Phytochem Rev (2021) 20:693–703
https://doi.org/10.1007/s11101-020-09730-4 (0123456789().,-volV)
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Molecular combination networks in medicinal plants:
understanding synergy by network pharmacology in Indian
traditional medicine
Pulok K. Mukherjee . Subhadip Banerjee . Amit Kar
Received: 11 April 2020 / Accepted: 26 October 2020 / Published online: 3 January 2021
Ó Springer Nature B.V. 2021
Abstract Translation of traditional medicine is
moving away from the reductionist approach towards
harnessing the synergy of poly-pharmacological
phyto-combinations that modulate the activity of
target networks of underlying disease phenotypes.
Metabolomics enabled the profiling of herbal extracts
and formulations in high resolution through LC-MS/
MS, GC-MS and NMR providing the possibility of
multi-component drug discovery. Phytopharmaceuti-
cals are multi-molecular combinations which follow a
combinatory treatment module which minimizes off-
target toxicity by synergistic potency and improve
outcomes by synergistic efficacy. Combination syn-
ergy is a multi-dimensional concept where synergy
can be observed from a network pharmacology
perspective. Network pharmacology uses systems-
level drug–response phenotypes originating from
various ‘omics’ platforms. This network pharmacol-
ogy with metabolomics has been proven to be
effective in elucidating the mechanisms of action of
P. K. Mukherjee (&)
An Autonomous Institute under Deptartment of
Biotechnology, Institute of Bioresources and Sustainable
Development, Government of India, Imphal 795001, India
e-mail: naturalproductm@gmail.com;
director.ibsd@nic.in
P. K. Mukherjee
S. Banerjee
A. Kar
Department of Pharmaceutical Technology, School of
Natural Product Studies, Jadavpur University,
Kolkata 700032, India
medicinal plants and complex traditional formula-
tions. LC-MS/MS of medicinal plants along with
network pharmacology unveil synergistic molecular
combinations which interact with neighbouring targets
in a combination synergy network. Case studies on
some traditional herbs mentioned in Ayurveda, has
been reported based on the combination of network-
based methods and metabolomics for establishing the
mechanism for their therapeutic potentials. The syn-
ergistic combination of molecules interacts with
target-disease networks which provide novel, mech-
anistic insights towards understanding their therapeu-
tic potentials. This review along with case-studies
illustrates various strategies to explore the mechanism
of action for different synergistic natural products
combinations through neighbourhood-based network
pharmacology approach.
Keywords Metabolomics
Synergy
Ayurveda

LC-MS/MS
Network pharmacology
Traditional
medicine
List of abbreviations
PPI protein-protein interact
LC-MS/ Liquid Chormatography Mass
MS Spectrometry
GC-MS Gas Chromatography Mass Spectrometry
NMR Nuclear Magnetic Resonance
DAVID Database for Annotation, Visualization
and Integrated Discovery
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KEGG Kyoto Encyclopedia of Genes and
Genomes
RNAi RNA interference
QED quantitative estimate of drug-likeness
EGFR Epidermal growth factor receptor
CAH7 carbonic anydrase 7
XDH xanthine oxidase
HSV Herpes Simplex Virus
MAPK mitogen-activated protein kinase
JAK- Janus kinases signal transducer and
STAT activator of transcription proteins
ESR1 Estrogen Receptor 1
AHR Aryl hydrocarbon receptor
MMP12 Matrix Metallopeptidase 12
FUT7 Fucosyltransferase 7
NR1H3 Nuclear Receptor Subfamily 1 Group H
Member 3
Introduction
Translation of traditional medicine by exploring
synergy is important to understand the mechanistic
pharmacology, safety and repurposing of medicinal
plants which present a multi-molecular matrix with an
array of target interactions (Mukherjee et al. 2017).
The concept of traditional drugs and formulations
finds its relevance in spite of changes in the environ-
ment, lifestyle, culture and disease patterns (Mukher-
jee et al. 2016). Medicinal plants used in traditional
medicine represent molecular combinations which
may achieve more stable ‘‘network responses’’ than
single drugs. Multi-component mixtures of known
compositions represent combined actions of single
substances. Network pharmacology has emerged as a
rational approach for traditional medicine studies to
understand ‘‘network-target, multiple-component-
therapeutics’’ by ‘‘compound-protein/gene-disease’’
network revealing the regulation principles of small
molecules in a high-throughput manner and analysis
of drug combinations, hence synergy (Zhang et al.
2019). Ethnopharmacology is an important way
towards the discovery of novel drugs and formulation
from the multiple bioactive components within the
herbs and herbal extracts (Mukherjee 2019). Systems
pharmacology is an important tool to provide impor-
tant perspectives in this aspect (Verpoorte et al. 2005).
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Phytochem Rev (2021) 20:693–703
Network pharmacology presents a systems perspec-
tive to drug discovery (Hopkins 2007). The rates of
attrition of new drug candidates into effective clinical
therapeutic agents are rising due to harmful off-target
effects causing adverse-effects and failing to show
favourable network responses in complex diseases.
The effective drugs are showing network responses,
and act on multiple rather than single targets, a
phenomenon known as polypharmacology (Hopkins
2008). Multi-component therapeutics is effective in
complex diseases, as it ensures novel therapeutics
and drug repositioning (Pujol et al. 2010).
Plant-derived herbal drugs or phytopharmaceuti-
cals can be considered as a natural ‘‘model’’ for
combinatory treatments with multi-molecular combi-
nations offering two objectives, i.e., minimizing off -
target toxicity by reducing doses (synergistic potency)
and improve outcomes by escalating effect (synergis-
tic efficacy). The traditional system is challenged by
the necessity to determine their complex composition
and their multi-target mode of action. A proper
guideline is needed to be explored to provide strategies
for effective evaluation of safety and toxicity of
botanical mixtures resulted with drug discovery
through natural product research (Caesar and Cech
2019). Concepts of functional annotation analysis
were reviewed based on databases like Binding DB,
DAVID, KEGG and STRING with combination
synergy using ‘‘neighbourhood approach’’.
Systems pharmacology and metabolomics
Systems biology follows a holistic approach to prove
efficacy and obtain information that might lead to
understanding the mode of action based on synergy,
prodrugs, novel targets (Verpoorte et al. 2005).
Metabolomics is an important tool for standardisation
and quality control in medicinal plants (Mukherjee
et al. 2016). This has been observed that concepts of
Ayurveda merge with systems biology to reveal the
pathways of active components which may lead to
novel drug discovery (Deocaris et al. 2008). There-
fore, comprehensive chemical analysis is required not
only to establish a correlation between complex
chemical mixtures and molecular pharmacology but
also to understand the combination synergy, complex
cellular processes and biochemical pathways via the
metabolite-to-gene network (Nakabayashi and Saito
Phytochem Rev (2021) 20:693–703
2013). In order to understand the mechanism of action,
we need to study the human metabolite network and its
possible effects in the realm of systems-polypharma-
cology (Kell and Goodacre 2014). Metabolomics
opens the possibility where we can study the effects
and mechanisms of complex mixtures which are
common in traditional medicine like Ayurveda. When
connected to network pharmacology we can under-
stand the synergy of components enabling the study of
systems from a holistic perspective (Wang et al. 2005).
Multi-target therapeutics
Drugs involving single targets cannot combat multi-
genic complex disorders like cancer, or diseases that
involve multiple tissues or cell types such as diabetes
and immune-inflammatory disorders. Combination
drugs that impact multiple targets simultaneously are
better in controlling complex disease systems which
are less prone to drug resistance and are the standard of
care in many important therapeutic areas (Zimmer-
mann et al. 2007). Drug discovery is now shifting
towards targeting multiple targets as drug combina-
tions shown phenotypic effect with knowledge of
transporters (Kell 2013). A number of studies have
been performed to analyse multi-target drug discovery
with a network-based approach (Xie et al. 2012;
Leung et al. 2013). Yildirim et al. applied network
analysis to drugs and their targets by integrating
publicly available drug data (Yıldırım et al. 2007). Li
et al. (2011) proposed an integrated network model,
which considers the correlation between the interac-
tions of drugs with their molecular targets. Multitarget
therapeutics considers pharmacogenomics may pro-
vide benefit to more patients than monotherapies
(Roses 2000). Understanding the polypharmacology
and potential network pharmacology of botanical
drugs is crucial for the rationalization of phytotherapy
(Gertsch 2011). Systems-oriented drug design based
on network pharmacology has shown to be a produc-
tive strategy (Keith et al. 2005; Csermely et al. 2005).
Computational methods like network pharmacology
explore the systems-level drug–response phenotypes
from genome-wide transcriptomic profiles, to under-
stand the mechanisms of action of multi-targeted
natural product pinosylvin (Kibble et al. 2016).Molec-
ular mechanisms of drug combinations involve poten-
tiative, synergistic, and antagonistic effects of drug
695
pairs which can help the discovery of novel efficient
combinations multi-target effects (Jia et al. 2009).
Molecular combinations at systems-level
Molecular combinations can be majorly synergistic,
antagonistic or additive. Synergy, in this context, is the
result of combining two or more chemical compounds
to produce an effect that is greater than additive effects
(where additive effects are computed from the indi-
vidual effects based on specific mathematical mod-
els) (Jia et al. 2009). Drug combinations show a
multitude of mechanisms which has been analysed by
various techniques. Despite the significance of com-
pound combinations in therapeutic and toxicity stud-
ies, the ability to mechanistically explain and model
compound combinations in a systematic fashion are
currently limited. Strategies for experimentally deter-
mining the possible effect of drug combinations can
range from screening several compounds at a time
from a large database to exhaustive pairwise screening
such that an effective combination can be found (Feala
et al. 2010; Cokol et al. 2011). The gene interaction
networks can be used to predict the effect of a
combination of knockouts on the resulting phenotype,
they could also provide insight into synergy, and have
been used as a context for exploring drug combina-
tions (Lehár et al. 2008). A synergistic drug-drug
surface not only encodes a genetic interaction but a
study found the surface itself provided deeper insights
into the relationship (Yeh and Kishony 2007). The
topological features of the related PPI network to
identify possible combinations of hit targets. The best
ranked combinations are subsequently computed on
the basis of a synergistic score (Vitali et al. 2013) In
fact, it was found that it was possible to classify the
local network topology of the drug targets according to
the relative shape of the response curve, paving the
way for more-detailed interaction networks (Lehár
et al. 2007). The development of RNAi yields another
biological perturbation the technique, because knock-
down of a gene also often has a similar effect to a drug
targeting the product of the same gene (although
variation in time and dosage elements can confound
this picture). Double or even higher-order knockdown
experiments, known as combinatorial RNAi (coR-
NAi), are RNAi equivalent to compound combina-
tions (Grimm and Kay 2007). Strategies like mRNA
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knockdown and gene knockout are related biological
perturbation methods to chemical-protein interaction.
According to this notion of potential equivalence, it
might be possible to consider a broader ‘biological
combination’, allowing data from these different
biological facets to be integrated into a single, more
descriptive model. Data integration of this nature has
proved fruitful in other data-driven fields such as
network biology, for example, the integration of gene
regulatory and protein-protein interaction networks,
and thus might also be conducive to progress in
modelling the effect of compound combinations
(Fazekas et al. 2013).
Systems biology techniques like protein-protein
interaction (PPI) network-based methods, pathway
dynamic simulations, synergy network motif recogni-
tions, integrative drug feature calculations, and
‘‘omic’’-supported analyses can be an effective tool
to understand drug combination mechanism (Chen
et al. 2015). The dynamics of networks of pathways
can be investigated through the use of mathematical
network models, and the outcome of potential target
inhibition within the model can be compared to assay
readouts to allow MOA hypothesis generation of a
combination (Lehár et al. 2007; Zhang et al. 2014).
These models could make use of large-scale datasets
of compound combination responses. Even though
limited in terms of availability, opportunities to train
and test predictive models can be provided. The ability
to integrate chemical bioactivity data from different
realms of the biological system, viz. gene expression,
gene–protein interaction networks and pathway anno-
tations, among others towards assessing compound
combinations across different disease areas have been
listed elsewhere (Bulusu et al. 2016). The ‘‘pathway–
pathway interaction’’ network-based synergy evalua-
tion method has been a success in predicting the
potential synergistic drug combinations (Chen et al.
2016). NIMS (Network target based Identification of
Multicomponent Synergy) studies the relationship
among agents with network interactions among the
molecular targets or responsive gene products
involved in the potential mechanisms of multicompo-
nent synergy (Li et al. 2011). Drug Synergy prediction
based on differential evolution based multinomial
random forest (Kaur et al. 2019), network topology
(Yin et al. 2014), chemogenomics knowledge (Zhang
et al. 2016), interaction potency model (Yadav et al.
2015) and neighbourhood approach (Zou et al. 2012)
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Phytochem Rev (2021) 20:693–703
has provided in depth understanding in traditional
medicine.
A key bottleneck in identifying and understanding
the molecular effects of compound mixtures on a
systems-level are the integration of biological and
chemical information (Searls 2005). The data that
needs to be integrated for a comprehensive systems-
level analysis to identify compound combinations
efficiently include chemical bioactivity data (Gaulton
et al. 2012), targets and pathway annotations (Fazekas
et al. 2013), gene expression data (Lamb et al.
2006) and protein interactions (Franceschini et al.
2013). In the area of medicinal plant and traditional
medicine combination therapies, a deeper understand-
ing of the underlying biological impact is required,
such as that on gene expression and regulatory
pathways. This can be achieved by integrating
metabolomics with bioactivity data from different
realms of the biological system, viz. gene expression,
gene–protein interaction networks and pathway anno-
tations, among others. This approach can also assist in
generating biologically meaningful MOA hypotheses
for combinations of phytomolecules and other
approved clinical alternatives which will help in our
understanding and applications of phyto-combination
therapies for the future.
Synergy and network pharmacology of medicinal
plants
Synergy of natural products is the holy chalice of
pharmacology and therapeutics. Network-based meth-
ods are emerging as tools for studies in new drug
discovery, systems pharmacology and toxicology (Wu
et al., 2018). It helps us to understand the novel
therapeutics using natural products as drugs with
multiple compounds responsible for the cumulative
activity and combination synergy (Yang et al.
2018a, b; Liu et al. 2018; Li et al. 2019; Ulrich-
Merzenich et al. 2019; Gong et al. 2020). These
methods have been shown to be effective in multiple
herb formulations used in traditional medicine (Yao
et al. 2013; Zhou et al. 2016; Zhong et al. 2017; Liu
et al. 2018).
Ayurveda is a traditional system of medicine for
disease management and health practiced in India for a
long time. However, the recurrent use of this medicine
faces challenges due to their composition and multi-
Phytochem Rev (2021) 20:693–703
target mode of action, which needs scientific valida-
tion. The term synergy or ‘samyoga’ is also scripted in
the ancient Ayurvedic text of Sarangadhar Samhita.
Research on network pharmacology protrudes the
understanding of system biology including protein-
protein interaction, expression of genome and mRNA
to establish the probable mechanism of action with its
proper validation (Mukherjee et al. 2018). The
synergy of curcumin was verified by an approach
towards target networks of drug pairs when combined
with multiple drugs like capsaicin, celecoxib, ralox-
ifene, silibinin, sulforaphane, tacrolimus, and tamox-
ifen followed by experimental verification in rat
against myocardial ischaemia (Wan et al. 2014).
Curcumin and piperine possess significant neuropro-
tective potential as they showed therapeutic potential-
ity as acetylcholine and amyloidogenic inhibitors
(Abdul Manap et al. 2019). Curcumin together in
combination with Artemisinin has shown synergistic
effect against malarial parasites (Nandakumar et al.
2006). Network pharmacological study showed the
potential of withanolide-phytosterol combination to
achieve effective immunomodulation (Chandran and
Patwardhan 2017). In recent scenario, network phar-
macology along with the multi-target approach serves
as the novel tool for searching novel therapeutics from
botanicals where multiple compounds are responsible
for cumulative activity and synergy (Wagner and
Ulrich-Merzenich 2009; Orland et al. 2014; Ulrich-
Merzenich et al. 2017, 2019).
Different computational methods explored differ-
ent strategies to investigate the complex mechanisms
of multi-component drug action in order to understand
their complex mechanism of action (Hopkins 2007;
Hutchinson and Kirk 2011). The application of
systems pharmacology for herbs may effectively
support the elucidation of the mechanism of drug
combinations and pharmacological networks (Kibble
et al. 2015). Using network pharmacology we showed
the combinatory modulation on the NF-jB and the
p38 MAPK pathway in the HSV-1 skin infection
model by the Boswellia serrata oleo-gum-resin
(Goswami et al. 2018). Similar strategies have been
a success in understanding the combination synergy of
Trigonella foenum-graecum plant from Ayurveda
against hyperlipidemia and hyperglycemia (Banerjee
et al. 2019).
697
Case study- combinatorial synergy of Trigonella
foenum-graecum seeds against hyperglycemia
and hyperlipidemia
The seeds of Trigonella foenum-graecum L. (Methika
in Sanskrit), is an Ayurvedic herb indicated in
Prameha or early diabetes mellitus together with
anti-obesity and also useful for lowering blood lipid
level. Our reported study based on the understanding
of the combinatory mechanism of action for validating
the therapeutic claims of Trigonella. In this study, we
have explored the metabolite fingerprinting and iden-
tification together with the network pharmacology
approach for better understanding of underlying
combination synergy. The LC-MS/MS analysis of
the ethanolic extract resulted in the identification of 13
phyto-compounds which were found to be bio-avail-
able and possess drug-like properties as per QED and
Veber drug-likeness parameters. The pathway analy-
sis showed enrichment for different pathways like
MAPK pathway (p- 4.69E-07), JAK-STAT pathway
(p-6.30E-05), adipocytokine (p-0.00179), Type 2
diabetes mellitus (0.00441), Insulin signalling path-
way (p-0.0121), mTOR signalling pathway (p-
0.000378), which are all connected to hyperlipidemia
and hyperglycemia. The combination synergy net-
work identified 23 targets interacting with these 13
compounds based on a network neighbourhood
approach. Most of the compounds interact with
carbonic anhydrase 7. Gallic acid was found to be
interacting with FUT7 which is a key protein that
activates EGFR/AKT/mTOR signalling cascade with
therapeutic implication in hyperglycemia and hyper-
lipidemia (Banerjee et al. 2019).
Screening of anti-hyperglycemic and lipid
lowering components in trigonella seeds
The target-compound network was constructed with
the interacting targets and compounds, in which many
targets found to interact with multiple compounds.
Higher degree distribution of hub nodes for P00533
epidermal growth factor receptor (EGFR), and P43166
carbonic anydrase 7 (CAH7) was found, while
peripheral nodes such as kaempferol interacts with
P47989- xanthine oxidase (XDH) suggesting a lower
distribution (Banerjee et al. 2019).
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Target gene interactions of Trigonella seed
molecules
The compound–target–disease network identified the
interaction between the compounds to hyperglycemia
and hyperlipidemia associated targets. This network
further elucidated target and compound combination
synergy. Quercetin and trigonelline was found to be
interacting with insulin-like growth factor (IGF)-1
receptor. Tricin has been found to be interacting with
pancreatic alpha-amylase (AMYP) which is reported
to play an important part in glucose metabolism
(Banerjee et al. 2019).
Target protein-protein interaction network
The protein-protein interaction (PPI) network was
used to elucidate significant biological processes,
cellular components, molecular functions and path-
ways of target proteins on the basis of the experimental
interaction curated from the database STRING inter-
actome (Szklarczyk et al. 2017). The PPI enrichment
p value was found to be less than unity which means
that proteins have more interactions among them-
selves than what would be expected for a random set of
proteins of similar size, drawn from the genome. Such
enrichment indicates that the proteins are at least
partially biologically connected, as a group. The
evidence were calculated on the basis of gene
ontologies (GO) and other databases like KEGG,
Pfam, InterPro. Module analysis of the PPI interaction
network was done to identify major hyperlipidemia
associated targets interaction. The PPI pathway anal-
ysis based on KEGG database showed significantly
lower p value, where false discovery rate (FDR) has
been observed in all connected hyperlipidemic and
hyperglycaemic pathways including MAPK pathway
(p- 4.69E-07), JAK-STAT pathway (p-6.30E-05),
adipocytokine (p-0.00179), Type 2 diabetes mellitus
(0.00441), Insulin signalling pathway (p-0.0121),
mTOR signalling pathway (p-0.000378). Functional
association clustering analysis of the protein targets
revealed involvement in different metabolic pathways
like alpha-linoleic acid metabolism, glycolipid meta-
bolism, glycolysis or gluconeogenesis etc. The com-
pound target network and metabolic cluster showed
apigenin interacts with an important target known as
beta-hydroxysteroid dehydrogenases (DHB1 and
DHB2). Kaempferol interacts with xanthine oxidase,
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Phytochem Rev (2021) 20:693–703
thereby providing protection against production of
free radicals together with protecting hyperlipidemia
associated endothelial dysfunction and the progression
of atherosclerosis. The other correlative mechanisms
are hypoxia-inducible factor (HIF) pathway, mTOR
pathway, insulin resistance which is involved in
hyperlipidemia and hyperglycaemic conditions. It
has been found that molecules of TGFE interact with
different proteins which are involved in these
pathways.
Combination synergy
The synergy mechanism of action of the molecular
combinations against interconnected diseases has been
analysed using a deductive approach, the combinatory
mechanism network showing molecules interacting
with the targets associated with hyperlipidemia and
hyperglycemia. Our network pharmacology guided
approach reported that 13 compounds together interact
with 23 targets connected with associated conditions
of hyperlipidemia and hyperglycemia as deduced from
the compound-target-disease networks which are only
‘‘first degree neighbours’’ of the disease terms forming
the drug combination network. We found molecules
working together on the targets (IGF1R, CA7, FUT7,
NR1H3) to combat insulin resistance. Most of the
compounds interact with carbonic anhydrase 7. Gallic
acid was found to interact with FUT7 which is a key
protein that activates EGFR/AKT/mTOR signal path-
way with therapeutic implication in hyperglycemia
and hyperlipidemia. Kaempferol and quercetin were
found to interact with IGFR1 which is involved in
glucose metabolism. Targets like ESR1, SERPINE1,
AHR, AXL, AHR, MMP12, FUT7, NR1H3 and ESR2
interact with most of the compounds providing
protection against the cardiovascular disease which
is associated with hyperglycemia and hyperlipi-
demia. The network pharmacology analysis strongly
suggests through multimode evidence that Trigonella
largely works on the insulin signalling pathway.
Therefore in totality, the work was based on LC-
MS/MS analyses which are a metabolomics technique
followed by network pharmacology investigation
elucidated a mechanistic landscape of Trigonella
seeds which are widely used in Ayurveda against
hyperglycemia and hyperlipidemia. The results indi-
cate that Trigonella contains various compounds like
apigenin, calycosin, gallic acid, kaempferol, luteolin,
Phytochem Rev (2021) 20:693–703
orientin, pratensin, quercetin, tricin, which are respon-
sible for its mechanism of action. The mechanism
involves insulin signalling pathway based on the
antioxidant potential, which interact with the protein
subunit of the carbonic anhydrase. It also showed
lipid-lowering and cardio-protective potential which
may be very much useful in the treatment of diabetic
and pre-diabetic patients as the targets show high
enrichment for the MAPK pathway, JAK-STAT
pathway, adipocytokine pathways. The results indi-
cate that EGFR/AKT/mTOR signalling cascade is also
involved in the mechanism of action of TGFE. The
following study will be helpful towards the develop-
ment of methods for better understanding of the
mechanism of action of similar multi-molecular
systems present in traditional medicine. The combi-
nation synergy network analysis based on neighbour-
hood approach can help us in further understanding of
the mechanism of multi-molecular fixed-dose
combinations. (Fig. 1).
Fig. 1 Combination synergy by network pharmacology and metabolomics
699
Case Study 2 Boswellia serrata oleo-gum-resin
against HSV-1 infection
Boswellia serrata oleo-gum-resin possesses a strong
traditional background of treating diverse skin ail-
ments including infection. A study was reported by our
group, elucidating the mechanism based antiviral
activity of B. serrata oleo-gum-resin methanol extract
(BSE) against Herpes Simplex Virus 1 (HSV-1)
(Goswami et al. 2018). The network pharmacology
analysis was undertaken on the compounds of B.
serrata, derived from UNPD. Our study involved a
multimode cross-validated network analysis to eluci-
date the possible molecular mechanism, underlying
pathways, genes and protein-protein interactions
based on chemical similarity of boswellic acid (BA)
to inherent target ligand (Fig. 2). Additionally, this
analysis resulted with the evaluation of synergy and
the possible mode of interaction of BSE with HSV-
infected host cell. The PPI network analyses reported
p38 MAP-kinase and NF-jB had a maximum degree
of interaction with other proteins. STRING 3.0 based
pathway enrichment analysis in network analyst
showed the enriched pathways against HSV-infection
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Fig. 2 Multimode cross-validated network analysis to elucidate the underlying pathways
which further validate the ethno-medicinal claim of
BSE and its phyto-constituents against HSV- 1
invasion. The protein–protein interaction network
with the module of the MAP-kinase signalling path-
way found that p38 MAP-kinase pathway proteins and
hub-proteins were responsible. The most important
targets were TNF-a, NFKB2, MAPK14, MAPK8,
MAPK9 indicated as hub proteins. The results also
suggest the importance of NOS2 and IL-1b in
signalling, as it shows the betweenness value of
1742.04 and 1485.50 in the PPI network. Thus, our
results clearly indicated that these hub-proteins are
associated with HSV-infection, as suggested by their
significant enrichment (Goswami et al. 2018).
Conclusions
Synergistic poly-pharmacological approach in tradi-
tional medicine reveals the combinatory therapeutic
efficacy of the constituent without producing any
potential adverse effect towards human health. Several
aspects on the existing synergy of traditional drugs are
very important and the integral part for deciphering the
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Phytochem Rev (2021) 20:693–703
traditional medicine and medicinal plant pharmacol-
ogy. The concept of network pharmacology is logi-
cally connected to pharmacognosy and traditional
medicine including Ayurveda, where the uses of
medicinal plants as ‘botanical drugs’ is very popular.
Metabolomics for chemo-profilling of these multi-
molecular herbs and herbal formulations is also very
important for translational research of traditional
medicine. LC-MS/MS, GC-MS, NMR are very crucial
hyphenated technologies in this regard which have
multiple applications in the field of targeted and
untargeted metabolomics. The ‘omics’ based systems
approach will promote translational research in tradi-
tional medicine. Deciphering the mechanism of multi-
molecular combinations network guided approach to
synergy research will not only be helpful for
researchers to discover new phytomedicines or drug
combinations but also help us to avoid the possible
negative synergy in case of herb-drug interaction.
Therefore drug discovery through this network neigh-
bour-based approach to understand combination syn-
ergy can leverage traditional medicine inspired drug
discovery. The future of drug discovery is based on
synergistic combinations of molecules which shows
Phytochem Rev (2021) 20:693–703
favourable network interactions against diseases and
also maintenance of health-promoting wellness.
Acknowledgements Authors wish to acknowledge the
financial support received from Science and Engineering
Research Board (SERB), Govt. of India, New Delhi and
Department of Biotechnology (DBT), Govt. of India, New
Delhi for financial support.
Compliance with ethical standards
Conflict of interest The authors confirm that there are no
known conflicts of interest associated with this publication.
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