In Vitro Development of Local Antiviral Formulations with Potent Virucidal Activity Against SARS-CoV-2 and Influenza Viruses
Juthaporn Ponphaiboon, Wantanwa Krongrawa, Sontaya Limmatvapirat, Sukannika Tubtimsri, Akanitt Jittmittraphap, Pornsawan Leaungwutiwong, Chulabhorn Mahidol, Somsak Ruchirawat, Prasat Kittakoop, Chutima Limmatvapirat
Pharmaceutics, doi:10.3390/pharmaceutics17030349
Background/Object: This study investigates the in vitro antiviral potential of D-limonene (DLM), monolaurin (ML), and cetylpyridinium chloride (CPC) in formulations targeting SARS-CoV-2 and influenza viruses. The aim was to develop oral and nasal formulations with optimized concentrations of these active ingredients to evaluate their efficacy, safety, and stability. Methods: Oral (formulation D) and nasal (formulation E) products were developed using specific concentrations of DLM (0.2-0.3% w/w), ML (0.1-0.2% w/w), and CPC (0.05-0.075% w/w). In vitro virucidal activity assays were conducted to assess the antiviral efficacy of the formulations against SARS-CoV-2 and influenza viruses. Stability testing was also performed under various storage conditions. Results: Formulation D (0.3% w/w DLM, 0.2% w/w ML, 0.05% w/w CPC, and 1.5% w/w Cremophor RH40) demonstrated a 3.875 ± 0.1021 log reduction and 99.99 ± 0.0032% efficacy against SARS-CoV-2 within 120 s. Formulation E (0.2% w/w DLM, 0.05% w/w CPC, and 0.75% w/w Cremophor RH40) showed a 2.9063 ± 0.1197 log reduction and 99.87 ± 0.0369% efficacy against SARS-CoV-2. Both formulations achieved >99.99% efficacy and log reductions exceeding 4.000 against various influenza strains. Stability testing confirmed optimal performance at 4 • C with no microbial contamination. Conclusions: The findings suggest that both formulations exhibit broad-spectrum antiviral activity against SARS-CoV-2 and influenza viruses in vitro. These results support their potential for further clinical evaluations and therapeutic applications, particularly in oral and nasal spray formulations.
Supplementary Materials: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/pharmaceutics17030349/s1 , Figure S1 : Physical characteristics (turbid or clear) of formulations containing DLM and surfactant at different ratios, measured 1 day after preparation and after a temperature cycling test (6 cycles); Table S1 : Cytotoxicity and neutralization validation control data for oral formulation D in MDCK cells; Table S2 : Cytotoxicity and neutralization validation control data for nasal formulation E in MDCK cells; Table S3 : Test results for oral formulation D and nasal formulation E at a dilution of 1:32 against FluA(H1N1pdm) (A/Thailand/104/2009); Table S4 : Virus recovery data for FluA(H1N1pdm) (A/Thailand/104/2009); Table S5 : Virucidal activity of oral formulation D at a dilution of 1:32 against FluA(H1N1pdm) (A/Thailand/104/2009); Table S6 : Virucidal activity of nasal formulation E at a dilution of 1:32 against FluA(H1N1pdm) (A/Thailand/104/2009); Table S7 : Test results for oral formulation D and nasal formulation E at a dilution of 1:32 against FluA(H3N2) (ATCC VR-1881™); Table S8 : Virus recovery data for FluA(H3N2) (ATCC VR-1881™); Table S9 : Virucidal activity of oral formulation D at a dilution of 1:32 against FluA(H3N2) (ATCC VR-1881™); Table S10 : Virucidal activity of nasal formulation E at a dilution of 1:32 against FluA(H3N2) (ATCC VR-1881™); Table S11 : Test results for oral formulation D and nasal..
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doi:10.1056/NEJMc2001737DOI record:
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"abstract": "<jats:p>Background/Object: This study investigates the in vitro antiviral potential of D-limonene (DLM), monolaurin (ML), and cetylpyridinium chloride (CPC) in formulations targeting SARS-CoV-2 and influenza viruses. The aim was to develop oral and nasal formulations with optimized concentrations of these active ingredients to evaluate their efficacy, safety, and stability. Methods: Oral (formulation D) and nasal (formulation E) products were developed using specific concentrations of DLM (0.2–0.3% w/w), ML (0.1–0.2% w/w), and CPC (0.05–0.075% w/w). In vitro virucidal activity assays were conducted to assess the antiviral efficacy of the formulations against SARS-CoV-2 and influenza viruses. Stability testing was also performed under various storage conditions. Results: Formulation D (0.3% w/w DLM, 0.2% w/w ML, 0.05% w/w CPC, and 1.5% w/w Cremophor RH40) demonstrated a 3.875 ± 0.1021 log reduction and 99.99 ± 0.0032% efficacy against SARS-CoV-2 within 120 s. Formulation E (0.2% w/w DLM, 0.05% w/w CPC, and 0.75% w/w Cremophor RH40) showed a 2.9063 ± 0.1197 log reduction and 99.87 ± 0.0369% efficacy against SARS-CoV-2. Both formulations achieved >99.99% efficacy and log reductions exceeding 4.000 against various influenza strains. Stability testing confirmed optimal performance at 4 °C with no microbial contamination. Conclusions: The findings suggest that both formulations exhibit broad-spectrum antiviral activity against SARS-CoV-2 and influenza viruses in vitro. These results support their potential for further clinical evaluations and therapeutic applications, particularly in oral and nasal spray formulations.</jats:p>",
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