vitamin B6 and ampicillin alone), a 2.84-fold increase in kanamycin sensitivity (survival rate decreased from 43.9% with kanamycin alone to 15.5% with vitamin B6 and kanamycin alone), and a 4.37-fold increase in ciprofloxacin sensitivity (survival rate decreased from 35.6% with ciprofloxacin alone to 8.14% with vitamin B6 and ciprofloxacin alone). The results show that after the addition of vitamin B6, the sensitivity of edwardsiella tarda to other antibiotics (ampicillin, kanamycin and ciprofloxacin) can be significantly improved.
The foregoing examples are provided to fully disclose and describe how to make and use the claimed embodiments by those skilled in the art, and are not intended to limit the scope of the disclosure herein. Modifications that are obvious to a person skilled in the art will be within the scope of the appended claims.

Claims (10)

1. The application of vitamin B6 in preparing medicines for improving the sensitivity of bacteria to antibiotics is characterized in that: the bacteria are Edwardsiella tarda; the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
2. The application of vitamin B6 combined antibiotics in preparing antibacterial drugs is characterized in that: the bacteria are Edwardsiella tarda; the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
3. Use according to claim 1 or 2, characterized in that: the effective concentration of the vitamin B6 is 1.25-20 mM.
4. A use according to claim 3, characterized in that: when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 100-2500 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM;
when the antibiotic is ampicillin, the effective concentration of the ampicillin is 200-400 mg/L, and the effective concentration of the vitamin B6 is 1.25-20 mM;
when the antibiotic is kanamycin, the effective concentration of kanamycin is 600-1000 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM;
when the antibiotic is ciprofloxacin, the effective concentration of the ciprofloxacin is 2-5 mg/L, and the effective concentration of the vitamin B6 is 1.25-20 mM.
5. The use according to claim 4, characterized in that: when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 2000 mg/L, and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is ampicillin, the effective concentration of ampicillin is 300 mg/L and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is kanamycin, the effective concentration of kanamycin is 800 mg/L, and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is ciprofloxacin, the effective concentration of ciprofloxacin is 3 mg/L, and the effective concentration of vitamin B6 is 2.5 mM.
6. An antibacterial agent characterized in that: the active ingredients are vitamin B6 and antibiotics; the bacteria are Edwardsiella tarda, and the antibiotics are selected from any one or more than two of chloramphenicol, ampicillin, kanamycin and ciprofloxacin.
7. An antibacterial agent according to claim 6, wherein: when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 100-2500 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM;
when the antibiotic is ampicillin, the effective concentration of the ampicillin is 200-400 mg/L, and the effective concentration of the vitamin B6 is 1.25-20 mM;
when the antibiotic is kanamycin, the effective concentration of kanamycin is 600-1000 mg/L, and the effective concentration of vitamin B6 is 1.25-20 mM;
when the antibiotic is ciprofloxacin, the effective concentration of the ciprofloxacin is 2-5 mg/L, and the effective concentration of the vitamin B6 is 1.25-20 mM.
8. An antibacterial agent according to claim 7, wherein: when the antibiotic is chloramphenicol, the effective concentration of chloramphenicol is 2000 mg/L, and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is ampicillin, the effective concentration of ampicillin is 300 mg/L and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is kanamycin, the effective concentration of kanamycin is 800 mg/L, and the effective concentration of vitamin B6 is 2.5 mM;
when the antibiotic is ciprofloxacin, the effective concentration of ciprofloxacin is 3 mg/L, and the effective concentration of vitamin B6 is 2.5 mM.
9. An antibacterial agent according to claim 6, wherein: the antibacterial drug also comprises a pharmaceutically acceptable carrier.
10. An antibacterial agent according to claim 6, wherein: the feed additive consists of vitamin B6, chloramphenicol and water, wherein the concentration of the chloramphenicol is 100-2500 mg/L, and the concentration of the vitamin B6 is 1.25-20 mM.
CN202310577199.3A 2023-05-22 2023-05-22 Application of vitamin B6 in preparation of medicines for improving sensitivity of bacteria to antibiotics Pending CN116549448A (en)

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