Chlorine dioxide aqueous solution can inactivate the binding of the variant spike proteins to the human ACE2 receptor protein

Proof that chlorine dioxide aqueous solution can inactivate the binding of the variant spike proteins to the human ACE2 receptor protein. This means that if you have had the VAX and you need to get rid of the spike proteins it will work.

http://www.remedypublications.com/open-access/inhibition-of-the-binding-of-variants-of-sars-cov-2-coronavirus-spike-7364.pdf

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The Time for Silence is Over

A unified pushback against the globalist agenda

It’s finally here, the Global Walkout begins September 4th at 8pm London time and continue every weeks. Next step january 29th.

One step at a time, hand in hand, we are walking out from the globalist society they are trying to enslave us into

ANYONE can participate
ANYWHERE in the world

JOIN or read about it here – https://globalwalkout.com

https://www.reignitefreedom.com/

The third step is to unsubscribe from all mainstream media outlets. Delete the apps from your phone, laptop, and tablet and unfollow all of their social media and YouTube channels. Try to avoid mainstream media for at least one week, even if the headline is intriguing.

In the same time why not removing all the big tech tracking/spying/social credit system around you: (Youtube, Facebook, Instagram, Twitter, Tik Tok, Google, Apple, Microsoft, Whatsapp, Zoom, Linkedln, Snapchat, Tumblr, Pinterest, Reddit, Myspace, etc.)

The fourth step of the global walkout is to move as many accounts as you can to a union or local bank.

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HERE

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https://childrenshealthdefense.org/child-health-topics/health-freedom/defender-days-sticker-gallery/

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Spike Protein Protocol 

Glutathione (most important for body detoxification) or better
NAC = N-Acetyl-Cysteine 600-750mg (causes the body to produce glutathione itself)
Zinc
Astaxantin 5mg (also improves vision)
Quercetin
vitamin D3
Milk thistle (also liver and stomach protection)
Melatonin 1mg to 10mg (against 5G)
Alternatively CDS/CDL and zeolite

Dr. Zelenko’s Protocol contains Ivermectin, Hydroxychloroquine (HCQ), Zinc, Vitamin D3, and Quercetin.

https://rightsfreedoms.wordpress.com/2021/09/13/summary-of-the-spike-protein-protocol-protection-against-spike-protein-and-vaccine-shedding-contagion-from-vaccinated-persons/

How to find the truth :

Search engine: https://presearch.org/, https://search.brave.com/, Searx (choose the server that you want) or https://metager.org/
Videos: www.odysee.com
www.bitchute.com
www.brandnewtube.com

Facebook style: www.gab.com or https://www.minds.com/

INTELLIGENCE ISN’T KNOWING EVERYTHING, IT’S THE ABILITY TO CHALLENGE EVERYTHING YOU KNOW

Annals of Pharmacology and Pharmaceutics
Remedy Publications LLC. 1 2021 | Volume 6 | Issue 1 | Article 1199
Introduction
Coronavirus Disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome
Coronavirus 2 (SARS-CoV-2) and has become a serious health issue since its emergence in late 2019
[1,2]. The outbreak of this disease originated in Wuhan, Hubei Province, China, before spreading
rapidly throughout the world [2]. The COVID-19 pandemic was declared an international
emergency in 2020 [3]. As of May 2021, global death toll due to COVID-19 is estimated to be
approximately 3.3 million [4]. SARS-CoV-2 is an enveloped virus and has a single-stranded
negative-sense RNA genome of 29.9 kilobases [5]. The virus attaches to the host cell surface of type
I and II pneumocytes, endothelial cells and ciliated bronchial epithelial cells [6]. Although several
vaccines against the disease have already been developed and administered [7], the pandemic has
not yet ended. Furthermore, currently variant types of SARS-CoV-2 virus have been reported
from England (B.1.1.7 variant) and South Africa (variant B.1.351) that appear to be spread more
readily than the original virus [8]. Both of these variants carry a common mutation (N501Y) in the
Receptor-Binding Domain (RBD) of the spike protein (S protein) [5]. This mutation is important
in binding of the spike protein to human cell surface receptor, Angiotensin-Converting Enzyme 2
(ACE2), a type I transmembrane glycoprotein with carboxypeptidase activity [5].
To control the spread of COVID-19, it is essential to develop and deploy safe but effective
disinfectants against the virus alongside novel antiviral drugs and vaccines [8]. While ethanol is
known to be effective against the virus [9], it cannot eradicate the virus present in the air. Chlorine
Dioxide (ClO2) (CD) is a water-soluble yellow gas at room temperature and is a stable free radical
[10]. CD can be used as a gas or an aqueous solution to inactivate both viruses and bacteria [11-15].
The powerful disinfection action of CD against microbes is due to its strong oxidizing activity against
proteins [16]. Moreover, safe and permissible concentrations of CD have been well documented
Inhibition of the Binding of Variants of SARS-CoV-2
Coronavirus Spike Protein to a Human Receptor by
Chlorine Dioxide
OPEN ACCESS
Correspondence: Norio Ogata, Department of R&D, Taiko Pharmaceutical Co., Ltd., 1-2-1 Hikaridai, Seikacho, 619-0237 Kyoto, Japan, Tel: 81 774 98 2716; Fax: 81 774 98 2737; E-mail: nogata7@yahoo.co.jp Received Date: 21 May 2021 Accepted Date: 16 Jun 2021 Published Date: 18 Jun 2021 Citation: Ogata N, Miura T. Inhibition of the Binding of Variants of SARS-CoV-2 Coronavirus Spike Protein to a Human Receptor by Chlorine Dioxide. Ann Pharmacol Pharm. 2021; 6(1): 1199. Copyright © 2021 Norio Ogata. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Research Article Published: 18 Jun, 2021 Abstract Aim: COVID-19 caused by a new coronavirus, SARS-CoV-2, has become an ongoing worldwide pandemic. A safe and potent virucidal disinfection system is urgently needed to protect the population from the virus. Chlorine Dioxide (ClO2) is a powerful disinfectant that is known to inactivate both viruses and bacteria. The aim of this study was to investigate whether chlorine dioxide inhibits the binding of the receptor-binding domain of the Spike protein (S protein) from variant coronavirus (British and South African variants) to human receptor, Angiotensin-Converting Enzyme 2 (ACE2). Materials and Methods: In vitro experiments to determine binding of the purified receptor-binding domain of spike protein to ACE2 were performed in the presence of various concentrations of chlorine dioxide. Purified spike proteins from the British and South African variants were used. Spike protein coated onto a microtiter plate was treated with chlorine dioxide aqueous solution or chlorine dioxide spray solution. Result: Binding of variant spike proteins was inhibited in a concentration-dependent manner (50% Inhibitory Concentration (IC50) of 7.6 µmol/L and 5.8 µmol/L for the British and the South African variants, respectively). Conclusion: These findings show that chlorine dioxide aqueous solution can inactivate the binding of the variant spike proteins to the human ACE2 receptor protein, indicating that this strategy may be useful in blocking the transmission of variant SARS-CoV-2 viruses. Keywords: Chlorine dioxide; COVID-19; SARS-CoV-2; Virus; Disinfection; IC50 Norio Ogata and Takanori Miura
Department of R&D, Taiko Pharmaceutical Co., Ltd, Japan
Norio Ogata, et al., Annals of Pharmacology and Pharmaceutics
Remedy Publications LLC. 2 2021 | Volume 6 | Issue 1 | Article 1199
[17]. Here, we demonstrate that CD aqueous solutions can inactivate
the binding of the spike proteins of both the British and South African
variant to human ACE2 in in vitro experiments, suggesting it may be
effective at inhibiting infection of the SARS-CoV-2 virus.
Materials and Methods
Chemicals
CD, prepared in our laboratory as described previously [16], was
dissolved in purified water and stored at 4ºC in a tightly capped amber
bottle. The stock CD was diluted in purified water to the desired
concentration immediately prior to use. A commercially available
CD spray solution sold under the brand name of Cleverin (Taiko
Pharmaceutical, Osaka, Japan) was also tested. The composition of
Cleverin was 1.48 mmol/L CD, 66.34 mmol/L sodium chlorite, 8.70
mmol/L sodium dihydrogen phosphates, 0.24 % (weight/weight)
decaglycerol monolaurate, 0.06 % silicone and 98.97 % water, pH
6.01. A tightly capped bottle of the spray solution made 3 years ago,
kept at room temperature and protected from the light was used. The
CD concentration of the spray solution at the time of the experiment
was 1.80 mmol/L (121 ppm (weight/weight)). The spray solution
was diluted as required with purified water immediately prior to the
experiment.
Binding assay
The spike protein binding assay kit was purchased from BPS
Bioscience (San Diego, California, USA). The kit codes for the British
and South African variant assay were 78140 and 78151, respectively.
The kits consisted of purified Receptor-Binding Domain (RBD) of
spike proteins derived from the corresponding variant virus. The kit
was used as recommended by the manufacturer with some
modifications. The spike protein RBD provided in the kit was first
coated onto a 96-well microtiter plate from the kit as indicated by the
manual. A 50 µL aliquot of CD or CD spray solution diluted to an
appropriate concentration was placed in each well and incubated at
25ºC for 5 min. Next, a 20 µL aliquot of 10 mmol/L sodium thiosulfate
in Blocking Buffer 2 provided in the kit was added to each well to
terminate the reaction of CD with the protein. Under this condition,
the CD is rapidly converted to ClO2- and becomes unreactive with
proteins [16]. Next, a 35 µL aliquot of biotin-labeled ACE2 diluted to
1.5 µg/mL by Blocking Buffer 2 was added to each well. Biotin-labeled
ACE2 was then bound to streptavidin-labeled horseradish peroxidase
and detected using a substrate of the horseradish peroxidase supplied
in the kit. The Chemiluminescence signal was measured after 1 min
using a luminometer (model SH-9000; Corona Electric, Hitachinaka,
Ibaraki, Japan). Each concentration of CD was assayed in four wells
(n=4). Because the chemiluminescence intensity fluctuated between
experiments, the final data was normalized (i.e. 0 µg/ml CD as 100%).
Results and Discussion
As shown in Figure 1, the intensity of the binding of human
ACE2 protein to the spike protein RBD decreased with increasing
concentrations of CD. The concentration of aqueous CD solution
required to give 50% binding inhibition (IC50) was 7.6 µmol/L and
5.8 µmol/L for the British and South African variant, respectively
(Table 1). An inhibitory effect of CD was also observed using the CD
spray solution (Figure 2). Specifically, IC50 for the binding of ACE2
to the spike protein RBD was 15.3 µmol/L and 4.7 µmol/L for the
British and South African variant, respectively (Table 1). Moreover,
binding of the Wuhan strain spike protein RBD to ACE2 was also
inhibited by the CD aqueous solution with an IC50 of 6.5 µmol/L
(Ogata N, unpublished data). There were slight differences in the
inhibitory effect between aqueous CD and the CD spray solution
in these experiments. The observed differences may be due to the
effect of other constituents in the spray solution, which are added to
prolong the shelf life of the product. These results suggest all the CD
solutions are effective in inactivating the binding of the SARS-CoV-2
virus to human cell receptor ACE2. As such, CD is likely to have an
inhibitory action on the infection of the virus.
The spike protein RBD of SARS-CoV-2 virus is located in the S1
Figure 1: Effects of chlorine dioxide (CD) aqueous solution on the binding
of the Receptor-Binding Domain (RBD) of variants of spike protein of SARSCoV-2 virus to human Angiotensin-Converting Enzyme 2 (ACE2) in in vitro
experiments. Binding of the spike protein of the British variant (filled circles)
and the South African variant (open circles) to ACE2 are shown. Each point
represents an average ± standard deviation of four experiments (n=4).
Figure 2: Effects of the Chlorine Dioxide (CD) spray solution on the binding
of the Receptor-Binding Domain (RBD) of variants of spike protein of SARSCoV-2 virus to human Angiotensin-Converting Enzyme 2 (ACE2) in in vitro
experiments. Binding of the spike protein of the British variant (filled circles)
and the South African variant (open circles) to ACE2 are shown. Each point
represents an average ± standard deviation of four experiments (n=4).
IC50 (µmol/L)
CD aqueous solution against
British variant 7.6
South African variant 5.8
CD spray solution against
British variant 15.3
South African variant 4.7
Table 1: The concentration of Chlorine Dioxide (CD) aqueous solution required
to inhibit binding of the receptor-binding domain of variant SARS-CoV-2 spike
proteins (S proteins) to human receptor protein ACE2 by 50% (IC50) is shown.
Variants of spike protein Receptor-Binding Domain (RBD) were treated with
either Chlorine Dioxide (CD) aqueous solution or CD spray solution at various
concentrations, and then assayed for its binding ability to the receptor protein
ACE2
Norio Ogata, et al., Annals of Pharmacology and Pharmaceutics
Remedy Publications LLC. 3 2021 | Volume 6 | Issue 1 | Article 1199
subunit of the spike protein, and the region of RBD that interacts with
ACE2 consists of a small patch of 25 amino acid residues [18].
The asparagine 501 (N501) residues, which is mutated to tyrosine
in both viral variants, forms part of the RBD patch region of the spike
protein [5]. The N501Y mutation is thought to result in enhanced
viral transmissibility by increasing the binding affinity to human
ACE2 [18]. Furthermore, it is worth noting that this mutation allows
the virus to escape from many antibodies raised against the virus [18],
suggesting poor effectiveness of vaccines against these variant strains
of SARS-CoV-2 virus. We previously demonstrated that the binding
of the spike protein RBD of the Wuhan strain to human ACE2 is
inhibited by CD aqueous solution [19]. These findings suggested CD
may be useful as a disinfectant against virus. Although the mechanism
of this inhibition has not been elucidated, we speculated that it may
involve oxidization of the tyrosine 453 residue of the RBD of the
spike protein, which forms a hydrogen bond with ACE2 [19]. Indeed,
CD is known to oxidize tyrosine residues in proteins [16]. Because
asparagine 501 is mutated to tyrosine (Y) in the variant strains,
treatment of the virus with CD may also oxidize 501Y. Given that
vaccines appear to have diminished effectiveness against the variant
viruses, disinfection systems employing CD may be invaluable. It
is also important to note that CD is a gas at room temperature and
can be used in this form to inactivate microbes floating in the air at
concentrations deemed safe to human health [17]. As such, gaseous
CD could be used to inactivate the SARS-CoV-2 virus floating in the
air of a crowded room.
Conclusion
CD solution, whether in water or as a formulated spray solution
in water, was found to potently inhibit the binding of spike protein
RBD to human receptor protein ACE2 in a concentration-dependent
manner. These findings suggest CD may be invaluable for inhibiting
the infection of SARS-CoV-2 virus to human.
Acknowledgement
The authors are employees of Taiko Pharmaceutical Co., Ltd. This
work was supported by the company.
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