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Review of Russian ezrin peptide treatment of acute viral respiratory disease and virus induced pneumonia; a potential treatment for covid-19

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1
Review of Russian ezrin peptide treatment of acute viral
respiratory disease and virus induced pneumonia; a potential
treatment for covid-19.
Holms, R.D., 1 2, Bessler W.G., 3, Konopleva, M.V., 4 Ataullakhanov R.I., 5
1 Nearmedic International Ltd, Nicosia, Cyprus
2 Newal R&D Ltd, London, UK
3 Institute of Molecular Medicine and Cell Research, University of Freiburg, Germany
4 Gamaleya Institute, Moscow, Russian Federation
5 Institute of Immunology, Moscow, Russian Federation
SUMMARY
In 2020, the world is gripped by a very infectious and
pathogenic novel coronavirus: Severe Acute Respiratory
Syndrome Coronavirus 2 (SARS-CoV-2), also known as COVID-
19 virus, which causes the disease named COVID-19. However,
Ezrin-peptides may be a prophylactic and treatment solution
especially for the elderly.
1
2
3
4
5
Details of three separate
successful post-registration clinical trials using Ezrin-peptide
TEKKRRETVEREKE, for the treatment of Acute Viral Respiratory
Infection (AVRI) with complications including pneumonia, were
performed in Moscow over the previous eighteen years: Trial 1 with
100 patients, Trial 2 with 48 patients and Trial 3, 135 patients, are
translated, summarized and presented in this review.
INTRODUCTION
COVID-19 virus, 96% identical to a bat coronavirus, has
affinity for the cells lining the mucus membranes of the airways,
where its spike-protein binds to the ACE2-receptor.
6
7
COVID-19
virus then uses human ezrin, a sub-membrane protein that
regulates cell-signalling, shape and motility in epithelial cells, to
fuse with the human cell membrane and gain entry.
8
9
Once inside
human cells, COVID-19 virus replicates rapidly and induces an
inflammation, mediated mainly by IL-1 IL-6 IL-8 and TNF, that
leads to viral pneumonia, ‘cytokine storm’, lung damage and other
organ damage.
10
2
COVID-19 patients display a spectrum of disease severity.
About 80% have Acute Viral Respiratory Disease (AVRI) with fever
around 38oC, dry cough and a mild pneumonia. About 15% have
severe disease with lung inflammation leading to Acute Respiratory
Distress Syndrome (ARDS): including dyspnoea (shortness of
breath), and hypoxia (low blood oxygen). About 5% have critical
disease: Acute Lung Injury (ALI) including respiratory failure,
shock, multi-organ dysfunction and in about 0.5% to 2% cases,
death.
11
Human Ezrin, Old-Age and COVID-19 fatality
In Chinese and Italian cohorts of COVID-19 virus infected
people, severe and critical disease was very common in people
over 65 years old. In contrast, symptomatic infection in children
with COVID-19 virus is rare and mild. In a Chinese CDC report,
less than 2% of all symptomatic infections were in individuals
younger than 20 years old. In a small study of 10 children in China
who did develop symptoms, clinical illness was mild; 80 per cent
had fever, which resolved within 24 hours, 60% had cough, 40%
had sore throat, and none required supplemental oxygen.
12
There is clearly a human factor related to aging that is
interacting with COVID-19 virus. In 2012, it was discovered that
human ezrin, a submembrane protein that is involved in cell shape,
motility, receptor organisation and cell signalling, specifically bound
to the carboxy-terminus of the SARS coronavirus spike protein,
using its FERM domain. The coronavirus was dependent on using
a specific conformation of human ezrin to fuse with the epithelial
cells of the airways and successfully infect them. The fully active
conformation of ezrin, restrains coronavirus infection at the cell-
entry stage.
13
Increased expression of non-functional ezrin is associated
with organism age and senescence, which accumulates on the
interior surface of the cell membrane.
14
In old rats, there is a
fourfold increase in membrane associated ezrin in old epithelial
cells, compared to young epithelial cells.
15
Old mice also have
3
defective CD4 T lymphocytes, which display age-related defects in
ezrin-mediated cytoskeletal signals
16
. The failure of Ezrin signalling
complexes over time with age, may in part explain the age-related
fatality observed with COVID-19 disease.
Ezrin Peptide Therapy
Ezrin peptides mimicking the trigger-hinge region of the alpha-
domain of human ezrin, are highly polar molecules, with alternating
negative and positive charges, which act locally on epithelial cells
and fibroblasts in mucus membranes.
The receptors for ezrin peptides are believed to be membrane
associated ezrin-protein signalling complexes. Ezrin peptides are
thought to have an allosteric effect, which results in changes of the
conformation of ezrin into various functional forms. These changes
can not only prevent viruses entering cells, but also can activate
specific signalling pathways. Ezrin-protein complexes are
associated with the regulation of the ras>raf>MEK>ERK and
PI3K>PKB>mTOR intracellular signalling pathways, and they are
involved in the control of cytokine and interferon expression. Ezrin
peptides also act on fibroblasts to stimulate tissue repair
processes.
17
Clinical studies in Russia over twenty-five years have shown
that ezrin peptides can safely and effectively treat viral infections
caused by HIV, HCV, HPV, Herpes Simplex 1 & 2, and the
spectrum of viruses that cause Acute Viral Respiratory Infection
(AVRI). Clinical trials of ezrin peptide TEKKRRETVEREKE
[Gepon], determined that the ezrin-peptides possess anti-viral,
immuno-modulating activity and anti-inflammatory activity, and
could be used for successful prevention and treatment of a wide
range of infectious diseases caused by viruses, bacteria,
chlamydia, mycoplasmas, and candida fungi.
18
19
20
21
22
23
24
25
26
27
28
29
30
Generally, clinical studies have demonstrated that ezrin
peptides are safe, reduce virally induced inflammation, and lead to
faster recovery from Acute Viral Respiratory Infection (AVRI). Ezrin
4
peptides have even successfully treated viral pneumonia by
shutting down non-specific inflammation, and symptoms of viral
infection, probably by amplifying specific anti-viral responses
selected by the immune system.
CLINICAL STUDY ONE
100 patient clinical study into intra-nasal Ezrin-peptide TEKKRRETVEREKE
[Gepon] solution, as a treatment of Acute Viral Respiratory Infection [AVRI],
inflammatory Laryngeal-Tracheal-Bronchitis with Stenosis [LTBS] and Recurrent
Croup [RC]
Introduction
In year 2000, V.F. Uchaikin, Member of The Russian
Academy of Medical Sciences, organized a post-registration
clinical study of fourteen amino-acid synthetic Ezrin-peptide
TEKKRRETVEREKE [Gepon], at the Morozov Moscow Children’s
Hospital, in collaboration with The Russian Government Medical
University, Moscow.
The Principal Clinical Investigators in the clinical study into the
safety and efficacy of Gepon in Recurrent Acute Respiratory
Diseases were: Kladova O.V., MD, Doctor of Medical Sciences,
Professor, Department of children’s infectious diseases of the
Russian State Medical University: Legkova T.P., Head of Moscow
Children’s Hospital No18 and Ovchinnikova G.S., Doctor of
Children’s Home No 5
The objective of the clinical study was to test the efficacy of
intra-nasal application of ezrin-peptide TEKKRRETVEREKE
[Gepon]
31
, in treating of recurrent Acute Viral Respiratory Infection
[AVRI], which results in inflammation, and laryngitis-tracheitis-
bronchitis complicated with laryngeal stenosis and/or croup
syndrome.
32
33
34
The clinical research was approved by a Decision of the
Committee on Ethics (Minutes No: 6 of 06 December 2000), and a
Decision of the Pharmacology Committee (Minutes No: 14 of 21
December 2000). Permission to conduct clinical trials was issued
by the Department of the State for control of quality, efficiency and
5
safety of the medical preparations and equipment (No: 183 of 31
January 2001).
Assessed Patient Population
125 child-patients between the ages of 1 year to 14 years old,
suffering Acute Respiratory Virus Infection [AVRI], Virus Induced
Inflammation, Laryngeal-Tracheal-Bronchitis with Stenosis [LTBS]
and Recurrent Croup [RC] syndrome, were assessed for the
clinical trial.
ARVI occurred in 85% of the children. 50 children suffered
Laryngeal-Tracheal-Bronchitis with Laryngeal Stenosis (LTBS) and
75 children suffered Recurrent Croup (RC). The recurrent croup
was caused by a chronic virus-induced non-specific inflammation
of the upper respiratory tract. The frequency of recurring croup
(RC) in child-patients was between 3 to 35 times per annum.
Recurrent croup syndrome was associated with 1st degree stenosis
of the larynx in 54 of the RC patients (72%), by 2nd degree stenosis
of the larynx in 21 of the RC patients (28%).
10% of the croup cases had no fever, 60% of the croup cases
suffered sub-febrile fever, 30% of the croup cases suffered febrile
fever. The average duration of fever was 3-4 days. Bacterial
complications were observed in 15%, including sore throats, acute
otitis media, eustachyitis, and pneumonia. Half of the child-patients
were under treatment in hospital, and half were treated as out-
patients at home.
Immune Status of Assessed Patient Population
The 125 children of the assessed patient population were
offered immune status analysis, which was then compared to the
average status of healthy children. Generally, a profound non-
specific inflammatory response was being induced and maintained
by the viral infection. In contrast, specific anti-viral immunity had
been disrupted.
Blood samples were assessed by flow-cytometry using
monoclonal antibody markers: CD3, CD4, CD8, CD16, CD20,
6
CD1b, CD25, CD38, CD54, CD71, CD95, and HLA-DR, to
determine the distribution of lymphocyte sub-populations. In
addition, co-expression of [CD4+, CD8+], [CD8, DR] and [CD16+,
CD8+] was also measured. The ex vivo phagocytic activities of
blood monocytes and neutrophils versus Staphylococcus Aureus
were also determined.
The levels of inflammatory cytokines IL-1, IL-6, IL-8 and
TNFwere analysed. The functional activity of Th1 and Th2 cells
were assessed and the level of expression of the interleukins IL-2
and IL-4 were measured. Interferon status of the children was
assessed by the functional 1988 Yershov method and by
expression of Interferon-gamma. The concentrations of serum IgA
IgE, IgG and IgM immunoglobulins were also analysed.
The viral respiratory infection had induced a marked
imbalance in T cell and B cell immunity, and also significantly
impaired the normal functionality of blood monocytes, neutrophils
and lymphocytes. For example, lymphocyte adhesion and
apoptosis were 4x to 5x above normal levels.
The most consistent differences between the assessed child-
patients and healthy children, were the large significant increases
in the concentrations of inflammatory cytokines: IL-1, IL-6, IL-8
and TNF IL-6 was significantly elevated 2.1x in vivo and 9x in
vitro, and hugely elevated 26x when induced in vitro. IL-8 and
TNF also showed a similar pattern of massive elevation.(Table 1)
In the Assessed Patient Population, levels of T cell activating
IL-2 and IL-4, were significantly increased. The level of IL-2 in vivo
was increased 24x, in vitro spontaneous production of IL-2 was
increased 12x, and in vitro induced production increased by 5.1x.
The level of IL-4 both in vivo and in vitro also showed similar
elevations.
The level of mature T-lymphocytes in children was increased
by 1.3x over healthy children. Activated [CD71+, CD38+, CDA+,
ADD+] lymphocytes were significantly reduced. The assessed
patient population had marked changes in the immuno-regulatory
7
population of cells. The Th1 cell population was 0.63x less than
normal, whereas the Th2 cell population was 1.49x more than
normal. There was a significant reduction in the expression of IL-2
Receptors (IL2R+).
There was also a significant reduction in the proportion of
cytotoxic T cells [CD16+, CD8+, HLA-DR+] and activated [CD8+,
HLA-DR+] cells. Macrophage function was disrupted; phagocytosis
was reduced 1.6x, phagocytic-index was reduced 1.2x, but
absolute-phagocyte-indicator was 1.6x above normal. Neutrophils
were significantly increased.
Serum IgG and IgE were significantly increased, but IgA was
decreased, while IgM stayed normal. Interferon production was
also disrupted: interferon- interferon- and interferon- were also
significantly reduced.
8
Table 1.
Immunological Status of Assessed Children
with AVRI, Virus Induced Inflammation, LTBS and RC,
vs healthy children
Less than healthy children: -, Same as healthy children: n, More than healthy children: +,
Immunological Virus Induced Change
Parameter
Leucocytes ++
Lymphocytes -
Mature T lymphocytes +
Th1 Subpopulation - -
Th2 Subpopulation +
IRI ++
Activated CD8+ - -
IL2R+ cells -
HLA DR+ cells - -
NK cells -
CD16+CD8+ - -
Mature B lymphocytes ++
IgG ++
IgA - -
IgM n
IgE ++
Neutrophils ++
Segmented nuclear +
Rod-like nuclear n/+
Phagocytes -
Phagocytic index - -
Absolute phagocytic index +
Production of interferon - -
IL-1 alpha (in vivo) ++
IL-1 alpha (in vitro) ++
IL-2 (in vitro) ++
IL-2 (in vivo) ++
IL-4 (in vitro) ++
IL-4 (in vivo) ++
IL-6 (in vitro) ++
IL-6 (in vivo) ++
IL-8 (in vitro) ++
IL-8 (in vivo) ++
TNF alpha (in vitro) ++
TNF alpha (in vivo) ++
________________________________________________________________________________________
9
Criteria for the Gepon Clinical Study
Child-patients aged between 1 and 14 years, with a verified
diagnosis of recurrent respiratory disease, with 5 or more incidents
per annum recorded in their medical documentation, and suffering
from Laryngeal-Tracheal-Bronchitis with Stenosis and / or
Recurrent Croup, were included in the study.
Child-patients were excluded from the study: if the child-
patient refused to take part in the clinical trials; if they were below
1 year, or over 14 years of age; they had received any immuno-
modulator therapy with the previous 4 months; if other diseases
were present, such as insulin-dependent diabetes, tuberculosis,
chronic kidney and liver diseases, oncological diseases or HIV-
infection. Patients were also excluded: if their doctor’s advice was
not followed, if side effects appeared which might require special
treatment, and if the child-patient’s doctor decided that it was in the
interest of the child-patient to terminate participation in the Clinical
Study.
Entry of Child-Patients to the Gepon Clinical Study.
Of the 125 child-patients who had been assessed, 100 were invited
to join the clinical study of Gepon. Child-patient’s voluntary
participation in the clinical research, was subject to informed written
agreement by their parents or guardians. Participation in the clinical
research was voluntary, free of charge, and free of incentive
payment.
Each child-patient was assessed for: body temperature, skin
condition, peripheral lymph nodes, fauces (the arched opening at
the back of the mouth leading to the pharynx), and tonsils; function
of the lungs, heart, nervous system, muscular system and other
evaluations. The time elapsed between receiving written
agreement and the start of the Gepon therapy was between 1 and
14 days.
10
Patient Allocation to Gepon Group and Control Group
There was an initial non-randomised selection of child-
patients into two groups of 50 patients each, in which the spectrum
of symptoms were matched as far as possible: Group 1 suffering
from Acute Viral Respiratory Infection [ARVI] and Laryngeal-
Tracheal-Bronchitis with Stenosis [LTBS]; and Group 2 suffering
from recurrent Acute Viral Respiratory Infection [ARVI] only.
25 patients from Group 1 and 25 patients from Group 2 were
then randomly assigned to the Gepon Group n=50. 25 patients
from Group 1 and 25 patients from Group 2 were then randomly
assigned to the Control Group n=50. (Table 2 & 3)
Table 2
Group 1: Associated pathology in ARVI+LTBS child-patients
suffering from recurrent Laryngeal-Tracheal-Bronchitis with Stenosis
[LTBS] assigned to the Gepon Group (Gepon + symptomatic therapy)
and Control Group (symptomatic therapy only)
Presence of associated pathologies
Gepon Group
AVRI + LTBS
(n=25)
Control Group
AVRI + LTBS
(n=25)
1st grade Stenosis of the larynx (n=20)
9
11
2nd grade Stenosis of the larynx (n=5)
2
3
Recurring laryngeal-trachea-bronchitis
(n=0)
-
-
Recurring obstructive bronchitis(n=0)
-
-
Monthly incidence of AVRI (n=0)
-
-
Obstruction of breathing passages
during AVRI (n=31)
15
16
Lymphadenopathy (n=8)
4
4
Hypertrophy of palatine tonsils (n=14)
7
7
Atopic dermatitis (n=21)
10
11
Recurring obstructive bronchitis in
children over 3 years of age (n=8)
4
4
Recurring Croup in children over 3 years
of age (n=6)
3
3
Chronic tonsillitis in children over 3
years of age (n=3)
2
1
11
Table 3
Group 2: Associated pathology in AVRI child-patients
suffering from recurrent disease only assigned to the Gepon Group
(Gepon + symptomatic therapy) and Control Group (symptomatic
therapy only)
Presence of associated pathologies
Gepon Group
AVRI only
(n=25)
Control Group
AVRI only
(n=25)
1-grade Stenosis of the larynx (n=0)
-
-
2-grade Stenosis of the larynx (n=0)
-
-
Recurring laryngeal-trachea-bronchitis
(n=3)
1
2
Recurring obstructive bronchitis (n=6)
3
3
Monthly incidence of AVRI (n=16)
7
9
Obstruction of breathing passages
during AVRI (n=3)
2
1
Lymphadenopathy (n=7)
4
3
Hypertrophy of palatine tonsils (n=9)
4
5
Atopic dermatitis (n=6)
3
3
Recurring obstructive bronchitis in
children over 3 years of age (n=5)
3
2
Recurring Croup in children over 3 years
of age (n=4)
2
2
Chronic tonsillitis in children over 3
years of age (n=2)
1
1
Clinical Trial
Therapy commenced in September 2001. During the trial the
child-patients visited the doctor not less than 7 times: prior to start
of the therapy; during the 5 days of therapy, following therapy; 1
month after the therapy, and 3 months after the therapy. All 100
children received standard symptomatic treatment, using
antihistamine, anti-pyretic, and mucolytic drugs, bronchodilators
and alkaline aerosol inhalations.
Control Group n=50
The 50 children selected for the Control Group received
standard symptomatic treatment only for viral inflammation of the
airways. 13 children of the control group also received antibiotics
for secondary bacterial infections.
12
Gepon Group n=50
The 50 children selected for the Gepon Group received
standard symptomatic treatment for viral inflammation of the
airways plus Gepon therapy: 2mg sterile lyophilised ezrin-peptide
TEKKRRETVEREKE Gepon produced by LLC Immapharma,
dissolved in 2ml water to make a solution of 1mg/ml. Gepon
solution was delivered intra-nasally as 5 (40 micro-litre) drops in
each nasal passage, twice a day, for a period of 5 days (total
administration 2mg Gepon in 2ml). In the case of 7 children, who
had AVRI with bacterial complications, antibiotic treatment was
also applied in parallel to Gepon treatment.
RESULTS
Fig 1: Duration of Clinical Symptoms (Days)
Gepon Group n=50 (dark) and Control Group n=50 (light)
Horizontal divisions in days.
a: Fever, b: difficulty in breathing, c: serous rhinitis, g: swelling of nasal mucous
membrane, d: hyperaemia of tissues, e: pharyngitis, zh: swollen palatine glands, 3:
hoarseness of voice, i: dry cough, k: moist cough, l: stenosis of larynx, m: swollen neck
lymph nodes, n: reduction in the appetite, o: weakness, p: sleepiness, r: reduction in
physical activity, s: conjunctivitis, t: complications.
13
Table 4
Clinical symptoms Duration in Days
Gepon Group Control Group
n=50 n=50
Fever 2 4
Difficulty in nasal breathing 4 6
Serious Rhinitis 3 7
Mucus from nose 3 6
Inflamed Throat 3 6
Perturbation of Pharynx 3 6
Enlarged tonsils 4 6
Laryngitis 2 4
Dry Cough 4 6
Wet Cough 4 10
Obstruction of larynx 2 4
Enlarged lymph nodes 4 6
Reduced appetite 2 3
Weakness 3 4
Low physical activity 2 4
Conjunctivitis 0 3
Complications 1 3
Bold means 2 or more times reduction in duration of symptoms
SAFETY OF GEPON
No child-patient displayed any adverse reaction, nor any
adverse drug interaction, when receiving Gepon. No side effects
were observed. No allergic reactions were detected with Gepon
therapy. No intestinal-dysbiosis (common with antibiotics) was
detected. No child-patient suffering from atopic dermatitis
displayed any aggravation of illness. There were no
hypersensitivities resulting from the intranasal introduction Gepon
and no evidence of any contra-indications at any patient age.
14
THERAPEUTIC EFFICACY OF GEPON
All child-patients who received Ezrin peptide
TEKKRRETVEREKE [Gepon], in addition to standard symptomatic
treatment, benefitted from a significant shortening in the duration
of the clinical symptoms, which was independent of the severity of
AVRI or LTBS. The child-patients experienced the rapid decrease
in the duration of symptoms of disease, regardless of the severity
of the Acute Viral Respiratory Infection (ARVI), the amount of
inflammation of the airways, stenosis of the larynx or upper-airway
obstruction.
Comparisons between the 50 child-patient Gepon Group and
50 child-patient Control Group, showed that the duration of the
fever and other manifestations of intoxication syndrome, including
malaise, reduced appetite, weakness, sleepiness, and decrease in
physical activity, were all reduced.
All child-patients presented with dry-cough and fever, but after
they received Gepon therapy, they benefitted from a reduction in
the duration of fever by 3.2x times to only two days, and the
duration of dry cough by 1.8x, so that it stopped in less than four
days. Duration of rhinitis was 2.2x less and laryngitis was reduced
by 2.3x. Croup-cough disappeared on Day-3 of treatment with
Gepon. In contrast, only 38% of the Control Group managed to
eliminate croup-cough in the same period.
As a result of Gepon therapy, dissolution of mucus and
appearance of productive wet cough, a sign of recovery occurred
on Day-2 of Gepon therapy. In the Control Group, productive wet-
cough only got established after Day-5. Regardless of the degree
of inflammation and SLTB in the Gepon Group, 67% of cases had
recovered by Day-2 of treatment. In the same two days, 72% of the
children in the Gepon Group increased sputum density, while only
46% of the Control Group improved.
Gepon treatment reduced the bacterial complications
requiring antibiotics. Gepon even reduced the manifestations of
atopic dermatitis in the child patients
15
In the child-patients who were suffering more severe
AVRI+LTBS, Ezrin peptide TEKKRRETVEREKE [Gepon] was
demonstrated to be a rapid acting therapy. (Table 5)
Fever was shut down after 30 hours of Gepon therapy,
whereas the Control AVRI+SLTB Sub-Group continued to suffer
fever for 3 days. Maximum clinical benefit with Gepon was
achieved in 67% of cases, within 48 hours of receiving the
treatment.
Dry-cough disappeared in less than 60 hours in three-quarters
of the child-patients in the Gepon AVRI+SLTB Sub-Group,
whereas only about a third of child-patients recovered from dry-
cough in the Control AVRI+SLTB Sub-Group, over the same
period.
Table 5
Comparison of duration of clinical symptoms in child-patients in
The ‘Gepon AVRI+SLTB Sub-Group vs ‘Control AVRI+SLTB Sub-Group
Clinical Symptoms
Control AVRI+SLTB
Sub-Group
Duration in Days
Fever
2,9+-0,08
Difficult nasal breathing
5,1+-1,2
Serous Rhinitis
3,68+-0,4
Mucous in nose
4,6+-1,4
Hypermia of fauces
5,1+-1,3
Appearance of pharyngitis
5,1+-1,3
Enlargement of palatine tonsils
5,4+-0,9
Hoarseness of voice
2,84+-0,06
Dry cough
3,4+-1,6
Wet cough
3,9+-1,4
Stenosis of larynx
2,4+-0,5
Swollen lymph nodes
5,3+-1,6
Loss of appetite
2,65+-0,1
Weakness
2,5+-0,1
Sleepiness
1,9+-0,2
Reduction of physical activities
1,9+-0,2
Conjunctivitis
5+-0,3
Complications
3+-0,2
Significance of differences: *-p<0,001; **-p<0,005
Bold means 2 or more times reduction in days of symptoms
16
In the first two days of therapy, 72% of the child-patients in the
Gepon AVRI+SLTB Sub-Group benefited from the alteration of the
mucus consistency towards dissolution. In contrast, in the Control
AVRI+SLTB Sub-Group, only 46% of the child-patients enjoyed
this improvement in symptoms.
Gepon therapy significantly benefitted all the very sick child-
patients who required antibiotics. In the sub-group of child-patients
who were suffering from both AVRI and SLTB, and who had been
be prescribed antibiotics to manage secondary bacterial infection,
Gepon therapy granted them a significant shortening of the clinical
symptoms and reduction in the duration of the antibiotics therapy.
(Table 6)
Table 6. Comparison of Duration of Clinical Symptoms, in Child-Patients requiring
antibiotics, in Gepon Group vs Control Group
Duration of clinical symptoms
(in days)
Control AVRI+SLTB
Sub-Group
Antibiotic Therapy
Subgroup (n=12)
Fever
4,34+-0,8
Difficult nasal breathing
5,9+-0,7
Rhinitis
7+-0,8
Mucous in nose
5,9+-1,3
Hypermia of fauces
5,8+-1,0
Pharyngitis
6,4+-1,1
Enlargement of palatine tonsils
6,1+-1,2
Hoarseness of voice
3,7+-0,07
Dry cough
5,3+-1,2
Wet cough
9,7+-1,1
Stenosis of larynx
3,2+-0,6
Swollen lymph nodes
5,1+-1,6
Decline in appetite
2,98+-0,1
Weakness
3,8+-0,3
Sleepiness
3,4+-0,2
Reduction of physical activities
3,9+-0,2
Conjunctivitis
3+-0,1
Complications
3+-0,1
Significance of differences: *-p<0,001; **-p<0,005
Bold means 2 or more times reduction in days of symptoms
In the sub-group of child-patients suffering from AVRI and
LTBS, and who were treated with antibiotics: Gepon therapy
17
eliminated conjunctivitis within hours, and reduced fever to under
two days, compared to over four days in the control sub-group.
Gepon more than halved the average duration of stenosis of the
larynx to around 34 hours.
In addition, Gepon therapy given to child-patients suffering
AVRI+SLTB who were on antibiotics, reduced the ten-day duration
of wet-cough observed in the control group, to around three days
LONG-TERM GEPON PROTECTION IN 3 MONTH FOLLOW-UP
Child-patients who had received Gepon therapy, had a
significant decline in recurrence of respiratory disease during the
3-month period of observation, which followed treatment. On the
rarer occasions when disease did re-occur, the illness progressed
in a much milder form, and for shorter duration. After the first Gepon
treatment for AVRI, only mild 3-day episodes of disease recurred,
if at all, and the child-patients did not need hospitalisation. Gepon
eliminated secondary bacterial complications requiring antibiotic
therapy in almost all child-patients. In the Control Group, there
were no such reductions in severity of recurring disease.
Fig 2 Duration of Clinical Symptoms (Days)
In the 3-month post-therapy follow-up Gepon Group n=50 (dark) and
Control Group n=50 (light). Horizontal divisions in days.
A: Fever, B: Rhinitis, C: Wet-
Cough, D: Antibiotics, E: AVRI
A B C D E
18
During the subsequent 3 months of observation in the Gepon
Group the number of episodes AVRI was 0.5 per patient, whereas
in the Control Group it was 1.6 per patient. The duration of one-
episode AVRI in the Gepon Group, was 3.2 +- 0.3 days, whereas
in the Control Group it was 6.9 +- 0.1 days. In those child-patients
who received Gepon therapy but then fell ill again in the following
3 months, the AVRI was very mild, the duration of fever was
reduced 3,2x, the duration of rhinitis 2,1x; and productive cough
appeared on the average, on Day-2, compared to Day 5 in controls.
In the Gepon Sub-Group with AVRI only, prior to therapy there
were 17 cases of AVRI registered in 3 months. However, during
the 3 months of observation following Gepon therapy, only 8 cases
of AVRI cases were recorded. In contrast in the Control Group,
prior to therapy there were 16 cases of AVRI registered in 3 months
and during the 3 months of observation following therapy, there
were 13 cases of AVRI.
Prior Gepon therapy reduced duration of fever from 2.5 days
to 0.7 days, reduced duration of Wet Cough from 5.1 days to 2.2
days, the number of AVRI episodes from 6.9 days to 3.2 days and
eliminated secondary bacterial infection and the need for
antibiotics. (Table 7)
Table 7. Clinical AVRI symptoms in children during the 3 months of the
observation period, following completion of Gepon prophylactic therapy
Clinical Symptoms
Control Group
AVRI only
(n=25)
Duration in days
Fever
2.5+-0,04
Rhinitis
4.1+-0,2
Wet Cough
5.1+-0,3
AVRI episode
6.9+-0,1
Incidence
Antibiotics therapy
17 of 25
Significance of difference: p<0,001
Bold means 2 or more times reduction in days of symptoms
In child-patients with AVRI+SLTB who had received Gepon
therapy, the frequency of respiratory diseases 3 months after
19
completion of therapy, were reduced over 60%. Gepon reduced the
frequency of respiratory disease in child-patients with AVRI + SLTB
from 1.8 in the three months prior to therapy to 0.69 in the three
months after completion of therapy. (Table 8)
Table 8. Frequency of respiratory diseases in child-patients with AVRI+ SLTB
3 months prior to therapy and 3 months after completion of therapy
AVRI frequency in AVRI + SLTB Patients
Prior to Gepon therapy
3 months follow-up after therapy
Gepon
AVRI+SLTB Sub-
Group
Control
AVRI+SLTB Sub-
Group
Gepon
AVRI+SLTB Sub-
Group
Control
AVRI+SLTB Sub-
Group
1.8+-0,1
2.0+-0,2
0.69+-0,2
1.2+-0.1
Significance of difference: p<0,005
Bold means 2 or more times reduction
In child-patients with Recurrent AVRI, who had received Gepon
therapy, the frequency of respiratory diseases 3 months after
completion of therapy were also reduced over 60%. Gepon
reduced frequency of respiratory diseases in children with
Recurrent AVRI (no SLTB), from 3.1 in the three months prior to
therapy, to only 1.1 in the three months after completion of therapy
(Table 9)
Table 9. Frequency of respiratory diseases in children with Recurrent AVRI
3 months prior to therapy and 3 months after completion of therapy
AVRI frequency in recurrent AVRI Patients
Prior to therapy
3 months follow-up after therapy
Gepon
AVRI only
Sub-Group
Control
AVRI only
Sub-Group
Gepon
AVRI only
Sub-Group
Control
AVRI only
Sub-Group
3,1+-0.2
2,9+-0.3
1,1+-0.1
3,4+-0.3
20
PROPHYLACTIC ADMINISTRATION OF GEPON
After the successful completion of the clinical study with intra-
nasal ezrin-peptide TEKKRRETVEREKE [Gepon] therapy, the
prophylactic efficiency of Gepon was assessed in child-patients
who regularly suffered from recurrent AVRI.
Child-patients attending hospital were assessed for the
frequency of recurrent AVRI, longevity of AVRI, the type of clinical
symptoms (fever, intoxication, sputum production sputum, and
rhinitis) and the associated increases in allergic reactions, duration
and enlargement of swollen lymph nodes, inflammation of the
pharynx and tonsils, and the development of obstructive bronchitis
or croup syndrome.
In the Prophylaxis Study, the prophylaxis treatment regime
was 1 drop of Gepon solution (1 mg/ml) into each nasal passage,
3 times in the day, for 4 weeks. The result was no AVRI cases being
registered over the following three months in the Gepon Group. In
contrast in the Control Group of 0.6 cases per child were
registered.
DISCUSSION OF CLINICAL STUDY ONE
The results of this clinical study show that Gepon treatment is
safe, without side effects and well tolerated; Gepon is an effective
prophylactic and treatment for viral diseases such as AVRI and
SLTB which have an inflammatory component.
Ezrin-peptide TEKKRRETVEREKE [Gepon] restores order to
the immune responses dysregulated by pathogenic respiratory
viruses, while suppressing non-specific inflammation. Gepon
inhibits the expression of the inflammatory cytokines IL-1, IL-6, IL-
8 and TNF triggered by viral replication, while at the same time
triggers tissue repair and recovery processes. Ezrin-peptide
TEKKRRETVEREKE [Gepon] stimulates fibroblasts to repair the
disturbed epithelial barrier to restore effective protection to
bacterial, fungus and virus infection in the mucus membranes of
the airways.
21
Earlier studies demonstrated that ezrin-peptides could
significantly enhance specific humoral immunity against infections,
even in AIDS patients, where they amplified antibody production
against opportunistic infections.
35
It is remarkable how such a complex disease process as AVRI
induced SLTB croup, is gently but effectively reversed by ezrin-
peptide TEKKRRETVEREKE [Gepon]. In children with Acute
Respiratory Viral Infection (AVRI) and Stenotic Laryngo-Tracheo-
Bronchitis (SLTB), simple intra-nasal therapy with Gepon solution,
reliably reduced the duration and intensity of fever, reduced the
concentrations of inflammatory cytokines, reduced the severity and
duration of stenosis, reduced the inflammation of the larynx, and
converted dry-cough into productive cough by liquefaction of
sputum.
Children suffering from recurrent inflammatory AVRI,
benefited from Ezrin-peptide TEKKRRETVEREKE [Gepon], which
reduced duration and severity of recurrent AVRI. Gepon decreased
morbidity of AVRI by almost 3 times, as well as reducing the annual
incidents of AVRI.
In children suffering from Acute Viral Respiratory Infection
(AVRI) combined with Laryngeal-Tracheal-Bronchitis with Stenosis
(SLTB), intra nasal Gepon therapy decreased fever duration, the
incidence and duration of stenosis of the larynx, terminated dry-
cough, as well as decreasing the period of time before appearance
of productive cough with sputum.
In addition, long term benefits have been observed with ezrin-
peptides. For three months after Gepon treatment, there was no
recurrence of respiratory obstruction, no re-hospitalisation was
required for normally chronic recurrent patients. Gepon also
reduced the need for the treatment of secondary bacterial infection
with antibiotics. In child-patients with AVRI + SLTB, intranasal
therapy with Gepon reliably shortened the duration of fever, cured
the stenosis of larynx, and also reduced period dry cough and
stimulated the appearance of a productive cough.
22
Intra-nasal administration of Gepon is remarkably non-toxic
and safe. No side effects or unfavourable drug interactions were
detected. There are no known contra-indications for Gepon for any
age of patient. Thus, Gepon was remarkably effective in eliminating
AVRI and SLTB Croup.
CLINICAL STUDY TWO
Post-registration clinical study into Ezrin-peptide
TEKKRRETVEREKE [Gepon] treatment of chronic inflammatory
diseases of throat.
A clinical study of Gepon therapy, was performed at Russian
Government Medical University, Moscow, on 48 adult patients who
suffered either chronic inflammatory pharyngitis or chronic
inflammatory tonsillitis, with durations from 5 years to 25 years. The
Principal Investigators were T. S. Polyakova, M. M. Magomedov,
M E Artyemev, E V Surikov, and V. T Palchun.
36
37
Ezrin peptide TEKKRRETVEREKE [Gepon] solution was
investigated as a new method of treatment for inflammatory chronic
disease of the throat. Gepon is a rapid-acting anti-inflammatory
peptide, which suppresses inflammatory cytokines IL-1, IL-6, IL-8
and TNF. Gepon also amplifies anti-viral immunity and possesses
interferon induction activity that increases the expression of Type I
interferons: -interferon and -interferon.
The open clinical investigation of Gepon therapy, was
performed on 48 adult patients (20 men, 28 women, aged between
15 and 75 years) who had been suffering inflammatory disease of
the throat triggered by viral infection, with durations from 5 years to
25 years. Of these patients, 28 were suffering chronic inflammatory
pharyngitis, and 20 were suffering chronic tonsillitis. Throat
inflammation was associated with chronic candida infection in 32
cases and cocci flora in 16 cases.
At the commencement of the clinical study, the inflamed
mucous membranes of throat were examined in both groups of
patients: sub-atrophic pharyngitis was diagnosed in 16 patients,
atrophic pharyngitis in 5 patients and hypertrophic pharyngitis in 7
23
patients. The 16 sub-atrophic and atrophic pharyngitis were all
women. All 20 chronic tonsillitis patients had clear manifestations
of the disease.
Patients complained of pain in the throat, tickling sensation,
dryness in the mouth and the sensation of foreign body obstruction.
Viscous mucus was detected on the fauces, the arched opening at
the back of the mouth leading to the pharynx, and on rear wall of
the pharynx.
After ultrasonic washing of the mouth, throat and nose with
saline, a solution of 2mg Gepon in 5ml water was applied to the
throat using an ultrasonic irrigator. Three doses of Gepon solution
were administered using an ultrasonic irrigator on Day-1, Day-3
and Day-5 of treatment
By Day-2, all signs of inflammation of the throat had
disappeared, in 46 of 48 patients. Only two patients still displayed
hyperemia of mucus membrane of rear wall of the pharynx, but this
resolved by Day-5. The anti-inflammatory effect was confirmed by
microscope examination. In 45 patients (94%) Candida infection
had disappeared and cocci flora were reduced to insignificant
levels.
The 30-day follow up examination, showed that 46 of 48
patients maintained a healthy pharynx and tonsils, after years of
chronic inflammation (2 patients relapsed). The rapid cure rate of
96% of chronic pharyngitis and tonsillitis was impressive. No side
effects or adverse reactions to Gepon were observed.
CLINICAL STUDY THREE
Ezrin-peptide TEKKRRETVEREKE [Gepon] Solution-Vapour
Treatment of Acute Viral Respiratory Infection (AVRI), and
complications (Pneumonia)
A post-registration clinical study of ezrin-peptide
TEKKRRETVEREKE [Gepon] solution-vapour, administered to the
airways to treat Acute Viral Respiratory Infection (AVRI), and
complications such as Pneumonia. The clinical study was
performed at Department of Infectious Diseases, Moscow Hospital
24
No 1, in collaboration with the Russian Ministry of Health Institute
of Immunology, during 2008. The Principal Investigators of the
study were O.A. Safonova, A B Pichukin, E Sh Kozhemyakina, N
A Malshev and R I Ataullakhanov.
38
151 adult Acute Viral Respiratory Disease patients were
assessed to participate in the clinical study of ezrin-peptide
TEKKRRETVEREKE [Gepon] therapy. 135 patients were recruited
and gave informed written consent to join the clinical study.
All patients received standard symptomatic therapy: anti-
inflammatory paracetamol, antihistamines, expectorants and
inhalation of vaporized 0.2% sodium bicarbonate solution.
On admission to hospital, all patients presented with Acute
Viral Respiratory Infection (AVRI) with the following symptoms:
sore throat, cough, runny nose, hoarseness of voice, together with
symptoms of systemic intoxication including headache and
weakness.
Some patients with AVRI presented evidence of serious
inflammation of the sinuses, bronchitis, obstruction of the pharynx,
together with hyperaemia of the mucous membrane of the pharynx,
swollen tonsils, sores on pharynx wall, and purulent deposits.
Some patients complained of severe congestion, mucopurulent
discharge from the nose and debilitating headache, which required
X-ray examination of the nose. Other patients with AVRI present
evidence of lung infection and pneumonia: dry or wet cough,
shortness of breath, dry or wet wheezing which required X-ray
examination of the lungs.
Patients were screened by blood tests to identify the infecting
agents. Diagnostic tests were performed for the antigens of
influenza virus, parainfluenza virus, adenovirus, respiratory
syncytial virus (RSV), and other viruses.
25
Table 10. Number of Patients in Clinical Study
Infection
GROUP A
AVRI
uncomplicated
GROUP B
AVRI +
Sinusitis
Laryngitis
Bronchitis
GROUP C
AVRI +
Pneumonia
TOTAL
Influenza
27
16
25
68
Parainfluenza
3
7
3
13
Adenovirus
5
12
2
19
RSV
4
4
3
11
Mixed Virus
5
6
3
14
Unknown ID
4
4
2
10
TOTAL
48
49
38
135
IgM, IgA, IgG and IgE were measured together with
concentrations of C-reactive protein in the blood. Bacteriological
analysis of the sputum was used to identify Mycobacterium
tuberculosis (if present), non-specific micro-flora and any antibiotic
drug-resistance.
Flow cytometry was used to count peripheral blood immune
cell subpopulations, their activation markers and functional
subtypes such as CD4+ T helper cells, CD8+ cytotoxic T
lymphocytes and NK cells. Chemo-luminescence was applied for
the functional study of granulocyte ex vivo response to zymosan, a
fungal glucan recognized by TLR2 receptors.
The patients were then allocated to three sub-groups,
depending on the type and severity of symptoms, for the following
clinical studies of the safety and efficacy of Gepon vapour therapy:
Clinical Study A; AVRI (uncomplicated)
Clinical Study B; AVRI + serious inflammation
Clinical Study C; AVRI + pneumonia
26
Clinical Study A: uncomplicated AVRI
The 48 adult patients presenting AVRI only, but no
pneumonia, were enrolled for a randomised Clinical Study A of
Gepon inhalation therapy. 26 patients were randomly assigned to
the Gepon Group A and 22 to the Control Group A. Both groups
received standard therapy of vitamins, antihistamines (calcium
gluconate, diazolin), as well as antipyretic and anti-inflammatory
treatment (paracetamol) or phenaca if body temperature exceeded
38.5oC.
In Gepon Group A, 22 patients received 1mg in 5 ml Ezrin-
peptide TEKKRRETVEREKE [Gepon] solution vapour inhalation
per treatment. Gepon was prepared for treatment in batches by
dissolving 2mg lyophilised Gepon in 10ml of isotonic NaCl solution,
resulting in a 0.02% Gepon solution. 5ml of solution was added to
an ultrasonic Beron inhaler and blown into the nasal cavity and
airways of the patient, once a day, for 5 consecutive days. Total
course of therapy 5mg of peptide.
Ezrin peptide TEKKRRETVEREKE [Gepon] significantly
accelerated recovery from of Acute Viral Respiratory Infection
(AVRI).
Table 11. Duration of symptoms: Gepon vs Control (Group A)
Symptom
Gepon Group A
Duration in Days
Control Group A
Duration in Days
Significance
Fever
2.68
4.05
p<0.003
Dry-Cough
2.50
5.50
p<0.0001
Intoxication
2.85
4.14
p<0.003
Headache
2.67
3.90
p<0.006
Rhinitis
2.56
4.05
p<0.003
Weakness
2.92
4.14
p<0.005
The application of Gepon inhalation significantly accelerated
recovery from Acute Viral Respiratory Infections without
complications. On average, normalisation of body temperature was
achieved in 2.68 days compared to 4.05 days in the control group
27
and dry cough was stopped in under 2.5 days, while it persisted for
5 to 6 days in the Control group.
Clinical Study B: AVRI + throat inflammation, sinusitis,
tonsillitis and bronchitis
49 patients in Clinical Study Group B, AVRI + throat
inflammation, sinusitis, tonsillitis and bronchitis, all received
standard therapy. Only 12 patients received additional Gepon
inhalation therapy. By the third day of hospitalisation, all 37 Control
Group B patients (n=37), suffered worsening symptoms and had to
receive antibiotics for 5 to 7 days.
In contrast, the 12 patients of Gepon Group B (n=12) steadily
improved and duration of illness was significantly less.
Table 12. Duration of symptoms: Gepon vs Control (Group B)
GEPON
GROUP B
CONTROL
GROUP B
Fever
2.9 days
3.5 days
p <0.05
Intoxication
3.0 days
4.2 days
p<0.0001
Headache
2.3 days
3.5 days
p<0.005
Weakness
3.0 days
4.2 days
p<0.0001
Loss of Voice
2.0 days
3.8 days
p<0.097
There was a rapid reduction of inflammation in patients who
received Gepon inhalation. Bronchitis, Laryngitis and Sinusitis
persisted for about 8 days in the Control Group B, while all 12
patients had recovered in the Gepon Group after 5 to 6 days.
Clinical Study C; AVRI + pneumonia
38 patients (aged 30+/-14 years) suffering Acute Viral
Respiratory Disease (AVRI), complicated with pneumonia. Patients
suffered fever up to 39oC and 82% presenting dry cough. All 38
patients with AVRI+pneumonia, gave their written informed
consent to the clinical study, in which immuno-modulators would
be added to the Gepon Group in addition to standard therapy.
28
Patients with secondary bacterial infection also received antibiotics
and Immunomax, a macromolecular peptidoglycan immuno-
stimulator.
The patients were randomised, and two subgroups were
created: 20 patients were allocated to Control Group C and 18
patients were allocated to Gepon Group C.
All patients received standard intra-venous anti-bacteria
therapy with both cephalosporin and aminoglycoside antibiotics,
anti-inflammatory therapy with antihistamines, expectorants and
inhalation of vaporized 0.2% sodium bicarbonate solution, after
developing symptoms of pneumonia with AVRI.
Gepon Group C received 1mg in 5 ml Gepon solution vapour
inhalation per treatment. Ezrin-peptide TEKKRRETVEREKE
[Gepon] was prepared for treatment in batches by dissolving 2mg
lyophilised Gepon in 10ml of isotonic NaCl solution, resulting in a
0.02% Gepon solution. 5ml of solution was added to an ultrasonic
Beron inhaler and blown into the nasal cavity and airways of the
patient, once a day, for 5 consecutive days. The total course of
therapy was 5mg of peptide.
Ezrin peptide TEKKRRETVEREKE [Gepon] significantly
accelerated recovery from of Acute Viral Respiratory Infection
(AVRI) complicated with Pneumonia.
39
In patients of the Gepon
Group C suffering pneumonia, the duration of fever, intoxication,
headache and weakness were significantly shorter.
Table 13. Duration of symptoms: Gepon vs Control (Group C)
GEPON
GROUP C
CONTROL
GROUP C
Fever
2.9 days
5.1 days
p <0.05
Intoxication
3.2 days
5.3 days
p<0.0001
Headache
2.6 days
4.1 days
p<0.005
Weakness
3.0 days
5.3 days
p<0.0001
Gepon was particularly effective at reversing high fever
temperatures (39oC), triggered by lung infection and inflammatory
29
pneumonia. Gepon reduced by twenty per cent, the duration
shortness of breath, hypoxemia (low arterial blood gas), the need
for supplemental oxygen and breathing support.
Fig 3. Duration of Symptoms in Days of Patients
with Acute Viral Respiratory Infection and Pneumonia
Control Group (Light) Gepon Group (Dark)
30
Protection by Gepon from Bacterial Infection in Hospital
On the first day of hospitalisation, there were 79 patients who
present ARVI without bacterial complications. During the studies,
48 of 79 patients received standard symptomatic therapy only,
while 31 of 79 patients received additional Gepon inhalation
therapy. Antibiotics had to be prescribed to 26 patients who
received standard therapy only. In contrast there were only 5 cases
with patients who were receiving Gepon, who also needed
antibiotic therapy. Gepon inhalation therapy had reduced the risk
of bacterial infection in hospital by more than three times.
DISCUSSION
Relevance of Ezrin Peptide therapy to COVID-19 Disease
Ezrin Peptide therapy may be a therapeutic approach to
COVID-19 Disease. The new acute viral respiratory disease was
first identified in mid-December 2019 in the city of Wuhan of Hubei
Province in the centre of China, which has a population of 11 million
people. A novel type of coronavirus was identified as being the
causal agent and named 2019-nCoV virus. In February 2020, WHO
renamed the virus as Severe Acute Respiratory Syndrome
Coronavirus 2 (SARS-CoV-2), which caused a new disease
renamed COVID-19.
By Friday 13th March 2020, COVID-19 disease was spreading
rapidly and exponentially around the globe. Over three months, the
cumulative number of cases world-wide was over 135,000 and
there had been almost 5,000 deaths. It is estimated that just over
70,000 people recovered from COVID-19. Only two weeks later the
cumulative number of cases world-wide was about 550,000 and
there had been almost 25,000 deaths.
40
The fatality rate in Europe appears higher than in China.
41
Italy
now has the highest number of deaths in the world from COVID-19
virus. The fatality rate is around 5% of the confirmed infected
population, much higher than the global average of 3.4%,
according to the World Health Organization.
42
31
The V-strain of COVID-19 infecting the Italian population, may
be more pathogenic. Another factor affecting the high death rate in
Italy may be the older average age of the infected population
infected with COVID-19-virus. In Italy, about a quarter of infected
people are 65 or older and many of Italy's deaths have been among
people in their 80s, and 90s. On the other hand, the number of non-
confirmed infections could be much higher in Italy.
43
Generally, Coronaviruses belong to a family of viruses which
can induce disease ranging from mild “common cold” symptoms,
to Severe Acute Respiratory Syndrome (SARS) and Middle East
Respiratory Syndrome (MERS).
44
During the 20022003 epidemic
of SARS, a highly pathogenic coronavirus-SCV infected
approximately 8,000 individuals, and there was overall mortality of
infected people of 10%. In 2012 MERS-CoV was first identified in
a patient in the Middle East, and infected 2374 individuals and
caused 823 deaths over the following eight years.
45
46
47
The pathogenic properties SARS-CoV and MERS-CoV
coronaviruses have been studied closely. The high pathogenicity
of SARS-CoV and MERS-CoV coronaviruses is due to their high
affinity for airway epithelial cells, type II pneumocytes, and
endothelial cells of human lung alveolar micro capillaries.
48
49
Infection with these types of coronaviruses can cause
systemic inflammation accompanied by persistent hypotension,
hyperthermia or hypothermia, leukocytosis or leukopenia,
thrombocytopenia, and Acute Lung Injury (known as ALI) causing
Acute Respiratory Distress Syndrome (known as ARDS). In ALI
cases, the mortality rate is in the range 2030%, with about 55% of
the cases progressing to ARDS within a few days. ARDS causes
significant morbidity and approximately 40% mortality.
50
The Chinese Centre for Disease Control and Prevention
issued a report on 72 314 COVID-19-virus infected cases, which
revealed that of the population of COVID-19-virus infected people,
about 80% had “Mild disease”: ~38oC fever and dry cough by mild
or no mild pneumonia. About 15% had “Severe Disease”: including
dyspnoea (shortness of breath), hypoxia (low blood oxygen), and
32
lung damage. About 5% had “Critical Disease”: including
respiratory failure, shock, and multi-organ dysfunction.
87% of patients were between 30 and 79 years old. Older age
was also associated with increased mortality, with a case fatality
rate of 8% among those aged 70 to 79 years old and 15% 80 years
or older. The overall case fatality rate was 2.3%, and no deaths
were reported among non-critical cases.
51
52
In several cohorts of hospitalized patients with confirmed
COVID-19, the median age of the infected population ranged from
49 to 56 years. A study describing 138 patients with COVID-19
pneumonia in Wuhan reported that the most common clinical
features at the onset of illness were: Fever in 99 % (above 37.5oC),
Fatigue in 70%, Dry cough in 59 %, Anorexia in 40 %, Myalgia in
35%, Dyspnoea in 31 %, Sputum production in 27 %. The
dyspnoea (shortness of breath) developed after a median of five
days of illness. Acute Respiratory Distress Syndrome (ARDS)
developed in 20%, and mechanical ventilation was implemented in
12.3%.
53
COVID-19: The Immune Response
The innate antiviral response, particularly production of Type
I Interferon: IFN-α and IFN-β, is the first line of defense against
multiple virus infections. Type I Interferon mediates antiviral effects
by directly inhibiting virus replication and indirectly modulating the
host immune response to virus infection, both of which are
mediated by induction of interferon-stimulated genes (ISGs).
However, SARS-CoV and MERS-CoV have developed specific
mechanisms to block the signaling pathways of interferons and
IFN-stimulated genes (ISGs) of the innate immune system, which
allows the virus to maintain infection and replication.
54
55
The dysregulated innate immune system of patients with
coronavirus infection, displays delayed expression of Type I
Interferons, which is critical for initiation of the anti-viral innate
immune response, together with elevated expression of IL-1, IL-6,
33
IL-8 pro-inflammatory cytokines and CXCL-10, and MCP-1
chemokines, leading to extensive lung damage.
56
57
58
59
In Severe Cases of COVID-19 an uncontrolled immune
response known as “Cytokine Storm”, is mediated by the pro-
inflammatory cytokines IL-1 and IL-6. The immune over-reaction
among COVID-19 patients, leads to Acute Respiratory Distress
Syndrome (ARDS) and potentially life-threatening damage to lung
tissue.
Symptoms of ARDS include shortness of breath, rapid
breathing, and a bluish low-oxygen skin coloration. ARDS is
respiratory failure, resulting from widespread inflammation in the
lungs, which impairs the exchange oxygen and carbon dioxide in
the alveoli of the lungs. There is no known effective treatment for
ARDS, so supportive provision of oxygen to the failing lungs is the
only option.
No Effective Treatment for COVID-19 Virus and Disease
There is no treatment available for the inflammation and
pneumonia induced by COVID-19-virus. Both WHO and US CDC
warn that glucocorticoids, a class of corticosteroids, should not be
used to control the inflammation and auto-immunity induced by
COVID-19 virus, because they have been associated with an
increased risk of death in patients with influenza, a delayed viral
clearance in patients with MERS coronavirus, and generally there
is significant evidence of both adverse short-term and long-term
harm to patients.
Actemra, an anti-IL-6 receptor therapy for rheumatoid arthritis
produced by Hoffman La Roche has been used to treat lung
damage in serious cases coronavirus patients but the efficacy is
still uncertain. The Russian Federal Medical-Biological Agency is
investigating mefloquine. No antiviral drug has been demonstrated
to stop COVID-19 virus replication. However, a combination of
antivirals lopinavir and ritonavir developed to treat HIV, called
Kaletra (Aluvia) are being tested. In addition, the Russian influenza
remedy called Arbidol (Umifenovir) is also being tested. In addition,
34
Gilead Sciences is promoting Remdesivir as a potential anti-viral
treatment. However, no effective treatment for COVID-19 disease
has been demonstrated.
60
Interferon-inducers trigger IFN-α and IFN-β and early use of
interferon-inducers may be useful for prophylaxis COVID-19
disease. The Caco2 human cell line derived from epithelial
colorectal adenocarcinoma cells, supports the replication of
coronavirus-SCV that caused SARS. In this experimental system,
IFN-, -, and - have been found effective in inhibiting this
replication.
61
In animal models, prophylactic or early therapeutic
administration of recombinant IFN-β (rIFN-β) completely protected
animals from lethal MERS-CoV and SARS-CoV infection by
inhibiting virus replication and inflammatory cytokine production.
62
63
64
65
The use of type I interferons can decrease the effects of
infection with SARS-CoV and MERS-CoV viruses in animals if
used early in the detection of symptoms.
However, the timing of IFN therapy is critical. Delay in starting
rIFN-β therapy led to a huge increase in inflammatory cytokine
levels, resulting in fatal disease in an otherwise sub-lethal infection.
These results suggest that the timing of IFN-αβ receptor (IFNAR)
signaling, relative to peak coronavirus replication, is a critical
determinant of either protective immunity or pathogenic immunity
in coronavirus disease.
66
67
68
69
70
71
72
73
74
75
The clinical results presented in this paper, suggest that oral,
nasal and vapor inhalation Ezrin Peptide therapy should amplify
antiviral immunity and reduce inflammatory events in COVID-19
patients. There are no known adverse side effects with Ezrin
Peptide therapy, and it should be especially helpful for the therapy
of the elderly population.
35
1
Holms Rupert Donald “Aids Prophylactics” International Application Number:
PCT/GB95/001285, (02.06.95), International Publication Number: WO 95/33768
2
Holms, Rupert Donald “Regulatory/Unfolding Peptides of Ezrin” International
Application Number: PCT/GB00/03566 (15.09.2000) International Publication Number:
WO 01/025275
3
Holms, Rupert Donald, Ataullakhanov Ravshan Inoyatov “HCV Combination Therapy”
International Application Number: PCT/GB2004/000330 (27.01.2004) International
Publication Number: WO 2004067024 A2
4
Holms, Rupert Donald, Ataullakhanov Ravshan Inoyatov “The Use Of Peptides In Anti-
Ulcer Therapy” International Application Number: PCT/GB2006/004390 (23.11.2006)
International Publication Number: WO/2007/060440
5
Holms, Rupert Donald, Ataullakhanov Ravshan Inoyatov, Ataullakhanov Rustam,
Sayadyan Khachik “Ezrin-Derived Peptides and Pharmaceutical Compositions Thereof”
International Application Number: PCT/EP2016/062336 (01.06.2016) International
Publication Number: WO/2016/193285A1
6
Zhou P, et al “A pneumonia outbreak associated with a new coronavirus of probable
bat origin.” Nature volume 579, pages270–273 (2020);
https://www.nature.com/articles/s41586-020-2012-7
7
Hoffmann M, Kleine-Weber H, Schroeder S, Krüger N, Herrler T, Erichsen S,
Schiergens TS, Herrler G, Wu NH, Nitsche A, Müller MA, Drosten C, Pöhlmann S.
SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a
Clinically Proven Protease Inhibitor. Cell. 2020 Mar 4. pii: S0092-8674(20)30229-4. doi:
10.1016/j.cell.2020.02.052.
8
Rouven B.B., Pietuch A., Nehls S., Rother J., Janshoff A. Ezrin is a Major Regulator of
Membrane Tension in Epithelial Cells. Sci Rep. 2015 Oct 5;5:14700. doi:
10.1038/srep14700.
9
Millet J K et al “Ezrin Interacts with the SARS Coronavirus Spike Protein and Restrains
Infection at the Entry Stage” Published: November 21,
2012https://doi.org/10.1371/journal.pone.0049566
10
Mehta P., et al. COVID-19: consider cytokine storm syndromes and
immunosuppression.” The Lancet VOLUME 395 (2020) p1033-1034,
DOI:https://doi.org/10.1016/S0140-6736(20)30628-0
11
Chen N, et al “Epidemiological and clinical characteristics of 99 cases of 2019 novel
coronavirus pneumonia in Wuhan, China: a descriptive study.” Lancet.
2020;395(10223):507. Epub 2020 Jan 30.
36
12
Wu Z, McGoogan JM, “Characteristics of and Important Lessons from the
Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of
72 314 Cases From the Chinese Centre for Disease Control and Prevention.” JAMA.
Published online February 24, 2020. doi:10.1001/jama.2020.2648
13
Millet J K et al Ezrin Interacts with the SARS Coronavirus Spike Protein and Restrains
Infection at the Entry Stage Published: November 21,
2012https://doi.org/10.1371/journal.pone.0049566
14
Yang HS, Hinds PW. Increased ezrin expression and activation by CDK5 coincident
with acquisition of the senescent phenotype. Mol Cell. 2003 May;11(5):1163-76.
15
Xiaorong Gu et al “Age-Related Changes in the Retinal Pigment Epithelium” PLoS
One. 2012;7(6):e38673. doi: 10.1371/journal.pone.0038673. Epub 2012 Jun 11.
16
Garcia G.G., Sadighi Akha A.A.,*, Miller R.A, “Age-related Defects in Moesin/Ezrin
Cytoskeletal Signals in Mouse CD4 T cells.” J Immunol. 2007 November 15; 179(10):
64036409.
17
Marina Chulkina M, Negmadjanov U, Lebedeva E, Pichugin A, Mazurova D,
Ataullakhanov R, Holmuhamedov E “Synthetic peptide TEKKRRETVEREKE derived
from ezrin induces differentiation of NIH/3T3 fibroblasts” European Journal of
Pharmacology 811 (2017) 249259
18
Tishchenko, A.L.: "A new approach in the treatment of recurrent urogenital
candidiasis", Attending Doctor, 2002, pages 46 47. Тищенко А.Л. Новый подход к
лечению рецидивирующего урогенитального кандидоза. // Лечащий врач, 2002
,№3, c.46-47.
19
Kladova, O. V.; Kharlamova, F.S.; Shcherbakova, A.A.; Legkova, T.P; Feldfix, L.I.;
Znamenskaya, A.A.; Ovchinnikova, G.S.; Uchaikin V.F.: "The first experience of Hepon
intranasal application in children with respiratory infections.", PEDIATRICS, 2002,
pages 86 88. Кладова О.В., Харламова Ф.С., Щербакова А.А., Легкова Т.П.,
Фельдфикс Л.И., Знаменская А.А., Овчинникова Г.С., Учайкин В.Ф. Первый опыт
интраназального применения Гепона у детей с респираторными инфекциями. //
Педиатрия, 2002, №2, c.86-88.
20
Dudchenko, M.A.; Lysenko, B.F.; Chelishvili, A.L.; Katlinsky, A. V.; Ataullakhanov,
R.R.: "Complex treatment of trophic ulcers", ATTENDING DOCTOR, 2002, pages 72
75. Дудченко М.А., Лысенко Б.Ф., Челишвили А.Л., Катлинский А.В., Атауллаханов
Р.Р. Комплексное лечение трофических язв.// Лечащий врач, 2002, №10, c.72-75.
21
Polyakova T.O.; Magomedov, M.M.; Artemyev, M.E.; Surikov, E.V.; Palchun, V.T.: "A
new approach in the treatment of chronic diseases of the pharynx", ATTENDING
DOCTOR, 2002, pages 64 65.. Полякова Т.О., Магомедов М.М., Артемьев М.Е.,
Суриков Е.В., Пальчун В.Т. Новый подход к лечению хронических заболеваний
глотки // Лечащий врач, 2002, №4, с.64-65.
37
22
Bardychev, M.S.: "Treatment of local radiation injuries with the activator of local
immunity", Russian Medical Journal, vol. 11, no. 11, 2003, pages 646 647. Бардычев
М.С. Лечение местных лучевых повреждений с помощью активатора местного
иммунитета.// Русский медицинский журнал, 2003, том 11, №11(183), c.646-647.
23
Cherednichenko, T. V.; Uchaikin, V.F.; Chaplygina, G. V.; Kurbanova, G.M.: "A new
efficient treatment of viral hepatitis", Attending Doctor, 2003, pages 82 83.
Чередниченко Т.В., Учайкин В.Ф., Чаплыгина Г.В., Курбанова Г.М. Новое
эффективное лечение вирусных гепатитов.// Лечащий врач, 2003, №3, с.82-83.
24
Gorbarets, I.P.; Voronkova, N.V.; Lopatina, T.V.; Ivanovskaya, V.N.; Braginsky, D.M.;
Blokhina, N.P.; Malyshev, N.A.: "The combined use of Hepon drug product and
recombinant interferon-alpha in the patients with chronic hepatitis C increases the
efficiency of antiviral treatment and reduces side effects of the therapy", Hepatology,
2003, pages 23 - 28 Горбарец И.П., Воронкова Н.В., Лопатина Т.В., Ивановская
В.Н., Брагинский Д.М., Блохина Н.П., Малышев Н.А. Сочетанное применение
препарата Гепон и рекомбинантного интерферона-альфа у больных хроническим
гепатитом С повышает эффективность противовирусного лечения и уменьшает
побочные эффекты терапии.// Гепатология, 2003, №4, с.23-28.
25
Lazebnik, L.B.; Zvenigorodskaya, L.A.; Firsakova, V. Yu.; Pichugin, A.V.;
Ataullakhanov, R.I: "The application of Hepon immunomodulator in the treatment of
erosive ulcerous lesions of gastroduodenal zone", EXPERIMENTAL AND CLINICAL
Gastroenterology, 2003, pages 17 20. Лазебник Л.Б., Звенигородская Л.А.,
Фирсакова В.Ю., Пичугин А.В., Атауллаханов Р.И. Применение иммуномодулятора
Гепон в лечении эрозивно-язвенных поражений гастродуоденальной зоны.//
Экспериментальная и клиническая гастроэнтерология, 2003, №3, c.17-20.
26
Parfenov, A.I.: "The activator of the local immunity Hepon in the complex treatment of
dysbiotic disorders of the intestine", Experimental And Clinical Gastroenterology, 2003,
pages 66 69. Парфенов А.И. Активатор местного иммунитета Гепон в
комплексной терапии дисбиотических нарушений кишечника. //
Экспериментальная и клиническая гастроэнтерология, 2003, №3, c.66-69.
27
Novokshonov, A.A.; Uchaikin, V.F.; Sokolova, N.V.; Tikhonova, O.N.; Portnykh,
O.Yu.: "Biocenosis-protecting therapy of intestinal infections in children", Russian
Medical Journal, vol. 6, no. 1, 2004. Новокшонов А.А., Учайкин В.Ф., Соколова Н.В.,
Тихонова О.Н., Портных О.Ю. Биоценоз-сберегающая терапия кишечных
инфекций у детей. // Российский медицинский журнал, специальное приложение
"Болезни органов пищеварения", 2004, том 6, №1.
28
Telunts, A. V.: "Treatment of candidiasis in infants", Questions Of Gynecology,
Obstetrics, And Perinatology, vol. 3, no. 4, 2004, pages 89 90. Телунц А.В. Лечение
кандидоза у детей раннего возраста. // Вопросы гинекологии, акушерства и
перинатологии, 2004, том 3, № 4, c.89-90.
38
29
Malakhova, N.S.; Pichugin, A.V.; Khalif, I.L.; Ataullakhanov, R.I.: "The application of
Hepon immunomodulator for the treatment of nonspecific ulcerative colitis", Farmateka,
no. 6, 2005, pages 105 108. Малахова Н.С., Пичугин А. В., Халиф И.Л.,
Атауллаханов Р.И. Применение иммуномодулятора Гепон для лечения
неспецифического язвенного колита.// Фарматека, 2005, № 6[101], c.105-108.
30
Salamov G.; Holms R D; Bessler W.; Ataullakhanov R.: "Treatment of Hepatitis C
Virus Infection with Human Ezrin Peptide One (HEP1) in HIV Infected Patients",
ARZNEIMITTEL FORSCHUNG. DRUG RESEARCH., vol. 57, no. 07, 1 January 2007
(2007-01-01), DE, pages 497 - 504, XP055288985, ISSN: 0004-4172, DOI: 10.1055/s-
0031-1296637
31
‘Gepon’ preparation is registered by the Russian Ministry of Health (registration
documents R No: 000015/01-2000 of 12.07.2000, FSP 42-0012-0015-00). The
instruction on the medical application of the ‘Gepon’ preparation was approved with the
decision of the Pharmacology Committee of the Ministry of Health of the Russian
Federation (Minutes No: 8 of 7 October 1999). The preparation belongs to the group of
immunomodulators and is prescribed to increase the immune defence mechanism, to
treat and to prevent bacterial, viral and fungal infections.
32
Kadova O V and Uchiakin V F, “Clinical Study Report on intranasal application of Gepon
in children with the respiratory infections: safety and efficacy of Gepon in Acute
Respiratory Virus Infection [AVRI], Virus Induced Inflammation [VII], Laryngo-Tracheo-
Bronchitis with Laryngeal Stenosis [LTBS] and Recurrent Croup [RC] syndrome.
Unpublished
33
Kladova O V et al. “Immunomodulator Gepon: An Effective Treatment for Croup
Syndrome” Russian Medical Journal, 2002 No3 [147] p138-141
34
Kladova O V et al. “First experiment on the intranasal administration of Gepon in
children with the respiratory infections” Pediatrics 2002 no2 1-2
35
Ataullakhanov, R.I.; Holms, R.D.; Katlinsky, A. V.; Pichugin A. V.; Papuashvili, M.N.;
Shishkova, N.M.: "Treatment with Gepon immunomodulator increases the efficacy of
the immune control of opportunistic infections in HIV-infected patients", ALLERGY,
ASTHMA, AND CLINICAL IMMUNOLOGY, 2002, pages 3 - 11 Атауллаханов Р.И.,
Холмс Р.Д., Катлинский А.В., Пичугин А.В., Папуашвили М.Н., Шишкова Н.М.
Лечение иммуномодулятором Гепон повышает эффективность иммунного
контроля оппортунистических инфекций у больных ВИЧ-инфекцией. // Аллергия,
астма и клиническая иммунология, 2002, №10, c.3-11.
36
Polyakova T.S., “New Approach to the Treatment of Chronic Diseases of the Throat
Attending Doctor 2002 no4 64-65
37
Polyakova T.S., “Application of Gepon in the Treatment of ENT Pathologies” Moscow
Medical Journal May 2003 p16-17 48 patients
39
38
Safonova O.A., Pichukin A. B., Kozhemyakina E. Sh., Malshev N. A., and
Ataullakhanov R. I., “Treatment of Acute Respiratory Disease and its Complications
(Pneumonia)” published in Chapter 7, “Immunotherapy of Respiratory Disease”, Section
2 “Immunotherapy of Diseases of the Respiratory Tract, in the book “Immunotherapia”,
p122-158, Editors: Khaitov R.M., and Ataullakhanov R.I., (2011) published by
Izdatekskaya Group, GEOTAR-Media.
39
Table 7-14 p148 fig7-8 p149 in Safonova O.A., Pichukin A. B., Kozhemyakina E. Sh.,
Malshev N. A., and Ataullakhanov R. I., “Treatment of Acute Respiratory Disease and
its Complications (Pneumonia)” published in Chapter 7, “Immunotherapy of Respiratory
Disease”, Section 2 “Immunotherapy of Diseases of the Respiratory Tract, in the book
“Immunotherapia”, p122-158, Editors: Khaitov R.M., and Ataullakhanov R.I., (2011)
published by Izdatekskaya Group, GEOTAR-Media.
40
https://www.worldometers.info/coronavirus/
41
https://www.ecdc.europa.eu/en/novel-coronavirus-china
42
https://www.who.int/emergencies/diseases/novel-coronavirus-2019/situation-reports
43
https://www.phc.ox.ac.uk/covid-19/evidence-service/reviews/global-covid-19-case-
fatality-rates
44
https://www.who.int/emergencies/diseases/novel-coronavirus-2019
45
Peiris, J.S., Guan, Y., and Yuen, K.Y. (2004). Severe acute respiratory syndrome.
Nat. Med. 10 (12, Suppl), S88S97.
46
Middle East respiratory syndrome coronavirus (MERS-CoV). World Health
Organization web site. http://www.who.int/emergencies/ mers-cov/en/. Updated
September 21, 2018. Accessed June 14, 2019.
47
Matsuno A K, Gagliardi T B, Paula F E, Luna L K S, Jesus B L S, Stein R T, Aragon D
C , Ana P. C. P. Carlott AP CP , Arruda E, “Human coronavirus alone or in co-infection
with rhinovirus C is a risk factor for severe respiratory disease and admission to the
pediatric intensive care unit: A one-year study in Southeast Brazil PLOS ONE |
https://doi.org/10.1371/journal.pone.0217744 June 3, 2019
48
Roberts A, Deming D, Paddock CD, et al. A mouse-adapted SARS-coronavirus
causes disease and mortality in BALB/c mice. PLoS Pathog. 2007;3:e5. [PMC free
article] [PubMed] [Google Scholar]
49
Gralinski LE, Bankhead A, 3rd, Jeng S, et al. Mechanisms of severe acute respiratory
syndrome coronavirus-induced acute lung injury. mBio. 2013;4:e0027113. [PMC free
article] [PubMed] [Google Scholar]
50
Reynolds HN, McCunn M, Borg U, et al. Acute respiratory distress syndrome:
estimated incidence and mortality rate in a 5 million-person population base. Crit Care.
1998;2:2934.
40
51
Wu Z, McGoogan JM, “Characteristics of and Important Lessons from the
Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of
72 314 Cases From the Chinese Center for Disease Control and Prevention.” JAMA.
Published online February 24, 2020. doi:10.1001/jama.2020.2648
52
Fei Zhou F et al, “Clinical course and risk factors for mortality of adult inpatients with
COVID-19 in Wuhan, China: a retrospective cohort study”, Published:March 11,
2020DOI:https://doi.org/10.1016/S0140-6736(20)30566-3
53
Wang D, et al “Clinical Characteristics of 138 Hospitalized Patients With 2019 Novel
Coronavirus-Infected Pneumonia in Wuhan, China.” JAMA. 2020;
54
Schneider WM, Chevillotte MD, Rice CM. 2Interferon-stimulated genes: a complex
web of host defenses.2 Annu Rev Immunol. 2014;32:513545.
55
Totura AL, Baric RS. 2SARS coronavirus pathogenesis: host innate immune
responses and viral antagonism of interferon.2 Curr Opin Virol. 2012;2(3):264275.
56
Law, H.K., Cheung, C.Y., Ng, H.Y., Sia, S.F., Chan, Y.O., Luk, W., Nicholls, J.M.,
Peiris, J.S., and Lau, Y.L. (2005). Chemokine up-regulation in SARS-coronavirus-
infected, monocyte-derived human dendritic cells. Blood 106, 23662374
57
Spiegel, M., Schneider, K., Weber, F., Weidmann, M., and Hufert, F.T. (2006).
Interaction of severe acute respiratory syndrome-associated coronavirus with dendritic
cells. J. Gen. Virol. 87, 19531960.
58
Yen, Y.T., Liao, F., Hsiao, C.H., Kao, C.L., Chen, Y.C., and Wu-Hsieh, B.A. (2006).
Modelling the early events of severe acute respiratory syndrome coronavirus infection
in vitro. J. Virol. 80, 26842693.
59
Channappanavar et al., 2016, Cell Host & Microbe 19, 181193 February 10, 2016
ª2016 Elsevier Inc. http://dx.doi.org/10.1016/j.chom.2016.01.007
60
Covid-19: what treatments are being investigated?BMJ 2020; 368 doi:
https://doi.org/10.1136/bmj.m1252 (Published 26 March 2020)
61
Cinatl J, Morgenstern B, Bauer G, Chandra P, Rabenau H, Doerr HW. Treatment of
SARS with human interferons. Lancet 2003; 362:2934.
62
Channappanavar R, Fehr AR, Zheng J, Wohlford-Lenane C, Abrahante JE, Mack M,
Sompallae R, McCray PB Jr, Meyerholz DK7, Perlman S. IFN-I response timing
relative to virus replication determines MERS coronavirus infection outcomes. J Clin
Invest. 2019 Jul 29;130:3625-3639. doi: 10.1172/JCI126363. eCollection 2019 Jul 29.
63
Falzarano D, et al. Treatment with interferon-α2b and ribavirin improves outcome in
MERS-CoV-infected rhesus macaques. Nat Med. 2013;19(10):13131317.
64
Cinatl J, Morgenstern B, Bauer G, Chandra P, Rabenau H, Doerr HW.
Treatment of SARS with human interferons. Lancet 2003; 362:2934.
41
65
Hamming, O. J., Terczyńska-Dyla, E., Vieyres, G., Dijkman, R., Jørgensen, S. E.,
Akhtar, H. Hartmann, R. “Interferon lambda 4 signals via the IFNλ receptor to regulate
antiviral activity against HCV and coronaviruses. (2013). The EMBO Journal, 32(23),
30553065. doi:10.1038/emboj.2013.232
66
Mayer-Barber K.D., Yan B. Clash of the cytokine titans: counter-regulation if
interleukin 1 and type I interferon-mediated inflammatory responses. Cellular &
Melecular Immunology. 2017; 14: 22-35;
67
Guarda G., Braun M., Staefli F., Tardivel A., Mattman Ch., Förster I., Farlik M., Decker
T., Du Pasquier R.A., Romero P., Tschopp J. Type I interferon inhibits interleukin-1
production and inflammasome activation. Immunity. 2011; 34: 213-223;
68
Coclet-Ninin J., Dayer J.-M., Burger D. Interferon-beta not only inhibits interleukin-
and tumor necrosis factor-α but stimulates interleukin-1 receptor antagonist production
in human peripheral blood mononuclear cells. Europ. Cytokine Netw. 1997; 8(4): 345-
349;
69
Schindler R., Ghezzi P., Dinarello C.A. Interferons as inhibitors of interleukin 1
induced interleukin 1 synthesis. Lymphokine Res. 1989; 8(3): 275-280).
70
Sharif M.N., Šošić D., Rothlin C.V., Kelly E., Lemke G., Olson E.N., Ivashkiv L.B.
Twist mediates suppression of inflammation by type I IFNs and Axl. J.Exp.Med. 2003;
203(8): 1891-1901;
71
Šošić D., Richardson J.A., Yu K., Ornitz D.M., Olson E.N. “Twist regulates cytokine
gene expression through negative feedback loop that represses NF-kB activity. Cell.
2003; 112: 169-180
72
Stifter S.A., et al. Functional interplay between type I and type II interferons is
essential to limit influenza A virus-induced tissue inflammation. PLoS Pathology. 2016;
12(1): e1005378
73
Jungo F., Dayer J.M., Modoux C., Hyka N, Burger D. IFN-beta inhibits the ability of T-
lymphocytes to induce TNF-alpha and IL-1 beta production in monocytes upon direct
cell-cell contact. Cytokine. 2001; 14(5): 272-282).
74
Kroetz D.N., Allen R.M., Schalter M.A., Cavallaro C., Ito T., Kunkel S.L. Type I
interferon induced epigenetic regulation of macrophages suppresses innate and
adaptive immunity in acute respiratory viral infection. PLoS Pathology. 2015; 11(12):
e10055338;
75
Stifter S., et al. Functional interplay between typeI and II interferons is essential to
limit influenza A virus-induced tissue inflammation. PLoS Pathogens. 2016; doi:
10.1371/journal.ppat.1005378, 20 pp.
... Burgeoning evidence is emerging from in vitro and in silico models, demonstrating interactions between S protein and immune receptors, including Neuropilin-1 (NRP1), Clectin type receptors (CLR) (mannose receptor (MR); dendritic cell-specific intracellular adhesion molecule-3-grabbing non-integrin (DC-SIGN); homologue dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin related (L-SIGN); and macrophage galactose-type lectin (MGL)) [34,35] and toll-like receptors (TLRs) (TLR1; TLR4; and TLR6) [2], and the non-immune receptor glucose regulated protein 78 (GRP78) [36]. Furthermore, Ezrin [37], and dipeptidyl peptidase-4 (DPP4) [38] have been postulated to be targets against SARS-CoV-2, but have yet to be confirmed in in vitro and in vivo models. ...
... As such, Ezrin enhances viral infectivity, through inhibition of unnecessary membrane fusion [55]. Contrary to this, in relation to SARS, previous studies noted that Ezrin interacts with the SARS-CoV spike protein through binding to the carboxy-terminus using its FERM domain [37], resulting in reduced viral entry [56]. This highlights a potential therapeutic option to prevent SARS-CoV-2 infection. ...
... In addition to inhibiting the key receptors involved in COVID-19, such as ACE2 and the newly suggested TLRs, an Ezrin agonist or molecule that increases Ezrin functionality could be a strategic approach to inhibiting SARS-CoV-2 viral entry. This hypothesis was investigated using Ezrin peptides, which have previously demonstrated effectiveness in treating a variety of viral infections, initiated by HIV-1, hepatitis C virus, human papillomavirus, herpes simplex I and II, and the causative viral agents in acute viral respiratory infection [37]. Specifically, it is particularly beneficial in inhibition of inflammation in viral pneumonia [37], a key pathophysiological complication observed in COVID-19. ...
Article
Full-text available
The occurrence of the novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), responsible for coronavirus disease 2019 (COVD-19), represents a catastrophic threat to global health. Protruding from the viral surface is a densely glycosylated spike (S) protein, which engages angiotensin-converting enzyme 2 (ACE2) to mediate host cell entry. However, studies have reported viral susceptibility in intra- and extrapulmonary immune and non-immune cells lacking ACE2, suggesting that the S protein may exploit additional receptors for infection. Studies have demonstrated interactions between S protein and innate immune system, including C-lectin type receptors (CLR), toll-like receptors (TLR) and neuropilin-1 (NRP1), and the non-immune receptor glucose regulated protein 78 (GRP78). Recognition of carbohydrate moieties clustered on the surface of the S protein may drive receptor-dependent internalization, accentuate severe immunopathological inflammation, and allow for systemic spread of infection, independent of ACE2. Furthermore, targeting TLRs, CLRs, and other receptors (Ezrin and dipeptidyl peptidase-4) that do not directly engage SARS-CoV-2 S protein, but may contribute to augmented anti-viral immunity and viral clearance, may represent therapeutic targets against COVID-19.
... Ezrin (EZR) is a protein encoded by the EZR gene and belongs to the family EZR-radixin moesin. The role of EZR has been described in the transmission of human immunodeficiency virus (HIV) but in SARS, it causes decreased entry of the virus by interacting with the SARS-CoV S protein [29], causing reduced viral entry. This can be considered as a potential therapeutic alternative to prevent SARS-CoV-2 infection. ...
... As EZR inhibits ACE2 and the Toll-like receptors (TLRs), the crucial receptors participating in the pathogenesis of COVID-19, an EZR agonist or molecule can be considered as a strategic therapeutic approach to affect SARS-CoV-2 viral entry [18]. It suppresses the inflammation seen in viral pneumonia [29], a significant pathophysiological consequence seen in COVID-19. As a result, more research is needed to fully comprehend its role in the SARS-CoV-2 virus infection. ...
Article
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The crown-like shaped viruses known as coronaviruses which were first reported in the 1960’s have caused three epidemics in the past two decades namely, coronavirus disease-19 (COVID-19), severe acute respiratory syndrome (SARS), and Middle East respiratory syndrome (MERS). SARS coronavirus 2 (SARS-CoV-2) was first reported in the latter half of December in Wuhan, a city of China, with people affected by deadly pneumonia with unknown etiology. Since then, the world has experienced two phases of virus spread with different symptoms and disease severity. This review embarks on the journey to investigate candidate molecules of this virus which can and are being investigated for various vaccine formulations and to discuss immunity developed against this virus.
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