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The Antiviral Activity of Lopinavir/Ritonavir and Clarythromycin Combination to SARS-CoV-2 from Local Isolate Indonesia ( In Vitro Study)
12 Pages Posted: 18 Sep 2020More...
Background: The evaluation of potential drugs is urgently needed against SARS-CoV2 and Lopinavir/ritonavir and Clarythromycin was considered as the drugs.
Methods: Cytotoxicity test using MTT assay was done using human mesenchymal stem cells and drugs concentration used: 0.2; 2; 10; 100; 400 ppm for each Lopinavir, Clarithromycin and combining Lopinavir+Clarithromycin. We prepared Vero cells in 12-well plate for viral inoculation then given drugs with concentration: 1 ppm, 15 ppm, and 37.5 ppm of Lopinavir/ritonavir (200/50 mg), either 0.5 ppm, 4 ppm and 8 ppm of Chlarithromycin (500 mg) and 0.5 ppm, 5 ppm and 15 ppm of combination Lopinavir+Chlarithromycin. The qRT-PCR was performed to observe viral load. The observation were done in 24, 48, 72 hours after inoculation.
Results: There is no cytotoxic effect in human mesenchymal stem cells. Testing with multilevel concentration 50% (CC50) showed cytotoxicity level (ppm) for each Lopinavir, Clarithromycin and Lopinavir+Clarithromycin are 3451.8 ppm, 2285.97 ppm and 12230.6 ppm. The morphology changes in Vero cells showed formation of giant cells, colony of detachment cells and cyto pathic effect. Antiviral activity or inhibition concentration of 50% to virus (IC50) value of each, Lopinavir, Clarithromycin and combination Lopinavir+Clarithromycin are 0.9757; 0.9246; and <0.5 ppm. The combination Lopinavir+Clarithromycin had the lowest value than others that indicated the most potential and has capability as an antiviral activity.
Conclusion: Using drugs combination would reducing the dosage than using single drugs, so it can reducing toxicity potency to cells and increasing effectiveness in reducing the number of copies of SARS-COV-2 virus.
Funding Statement: The authors received no specific funding for this work.
Declaration of Interests: The authors report no conflicts of interest in this work.
Ethics Approval Statement: The patient's sputum sampling requires an ethics test that was submitted to the Rumah Sakit Universitas Airlangga (RSUA) Ethics Commission and has been approved as evidenced by the issuance of the ethics-worthy certificate number 136/KEP/2020 on April 20, 2020, following the regulatory guidelines of the country.
Keywords: Lopinavir; Clarythromycin; SARS-CoV2; in vitro; qRT-PCR
Suggested Citation: Suggested Citation