Small molecule inhibitors have previously been investigated in different studies aspossible therapeutics in the treatment of SARS-CoV-2. In the current drug repurposingstudy, the investigators identified the leukotriene (D4) receptor antagonist Montelukastas a novel agent that simultaneously targets two important drug targets of SARS-CoV-2.The investigators initially demonstrated the dual inhibition (main protease andSpike/ACE2) profile of Montelukast through multiscale molecular modeling studies. Next,the investigators characterized its effect on both targets by different in vitroexperiments including the Fluorescent Resonance Energy Transfer (FRET)-based mainprotease enzyme inhibition assay, surface plasmon resonance (SPR) spectroscopy,pseudovirus neutralization on HEK293T / hACE2, and virus neutralization assay usingxCELLigence MP real time cell analyzer.
The 2019 new coronavirus (SARS-CoV-2), was first reported in December 2019 in Wuhan
(Hubei, China). It has quickly spread to other countries all around the world and
effected more than 67 million people worldwide becoming an urgent global pandemic.
Coronaviruses are enveloped, non-segmented positive-sense RNA viruses belonging to the
family of Coronaviridae, the largest family in Nidovirales and widely distributed in
humans, other mammals and birds, causing respiratory, enteric, hepatic and neurological
diseases. Seven species of coronavirus are known to cause disease in humans. Four of them
(229E, OC43, NL63, and HKU1) are common and they mostly cause common cold symptoms in
immunocompetent individuals while the other three, SARS-CoV, MERS-CoV, and SARSCoV-2
cause serious symptoms and death.
SARS-CoV-2 has four structural proteins which are nucleocapsid, envelope, membrane and
spike. These four proteins play a vital role during the viral infection. The Spike
glycoprotein (S protein) located on the external surface of coronaviruses are responsible
for the connection and entry of the virus to host cells. The S protein mediates receptor
recognition, cell attachment, and fusion during viral infection. While the virus is in
its natural environment, S protein of coronavirus is inactive. During viral infection,
target cell proteases activate the S protein by cleaving it into S1 and S2 subunits,
which are required to activate the membrane fusion domain after viral entry into target
cells. The S1 subunit includes the receptor binding domain (RBD). This domain binds
directly to the peptidase domain angiotensin converting enzyme 2 (ACE-2). S2 functions
during membrane fusion. The chymotrypsin-like cysteine protease called 3C-like protease
(3CLpro) aka main protease (Mpro) in SARS-CoV-2 is a vital enzyme involved in processes
such as the processing, assembly, and replication of the virus.
One of the key characteristics of severe COVID-19 is increased cytokine production. It is
thought that the severity of the disease is primarily associated with the cytokine storm,
which is an aggressive immune response to the virus. The number of white blood cells,
neutrophils, and levels of procalcitonin, C-reactive protein and other inflammatory
indices like IL2, IL7, IL10, granulocyte-colony stimulating factor (GSCF), interferon
inducible protein -10 (IP10), monocyte chemotactic protein-1 (MCP1), macrophage
inflammatory protein-1α (MIP1A), and TNF are significantly higher in severe cases in
patients with COVID-19. Specifically, IL-1β, IL-6, and IL-10 are the three most elevated
cytokines in serious cases. One result of the cytokine storm is lung injury that can
develop into acute lung injury or its more severe type (acute respiratory distress
syndrome, ARDS). Studies have shown the relation between COVID-19 and the most common
chronic conditions such as diabetes, cardiovascular diseases, respiratory system
diseases, immune system disorders, etc. Asthma and chronic obstructive pulmonary disease
(COPD) are among the diseases of the respiratory system that are most emphasized. Asthma
is a chronic inflammatory airway condition. There is significant evidence that represents
the relation of asthmatic patients in the population with viral infections like
rhinoviruses. Virus infections cause upper respiratory tract infection, like influenza A,
rhinovirus, and respiratory syncytial virus (RSV) elevate local leukotriene levels.
Leukotrienes, which play a role in the contraction of bronchial muscles, are effective in
initiating and amplifying many biological responses, including mast cell cytokine
secretion, macrophage activation, and dendritic cell maturation and migration.
Leukotrienes (LTC4, LTD4 and LTE4), activated basophils, eosinophils, macrophages, and
products of mast cells are types of lipids conjugated with peptides. LTD4 receptors
belong to G protein-coupled receptor (GPCR) family. Montelukast is a selective
leukotriene (D4) receptor antagonist which is a member of quinolines and it was approved
by FDA as an oral tablet in 1998. It is a licensed drug used for allergic rhinitis,
exercise-induced bronchospasm and especially prophylaxis and chronic treatment of asthma.
As a result of LTD4 blockage, NF-kB pathway activation and release of the proinflammatory
mediators (i.e., IL-6,8 and 10, TNF-a and MCP-1) decrease. Considering these
anti-inflammatory effects by leukotriene receptor inhibition and possible antiviral
effects, Montelukast maybe considered for the effective medication against SARS CoV-2.
Here, initially the investigators explored the potential role of Montelukast in the
management of SARS-CoV-2 infection with multiscale molecular modeling approaches and its
promising results both in main protease and Spike/ACE2 interface encouraged the
investigators to perform further detailed in vitro experiments. The results of FRET-based
biochemical assays, surface plasmon resonance (SPR), pseudovirus neutralization and virus
neutralization experiments demonstrated the effect of Montelukast on SARS-CoV-2.
This study was designed as a national, multi-center, open-label, randomized, parallel,
three-arm, phase-II study.
Drug: Montelukast Oral Tablet
3x10 mg oral montelukast first day and other 13 days 1 x 10 mg montelukast
Drug: Montelukast plus Favicovir (Favipiravir)
200 mg oral favicovir for 5 days in a regimen of 2x1600 mg (oral) loading dose on day-1
(eight tablets in the morning and eight tablets in the evening) followed by 2x600 mg
maintenance dose (three tablets in the morning and three tablets in the evening) on day-2
to day-5 and 3x10 mg oral montelukast first day and rest of 13 days 1 x 10mg,
concurrently.
Drug: Favicovir (Standard Treatment)
200 mg oral favicovir for 5 days in a regimen of 2x1600 mg (oral) loading dose on day-1
(eight tablets in the morning and eight tablets in the evening) followed by 2x600 mg
maintenance dose (three tablets in the morning and three tablets in the evening) on day-2
to day-5.
Inclusion Criteria:
- Male or female patients aged 18 years and older infected with the SARS-CoV-2
infection
- Patients with COVID-19 symptoms and have a positive PCR test result
- Patients in a stable clinical condition and basically in an outpatient condition
- Patients who sign the informed consent
Exclusion Criteria:
- Patients with a partial oxygen pressure < 90% and who have required hospitalization
- Patients who have required intensive care
- Any condition which, in the opinion of the Principal Investigator, would prevent
full participation in and compliance with the trial protocol
- Patients who have been involved in any other interventional studies
- Patients with uncontrolled Type I or Type II diabetes mellitus (DM)
- Patients with severe liver failure (Child Pugh score ≥ C, AST> 5 times the upper
limit of normal (ULN)
- Patients with severe renal failure (GFR ≤30 mL/min/1.73 m2) or continuous dialysis
(hemodialysis, peritoneal dialysis) or continuous renal replacement therapy
- Patients with serious cardiac problems such as heart failure
- Patients with hypersensitivity to montelukast or other drugs in the study
- Patients with rare hereditary problems of galactose / fructose intolerance, glucose-
galactose malabsorption or sucrase-isomaltase insufficiency
- Pregnant and lactating women
- Patients who cannot use sexual abstinence or appropriate contraceptive method during
the study
- Patients who are treated with any other antiviral drugs for COVID-19 in the last 30
days
Bahcesehir University, School of Medicine, Department of Biophysics,
Istanbul, Turkey
Istanbul University, Cerrahpaşa School of Medicine
Istanbul, Turkey
Prof. Serdar Durdagi, Ph.D.
+90-216-579-8217
serdar.durdagi@med.bau.edu.tr