Individual ingredients of NP-101 (Thymoquinone formula) inhibit SARS-CoV-2 pseudovirus infection
Abdelrahim Maen, Betul Gok Yavuz, Yehia I Mohamed, Abdullah Esmail, Jianming Lu, Amr Mohamed, Asfar S Azmi, Mohamed Kaseb, Osama Kasseb, Dan Li, Michelle Gocio, Mehmet Kocak, Abdelhafez Selim, Qing Ma, Ahmed O Kaseb
Frontiers in Pharmacology, doi:10.3389/fphar.2024.1291212
Thymoquinone TQ, an active ingredient of Nigella Sativa, has been shown to inhibit COVID-19 symptoms in clinical trials. Thymoquinone Formulation (TQF or NP-101) is developed as a novel enteric-coated medication derivative from Nigella Sativa. TQF consists of TQ with a favorable concentration and fatty acids, including palmitic, oleic, and linoleic acids. In this study, we aimed to investigate the roles of individual ingredients of TQF on infection of SARS-CoV-2 variants in-vitro, by utilizing Murine Leukemia Virus (MLV) based pseudovirus particles. We demonstrated that NP-101, TQ, and other individual ingredients, including oleic, linoleic, and palmitic acids inhibited SARS-CoV-2 infection in the MLV-based pseudovirus model. A large, randomized phase 2 study of NP-101 is planned in outpatient COVID-19 patients.
Author contributions AbM: Conceptualization, Data curation, Formal Analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Software, Supervision, Validation, Visualization, Writing-original draft, Writing-review and editing. BG: Conceptualization, Investigation, Software, Writing-original draft, Writing-review and editing. YM: Conceptualization, Data
Conflict of interest Author JL was employed by Codex BioSolutions Inc. Authors MoK, OK, and MG were employed by Novatek Pharmaceuticals, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.
Publisher's note All claims expressed in this article are solely those of the authors and do not necessarily represent those of their
Supplementary material The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fphar.2024.1291212/ full#supplementary-material
References
Abdelrahim, Esmail, Al Saadi, Zsigmond, Al Najjar et al., Thymoquinone's antiviral effects: it is time to be proven in the covid-19 pandemic era and its Omicron variant surge, Front. Pharmacol,
doi:10.3389/fphar.2022.848676Ashraf, Ashraf, Ashraf, Imran, Kalsoom et al., Therapeutic efficacy of Honey and Nigella sativa against COVID-19: a multicenter randomized controlled clinical trial (HNS-COVID-PK), medRxiv,
doi:10.1101/2020.10.30.20217364Barakat, El Wakeel, Hagag, Effects of Nigella sativa on outcome of hepatitis C in Egypt, World J. Gastroenterol,
doi:10.3748/wjg.v19.i16.2529Bencheqroun, Ahmed, Kocak, Villa, Barrera et al., A randomized, double-blind, placebo-controlled, multicenter study to evaluate the safety and efficacy of ThymoQuinone formula (TQF) for treating outpatient SARS-CoV-2, Pathogens,
doi:10.3390/pathogens11050551Chen, Xu, Pradhan, Gorshkov, Petersen et al., Identifying SARS-CoV-2 entry inhibitors through drug repurposing screens of SARS-S and MERS-S pseudotyped particles, ACS Pharmacol. Transl. Sci,
doi:10.1101/2020.07.10.197988Chen, Zhang, Li, Li, Liu et al., The impact of ACE2 polymorphisms on COVID-19 disease: susceptibility, severity, and therapy, Front. Cell. Infect. Microbiol,
doi:10.3389/fcimb.2021.753721Das, Arachidonic acid and other unsaturated fatty acids and some of their metabolites function as endogenous antimicrobial molecules: a review, J. Adv. Res,
doi:10.1016/j.jare.2018.01.001Goc, Niedzwiecki, Rath, Polyunsaturated ω-3 fatty acids inhibit ACE2-controlled SARS-CoV-2 binding and cellular entry, Sci. Rep,
doi:10.1038/s41598-021-84850-1Gu, Chen, Yang, He, Fan et al., Adaptation of SARS-CoV-2 in BALB/c mice for testing vaccine efficacy, Science,
doi:10.1126/science.abc4730Kaseb, Mohamed, Malek, Raad, Ii et al., The impact of angiotensin-converting enzyme 2 (ACE2) expression on the incidence and severity of COVID-19 infection, Pathogens,
doi:10.3390/pathogens10030379Kohn, Gitelman, Inbar, Interaction of polyunsaturated fatty acids with animal cells and enveloped viruses, Antimicrob. Agents Chemother,
doi:10.1128/AAC.18.6.962Kohn, Gitelman, Inbar, Unsaturated free fatty acids inactivate animal enveloped viruses, Arch. Virol,
doi:10.1007/bf01320626Koshak, Koshak, Mobeireek, Badawi, Wali et al., Nigella sativa for the treatment of COVID-19: an open-label randomized controlled clinical trial, Complement. Ther. Med,
doi:10.1016/j.ctim.2021.102769Koshak, Koshak, Mobeireek, Badawi, Wali et al., Nigella sativa for the treatment of COVID-19: an open-label randomized controlled clinical trial, Complementary Ther. Med,
doi:10.1016/j.ctim.2021.102769Liu, Wei, Kappler, Marrack, Zhang, SARS-CoV-2 variants of concern and variants of interest receptor binding domain mutations and virus infectivity, Front. Immunol,
doi:10.3389/fimmu.2022.825256Millet, Whittaker, Murine leukemia virus (MLV)-based Coronavirus spike-pseudotyped particle production and infection, Bio Protoc,
doi:10.21769/BioProtoc.2035Mostofa, Hossain, Basak, Sayeed, Thymoquinone as a potential adjuvant therapy for cancer treatment: evidence from preclinical studies, Front. Pharmacol,
doi:10.3389/fphar.2017.00295Obukhov, Stevens, Prasad, Li Calzi, Boulton et al., SARS-CoV-2 infections and ACE2: clinical outcomes linked with increased morbidity and mortality in individuals with diabetes, Diabetes,
doi:10.2337/dbi20-0019Onifade, Jewell, Adedeji, Nigella sativa concoction induced sustained seroreversion in HIV patient, Afr. J. traditional, complementary, Altern. Med,
doi:10.4314/ajtcam.v10i5.18Onifade, Jewell, Okesina, Seronegative conversion of an HIV positive subject treated with Nigella sativa and honey, Afr. J. Infect. Dis,
doi:10.4314/ajid.v9i2.6Pop, Sabin, Suciu, Vesa, Socaci et al., Nigella sativa's anti-inflammatory and antioxidative effects in experimental inflammation, Antioxidants,
doi:10.3390/antiox9100921Starr, Greaney, Hilton, Ellis, Crawford et al., Deep mutational scanning of SARS-CoV-2 receptor binding domain reveals constraints on folding and ACE2 binding, Cell,
doi:10.1016/j.cell.2020.08.012Villoutreix, Calvez, Marcelin, Khatib, In silico investigation of the new UK (B.1.1.7) and South African (501Y.V2) SARS-CoV-2 variants with a focus at the ACE2-spike RBD interface,
doi:10.3390/ijms22041695Xiao, Lu, Zhang, Johnson, Mckay et al., A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent, Nat. Struct. Mol. Biol,
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'abstract': '<jats:p>Thymoquinone TQ, an active ingredient of Nigella Sativa, has been shown to inhibit '
'COVID-19 symptoms in clinical trials. Thymoquinone Formulation (TQF or NP-101) is developed '
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