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Advances in Pharmacology & Clinical Trials Research Article 5 min read

DNA Binding Potential and Antibacterial Activity of Metamizole

Mostafa Norizadeh Tazehkand*
* Corresponding author
ISSN: 2474-9214  10.23880/apct-16000172  Received: December 09, 2019  Published: January 08, 2020
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Keywords
Metamizole Antibacterial DNA Binding PUC18
Abstract

Metamizole is an anti-inflammatory medicine that is using in spasm, painkiller, reliever. The drug is using from a lot of people but there is a paucity of study about the antimicrobial and DNA binding activity of Metamizole. This study was aimed to investigate the antimicrobial (against Escherichia coli and Enterococcus faecalis) and DNA binding activity (on PUC18 Plasmid DNA) of Metamizole. The result of this study showed that Metamizole does not have antibacterial effect on Escherichia coli and Enterococcus faecalis, because both bacterial strain were grown at all concentrations (0.049 to 12.5 mg/ml). The electrophoretic pattern of DNA treated with Metamizole (concentrations of 250, 125 and 62.5 μg) showed same bands on agarose gel as negative control. Therefore Metamizole did not break the PUC18 vector DNA. It can be concluded that Metamizole might not pose a potential risk for our life. However, it must be investigated in other test systems.

Bacterial Strains

The test bacteria used in this study were from Bulent Ecevit University Medical Microbiology lab. Enterococcus faecalis ATCC 51299 as a gram positive bacteria and Escherichia coli ATCC 25922 as a gram negative bacteria. The bacterial strains were cultured in viable state via inoculation on Mueller-Hinton Broth and overnight incubation at 37°C.

Introduction
Material and Methods

Antimicrobial Activity of Metamizole

Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Metamizole were determined by dilution method.

Different concentrations of Metamizole (final concentrations of 0.049 to 12.5 mg/ml) were prepared and added to different tubes with 4 ml Mueller-Hinton Broth medium. So, a microbial suspension of 0.5 McFarland was obtained from two bacterial strains (Enterococcus faecalis ATCC 51299 as a gram positive bacteria and Escherichia coli ATCC 25922 as a gram negative bacterium) and the bacterial strains were cultured in prepared Mueller-Hinton Broth medium containing different Metamizole concentrations tubes. The tubes were then incubated at 37°C for overnight. The tube with the lowest concentration of the Metamizole at which no visible growth was reported as the MIC of the tested bacteria. The tubes were shaken to homogenize the contents and 100 μL of the contents of each tube was sub cultured by spreading Mueller-Hinton agar plastic petri dishes. The plates were incubated for 24 hours and then observed for any growth of colonies. Minimum bactericidal concentration was determined as the lowest concentration of Metamizole at which no colony observed following the sub culturing onto Mueller Hinton agar mediums [9, 10, 11].

DNA Binding Activity of Metamizole

DNA breaking potential of Metamizole was evaluated on PUC18 plasmid vector. The experiments were done in a volume of 8 μl in a microfuge tube containing 5 μl PUC18 (50μg/ml) plasmid DNA, and 5 μl of Metamizole in the concentrations of 250, 125 and 62.5 μg, respectively. In this research, untreated controls (PUC18 DNA) were also used. The reactions were incubated at 37°C temperature for 30 min. After incubation, the reaction mixture along with gel loading dye (6×) was loaded on a 2 % agarose gel for electrophoresis [12].

Result and Discussion

Antibacterial Activity of Metamizole

Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Metamizole were determined by dilution method. In this research we tested Enterococcus faecalis ATCC 51299 as a gram positive bacteria and Escherichia coli ATCC 25922 as gram negative bacteria. The result obtained from this research showed that Metamizole has no antibacterial effect on Enterococcus faecalis and Escherichia coli, because both bacterial strains were grown at all tested concentrations. For this reason we cannot report MIC and MBC for Metamizole against Enterococcus faecalis and Escherichia coli (Figure 2 & 3).

Figure 1: **CAS ID:** 68-89-3 **Molar mass:** 333.341 g/mol **Formula:** C13H17N3O4S **Excretion:** Urine (96%, IV; 85%, oral), faeces (4%, IV). **Bioavailability:** 100% (active metabolites) **Trade name:** Metamizole, dipyrone, Novalgin, others
Click to enlarge
Figure 1: CAS ID: 68-89-3 Molar mass: 333.341 g/mol Formula: C13H17N3O4S Excretion: Urine (96%, IV; 85%, oral), faeces (4%, IV). Bioavailability: 100% (active metabolites) Trade name: Metamizole, dipyrone, Novalgin, others
Figure 2: MIC result of _Enterococcus faecalis_ ATCC 51299_._ 1: 12.5 mg/ml, 2: 6.25 mg/ml, 3: 3.125 mg/ml, 4: 1.56 mg/ml, 5:0.781 mg/ml. Metamizole could not inhibited the growing of _Enterococcus faecalis._
Click to enlarge
Figure 2: MIC result of Enterococcus faecalis ATCC 51299_. 1: 12.5 mg/ml, 2: 6.25 mg/ml, 3: 3.125 mg/ml, 4: 1.56 mg/ml, 5:0.781 mg/ml. Metamizole could not inhibited the growing of _Enterococcus faecalis.

DNA Breaking Activity of Metamizole

As it was shown in Figure 4, the electrophoretic pattern of DNA treated with Metamizole (concentrations of 250, 125 and 62.5 μg) showed same bands on agarose gel electrophoresis as untreated control. Therefore Metamizole did not break the plasmid DNA.

The results of this study showed that Metamizole has not antibacterial effect on Enterococcus faecalis and Escherichia coli. According to our knowledge, it was the first study that addresses the antibacterial effects Metamizole against Enterococcus faecalis and Escherichia coli. The results obtained from PUC18 plasmid DNA damaging assay showed that Metamizole has no genetoxic effect on PUC18 DNA, because the electrophoretic pattern of PUC18 DNA after Metamizole treatment showed same bands on agarose gel electrophoresis as untreated control. In our life some time we use different drugs for pain relief. Some of these drugs have genotoxic or antibacterial effects, for this reason we need to study about these substances.

Figure 3: MIC result of _E. coli_ ATCC 25922. 1: 12.5 mg/ml, 2: 6.25 mg/ml, 3: 3.125 mg/ml, 4: 1.56 mg/ml, 5:0.781 mg/ml. Metamizole could not inhibited the growing of _E. coli._
Click to enlarge
Figure 3: MIC result of E. coli ATCC 25922. 1: 12.5 mg/ml, 2: 6.25 mg/ml, 3: 3.125 mg/ml, 4: 1.56 mg/ml, 5:0.781 mg/ml. Metamizole could not inhibited the growing of E. coli.

In contrast to Metamizole, number of reviews concerning the genotoxic property of some other pain relief drugs has been published. For example Oliván, et al. [13] showed that Diclofenac, ibuprofen and naproxen were responsible of alterations in biochemical biomarkers evaluated and DNA damage.

P.
aeruginosa ATCC 9027 and E. coli ATCC 8739.Milani, et
al. [16]investigated the antimicrobial activity of
diclofenac and ibuprofen on E. faecalis.In accordance with
this result,Milani, etal. [16]diclofenac and ibuprofen
have significantly more pronounced antibacterial activity
against E. faecalis.

Conclusion

The result of our research showed that Metamizole don’t have antimicrobial and genotoxic effect. It can be concluded that Metamizole might not pose a potential risk for our life. However, it must be investigated in other test systems.

Acknowledgments

This study was funded by Zonguldak Bulent Ecevit University Research Fund (Zonguldak Bulent Ecevit University- BAP) 2018-27095600-02.

References

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@article{mostafa2020,
  title   = {DNA Binding Potential and Antibacterial Activity of
Metamizole},
  author  = {Mostafa Norizadeh Tazehkand},
  journal = {Advances in Pharmacology & Clinical Trials},
  year    = {2020},
  volume  = {5},
  number  = {1},
  doi     = {10.23880/apct-16000172}
}
Mostafa Norizadeh Tazehkand (2020). DNA Binding Potential and Antibacterial Activity of
Metamizole. Advances in Pharmacology & Clinical Trials, 5(1). https://doi.org/10.23880/apct-16000172
TY  - JOUR
TI  - DNA Binding Potential and Antibacterial Activity of
Metamizole
AU  - Mostafa Norizadeh Tazehkand
JO  - Advances in Pharmacology & Clinical Trials
PY  - 2020
VL  - 5
IS  - 1
DO  - 10.23880/apct-16000172
ER  -