CN112999220A - Application of alpha-lipoic acid as and/or preparing metallo-beta-lactamase inhibitor - Google Patents
Application of alpha-lipoic acid as and/or preparing metallo-beta-lactamase inhibitor Download PDFInfo
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Abstract
The invention belongs to the field of medicines, and discloses application of alpha-lipoic acid as and/or in preparation of a metal beta-lactamase inhibitor. The invention discloses the application of alpha-lipoic acid and/or salt thereof as and/or for preparing a metallo-beta-lactamase inhibitor for the first time, wherein the alpha-lipoic acid and/or salt thereof has a good inhibition effect on the metallo-beta-lactamase, can protect antibiotics from being degraded by bacteria, improves the sensitivity of the bacteria to the antibiotics and reverses the drug resistance of the bacteria to the antibiotics; meanwhile, the alpha-lipoic acid and/or the salt thereof has good synergistic effect when being combined with antibiotics, and can be used as a medicament for inhibiting bacteria. The alpha-lipoic acid is a natural compound, basically has no toxic or side effect on a human body under normal clinical dosage, has been used as a medicament for treating the abnormal sensation caused by the diabetic peripheral neuropathy for many years in clinic, and has good safety.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to application of alpha-lipoic acid as and/or in preparation of a metal beta-lactamase inhibitor.
Background
Beta-lactam antibiotics become the most common clinical antibiotics due to the characteristics of high efficiency and low toxicity, and play an important role in controlling and treating gram-negative bacterial infection. However, long term unjustified use and even abuse of antibiotics has caused serious resistance to bacteria, which poses a serious threat to public health problems worldwide.
The most prominent resistance mechanism of bacteria is the production of beta-lactamases, which are resistant to antibiotics by hydrolyzing the lactam ring of beta-lactam antibiotics, rendering the antibiotics ineffective. Beta-lactamases can be classified into four classes a-D, based on their amino acid sequence, wherein the active site of class B beta-lactamases comprises 1-2 zinc ions and are therefore also known as metallo beta-lactamases (MBLs).
Based on the amino acid sequence, MBLs are further divided into 3 subclasses, B1, B2 and B3, while class B1 class of New Delhi metallo-beta-lactamase (NDM-1) has a broader substrate spectrum than the other subclasses of metallo-beta-lactamase, and can hydrolyze almost all beta-lactam antibiotics, including the carbapenem antibiotics known as the "last line of defense". The gene can not only rapidly carry out cross-strain propagation through plasmids, but also is very easy to generate variation due to the action of rearrangement of the plasmids, and the like, and can be integrated on the chromosome of bacteria to ensure that the heredity and the drug resistance of the bacteria are more stable. At present, no effective metallo-beta-lactamase inhibitor is available on the market clinically. Therefore, the development of metallo-beta-lactamase inhibitors is imminent.
alpha-Lipoic Acid (LA) is a natural product with biological activity and is a coenzyme for pyruvate dehydrogenase and glycine decarboxylase. The disulfide five-membered ring structure of the alpha-lipoic acid ensures that the alpha-lipoic acid has strong antioxidation, can eliminate active oxygen free radicals in vivo, simultaneously improves the levels of antioxidants such as glutathione and the like in cells, and greatly enhances the antioxidation. The alpha-lipoic acid can also improve the blood sugar control of the diabetic patients and reduce the dependence of the patients on insulin and hypoglycemic drugs; protecting nervous tissues of patients with diabetes, and helping to treat peripheral neuropathy; it also has positive effects on other chronic diseases, such as cardiovascular diseases, and nephropathy of liver and kidney. In recent years, alpha-lipoic acid has received high attention from the biological world at home and abroad because of playing an important role in the treatment of various diseases, but at present, no report on the aspect of the alpha-lipoic acid as a metallo-beta-lactamase inhibitor is found.
Disclosure of Invention
The object of the first aspect of the present invention is to provide the use of alpha-lipoic acid and/or salts thereof as inhibitors of metallo-beta-lactamases.
The object of the second aspect of the present invention is to provide the use of alpha-lipoic acid and/or its salts as and/or in the preparation of a medicament for increasing the sensitivity of bacteria to antibiotics.
The object of a third aspect of the invention is to provide the use of an antibiotic and alpha-lipoic acid and/or a salt thereof in the manufacture of a medicament for inhibiting bacteria.
The fourth aspect of the present invention is directed to a medicament comprising an antibiotic and alpha-lipoic acid and/or a salt thereof.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
in a first aspect of the invention, there is provided the use of alpha-lipoic acid and/or a salt thereof as and/or in the manufacture of a metallo-beta-lactamase inhibitor.
Alpha-lipoic acid is a natural product with biological activity, and is prepared from pyruvate dehydrogenase and glycine decarboxylaseCoenzyme of formula C8H14O2S2The CAS number is 1077-28-7, and the structural formula is shown as the formula (I).
Preferably, the metallo-beta-lactamase is at least one of an IMP-7 type metallo-beta-lactamase, an NDM-1 type metallo-beta-lactamase and a VIM-2 type metallo-beta-lactamase; further preferably, the metallo-beta-lactamase is an NDM-type 1 metallo-beta-lactamase.
Preferably, the source of the metallo-beta-lactamase comprises extraction in nature or preparation from a genetically engineered strain.
In a second aspect of the invention, there is provided the use of alpha-lipoic acid and/or a salt thereof as and/or in the manufacture of a medicament for increasing the sensitivity of a bacterium to an antibiotic.
Alpha-lipoic acid is a natural product with biological activity, and is coenzyme of pyruvate dehydrogenase and glycine decarboxylase, and its molecular formula is C8H14O2S2The CAS number is 1077-28-7, and the structural formula is shown as the formula (I).
Preferably, the bacterium is a drug-resistant bacterium that expresses a metallo-beta-lactamase; further preferably, the bacteria are Escherichia coli (Escherichia coli), Klebsiella pneumoniae (Klebsiella pneumoniae), Pseudomonas aeruginosa (Pseudomonas aeruginosa).
Preferably, the antibiotic is a beta-lactam antibiotic, and the beta-lactam antibiotic is an antibiotic with a beta-lactam ring in the molecular structure; further preferably, the antibiotic is at least one of penicillin, cephalosporin, cephamycin, thiomycin and carbapenem antibiotics; still more preferably, the antibiotic is at least one of meropenem, imipenem, ertapenem, cephalexin, cefuroxime, cefdinir, ceftriaxone, ceftazidime, ampicillin, and amoxicillin.
Preferably, the metallo-beta-lactamase is at least one of an IMP-7 type metallo-beta-lactamase, an NDM-1 type metallo-beta-lactamase and a VIM-2 type metallo-beta-lactamase.
In a third aspect of the invention, there is provided the use of an antibiotic and alpha-lipoic acid and/or a salt thereof in the manufacture of a medicament for inhibiting bacteria.
Alpha-lipoic acid is a natural product with biological activity, and is coenzyme of pyruvate dehydrogenase and glycine decarboxylase, and its molecular formula is C8H14O2S2The CAS number is 1077-28-7, and the structural formula is shown as the formula (I).
Preferably, the antibiotic is a beta-lactam antibiotic, and the beta-lactam antibiotic is an antibiotic with a beta-lactam ring in the molecular structure; further preferably, the antibiotic is at least one of penicillin, cephalosporin, cephamycin, thiomycin and carbapenem antibiotics; still more preferably, the antibiotic is at least one of meropenem, imipenem, ertapenem, cephalexin, cefuroxime, cefdinir, ceftriaxone, ceftazidime, ampicillin, and amoxicillin.
Preferably, the bacterium is a drug-resistant bacterium that expresses a metallo-beta-lactamase; further preferably, the bacteria are Escherichia coli (Escherichia coli), Klebsiella pneumoniae (Klebsiella pneumoniae), Pseudomonas aeruginosa (Pseudomonas aeruginosa).
Preferably, the metallo-beta-lactamase is at least one of an IMP-7 type metallo-beta-lactamase, an NDM-1 type metallo-beta-lactamase and a VIM-2 type metallo-beta-lactamase.
In a fourth aspect of the invention, there is provided a medicament comprising:
(1) (ii) an antibiotic; and
(2) alpha-lipoic acid and/or salts thereof.
Alpha-lipoic acid is a natural product with biological activity, and is coenzyme of pyruvate dehydrogenase and glycine decarboxylase, and its molecular formula is C8H14O2S2The CAS number is 1077-28-7, and the structural formula is shown as the formula (I).
Preferably, the antibiotic is a beta-lactam antibiotic, and the beta-lactam antibiotic is an antibiotic with a beta-lactam ring in the molecular structure; further preferably, the antibiotic is at least one of penicillin, cephalosporin, cephamycin, thiomycin and carbapenem antibiotics; still more preferably, the antibiotic is at least one of meropenem, imipenem, ertapenem, cephalexin, cefuroxime, cefdinir, ceftriaxone, ceftazidime, ampicillin, amoxicillin.
Preferably, the medicament further comprises pharmaceutically acceptable auxiliary materials.
Preferably, the adjuvant preferably includes at least one of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, a surfactant, an adsorption carrier, and a lubricant.
Preferably, the formulation types of the drug include solid formulations, liquid formulations, and semi-solid formulations.
Preferably, the solid formulation includes tablets, granules, powders and capsules.
Preferably, the liquid formulation comprises an injection.
Preferably, the semi-solid formulation comprises an ointment and a cream.
The invention has the beneficial effects that:
the invention discloses the application of alpha-lipoic acid and/or salt thereof as and/or for preparing a metallo-beta-lactamase inhibitor for the first time, wherein the alpha-lipoic acid and/or salt thereof has a good inhibition effect on the metallo-beta-lactamase, can protect antibiotics from being degraded by bacteria, improves the sensitivity of the bacteria to the antibiotics and reverses the drug resistance of the bacteria to the antibiotics; meanwhile, the alpha-lipoic acid and/or the salt thereof has good synergistic effect when being combined with antibiotics, and can be used as a compound medicine for inhibiting bacteria. Alpha-lipoic acid is a natural compound, basically has no toxic or side effect on a human body under normal clinical dosage, has been used as a medicament for treating abnormal sensations caused by diabetic peripheral neuropathy for many years in clinic, and has good safety.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. The materials, reagents and the like used in the present examples are commercially available reagents and materials unless otherwise specified.
Example 1 determination of the inhibitory Activity of alpha-lipoic acid on metallo beta-lactamases
The substrate is hydrolyzed by the enzyme to cause the decrease of the absorbance value, so that the degree of hydrolysis of the substrate can be characterized by the change of the absorbance, thereby judging the activity of the enzyme. The change in absorbance of the substrate after hydrolysis by metallo-beta-lactamase was measured at a wavelength of 300nm using meropenem (50. mu.M) as a reporter substrate. The concentration of metallo-beta-lactamase was 2nM, the buffer was 50mM HEPES (4-hydroxyethylpiperazineethanesulfonic acid), 0.1mM ZnSO4The pH is 7.2, the reaction temperature is 25 ℃, and the specific experimental method is as follows:
1. determination of alpha-lipoic acid inhibitory Activity on IMP-7 type metallo beta-lactamase
(1) Dissolving alpha-lipoic acid in HEPES buffer solution, preparing into different concentrations (0.1, 1, 10, 50, 100, 200, 300 and 500 mu M respectively), setting three multiple holes for each concentration, adding 10 mu L of IMP-7 type metallo-beta-lactamase solution (the final concentration is 2nM), and incubating at 25 ℃ for 15min to ensure that the alpha-lipoic acid and the enzyme are fully combined.
(2) The system is transferred into a quartz cuvette, 50 mu L of meropenem (the final concentration is 50 mu M) is added, the change of the absorbance value is rapidly measured, and the data is recorded.
(3) Calculating the inhibition rate of alpha-lipoic acid with different concentrations on IMP-7 type metallo-beta-lactamase, plotting the concentration of the compound on the inhibition rate, and calculating the IC by fitting a curve50The value is obtained.
2. Determination of the inhibitory Activity of alpha-lipoic acid on NDM-1 type metallo-beta-lactamase
(1) Dissolving alpha-lipoic acid in HEPES buffer solution, preparing into different concentrations (0.1, 1, 10, 50, 100, 200, 300 and 500 mu M respectively), setting three wells for each concentration, adding 10 mu L of NDM-1 type metallo-beta-lactamase solution (final concentration is 2nM), and incubating at 25 ℃ for 15min to fully combine the alpha-lipoic acid with the enzyme.
(2) The system is transferred into a quartz cuvette, 50 mu L of meropenem (the final concentration is 50 mu M) is added, the change of the absorbance value is rapidly measured, and the data is recorded.
(3) Calculating the inhibition rate of alpha-lipoic acid with different concentrations on NDM-1 type metallo-beta-lactamase, plotting the concentration of the compound on the inhibition rate, and calculating the IC by fitting a curve50The value is obtained.
3. Determination of the Activity of alpha-lipoic acid on the inhibition of VIM-2 metallo-beta-lactamase
(1) Dissolving alpha-lipoic acid in HEPES buffer solution, preparing into different concentrations (0.1, 1, 10, 50, 100, 200, 300 and 500 mu M respectively), setting three wells for each concentration, adding 10 mu L of VIM-2 type metallo-beta-lactamase solution (final concentration is 2nM), and incubating at 25 ℃ for 15min to fully combine the alpha-lipoic acid with the enzyme.
(2) The system is transferred into a quartz cuvette, 50 mu L of meropenem (the final concentration is 50 mu M) is added, the change of the absorbance value is rapidly measured, and the data is recorded.
(3) Calculating the inhibition rate of alpha-lipoic acid with different concentrations on VIM-2 type metallo-beta-lactamase, plotting the concentration of the compound on the inhibition rate, and calculating the IC by fitting a curve50The value is obtained.
The results of the inhibitory activity of alpha-lipoic acid against metallic beta-lactamase of IMP-7 type, NDM-1 type and VIM-2 type are shown in tables 1 and 2: IC of alpha-lipoic acid for NDM-1 type metallo beta-lactamase5062.75 +/-15.53 mu M and 36.29 +/-8.98 mu M of Ki value; IC for VIM-2 and IMP-7 metallo beta-lactamases50Are all more than 500 mu M; as can be seen, the alpha-lipoic acid has good inhibition effect on NDM-1 type metallo-beta-lactamase.
TABLE 1 IC of alpha-lipoic acid on MBLs50Value (μ M)
TABLE 2 inhibition constants (Ki, μ M) of alpha-lipoic acid to MBLs
Example 2 evaluation of Effect of alpha-lipoic acid in combination with Meropenem in inhibiting MBLs drug-resistant bacteria
The Minimum Inhibitory Concentration (MIC) of alpha-lipoic acid and meropenem combined to MBLs drug-resistant strains is determined by a broth dilution method. The strains for producing MBLs used in the experiment are genetically engineered bacteria E.coli BL21(DE3)/pET28a-IMP-7, E.coli BL21(DE3)/pET28a-VIM-2 and E.coli BL21(DE3)/pET28a-NDM-1, which are purchased from Shanghai Biotech Co., Ltd.
FICI is used to judge the interaction of two drugs when used in combination and is defined according to the following equationA+FICB＝CA/MICA+CB/MICBWherein MICAAnd MICBMIC values for Compounds A and B, respectively, alone, and CAAnd CBIs the drug concentration of compound a and B in the effective combination. If the FICI is less than or equal to 0.5, the two medicines are considered to have synergistic action, if the FICI is more than 0.5 and less than or equal to 4, the two medicines are considered to have weak synergistic action or no related action, and if the FICI is more than or equal to 4, the two medicines are considered to have antagonistic action. The smaller the FICI, the stronger the drug synergy.
The specific experimental method is as follows:
1. evaluation of Effect of alpha-lipoic acid and meropenem in combination on inhibition of IMP-7-producing type drug-resistant bacteria
(1) Inoculating a strain (E.coli BL21(DE3)/pET28a-IMP-7 strain) preserved at ultralow temperature into a sterile LB solid culture medium under the aseptic operating condition, putting the strain into a constant temperature incubator at 37 ℃ for overnight culture, picking a single colony, transferring the single colony into 3mL of LB liquid culture medium (containing 50mg/mL of kanamycin), and culturing the single colony in the constant temperature incubator at 37 ℃ until the logarithmic growth phase to obtain a bacterial suspension; adjusting bacteria liquid by using McLeod turbidimeterThe concentration is 0.5 McLee's concentration, LB liquid medium is diluted 100 times, and the bacterial number is about 1 × 106CFU/mL。
(2) Adding 100 mu L of LB liquid culture medium into the 2 nd to 12 th rows of the 96-well plate, adding 100 mu L of meropenem solution (256 mu g/mL) or inhibitor (alpha-lipoic acid, 256 mu g/mL) into the 1 st row, sucking 100 mu L of the liquid medicine in the 2 nd row after fully mixing the liquid medicine, adding the liquid medicine into the 3 rd row, mixing the liquid medicine again, and sequentially diluting the liquid medicine by the multiple dilution method to obtain the medicine concentration of 0.0625 to 128 mu g/mL; the MIC of IMP-7 type drug-resistant bacteria with meropenem or alpha-lipoic acid alone was determined by adding 100. mu.L of diluted bacterial solution to each well, and each experiment was repeated three times.
(3) Jointly diluting the liquid medicine on a 96-well plate according to the horizontal direction and the vertical direction, and carrying out gradient dilution on meropenem in the horizontal row, wherein the method is the same as the step 2, but the volumes of the added LB liquid culture medium and the meropenem are both 50 mu L (the final concentration of the meropenem is 0.0625-128 mu g/mL), and 50 mu L of inhibitor (alpha-lipoic acid) with different concentrations diluted in a multiple ratio in advance is added in the vertical row, and the final concentration is 2-128 mu g/mL; 100 μ L of diluted bacterial solution was added to each well to determine the MIC of meropenem in combination with an inhibitor (α -lipoic acid) against IMP-7 type MBLs-producing drug-resistant bacteria, and each experiment was repeated three times.
(4) Each set of experiments was set up with three parallel controls: taking Escherichia coli ATCC25922 as a quality control standard, taking captopril as a positive control, and simultaneously arranging a sterile hole and a drug-free hole; the 96-well plate was incubated in a 37 ℃ incubator for 24 hours, and the results were observed and MIC values were recorded.
2. Evaluation of effect of alpha-lipoic acid and meropenem in combination on inhibiting VIM-2-producing type drug-resistant bacteria
(1) Inoculating a strain (E.coli BL21(DE3)/pET28a-VIM-2 strain) preserved at ultralow temperature into a sterile LB solid culture medium under the aseptic operating condition, placing the strain in a constant temperature incubator at 37 ℃ for overnight culture, picking a single colony, transferring the single colony into 3mL of LB liquid culture medium (containing 50mg/mL of kanamycin), and culturing the single colony in the constant temperature incubator at 37 ℃ until the logarithmic growth phase to obtain a bacterial suspension; adjusting the concentration of the bacterial liquid to 0.5 McLeod concentration by a McLeod turbidimeter, diluting the LB liquid culture medium by 100 times, and obtaining the bacterial number of about 1 × 106CFU/mL。
(2) Adding 100 mu L of LB liquid culture medium into the 2 nd to 12 th rows of the 96-well plate, adding 100 mu L of meropenem solution (256 mu g/mL) or inhibitor (alpha-lipoic acid, 256 mu g/mL) into the 1 st row, sucking 100 mu L of the liquid medicine in the 2 nd row after fully mixing the liquid medicine, adding the liquid medicine into the 3 rd row, mixing the liquid medicine again, and sequentially diluting the liquid medicine by the multiple dilution method to obtain the medicine concentration of 0.0625 to 128 mu g/mL; the MIC of ViM-2-type drug-resistant bacteria with meropenem or alpha-lipoic acid alone was determined by adding 100. mu.L of diluted bacterial solution to each well, and each experiment was repeated three times.
(3) Jointly diluting the liquid medicine on a 96-well plate according to the horizontal direction and the vertical direction, and carrying out gradient dilution on meropenem in the horizontal row, wherein the method is the same as the step 2, but the volumes of the added LB liquid culture medium and the meropenem are both 50 mu L (the final concentration of the meropenem is 0.0625-128 mu g/mL), and 50 mu L of inhibitor (alpha-lipoic acid) with different concentrations diluted in a multiple ratio in advance is added in the vertical row, and the final concentration is 2-128 mu g/mL; the MIC of Meropenem in combination with an inhibitor (alpha-lipoic acid) against VIM-2 type MBLs-producing drug-resistant bacteria was determined by adding 100. mu.L of diluted bacterial solution to each well.
(4) Each set of experiments was set up with three parallel controls: taking Escherichia coli ATCC25922 as a quality control standard, taking captopril as a positive control, and simultaneously arranging a sterile hole and a drug-free hole; the 96-well plate was incubated in a 37 ℃ incubator for 24 hours, and the results were observed and MIC values were recorded.
3. Evaluation of effect of alpha-lipoic acid and meropenem in combination on inhibiting NDM-1-type drug-resistant bacteria
(1) Inoculating a strain (E.coli BL21(DE3)/pET28a-NDM-1 strain) preserved at ultralow temperature into a sterile LB solid culture medium under the aseptic operating condition, placing the strain in a constant temperature incubator at 37 ℃ for overnight culture, picking a single colony, transferring the single colony into 3mL of LB liquid culture medium (containing 50mg/mL of kanamycin), and culturing the single colony in the constant temperature incubator at 37 ℃ until the logarithmic growth phase to obtain a bacterial suspension; adjusting the concentration of the bacterial liquid to 0.5 McLeod concentration by a McLeod turbidimeter, diluting the LB liquid culture medium by 100 times, and obtaining the bacterial number of about 1 × 106CFU/mL。
(2) Adding 100 mu L of LB liquid culture medium into the 2 nd to 12 th rows of the 96-well plate, adding 100 mu L of meropenem solution (256 mu g/mL) or inhibitor (alpha-lipoic acid, 256 mu g/mL) into the 1 st row, sucking 100 mu L of the liquid medicine in the 2 nd row after fully mixing the liquid medicine, adding the liquid medicine into the 3 rd row, mixing the liquid medicine again, and sequentially diluting the liquid medicine by the multiple dilution method to obtain the medicine concentration of 0.0625 to 128 mu g/mL; the MIC of NDM-1 type drug-resistant bacteria with meropenem or alpha-lipoic acid alone was determined by adding 100. mu.L of diluted bacterial solution to each well, and each experiment was repeated three times.
(3) Jointly diluting the liquid medicine on a 96-well plate according to the horizontal direction and the vertical direction, and carrying out gradient dilution on meropenem in the horizontal row, wherein the method is the same as the step 2, but the volumes of the added LB liquid culture medium and the meropenem are both 50 mu L (the final concentration of the meropenem is 0.0625-128 mu g/mL), and 50 mu L of inhibitor (alpha-lipoic acid) with different concentrations diluted in a multiple ratio in advance is added in the vertical row, and the final concentration is 2-128 mu g/mL; 100 μ L of diluted bacterial solution was added to each well to determine the MIC of meropenem in combination with an inhibitor (α -lipoic acid) against NDM-1 type MBLs-producing drug-resistant bacteria, and each experiment was repeated three times.
(4) Each set of experiments was set up with three parallel controls: taking Escherichia coli ATCC25922 as a quality control standard, taking captopril as a positive control, and simultaneously arranging a sterile hole and a drug-free hole; the 96-well plate was incubated in a 37 ℃ incubator for 24 hours, and the results were observed and MIC values were recorded.
The results of the antibacterial activity of meropenem in combination with alpha-lipoic acid against drug-resistant bacteria expressing NDM-1, VIM-2 or IMP-1 are shown in Table 3: the inhibitor (alpha-lipoic acid, captopril) can improve the antibacterial activity of meropenem: when the concentration of the inhibitor is 128 mug/mL, the alpha-lipoic acid and the meropenem are used together, the bacteriostatic effect of the meropenem on various MBLs drug-resistant bacteria can be improved, and compared with the single use of the meropenem, the combined use can effectively reduce the MIC value of the meropenem on the drug-resistant strains, the MIC value can be reduced by 8 times to the maximum, and the effect is better than that of captopril.
The synergistic antimicrobial index of alpha-lipoic acid or captopril in combination with meropenem against MBLs-expressing resistant bacteria is shown in Table 4: the FICI of the alpha-lipoic acid and the Meropenem used together for the drug-resistant bacteria expressing the MBLs is less than or equal to 0.5, which shows that the alpha-lipoic acid and the Meropenem have good synergistic effect; the FICI of the captopril and the meropenem which are combined to the drug-resistant bacteria expressing MBLs is larger than that of alpha-lipoic acid, which shows that the synergistic effect of the captopril and the meropenem is not as good as that of the alpha-lipoic acid and the meropenem.
The above results show that: when the alpha-lipoic acid is combined with meropenem, the alpha-lipoic acid has effective synergistic antibacterial activity on various drug-resistant bacteria expressing metallo-beta-lactamase, which shows that the alpha-lipoic acid can be used as an MBLs inhibitor, can reverse the drug resistance of carbapenem drug-resistant bacteria, effectively protects meropenem from being hydrolyzed by MBLs, improves the antibacterial activity of the meropenem on the drug-resistant bacteria producing the MBLs, and further maintains the sensitivity to the meropenem. Therefore, the alpha-lipoic acid can be used as a metallo-beta-lactamase inhibitor and a beta-lactam antibiotic to prepare a composite preparation.
TABLE 3 MIC values (μ g/mL) for MBLs-expressing drug-resistant bacteria for alpha-lipoic acid or the positive control drug captopril in combination with meropenem
TABLE 4 synergistic antimicrobial index (FICI) of alpha-lipoic acid or the positive control drug captopril in combination with meropenem against MBLs-expressing resistant bacteria
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. Use of alpha-lipoic acid and/or salts thereof as inhibitors of metallo-beta-lactamases.
2. Use according to claim 1, characterized in that:
the metallo-beta-lactamase is at least one of IMP-7 type metallo-beta-lactamase, NDM-1 type metallo-beta-lactamase and VIM-2 type metallo-beta-lactamase.
3. Use of alpha-lipoic acid and/or salts thereof as and/or in the manufacture of a medicament for increasing the sensitivity of bacteria to antibiotics.
4. Use according to claim 3, characterized in that:
the antibiotic is beta-lactam antibiotic.
5. Use according to claim 4, characterized in that:
the bacteria are drug-resistant bacteria that express metallo-beta-lactamases.
6. Use of an antibiotic and alpha-lipoic acid and/or a salt thereof for the preparation of a medicament for inhibiting bacteria.
7. Use according to claim 6, characterized in that:
the antibiotic is beta-lactam antibiotic.
8. Use according to claim 7, characterized in that:
the bacteria are drug-resistant bacteria that express metallo-beta-lactamases.
9. A medicament, comprising:
(1) (ii) an antibiotic; and
(2) alpha-lipoic acid and/or salts thereof.
10. The medicament of claim 9, wherein:
the antibiotic is beta-lactam antibiotic.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110371383.3A CN112999220B (en) | 2021-04-07 | 2021-04-07 | Application of alpha-lipoic acid as and/or preparing metallo-beta-lactamase inhibitor |
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| CN113521063A (en) * | 2021-07-12 | 2021-10-22 | 中山大学 | Application of alpha-lipoic acid in resisting fish viral hemorrhagic septicemia virus infection |
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| CN113521063B (en) * | 2021-07-12 | 2023-03-10 | 中山大学 | Application of alpha-lipoic acid in resisting fish viral hemorrhagic septicemia virus infection |
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