Nanoparticles of ZnO/Berberine complex contract COVID-19 and respiratory co-bacterial infection in addition to elimination of hydroxychloroquine toxicity

Doaa A Ghareeb; Samar R Saleh; Mohamed G Seadawy; Mohammed S Nofal; Shaymaa A Abdulmalek; Salma F Hassan; Shaimaa M Khedr; Miral G AbdElwahab; Ahmed A Sobhy; Ali Saber Ali Abdel-Hamid; Abdelrahman Mohamed Yassin; Alshimaa A Abd Elmoneam; Aliaa A Masoud; Mohamed M Y Kaddah; Sally A El-Zahaby; Abdulaziz Mohsen Al-Mahallawi; Alaa M El-Gharbawy; Ahmed Zaki; Inas K Seif; Marwa Y Kenawy; Magdy Amin; Khaled Amer; Maha Adel El Demellawy

doi:10.1007/s40005-021-00544-w

Nanoparticles of ZnO/Berberine complex contract COVID-19 and respiratory co-bacterial infection in addition to elimination of hydroxychloroquine toxicity

J Pharm Investig. 2021;51(6):735-757. doi: 10.1007/s40005-021-00544-w. Epub 2021 Sep 6.

Authors

Doaa A Ghareeb^{1

2

3}, Samar R Saleh^{1

2

3}, Mohamed G Seadawy⁴, Mohammed S Nofal¹, Shaymaa A Abdulmalek^{1

2

3}, Salma F Hassan¹, Shaimaa M Khedr¹, Miral G AbdElwahab¹, Ahmed A Sobhy^{1

2

5}, Ali Saber Ali Abdel-Hamid¹, Abdelrahman Mohamed Yassin¹, Alshimaa A Abd Elmoneam^{2

3}, Aliaa A Masoud^{2

3}, Mohamed M Y Kaddah¹, Sally A El-Zahaby⁶, Abdulaziz Mohsen Al-Mahallawi^{7

8}, Alaa M El-Gharbawy¹, Ahmed Zaki¹, Inas K Seif^{1

2

3}, Marwa Y Kenawy^{2

3

9}, Magdy Amin¹⁰, Khaled Amer¹¹, Maha Adel El Demellawy^{1

12}

Affiliations

¹ Center of Excellence for Drug Preclinical Studies (CE-DPS), Pharmaceutical and Fermentation Industry Development Center, City of Scientific Research & Technological Applications, New Borg El Arab, Alexandria, Egypt.
² Bio-Screening and Preclinical Trial Lab, Biochemistry Department, Faculty of Science, Alexandria University, Alexandria, Egypt.
³ Biochemistry Department, Faculty of Science, Alexandria University, Alexandria, Egypt.
⁴ Chemical Warfare Department, MCL Almaza, Cairo, Egypt.
⁵ Clinical Pharmacy Program, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.
⁶ Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Pharos University in Alexandria, Alexandria, Egypt.
⁷ Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
⁸ School of Life and Medical Sciences, University of Hertfordshire Hosted by Global Academic Foundation, New Administrative Capital, Cairo, Egypt.
⁹ Fabrication Technology Research Department, Advanced Technology and New Materials Research Institute (ATNMRI), City of Scientific Research and Technological Applications (SRTA-City), New Borg El-Arab City, Alexandria, 21934 Egypt.
¹⁰ Military Medical Services, Cairo, Egypt.
¹¹ Egypt Center for Research and Regenerative Medicine, Cairo, Egypt.
¹² Medical Biotechnology Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research & Technological Applications, New Borg El Arab, Alexandria, Egypt.

Abstract

Purpose: A novel coronavirus (COVID-19) that has not been previously identified in humans and has no specific treatment has recently spread. Treatment trials using antiviral and immune-modulating drugs such as hydroxychloroquine (HCQ) were used to control this viral outbreak however several side effects have emerged. Berberine (BER) is an alkaloid that has been reported to reveal some pharmacological properties including antioxidant and antimicrobial activities. Additionally, Zinc oxide nanoparticles (ZnO-NPs) possess potent antioxidant and anti-inflammatory properties. Therefore, this study was undertaken to estimate the efficiency of both BER and synthetic ZnO/BER complex as an anti-COVID-19 therapy.

Methods: First, the ZnO/BER complex was prepared by the facile mixing method. Then in vitro studies on the two compounds were conducted including VeroE6 toxicity, anti-COVID-19 activity, determination of inhibitory activity towards papain-like proteinase (PL pro) and spike protein- and receptor- binding domain (RBD) as well as assessment of drug toxicity on RBCs.

Results: The results showed that ZnO/BER complex acts as an anti-COVID-19 by inhibiting spike protein binding with angiotensin-converting enzyme II (ACE II), PL pro activity, spike protein and E protein levels, and expression of both E-gene and RNA dependent RNA polymerase (RdRp) at a concentration lower than that of BER or ZnO-NPs alone. Furthermore, ZnO/BER complex had antioxidant and antimicrobial properties where it prevents the auto oxidation of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and the culture of lower respiratory system bacteria that affected Covid 19 patients. The ZnO/BER complex prevented as well the HCQ cytotoxic effect on both RBC and WBC (in vitro) and hepatotoxicity, nephrotoxicity and anemia that occurred after HCQ long administration in vivo.

Conclusion: The ZnO/BER complex can be accounted as promising anti-COVID 19 candidate because it inhibited the virus entry, replication, and assembly. Furthermore, it could be used to treat a second bacterial infection that took place in hospitalized COVID 19 patients. Moreover, ZnO/BER complex was found to eliminate the toxicity of long-term administration of HCQ in vivo.

Keywords: ACE2; Berberine; COVID-19; Molecular docking analysis; Papain-like proteinase; Spike protein RBD; Vero E6 toxicity; ZnO nanoparticles.