TaffiX® nasal powder spray forms an effective barrier against infectious new variants of SARS-CoV-2 (COVID-19)

doi:10.21203/rs.3.rs-420365/v1

Download PDF

Short Report

TaffiX^® nasal powder spray forms an effective barrier against infectious new variants of SARS-CoV-2 (COVID-19)

https://doi.org/10.21203/rs.3.rs-420365/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 10 Oct, 2021

Read the published version in Archives of Clinical and Biomedical Research →

Version 1

posted

You are reading this older preprint version

Read the latest preprint version →

Introduction: While vaccination efforts against SARS-CoV-2 around the world are ongoing -, new high-infectious variants of the virus are being detected. The protection of the available vaccines against some of the new variants is weaker, and experts are concerned that newer as yet undescribed variants of this mutated RNA virus will eventually prove stable against the current vaccines. Additional preventive measures will therefore be needed to protect the population until effective vaccinations are widely available.

TaffiX^® is a personal, anti-viral nasal powder spray comprised of low pH Hypromellose that upon insufflation into the nose creates a thin gel layer covering the nasal mucosa and forming a protective mechanical barrier that prevents viruses from engaging with nasal cells- the main portal of entry for viruses. Taffix is commercially available in many countries across Europe, Asia America and Africa. In a prior preclinical study, TaffiX^®was found to be effective against SARS-CoV-2 Hong Kong/VM20001061/2020 in experimental in vitro conditions. A real-life clinical survey demonstrated that TaffiX^®nasal spray significantly reduced the SARS-CoV-2 infection rate post mass-gathering event in a highly endemic community.

Objective: The current study aimed to test the protective effect of Taffix against new pathogenic, highly infectious SARS-CoV-2 variants in vitro: the “British” B.1.1.7 (hCoV-19/Israel/CVL-46879-ngs/2020) and the “South African” B.1.351 (hCoV-19/Israel/CVL-2557-ngs/2020) variants.

Study design: A TaffiX® gel was formed on a nylon filter, using an amount equivalent to a clinical dose of Taffix . Filters were then seeded with SARS-CoV-2 B.1.1.7 (“British”) and B.1.351 (“South African”) variants. After a 10 -minute incubation at room temperature, the bottom of each filter was washed, and the resulting flow-through was collected and seeded into 24 -well plates containing Vero-E6 cells. After 5 days of incubation, a 200 µl sample from each well was taken for viral RNA extraction followed by SARS-CoV 2 RT-PCR analysis.

Results: The TaffiX^® gel completely blocked SARS-CoV-2 highly infectious variants B.1.1.7 and B.1.351 in vitro, reducing the titer of recoverable infectious virus as well as viral RNA by 100%.

Conclusions: Under in vitro conditions, TaffiX® formed an effective protective barrier against SARS-COV-2 variants (British variant and South African Variant). These results are consistent with prior findings demonstrating the in vitro high efficacy of Taffix gel in preventing viruses from reaching cells and infecting them. These results, added to clinical real-life studies performed with Taffix , support its use as an effective barrier against new variants of SARS-CoV-2 in conjunction with other protective measures.

Virology

CoVID-19

SARS-CoV-2

Variant B.1.1.7

Variant B.1.351

Taffix

Nasal spray

While vaccination efforts against SARS-CoV-2 around the world are ongoing, new high-infectious variants of the virus are being detected. The protection of the available vaccines against some of the new variants is weaker, and experts are concerned that newer as yet undescribed variants of this rapidly mutating RNA virus will eventually prove stable against the current vaccines .Additional preventive measures will therefore be needed to protect the population until effective vaccinations are widely available.

It is now well established that the nasal epithelium, specifically goblet and cilia cells, is the primary portal of entry of the SARS-CoV 2 virus into the body, and protection of nasal cells could therefore be meaningful in reducing the risk of infection .

TaffiX® is a personal, anti-viral nasal powder spray comprised of low pH Hypromellose that upon insufflation into the nose creates a thin gel layer covering the nasal mucosa forming a protective mechanical barrier that prevents viruses from engaging with nasal cells. The acidity of the gel enhances the barrier by inactivating the viruses trapped in it. Taffix is commercially available in many countries across Europe, Asia America and Africa. In a prior pre-clinical study TaffiX® was found effective against SARS-CoV-2 Hong Kong/VM20001061/2020 using experimental in vitro conditions. In a real-life clinical survey TaffiX® nasal spray significantly reduced SARS-CoV-2 infection rate post mass-gathering event at a highly endemic community

In this study we present in vitro data on the protective effect of Taffix against the new pathogenic, highly infectious variants of the SARS- CoV-2 : the “British” B.1.1.7 (hCoV-19/Israel/CVL-46879-ngs/2020) and the “South African” B.1.351 (hCoV-19/Israel/CVL-2557-ngs/2020) variants.

Virus B.1.1.7 stock was isolated from patients in Israel on 20 December 2020; virus B.1.351 stock was isolated from patients in Israel on 15 January 2021; cell strainers (40 µm Cat# SO SCS402) were obtained from AlexRed; qRT-PCR is described below; cell culture media (MEM-EAGLE) was obtained from BI Beit Haemek; Vero-E6 cells were obtained from HTCC; TaffiX® was supplied by Nasus Pharma.

TaffiX® Assay

One hundred fifty (150) µl of sterile water was added to individual sterile 40 µm cell strainers placed on a 50 ml tube. Twenty milligrams of TaffiX® was added and mixed with the water with a pipet tip until homogeneous across the cell strainer surface and incubated at room temperature for 10 minutes. Control filters were incubated with 150 µl of sterile water. Filters were then seeded with 10 µl of SARS-CoV-2 British or South African variants (each variant separately) at a concentration of Cycle Threshold (CT) 16.5 (approximately 5,000,000 copies/ml) or CT 20 (approximately ~ 600,000 copies/ml), which are parallel to the clinical concentrations found in patients’ nasal swabs diagnosed positive for SARS-CoV-2 (COVID-19).

After a 10-minute incubation at room temperature, the bottom of each filter was washed with 500 µl cell culture media, and the resulting flow-through was collected and diluted 1:10 with culture media (to prevent cytotoxicity to the Verro-E6 cells due to the acidic pH of TaffiX®). Then, 50 µl of the diluted flow-through was seeded into 24-well plates containing Vero-E6 cells (2x10⁵ cells/well, in 2% fetal calf serum). After 5 days of incubation at 33°C (5% CO₂), a 200 µl sample from each well was taken for viral RNA extraction followed by qRT-PCR analysis. Each treatment was repeated twice.

Quantitative Reverse Transcriptase PCR (qRT-PCR)

Nucleic acids were extracted using the MagLead extractor (PSS, Japan) according to the manufacturer’s instructions. RT-PCR reactions using primers corresponding to the SARS-CoV-2 envelope (E) gene were performed as previously described (Corman et al., Eurosurveillance, 2020) . qRT-PCRs were performed in 25 µl SensiFast reaction mix (Bioline, USA ) using TaqMan Chemistry on a CFX-96 instrument.

Using an amount equivalent to a clinical dose, the TaffiX® layer completely (100%) protected Vero-E6 cells from infection by two variants of SARS-CoV-2, B.1.1.7 (British) and B.1.351 (South African). For both variants, virus solution at an initial concentration of approximately 5,000,000 copies/ml (parallel to a clinical PCR result of CT 16.5) and 600,000 copies/ml (parallel to a clinical PCR result of CT 20) was applied to filter with and without TaffiX®. After 5 days of incubation of Vero- E6 cells with the flow-through solution eluted from the filter, no infection (zero) was diagnosed in the cells by qRT-PCR following the TaffiX® gel protection, in comparison with a control of filters without TaffiX®, were very high number of virus copies were detected in the cells: 2,236,550 copies/ml (filters inoculated with CT 20) and 25,027,670 copies/ml of B.1.1.7 variant (filters inoculated with CT 16.5, Fig. 1); and 506,578,602 copies/ml of the B.1.351 (filters inoculated with CT 20, Fig. 2).

In this series of experiments, the initial virus concentration used was equivalent to the clinical concentration found in nasal swabs taken from positive COVID-19 patients (CT 16.5, CT20), and they were all effectively blocked by the TaffiX® gel layer, which was applied at a clinical equivalent dose.

In a prior study, TaffiX® nasal spray powder formed a protective barrier against SARS-COV-2 Hong Kong/VM20001061/2020 under in vitro conditions preventing > 99.9% of viruses from infecting cells ⁸. The protective effect of TaffiX® in vitro was further verified in a human real-life survey conducted in Israel during September 2020 in a “superspreader” event where a reduction of 78% in COVID-19 cases was recorded among TaffiX® users with no reported side effects ⁹ .

Since then, new variants of the SARS-COV-2 infectious virus have emerged. According to the American Centers of Disease Control and Prevention (CDC) and the United Kingdom (UK) government publication , a newly identified variant called B.1.1.7 (“British”) with a large number of mutations was detected in the fall of 2020. This variant spreads more easily and quickly than other variants. Recent data from the UK reported that this variant may be associated with an increased risk of death compared to other variant viruses. It has since been detected in many countries around the world. This variant was first detected in the US at the end of December 2020 and is now the dominant variant found in new cases.

In South Africa, another variant called B.1.351 emerged independently of B.1.1.7. Originally detected in early October 2020, B.1.351 shares some mutations with B.1.1.7. Cases caused by this variant have been reported in the US at the end of January 2021

In Brazil, a variant called P.1 emerged that was first identified in travelers from Brazil who were tested during routine screening at an airport in Japan in early January 2021. This variant contains a set of additional mutations that may affect its ability to be recognized by antibodies. This variant was first detected in the US at the end of January 2021. As of March 2021, the “British” variant was the most abundant virus found in clinical samples of COVID-19 patients around the world, and the “South African” variant was the second most abundant (according to the CDC data).

Taffix is used worldwide as an additional layer of protection to help reduce the risk of contracting viral diseases caused by viruses whose first portal of entry to the body passes through the nasal cavity, such as influenza rhinoviruses and COVID-19. Its efficacy against the newly emerging variants of SARS-CoV 2 is therefore clinically relevant.

The concept of nonspecific protection against upper respiratory infectious viruses was first described. Hull et al. emphasized the potential value of creating a hostile microenvironment in the nasal cavity as an effective means of activating upper respiratory infectious pathogens. Indeed, many upper respiratory viruses are sensitive to low pH, including rhinoviruses , influenza , respiratory syncytial virus (RSV), and coronavirus . Hull et al. demonstrated in a randomized, double blind, placebo-controlled clinical study that irrigation with an acidic nasal hydrogel spray reduced the severity and duration of the common cold symptoms ¹⁹. Sungnak et al ^{5 9} found that SARS-CoV-2 entry factors are highly expressed in goblet and ciliated nasal epithelial cells together with innate immune genes. These factors make the nose likely a COVID virus entry point.

TaffiX® low pH gel is formed within 1 minute from spray and an acidic film is maintained on the nasal tissue for about 5 hours. The human nasal mucosal pH is approximately 5.5–6.5 ,which is a comfortable environment for most viral pathogens including the SARS-COV-2 virus.

In this study, we have shown that TaffiX® is very efficient in blocking the new variants under in vitro conditions.

Under in vitro conditions, TaffiX® formed an highly effective protective barrier against SARS-COV-2 variants (British variant and South African Variant). These results are consistent with prior findings demonstrating the in vitro high efficacy of Taffix gel in preventing viruses from reaching cells and infecting them. The body of work, in addition to clinical real-life studies performed with Taffix, support the use of nasal spray as an effective barrier against new variants of SARS-CoV-2 in conjunction with other protective measures.,

Since the development and distribution of new vaccines against novel variants is a long and complex process, additional protective measures are required. Taffix may provide a simple and safe way to reduce infections and thus the spread of COVID-19.

Funding Statement

This work was supported by Nasus Pharma, Tel Aviv, Israel.

Declaration of Competing Interest

T. Lapidot, PhD and D. Megiddo, MD are employees of Nasus Pharma.

Abdool Karim SS, de Oliveira T. New SARS-CoV-2 Variants - Clinical, Public Health, and Vaccine Implications. N Engl J Med. 2021 Mar 24:NEJMc2100362. doi: 10.1056/NEJMc2100362. Epub ahead of print. PMID: 33761203; PMCID: PMC8008749.
Rubin R. COVID-19 Vaccines vs Variants—Determining How Much Immunity Is Enough. JAMA. 2021;325(13):1241–1243. doi:10.1001/jama.2021.3370
https://www.medicalnewstoday.com/articles/new-sars-cov-2-variants-how-can-vaccines-be-adapted ( accessed April 12 2021)
Wang W, Xu Y, Gao R, et al. Detection of SARS-CoV-2 in Different Types of Clinical Specimens. JAMA. Published online March 11, 2020. doi:10.1001/jama.2020.3786
Sungnak W, Huang N, Bécavin C, et al. SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes. Nat Med. 2020;26(5):681‐687. doi:10.1038/s41591-020-0868-6
Hou YJ, Okuda K, Edwards CE, et al. SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract [published online ahead of print, 2020 May 27]. Cell. 2020;S0092-8674(20)30675-9. doi:10.1016/j.cell.2020.05.042
Gallo, O., Locatello, L.G., Mazzoni, A. et al. The central role of the nasal microenvironment in the transmission, modulation, and clinical progression of SARS-CoV-2 infection. Mucosal Immunol (2020). https://doi.org/10.1038/s41385-020-00359-2
Mann, Barbara J., et al. "TaffiX® Nasal Powder forms an Effective Barrier against SARS-CoV-2." Biomedical Journal of Scientific & Technical Research 33.3 (2021): 25843-25845.
Klang Shmuel, Megiddo Dalia, Lapidot Tair & Naparstek Yaakov (2021) Low pH Hypromellose (Taffix) nasal powder spray could reduce SARS-CoV-2 infection rate post mass-gathering event at a highly endemic community: an observational prospective open label user survey, Expert Review of Anti-infective Therapy, DOI: 10.1080/14787210.2021.1908127
https://www.cdc.gov/mmwr/volumes/70/wr/mm7003e2.htm
https://www.cdc.gov/mmwr/volumes/70/wr/mm7008e2.htm
Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DK, Bleicker T, Brünink S, Schneider J, Schmidt ML, Mulders DG, Haagmans BL, van der Veer B, van den Brink S, Wijsman L, Goderski G, Romette JL, Ellis J, Zambon M, Peiris M, Goossens H, Reusken C, Koopmans MP, Drosten C.Corman VM, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020 Jan;25(3):2000045. doi: 10.2807/1560-7917.ES.2020.25.3.2000045]
https://www.cdc.gov/mmwr/volumes/70/wr/mm7003e2.htm ( accessed April 12 2021)
https://www.gov.uk/government/news/phe-investigating-a-novel-variant-of-covid-19
Challen R, Brooks-Pollock E, Read J M, Dyson L, Tsaneva-Atanasova K, Danon L et al. Risk of mortality in patients infected with SARS-CoV-2 variant of concern 202012/1: matched cohort study BMJ 2021; 372 :n579 doi:10.1136/bmj.n579
https://www.nbcnews.com/science/science-news/uk-coronavirus-variant-now-dominant-strain-us-rcna606 (accessed April 12 2021)
Houriiyah Tegally; et al. (22 December 2020). "Emergence and rapid spread of a new severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) lineage with multiple spike mutations in South Africa". medrxiv. doi:10.1101/2020.12.21.20248640. Retrieved 10 March 2021
https://www.cdc.gov/coronavirus/2019-ncov/cases-updates/variant-surveillance/global-variant-map.html
Hull, D., et al. "Effects of creating a non-specific, virus-hostile environment in the nasopharynx on symptoms and duration of common cold." Acta otorhinolaryngologica italica 27.2 (2007): 73.
James E. Gern, Anne G. Mosser, Cheri A. Swenson, Paul J. Rennie, R. James England, Jacqueline Shaffer, Haruko Mizoguchi, Inhibition of Rhinovirus Replication In Vitro and In Vivo by Acid-Buffered Saline, The Journal of Infectious Diseases, Volume 195, Issue 8, 15 April 2007, Pages 1137–1143, https://doi.org/10.1086/512858
Hughes JH, Thomas DC, Hamparian VV: Acid lability of rhinovirus type 14 : effect of pH, time and temperature. Proc Soc Exp Biol Med 1973, 144:555–560.
Rennie, P., Bowtell, P., Hull, D. et al. Low pH gel intranasal sprays inactivate influenza viruses in vitro and protect ferrets against influenza infection. Respir Res 8, 38 (2007). https://doi.org/10.1186/1465-9921-8-38
Ausar SF, Rexroad J, Frolov VG, Look JL, Konar N, Middaugh CR. Analysis of the thermal and pH stability of human respiratory syncytial virus. Mol Pharm. 2005 Nov-Dec;2(6):491-9. doi: 10.1021/mp0500465. PMID: 16323956.
Alain Lamarre and, Pierre J. Talbot Effect of pH and temperature on the infectivity of human coronavirus 229E; Canadian Journal of Microbiology, 1989, 35(10): 972-974
England, R.J.A., Homer, J.J., Knight, L.C. and Ell, S.R., Nasal pH measurement: a reliable and repeatable parameter. Clinical Otolaryngology & Allied Sciences, (1999) 24: 67-68. doi:10.1046/j.1365-2273.1999.00223.x

Download PDF

Journal Publication

published 10 Oct, 2021

Read the published version in Archives of Clinical and Biomedical Research →

Version 1

posted

You are reading this older preprint version

Read the latest preprint version →

TaffiX^® nasal powder spray forms an effective barrier against infectious new variants of SARS-CoV-2 (COVID-19)

Status:

Journal Publication

Version 1

Abstract

Figures

Introduction

Materials And Methods

Results

Discussion

Conclusion

Declarations

References

Status:

Journal Publication

Version 1