Introduction
Microalgae are defined as photosynthetic and unicellular organisms that demonstrate
a wide range of adaptability to adverse environmental conditions like temperature
extremes, photooxidation, high or low salinity, and osmotic stress (Holzinger and
Karsten, 2013; Singh et al., 2019). Alternatively, macroalgae or seaweed includes
multicellular, macroscopic, and marine algae belonging mostly to two phyla, namely,
Rhodophyta and Phaeophyta (Peng et al., 2015). The micro/macro-algae have recently
emerged as a source of various bioactive compounds like phycocyanin, lutein, vitamin
E, B12 and K1, polyunsaturated fatty acids, polysaccharides and phenolics (Peng et
al., 2015; Costa et al., 2020). These secondary metabolites have been studied for
their anti-microbial, anti-inflammatory, immunosuppressive, anti-cancer and other
pharmacologically important activities (Sathasivam et al., 2019). Thus, the algal
metabolites find wide applicability in a vast array of biotechnological and pharmaceutical
fields.
In view of the ongoing COVID-19 pandemic caused by a novel coronavirus, designated
as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), increased efforts
are being made for developing efficient treatment options to tackle the disease. The
SARS-CoV-2 has been identified as a single-stranded, positive-sense RNA virus belonging
to the Betacoronavirus family (Yeo et al., 2020). Further, various structural (spike
glycoprotein), non-structural (3-chymotrypsin-like protease, helicase, papain-like
protease, and RNA-dependent RNA polymerase), and accessory proteins are encoded by
SARS-CoV-2 genome (Li and De Clercq, 2020). The spike glycoprotein has been considered
to be involved in the interaction between viruses and receptors present on the host
cell (Li and De Clercq, 2020). Since this glycoprotein is an essential requirement
for the entry of virus in host cells, many recent studies are focused on this structural
protein (Zumla et al., 2016). It has been further concluded that the above mentioned
five proteins also emerged as attractive targets for antiviral studies against SARS
(Severe Acute Respiratory Syndrome) and MERS (Middle East respiratory syndrome) (Zumla
et al., 2016).
Considering all the facts related to exploration of algae for bioactive molecules,
the present study provides an insight into the utilization of micro/macro-algal metabolites
as therapeutic compounds against SARS-CoV-2 and like viruses. The key antiviral metabolites,
namely, phycocyanobilins, lectins, and, sulphated polysaccharides have been discussed.
Algae; A Treasure-trove of Bioactive Metabolites
Phycocyanobilins; The Antiviral Chromophores
Phycocyanobilins (PCBs) are tetrapyrrole chromophores present in certain cyanobacteria,
rhodophytes, and are classified as blue phycobilinis (Figure 1) (Guedes et al., 2019).
These light-capturing pigments are now widely studied for their antioxidative, antiviral
(Hirata et al., 2000; Ramakrishnan, 2013) and NADPH-oxidase inhibitory activity (McCarty,
2007). Recently, Pendyala and Patras (2020) discussed the possible utilization of
PCBs (source—Spirulina sp.) as inhibitors for the SARS-CoV-2 infection. The study
involved in-silico screening (by the COVID-19 Docking Server) of the bioactive compounds
for their activity against SARS-CoV-2. It was observed that the phycocyanobilin demonstrates
a high binding affinity toward the potential targets, namely, the Main protease (Mpro)
and RNA-dependent RNA polymerase (RdRp). The Main protease is involved in the processing
of polyproteins (translated from SARS-CoV-2 RNA) while the replication of viral RNA
is catalyzed by the polymerase. High binding energy of −8.6 kcal/mol was observed
for PCB-Mpro while −9.3 kcal/mol for PCB-RdRp. Noteworthy, the PCB demonstrated a
superior binding to target enzymes as compared to antiviral drugs like remdesivir
(−8.1 kcal/mol for Mpro, −9.0 kcal/mol for RdRp), lopinavir (−7.9 kcal/mol) and nelfinavir
(−7.9 kcal/mol for Mpro, −9.3 kcal/mol for RdRp). Thus, the study highlighted the
significant potential of PCB as antiviral. However, as recommended by Pendyala and
Patras (2020), further in-vitro and/or in-vivo studies will be crucially needed to
support the obtained docking results and unravel the underlying potential of PCB as
therapeutic for COVID-19. Additionally, the purified allophycocyanin obtained from
Spirulina platensis has been demonstrated to exhibit significant activity against
enterovirus 71 (Singh et al., 2020). It was observed that the cytopathic effects of
the viral infection were neutralized and the viral RNA synthesis was delayed by the
microalgal pigment allophycocyanin. Likewise, results of an in-silico study reported
that the PCB expressed by Arthrospira sp. could serve as a potent antiviral against
SARS-CoV-2 (Petit et al., 2020). The study evaluated the interaction between the Arthrospira
sp. PCB and the receptor binding domain (RBD) of SARS-CoV-2 spike glycoprotein. It
was observed that five Van der Waals interactions (involving residues ARG403, TYR453,
LEU492, GLN493, and ASN501) contributed to the PCB/Spike RBD complex. The five π-alkyl
bonds between the PCB and spike RBD involved the residues TYR449, TYR495, PHE497,
and TYR505 with a hydrogen-bond on TYR449. The other residues involving the hydrogen-bond
were SER494, GLY496, and GLN498 with the GLY496 linked to PCB by a π-donor hydrogen
bond. Finally, a competitive binding energy (−7.2 kcal/mol) demonstrated the possibility
to employ PCB as a potential antiviral agent (Petit et al., 2020). A recent study
also reported the probability to utilize phycocyanobilin containing cyanobacteria
like Spirulina sp. to control the RNA virus infections (Nikhra, 2020). A decrease
in mortality rate in influenza-infected mice has been observed when administered orally
with phycocyanin rich cold-water Spirulina sp. extracts in animal experimentation
studies. The cold-water extract was well-tolerated even at high concentrations of
3,000 mg/kg/day in animal models for a period of 14 days (Chen et al., 2016). The
PCB extracts thus demonstrated a substantial reduction in the survival of zoonotic
RNA viruses by enhancing the type 1 interferon response of host immune system (Nikhra,
2020). Hence, it is likely possible that PCB producing microalgae may demonstrate
substantial activity against SARS-CoV-2 as well (Cascella et al., 2020; Zhou et al.,
2020). Moreover, further research along with in-vivo studies is necessary to understand
the specific bioactivity of PCBs for the development of therapeutic strategies against
human pathogenic viruses, including SARS-CoV-2.
Figure 1
Algal antiviral metabolites (a) phycocyanobilin; (b) lectin; (c) fucoidan (a representative
SP); (d) SPs-mediated humoral activation; (e) activation of host cellular immune response
by SPs; (f) SPs-driven inhibition of virus entry/attachment to the host cell receptor.
Lectins; Potential Therapeutic Against SARS-CoV-2
The macroalgae are rich in certain carbohydrate-binding proteins called lectins that
demonstrate high specificity for sugar groups of other molecules like the oligosaccharide
chains of the viral glycoproteins (Figure 1). Thus, lectins have been widely employed
in various pharmacological and medical applications (Breitenbach Barroso Coelho et
al., 2018). The mannose-binding lectins (MBL) are the predominant proteins to be studied
in the viral infection pathways (Mitchell et al., 2017). The self-assembly of viruses
during replication is interrupted by MBLs (Liu et al., 2015); thus, they have also
emerged as a potential therapy against Ebola (Michelow et al., 2011).
The red algae-derived lectins were initially brought to the limelight when griffithsin
was discovered by Watson and Waaland (1983) from Griffithsia sp. Since then, it has
been widely studied for various applications (Mori et al., 2004). It has been observed
to possess high specificity for mannose residues present on viral glycoproteins. Some
studies have demonstrated its antiviral activity against HIV-1 (Lusvarghi et al.,
2016), Hepatitis C (Meuleman et al., 2011), and SARS-CoV glycoprotein (Zumla et al.,
2016). A recent study analyzed the anti MERS-CoV activity of griffithsin and concluded
that the lectin inhibits the entry of the virus while imparting negligible cellular
toxicity (Millet et al., 2016). The inhibitory effect of griffithsin at the binding
step during virus infection was assayed by time-course experiments. Thus, the study
by Millet et al. (2016) demonstrated the griffithsin-mediated inhibition of MERS-CoV
infectivity in-vitro. Additionally, various studies have reported the in-vivo antiviral
activity of griffithsin against Japanese encephalitis virus (Ishag et al., 2013),
herpes simplex virus 2 (Nixon et al., 2013) and human papillomavirus (Levendosky et
al., 2015). For instance, the impact of an anti-HIV griffithsin containing microbicide
on the rectal microbiome was assessed in the non-human primates (Rhesus macaques)
(Girard et al., 2018). It was observed that 0.1% of griffithsin gel did not negatively
impact the rectal mucosal proteome or microbiome. Further, O'Keefe et al. (2010) reported
a 100% survival of model mice infected with a high dose of SARS-CoV upon providing
a griffithsin dose of 10 mg/kg(b.w.)/day. Based on griffithsin activity against SARS-CoV,
it may be investigated as a therapeutic option for SARS-CoV-2.
Likewise, a novel D-mannose-binding lectin was identified from the red macroalgae
Grateloupia chianggi and designated as GCL (Grateloupia chianggi lectin) (Hwang et
al., 2020). The study focussed on GCL purification, its molecular and functional characterization,
and subsequent analysis of its antiviral activity against influenza virus, herpes
simplex virus and HIV. A quantity of 1–20 nM GCL was required for effective inhibition
of HSV. Thus, it may be concluded that GCL also holds the potential to be utilized
in virology and biomedical research. It is significant to note here that the SARS-CoV-2
is similar to the influenza virus as both are characterized as enveloped RNA viruses
(Noda, 2012; Yeo et al., 2020). Based on the activity of GCL against the influenza
virus, its activity may be explored against SARS-CoV-2 as well.
Sulphated Polysaccharides; Favorable Antiviral Candidates
Various researchers have demonstrated the beneficial effects of algal sulphated polysaccharides
(SPs) under defined in-vitro and/or in-vivo conditions. Both the cellular and/or the
humoral response of the immune system can be activated by these compounds (de Paniagua-Michel
et al., 2014) (Figure 1).
A recent study emphasized on the purification and structural characterization of two
fucoidans from the brown macroalgae Sargassum henslowianum (Sun et al., 2020). These
fucoidans designated as SHAP-1 and SHAP-2 were studied for their activity against
two strains of herpes simplex virus, i.e., HSV-1 and HSV-2. It was observed that both
compounds possessed significant anti-HSV activity with the IC50 value estimated to
be 0.89 and 0.82 μg/mL for SHAP-1 and SHAP-2, respectively, against HSV-1 strain.
Surprisingly, the IC50 values for both polysaccharides against HSV-2 were very low,
i.e., 0.48 μg/mL. Also, time-of-addition experiments revealed that more efficient
anti-HSV activities were obtained when fucoidans were added during the infection stage,
thereby signifying their role at the early stages of viral infection. The adsorption
and penetration assays further demonstrated that the fucoidans were involved in interruption
of HSV adsorption to the host cell. Hence, it may be concluded that fucoidans could
serve as promising candidates for inhibition of HSV-2 viruses and may be successfully
utilized for various clinical applications. Similarly, a sulphated polysaccharide
was isolated from the green macroalgae Monostroma nitidum (Wang et al., 2020). The
compound isolated from M. nitidum was identified as a water-soluble sulphated glucuronorhamnan
and thus designated as MWS. Various cytotoxicity and antiviral assays were performed
to estimate the activity of MWS against EV71, a strain of human pathogenic enterovirus.
It was observed that MWS was not toxic to the used cell lines and demonstrated a broad-spectrum
of antiviral activity, especially against EV71 under defined in-vitro conditions.
Further, it was concluded that MWS inhibits the EV71 infection by either targeting
the host signaling pathway (down-regulation of host phosphoinositide 3-kinase/protein
kinase B signaling pathway) in EV71 early life cycle and/or interrupting adsorption
of virus to the host cell. The former mechanism has been concerned with the suppression
of viral infection. The study also involved animal experiments, and a significant
reduction in the viral titers was observed upon intramuscular administration of MWS
in EV71 infected mice (Wang et al., 2020). Additionally, the SPs obtained from macroalgae
Cladosiphon okamuranus and Ulva clathrata were also observed to demonstrate significant
antiviral activity against the Newcastle disease virus under defined in-vitro conditions
(Aguilar-Briseño et al., 2015). Another study elaborated the antiviral activity of
SPs obtained from Ulva pertusa, Grateloupia filicina, and Sargassum qingdaoense against
the avian influenza virus under in-vitro and in-vivo conditions (Song et al., 2016).
A recent review highlighted the possibility of utilizing the SPs obtained from Porphyridium
sp. (red microalga) as a potential therapeutic to combat COVID-19 disease (Gaikwad
et al., 2020). Based on the antiviral activity of Porphyridium polysaccharides against
a wide range of viruses including HSV (Huheihel et al., 2002), varicella zoster virus
(Raposo et al., 2013), hepatitis B virus, vaccinia virus (Radonić et al., 2010) and
retroviruses (Xiao and Zheng, 2016), this microalga has been considered to hold immense
potential for the development of an antiviral pharmaceutical composition against SARS-CoV-2
as well (Gaikwad et al., 2020). Also, the effective inhibition (in-vitro) of SARS-CoV-2
by SPs (fucoidans) obtained from macroalgae Saccharina japonica was reported by Kwon
et al. (2020). The fucoidans labeled as RPI-27 and RPI-28 demonstrated significant
activity against SARS-CoV-2 with RPI-27 being more potent than the antiviral drug
remdesivir. These highly branched fucoidans were observed to interfere with the binding
of viral S protein to the heparan sulfate co-receptor of the host cells, thereby,
inhibiting the viral infection. Thus, the study suggested the possibility of utilizing
fucoidans alone or in combination with other antivirals as a promising therapeutic
strategy against SARS-CoV-2 infection (Kwon et al., 2020). These studies indicate
the potential therapeutic role of algal sulphated polysaccharides.
Discussion and Conclusion
There has been a substantial increase in evidence that reveals the antiviral activity
of various microalgal and macroalgal metabolites like lectins, sulphated polysaccharides,
and phycocyanobilins. Recent studies have reported that these compounds demonstrate
substantial activity against a wide array of DNA and RNA viruses, including the influenza
virus known to be associated with respiratory illnesses. As discussed, the bioactive
molecules could serve as a novel therapeutic option to tackle SARS-CoV-2 and alike
viruses. Considering the dire need for the development of therapeutics against SARS-CoV-2,
there is a necessity to screen through the myriad of algae-derived potential antivirals
which demands further evaluation and research.
Author Contributions
AB: conceptualization, data curation, visualization, and writing - original draft.
PA: validation, writing - review & editing. SP: conceptualization, writing - review
& editing, and supervision. All authors contributed to the article and approved the
submitted version.
Conflict of Interest
The authors declare that the research was conducted in the absence of any commercial
or financial relationships that could be construed as a potential conflict of interest.