The Role of Folic Acid in the Management of Respiratory Disease Caused by COVID-19
The Role of Folic Acid in the Management of Respiratory Disease Caused by COVID-19
Submitted date: 26/03/2020 • Posted date: 30/03/2020 Licence: CC BY-NC-ND 4.0
Citation information: Sheybani, Zahra; Dokoohaki, Maryam Heydari; Negahdaripour, Manica; Dehdashti, Mehdi; Zolghadr, Hassan; Moghadami, Mohsen; et al. (2020): The Role of Folic Acid in the Management of Respiratory Disease Caused by COVID-19. ChemRxiv. Preprint. https://doi.org/10.26434/chemrxiv.12034980.v1
Entrance of coronavirus into cells happens through the spike proteins on the virus surface, for which the spike protein should be cleaved into S1 and S2 domains. This cleavage is mediated by furin, which can specifically cleave Arg-X-X-Arg↓ sites of the substrates. Furin, a member of proprotein convertases family, is moved from the trans-Golgi network to the cell membrane and activates many precursor proteins. A number of pathological conditions such as atherosclerosis, cancer, and viral infectious diseases, are linked with the impaired activity of this enzyme. Despite the urgent need to control COVID-19, no approved treatment is currently known. Here, folic acid (folate), a water-soluble B vitamin, is introduced for the first time for the inhibition of furin activity. As such, folic acid, as a safe drug, may help to prevent or alleviate the respiratory involvement associated with COVID-19.
The role of folic acid in the management of respiratory disease caused by COVID-19
Zahra Sheybani1,9, Maryam Heydari Dokoohaki2,9, Manica Negahdaripour3,4, Mehdi Dehdashti5, Hassan Zolghadr6, Mohsen Moghadami7, Seyed Masoom Masoompour7, and Amin Reza Zolghadr2,8*
- Department of Internal Medicine, Aliasghar Hospital, Shiraz University of Medical Sciences, Shiraz, Iran.
- Department of Chemistry, Shiraz University, Shiraz, 71946-84795, Iran.
- Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
- Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
- Microbiology laboratory, Moslemin Hospital, Shiraz, Iran.
- Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.
- Non‐Communicable Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Fars Science and technology Park, Shiraz, Iran.
- Sh and M.H.D. are co-first authors.
0788, ORCID: 0000-0002-6289-3794 (A.R.Z).
Entrance of coronavirus into cells happens through the spike proteins on the virus surface, for which the spike protein should be cleaved into S1 and S2 domains. This cleavage is mediated by furin, which can specifically cleave Arg-X-X-Arg↓ sites of the substrates. Furin, a member of proprotein convertases family, is moved from the trans-Golgi network to the cell membrane and activates many precursor proteins. A number of pathological conditions such as atherosclerosis, cancer, and viral infectious diseases, are linked with the impaired activity of this enzyme.
Keywords: Coronavirus, 2019 novel coronavirus, Furin, Folic acid, Spike protein
Coronaviruses, a family of Coronaviridae, can cause significant human pathologies such as respiratory tract infections in humans and other mammals.1 Coronavirus infections are usually mild, but some beta coronaviruses including Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus (SARS-CoV) may induce critical symptoms.2,3
In December 2019, an outbreak of lower respiratory tract infections was reported in Wuhan, China.4 The pathogen was recognized as a novel RNA beta coronavirus, later named as SARS-CoV-2.5 The infection caused by this virus, COVID-19, is declared by the World Health Organization (WHO) as a pandemic.6 In view of SARS-CoV-2 novelty, further researches are required to obtain more insights about its pathogenesis.
Coronavirus (CoV) genome encodes four structural proteins, comprising spike (S), membrane (M), envelope (E), and nucleocapsid (N). The spike (S) protein of coronaviruses mediates receptor binding and fusion of the virus with the target cells.7 Each class of coronavirus attaches to a specific cellular receptor to facilitate virus entrance into cells. Angiotensin- converting enzyme 2 (ACE2) and CD209L are shown responsible for SARS-CoV entrance.8-9 It is reported that SARS-CoV-2 enters the respiratory tract by interacting with ACE2 receptor.10
The spike protein comprises an amino (N)-terminal S1 subunit and a carboxyl (C)-terminal S2 subunit. The entrance of the virus is facilitated by cleavage of S protein to S1/S2 subunits. The S1 subunit binds to the ACE2 receptor, while the S2 site interacts with the cell membrane to mediate receptor-dependent endocytosis11, as shown in Fig1a. The coronavirus spike protein is cleaved into S1 (receptor binding subunit) and S2 (membrane fusion subunit) by a proteolytic activation at the furin consensus motif RRRR537↓S (R=arginine, ↓: cleavage site) in virus-infected cells. Additionally, the S2 subunit of the S protein is further cleaved at the second furin site (RRRR690↓S) in the infected cells expressing S constructs.12-16 Mutations of one basic residue in the RRRR690↓S motif and use of non-furin cleavable PRRR↓S sequence demonstrated that furin may play an important role in furin-dependent entry.17 The working protease is a cellular proprotein convertase that circulates between plasma membrane, early endosome, and trans- Golgi network (TGN), by participation in endocytic and exocytic paths.18,19 This proprotein convertase is a major candidate for processing the surface glycoproteins of pathogenic
viruses.20,21 Furin can cleave precursor proteins with specific motifs to produce mature proteins with biological activity. The first (P1) and fourth (P4) amino acids at the N-terminus of the substrate cleavage site must be arginine "Arg-X-X-Arg ↓" (R-X-X-R, X= any amino acid, ↓: cleavage site). If the P2 position is basic lysine or arginine, the cleavage efficiency could be improved by about 10 times.22 The results of a series of analyses have proposed that one of the important reasons for the high infectivity of COVID-19 is a redundant furin cut site in the virus spike protein.23
Our aim is to suggest folic acid as a potential inexpensive, safe, and non-immunogenic drug candidate for the prevention or treatment of early stages of respiratory disease associated with COVID-19 (Fig. 1). Folic acid is a type of B vitamin normally found in foods such as spinach, broccoli, asparagus, dried beans, lentils, peas, and oranges. Folic acid helps the body produce and maintain new cells and also prevent changes to DNA that may lead to cancer. Noticeably, folic acid deficiency is associated with a variety of human malignancies, including colorectal cancer. The over-expression of folate receptors in the early stages of malignant cell formation can be due to folic acid deficiency. Besides, folate malnutrition can cause a high incidence of adenomatous polyps and premalignant lesions of the colon.24 To this aim, the molecular dynamics (MD) simulations of the interactions of furin enzyme with folic acid and one of its active metabolites, folinic acid, was performed here for the first time to evaluate the interplay of these molecules with furin.
Fig. 1- A schematic representation of inhibitory action of folic acid. a, The mechanism of fusion and replication of COVID-19 virus. b, Inhibition of furin protein by folic acid.
were -161.6, and -159.2 kcal/mol for folic acid and folinic acid, respectively. The results showed that folic acid and folinic acid molecules interacted well with the active site residues of furin by formation of hydrogen bonds. Different atom sites of the two drug molecules established hydrogen bonding interactions with various amino acids of furin as shown in Fig. 2a. Interestingly, the binding sites of folinic acid and furin were clearly different. The interactions of Gly307, Glu271, Tyr313, Gln488, Ala532, Arg490, and Asp530 residues with hydrogen, oxygen, and nitrogen atoms of folic acid were dominant; whereas, strong hydrogen bonds were established through Ser311, Glu271, Arg490, Lys449, and Gln488 residues of folinic acid molecules. Moreover, while the H19 and H12 atoms of folic acid interacted substantially with Glu271, the H2 and H4 atoms of folinic acid formed hydrogen bonds with the Glu271 residue of furin. These findings proposed different orientation preferences of folic acid and folinic acid molecules in the binding site of furin.
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