Journal of Herbal Medicine 14 (2018) 1–16

Contents lists available at ScienceDirect

Journal of Herbal Medicine

journal homepage: www.elsevier.com/locate/hermed

Review article

Validation of medicinal herbs for anti-tyrosinase potential T

a,⁎ a a a a
Pulok K. Mukherjee , Rajarshi Biswas , Akanksha Sharma , Subhodip Banerjee , Sayan Biswas ,
C.K. Katiyarb
a
School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700 032, India
b
Health Care Division, Emami Limited, 13, BT Road, Kolkata, 700056, India

ARTICLE INFO ABSTRACT

Keywords: Tyrosinase is a key regulatory multifunctional enzyme containing copper that is responsible for the biosynthesis
Tyrosinase inhibitor of melanin that determines the color of the skin. Accumulation of excessive melanin causes various dermato-
Medicinal plant logical disorders including melasma and age spots. Tyrosinase is also responsible for enzymatic browning re-
Anti-melanogenesis actions in damaged fruits and vegetables. It deteriorates the color clarity of plant-derived food product which
Skin whitening
results in loss of nutritional quality. The study of Tyrosinase inhibition is an active field of research in derma-
Phytoconstituents
tological, biomedical, food and agricultural science and also has potential impact in the domain of insect
physiology. Despite several developments on tyrosinase inhibitors, their safety concern still requires investiga-
tions due to their undesirable side-effects. Research in this context is being carried out to find potent and safe
leads as tyrosinase inhibitors from medicinal plants. This review provides comprehensive overviews of various
tyrosinase inhibitors obtained from medicinal plants with their mechanism of action. Several medicinal plants as
tyrosinase inhibitors have been extensively studied and successfully marketed. The polyphenol and stilbenes
group of phyto-molecules have been established as effective tyrosinase inhibitors. Few of these molecules
however have been clinically investigated in the context of potential anti-melanogenic medicinal plants.
Tyrosinase inhibitors are commercially available for cosmetic purposes to lighten the complexion. Clinically,
they are also used in the treatment of hyper-pigmentary disorders. They are equally applicable for the anti-
browning agents in the food-processing industry. This review will be useful for the development and evaluation
of tyrosinase inhibitors from medicinal plants.

1. Introduction is used further for the synthesis of melanin pigment. Overproduction

Tyrosinase or polyphenol oxidase is a multifunctional copper-con-
taining enzyme prevalent in the plant and animal kingdom. It is a key
regulatory enzyme that greatly influences the process of melanogenesis
within melanocytes. Melanin, a mixture of different biopolymers de-
termines the color of the skin and hair as well as gives protection from
harmful UV radiation. Tyrosinase catalyzed standard quinone precursor
and accumulation of melanin pigment in the skin leads to the devel-
opment of dermatological ‘hyperpigmentation’ manifesting in clinical
conditions such as solar lentigo, melasma, post-inflammatory hy-
perpigmentation (PIH), and linea nigra. It may also occur due to hor-
monal imbalance in the body, i.e. Cushing's disease, Addison's disease
and Nelson's syndrome. Arbutin and kojic acid are known tyrosinase
inhibitors commonly used in cosmetic products for skin whitening.

Abbreviation: UV, ultraviolet; PIH, post-inflammatory hyperpigmentation; ROS, reactive oxygen species; L-dopa, L-3,4-dihydroxyphenylalanine; IQ, indole-5, 6-
quinone; Tyrp-2, tyrosinase-related protein-2; DHICA, 5, 6-dihydroxyindole-2-carboxylic acid; Tyrp-1, tyrosinase-related protein-1; IQCA, indole-2-carboxylic acid-5,
6-quinone; MITF, microphthalmia-associated transcription factor; DAG, diacylglycerol; PKC, protein kinase-C; DGK-ζ, diacylglycerol kinase-ζ; ACTH, adrenocorti-
cotropic hormone; MC1R, melanocortin receptor-1; cAMP, adenosine monophosphate; MEK, mitogen-activated orotein kinase; ERK, extracellular signal-regulated
kinase; PI3K, phosphatidylinositol 3-kinase; Akt, serine-threonine protein kinase; EGF, epidermal growth factor; PPO, polyphenol oxidase; IC50, 50% inhibitory
concentration; α-MSH, α-melanocyte-stimulating hormone; EC, (-) epicatechin; C, (+) catechin; EGC, epigallocatechin; ECG, (-)-epicatechin 3-0-gallate; GCG,
(-)-gallocatechin 3-0-gallate; EGCG, (-)-epigallocatechin 3-0-gallate; mRNA, messenger ribonucleic acid; DNA, deoxyribonucleic acid; GR, glycyrrhizin; SCF, stem cell
factor; GC analysis, gas chromatography; GSK-3β, glycogen synthase kinase-3β; HQ, hydroquinone; HHQ, hydroxyhydroquinone; BQ, benzoquinone; RH, rhodo-
dendrol; TCM, traditional Chinese medicines; Cd, cadmium; As, arsenic; Hg, mercury; Pb, lead
⁎
Corresponding author.
E-mail addresses: naturalproductm@gmail.com (P.K. Mukherjee), rajarshi_mpharm@rediff.com (R. Biswas), akanksha2431@gmail.com (A. Sharma),
ami.subhadipbanerjee@gmail.com (S. Banerjee), sayanb0@gmail.com (S. Biswas), ck.katiyar@emamigroup.com (C.K. Katiyar).

https://doi.org/10.1016/j.hermed.2018.09.002
Received 14 October 2017; Received in revised form 22 June 2018; Accepted 14 September 2018
Available online 24 September 2018
2210-8033/ © 2018 Published by Elsevier GmbH.
P.K. Mukherjee et al.
Clinically, these de-pigmenting agents are applied for hyperpigmenta-
tion therapy. However, Kojic acid causes dermal sensitization at a
therapeutic concentration whereas arbutin has potential cytotoxicity
(Sarkar et al., 2013; Burnett et al., 2010; Zhu and Gao, 2008).
Tyrosinase inhibitors are also widely used in the cosmetic field as
fairness enhancers. Approximately 15% of the world population uses
skin whitening agents. India, Japan and China are the global hub for
consuming skin whitening agents. A recent survey indicated that a large
number of Indian men (< 80%) use fairness creams (Pillaiyar et al.,
2017).
Utilization of plant extracts and their derived phytoconstituents has
a likely future for controlling hyperpigmentation. Much evidence has
been explored to support the use of herbs as an alternative way for its
management (Mukherjee and Wahile, 2006; Sarkar et al., 2013; Zhu
and Gao, 2008). Tyrosinase is an essential enzyme in an insect’s de-
fensive mechanisms. It has a broad range of functions in insects, in-
cluding wound healing, sclerotization, molting and encapsulation.
Thus, some insecticides have been developed targeting the inactivation
of tyrosinase activity (Kim and Uyama, 2005). Tyrosinase is also re-
sponsible for enzymatic browning reactions in fruits and vegetables.
Browning usually damages the color of plant-derived food products,
which may indicate spoilage of its nutritional quality. This excessive
tyrosinase activity in food can be prevented by using tyrosinase in-
hibitors. Additionally, tyrosinase inhibitors are equally valuable in
scavenging reactive oxygen species (ROS), and protect from UV-radia-
tion induced skin cancer, e.g. melanoma (Mukherjee et al., 2011; Rao
et al., 2013). Several review articles have summarized the currently
available tyrosinase inhibitors from synthetic, semi-synthetic and nat-
ural origin (Chang, 2009; Loizzo et al., 2012). Several available review
articles have described the tyrosinase inhibitory activities of phyto-
molecules and their current status however none of them have dis-
cussed the commercialization of medicinal plants as a tyrosinase in-
hibitor.
In this context, this review focusses on tyrosinase inhibitors from
medicinal plant resources particularly on the systematic study of
medicinal plants and its phytoconstituents with wide-spread utility,
mechanism of action, adverse effects and prospects. The information
outlined in this review will provide a new dimension to the research for
the development of safe and potent tyrosinase inhibitors from medic-
inal plants.
2. Methodology
Scientific information on the topic was collected from the literature
available in the books on medicinal plants as well as from databases
such as PubMed, Google Scholar, Springer, Scopus, and Science Direct.
Different combination of keywords i.e.; “Tyrosinase inhbition/medic-
inal plants”; “Phytochemicals/anti-tyrosinase”; “Plants as anti-tyr-
osinase”; and “Plants secondary metabolites/anti-tyrosinase”; were
applied to gather information available on the topic. Moreover, refer-
ences of selected articles were also screened manually for additional
information. The authors also searched key articles from journals such
as Industrial crops and products; Archives of Pharmacal Research; Food
and Chemical Toxicology; Journal of Agricultural and Food Chemistry;
Food Chemistry; Evidence-Based Complementary and Alternative
Medicine; Phytomedicine; International Journal of Biological
Macromolecules; Process biochemistry; Fitoterapia; Journal of
Ethnopharmacology; PLoS One; Bioorganic and Medicinal chemistry;
Phytochemistry; Journal of Dermatological Science; Journal of Natural
Products; Planta Medica; Phytotherapy Research and Journal of
Bioscience and Bioengineering to collect as much information as pos-
sible which was relevant to the topic.
3. Role of Tyrosinase in melanogenesis
The name “melanin” comes from the ancient Greek word “melanos,”
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Journal of Herbal Medicine 14 (2018) 1–16
meaning dark. Chemically melanins are of variable structures, but its
parent structure is derived from the oxidation of phenolic precursors.
The primary biosynthesis pathway of melanin is called the Raper-Mason
pathway. Oxidation of L-tyrosine and L-dopa is the first step of mela-
nogenesis catalyzed by tyrosinase. This first step is the rate-limiting step
in melanin synthesis (Chang, 2009). Thus, tyrosinase inhibitors have
emerged as inhibitors of melanin synthesis, thus inhibiting melano-
genesis. The primary action of this enzyme is oxidation of L-tyrosine and
L-dopa to dopaquinone and subsequent cyclization of quinine precursor
to leukodopachrome. In subsequent steps leukodopachrome (cyclo-
dopa) is oxidized by redox exchange to dopachrome. Further, dopa-
chrome is decarboxylated and with the help of tyrosinase, it is con-
verted to indole-5, 6-quinone (IQ). Simultaneously, dopachrome is
reduced by Tyrosinase related protein (Tyrp-2) proteins to 5, 6-dihy-
droxyindole-2-carboxylic acid (DHICA) and oxidized by tyrosinase or
Tyrosinase related protein 1(Tyrp-1) to indole-2-carboxylic acid-5, 6-
quinone (IQCA). DHICA, Indole 2 carboxylic acid -5, 6-quinone IQCA,
and IQ again polymerized to form eumelanin (black/brown in color). In
another sub-pathway, a thio group containing amino acid cysteine is
added to dopaquinone and further following a series of redox reactions
pheomelanin (reddish/yellow in color) is produced. Formation of
neuromelanin is formed by mixing eumelanin and pheomelanin to-
gether. Neuromelanin is found within the catecholaminergic neuron
(Solano, 2014).
Regulation of melanogenesis in mammals is a complex process. Its
regulation at the cellular level is controlled through formation and
destruction of melanocyte organelle ‘melanosomes’. Melanogenesis is
regulated at the sub-cellular level by expression of the melanogenesis-
related enzymes, including tyrosinase, Tyrp-1, and Tyrp-2. The sche-
matic representation of melanin synthesis and its cellular and sub-cel-
lular regulation is shown in Fig. 1. Regulation of melanogenesis is
closely associated with a variety of hormones, growth factors, inter-
leukins, prostaglandins, and interferons, which determine not only the
quantity but also the quality of the synthesized melanin. Ultraviolet
(UV) exposure and/or environmental stimulations also plays a key role
in Melanogenesis (Sardana and Ghunawat, 2015; Solano, 2014).
Fig. 1 shows the two commonly known signalling pathways in-
volved in the up-regulation of melanogenesis. In contrast, two signal-
ling pathways are down-regulating the melanogenesis. Micro-
phthalmia-associated transcription factor (MITF) is a class E basic helix-
loop-helix transcription factor, which is regulated by all signalling
pathways. MITF has an M-box as a promoter region and regulates the
gene expression of tyrosinase, Tyrp-1, and Tyrp-2. The up-regulation of
MITF activity increases the expression of the melanogenesis-related
enzymes, thus stimulating melanogenesis. In contrast the down-reg-
ulation of MITF activity depresses the expression of the related en-
zymes, thereby inhibiting melanogenesis (Chang, 2012). Diacylglycerol
(DAG) is a plasma membrane-bound component activated by UV irra-
diation or other receptors. It acts on the protein kinase-C (PKC) which
phosphorylate the serine/threonine residues of tyrosinase and activates
the enzyme. It has been reported that phosphorylated isoform dia-
cylglycerol kinase-ζ (DGK-ζ) regulates the melanin synthesis without
affecting tyrosinase and MITF mRNA levels (Bae-Harboe and Park,
2012).
Adrenocorticotropic hormone (ACTH) and Alpha-melanocyte-sti-
mulating hormone (α-MSH) act as an agonist on melanocortin receptor-
1 (MC1R) and activates the cyclic adenosine monophosphate (cAMP)-
dependent pathway. Then cAMP activates protein kinase A (PKA),
which then activates the gene expression of MITF and stimulates mel-
anogenesis. Another signalling pathway also targeting the gene ex-
pression of MITF is the Wnt signal pathway. A key control in this
pathway is the increase in the level of intracellular β-catenin. In con-
trast to up-regulation, the down-regulation of melanogenesis takes
place by the mitogen-activated protein kinase (MEK) and extracellular
signal-regulated kinase (ERK) pathway via the degradation of the MITF
protein (Chang, 2012). Furthermore, phosphatidylinositol 3-kinase
P.K. Mukherjee et al.
Fig. 1. Melanogenesis in melanocyte.
(PI3K) activates the serine-threonine protein kinase (Akt). Stimulation
of Akt pathway causes down-regulation of melanogenesis (Oka et al.,
2000).
3.1. Tyrosinase and its mode of inhibition
The active site of the tyrosinase contains type-3 (two) copper metals
coordinated by three histidine residues to which two oxygen atoms are
bound in a peroxy configuration. Tyrosinase exhibits both mono-
oxygenase or monophenolase and oxidase or diphenolase activity, and
both activities arise from the binding of di-oxygen to the two copper
ions (usually identified as CuA and CuB) located in the active site. In
mammalian tyrosinase, there is evidence that catechols bind to CuB,
whereas monohydric phenols bind to CuA. Tyrosinase (oxy-, met-, and
deoxy-tyrosinase) is present in three different states which is involved
in the melanin bio-synthetic pathway (Ismaya et al., 2011). Tyrosinase
is used to catalyze two distinct oxidation reactions as shown in Fig. 2.
Enzyme’s monophenolase activity is initiated by oxidation of phenols
(L-tyrosinase) to ortho-quinones by oxy-tyrosinase and which is then
converted to dopaquinone. The phenolic oxygen is initially bound to
the CuA of oxy-tyrosinase followed by binding with another copper ion
and subsequent homolytic dissociation of the complex take place to give
the ortho-quinone and deoxy-tyrosinase. This process continuously
takes place until the substrates (phenol and oxygen) are depleted. A
characteristic feature of monophenolase activity of the enzyme is re-
ferred to as the ‘lag phase’. In this stage, phenolic substrates are oxi-
dized very slowly and reach to maximum velocity after a certain in-
duction period. It is important to understand the necessities for
conversion of met-tyrosinase to deoxy-tyrosinase for comprehending
the lag period. Met-tyrosinase is the stable form (resting state) of the
enzyme because a copper ion is in the oxidized state, Cu(II)2. In this
form, phenolic oxygen cannot bind to the enzyme. As a result,
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Journal of Herbal Medicine 14 (2018) 1–16
monophenolase activity cannot be expressed (Ramsden and Riley,
2014).
It has been experimentally proven that catechols (L-dopa) are not
formed from phenols (L-tyrosinase). Catechols are the by-product of
leukodopachrome (cyclodopa) which is formed during dopachrome
generation. Native tyrosinase contains small amounts of oxy-tyrosinase
which can generate a correspondingly little amount of ortho-quinone.
Ortho-quinones then produce catechols that can activate more met-
tyrosinase. This process continues until all the enzymes are converted to
the oxy form. Therefore, it is clear that small amounts of L-dopa are
required to generate more of the met-enzyme. The lag period of the
monophenolase activity is the result of this process (Ramsden and Riley,
2014).
Diphenolase activity of tyrosinase is accomplished by binding two
adjacent hydroxyl groups of the catechols with two copper ions of the
active site. The hydrophobic ‘pocket’ of the tyrosinase active site is
sterically favorable for Catechol's bindings. This binding pattern is
different from the phenol (L-tyrosinase) binding model; catechols in-
itially bind to the active site through the CuB. The oxidation cycle of
catechols involves two steps. In the first step, oxy-tyrosinase is con-
verted to meta-tyrosinase. Catechol reduces the peroxy-bridge of the
active site of oxy-tyrosinase. This step is yielding the corresponding
ortho-quinone and water. The resultant copper ions are an oxidized
state in met-tyrosinase form, probably in a protonated form. In the
second stage, catechols again reduce the active site copper ions of met-
tyrosinase and give deoxy-tyrosinase [Cu(I)] and a molecule of ortho-
quinone. Binding di-oxygen restores the oxidation states of the active
site copper ions [Cu(II)] (Ramsden and Riley, 2014).
Many tyrosinase inhibitors from both natural and synthetic sources
have been identified. Tyrosinase inhibitors are examined in the pre-
sence of L-tyrosine or L-dopa as the enzyme substrate, and activity is
assessed regarding dopachrome formation. True tyrosinase inhibitors
P.K. Mukherjee et al. Journal of Herbal Medicine 14 (2018) 1–16

Fig. 2. Catalytic cycles of the monophenol and o-diphenol to o-quinone by tyrosinase [Eoxy, Emet, and Edeoxy are the three types of tyrosinase, respectively].

are classified into four types, including competitive inhibitors, un- 4. Medicinal plants with tyrosinase inhibition potential
competitive inhibitors, mixed type (competitive/uncompetitive) in-
hibitors, and non-competitive inhibitors. In most studies conducted to Several tyrosinase inhibitors have been developed from medicinal
discover new tyrosinase inhibitors, scientifically validated tyrosinase plants, but there is still a long way to go. Various medicinal plants have
inhibitors are often used as a positive standard. Kojic acid and arbutin been reported as being used as potential de-pigmenting agents re-
are intensively studied tyrosinase inhibitors and widely used as positive presented as follows. Phyto-constituents with tyrosinase inhibitory
controls in a tyrosinase inhibitory assay. Kojic acid is a fungal meta- potential are shown in Fig. 3.
bolite, currently functional as a cosmetic skin-whitening agent and as a
food additive for preventing enzymatic browning. Its ability to chelate
4.1. Aloe
copper in the enzyme active site may well explain the observed com-
petitive inhibitory effect. Kojic acid showed a competitive mode of in-
Aloe (Family-Liliaceae) is extensively used in the cosmetic industry
hibition on monophenolase activity and a mixed inhibitory effect on the
as a skin whitening agent. Approximate 350 species of aloe have been
diphenolase activity of mushroom tyrosinase (Chang, 2009). β-arbutin
identified. It has a long history of use as a traditional skin care herb thus
is a naturally-occurring hydroquinone-β-D-glucopyranoside of hydro-
inspiring researchers to research and develop de-pigmenting agents
quinone, commercially extracted from the Arctostaphylos uva-ursi
from them. Aloesin (1) and 2′'-o-feruloylaloesin (2) isolated from Aloe
(bearberry) plant. It showed competitive inhibition of monophenolase
arborescens Mill. (Candelabra aloe) showed mushroom-tyrosinase in-
activity of tyrosinase (Tomita et al., 1990). In 2004, Hori et al. (2004)
hibition activity. Enzyme kinetic study revealed 2′'-o-feruloylaloesin
found β-arbutin itself exhibited slow oxidizing potential upon L-tyr-
was a non-competitive inhibitor; Ki was 0.85 μM (Yagi et al., 1987). In
osinase when L-dopa is present as a cofactor.
1999, Jin and co-workers also found aloesin to be a non-competitive
inhibitor of mushroom tyrosinase (Ki 5.3 mM) (Jin et al., 1999). It was
found to interact synergistically with arbutin thus effectively inhibiting
melanin production (Wu et al., 2012a,b). Its synergistic interaction with

4
P.K. Mukherjee et al.
Fig. 3. Different phyto-molecules as tyrosinase inhibitors.
arbutin can effectively inhibit melanin production (Jin et al., 1999; Wu
et al., 2012a,b). Furthermore, a separate experiment was performed by
Choi and colleagues on UV radiated human skin. This work revealed
that aloesin and arbutin can also be useful as combination therapy for
UV mediated hyper-pigmentation (Choi et al., 2002). Later in 2002,
aloesin was identified as a competitive inhibitor with respect to human
tyrosinase (ex-vivo). Due to its higher molecular weight and poor per-
meability and/or solubility, it showed moderate human tyrosinase in-
hibition in comparison to mushroom tyrosinase inhibition in vitro
(Jones et al., 2002). Two chromone derivatives, isoaloeresin D and
aloeresin E obtained from the leaves of A. vera L. also showed potent
tyrosinase inhibition. Besides isolation from aloe several chromone
derivatives have been synthesized, but none of them are as active as
aloesin (Piao et al., 2002). Notably, a recent study suggests A. vera gel
extracted from different germplasms have promising mushroom tyr-
osinase inhibitory activity (Gupta and Masakapalli, 2013).
Aloin (3), an anthraquinone glycoside, also known as barbaloin has
been found to be a potent mixed type of mushroom-tyrosinase inhibitor.
The principal constituent of A. vera leaves and its extract has shown
depigmentory activity on isolated melanophores of tadpoles (Zhu and
Gao, 2008; Ali et al., 2012). Contrary to popular belief, it is evidenced
that aloin induced melanogenesis triggered by tyrosinase activity in
murine melanoma B16 cell line after three-day treatment (Tabolacci
et al., 2013). Besides, several species of aloe have been tested as tyr-
osinase inhibitors. Among them A. ferox showed strong monophenolase
inhibition (Mapunya et al., 2012).
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Journal of Herbal Medicine 14 (2018) 1–16
4.2. Crocus sativus L
C. sativus (Family-Iridaceae), commonly known as “spice saffron” is
used extensively in commercial skin whitening formulations.
Kaempferol (4) and its 3-O-glycoside derivative were isolated from
fresh flower petals of C. sativus and investigated on mushroom tyr-
osinase. The 50% enzyme inhibition concentration (IC50) of kaempferol
was estimated to be 67 μg/mL and acted as a competitive inhibitor,
whereas kaempferol-3-O-glycoside did not produce any activity. A si-
milar trend was also observed with flavonols such as quercetin 3-O-
glucoside (Kubo and Kinst-Hori, 1999). In another finding, isolated
flavonoid glycoside, isorhamnetin-3-O-robinobioside (5) from saffron
showed potent tyrosinase inhibition (IC50 ≈ 1.8 mM) as compared to
arbutin. Crocusatin H (6), crocin-1 (7), and crocin-3 (8) obtained from
stigmas of C. sativus exhibited significant tyrosinase inhibition. IC50 of
crocin-1 (≈0.15 mM) was found to be lower than kojic acid. In a recent
study, it was shown that crocin has diphenolase inhibition, but affinity
to mushroom tyrosinase was low (Patil et al., 2014). Furthermore,
Anantharaman and co-authors have demonstrated that crocin inhibits
the diphenolase activity of tyrosinase in a mixed competitive manner.
However, it did not minimize melanin production significantly in
comparison to kojic acid when tested on mouse melanoma cell-line
(Anantharaman et al., 2016). In 2004, Li and associates isolated and
identified 35 compounds from the petals of saffron and out of them
crocusatin-K (9) showed strong tyrosinase inhibition in a competitive
manner (Li et al., 2004).
P.K. Mukherjee et al.
Fig. 3. (continued)
4.3. Curcuma longa L
Ample evidence suggests that Curcuma longa (Family-Zingiberaceae)
present in a traditional semisolid preparation of powdered herbs called
‘Ubtan’ enhance the skin color (Mukherjee, 2015; Dixit and Goyal, 2011).
The rhizome of C. longa contains curcumin (10) as major phyto-con-
stituent. Du and co-worker have demonstrated mushroom tyrosinase in-
hibition potential of curcuminoids isolated from turmeric. The percentages
of tyrosinase inhibition (IC50 in μM) of curcuminoids are ranked as fol-
lows: curcumin < demethylcurcumin (11) < bis-demethylcurcumin
(12). Curcumin-bis-β-D-glycoside also displayed the potential for better
tyrosinase inhibition compared to curcumin (Du et al., 2011; Prasad et al.,
2014). Semi-purified fraction of C. longa was found to suppress melanin
biosynthesis in α-MSH-stimulated B16F10 melanoma cells by activation of
MEK ⁄ERK signalling pathway (Jang et al., 2009). The anti-melanogenic
potential of curcuminoids was further supported by UV-A mediated mel-
anogenesis inhibition in G361 melanoma cells by C. aromatica extract
(Panich et al., 2010). Several attempts have been made to synthesize
curcumin derivatives for tyrosinase inhibition, but none of them emerge as
an effective inhibitor. These may be due to the high degree of cytotoxicity
and low affinity to the active site of tyrosinase (Prasad et al., 2014).
4.4. Camellia sinensis (L.) Kuntze
C. sinensis (Family- Theaceae) commonly known as green tea is a
popular ingredient in topical skin care preparations. Green tea extract is
enriched with polyphenolic compounds that have significant antioxidant
and anti-inflammatory and UV-protection activities. Tea polyphenols, i.e.
(-) epicatechin (EC) (13), (+) catechin (C) (14), epigallocatechin (EGC)
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Journal of Herbal Medicine 14 (2018) 1–16
(15), (-)-epicatechin 3-0-gallate (ECG) (16), (-)-gallocatechin 3-0-gallate
(GCG) (17), and (-)-epigallocatechin 3-0-gallate (EGCG) (18) were in-
vestigated on mushroom tyrosinase. ECG, GCG and EGCG showed a high
degree of a competitive mode of monophenolase inhibition. Moreover,
EGCG, EGC and Catechin were tested in B16 melanoma cells along with
another green tea polyphenol (gallic acid). Except Catechin, all of them
inhibited melanogenesis by decreasing expression of tyrosinase protein.
The most abundant polyphenol EGCG in green tea works via down-
regulation of MITF protein. Enzyme inhibition study and murine cell-line
analysis indicated that the structure of flavones containing a 3-position
gallic acid moiety is responsible for depigmentation activity (Sato and
Toriyama, 2009). Therefore, they do not inhibit the melanogenesis
pathway by acting on tyrosinase. The present finding is contrary to the
previous study that claims EC-enriched extract significantly improves the
tyrosinase inhibition capacity (Hong et al., 2014).The traditional Chinese
preparation ‘Oolong tea’ consists of partially fermented leaves of C. si-
nensis, which contain polymerized polyphenols. Oolong tea did not in-
hibit cell-free tyrosinase activity but decreased its action by reducing the
intracellular tyrosinase mRNA level in B16 cell-line. A similar trend was
also observed with theaflavin-3, 3′-digallate (19). This is a polymerized
polyphenol present in black tea which is produced by a PPO-catalyzed
reaction during the fermentation process of tea. Recent experiments have
indicated that water extract of black tea significantly decreases tyr-
osinase activity by reducing the levels of melanogenesis-related proteins
(Tyrosinase, Tyrp-1, and Tyrp-2) in melan-A cell without affecting mRNA
level (Yamaoka et al., 2009; Kim et al., 2015). However, major limita-
tions of green tea polyphenols are poor bioavailability and transient
stability.
P.K. Mukherjee et al. Journal of Herbal Medicine 14 (2018) 1–16

Fig. 3. (continued)

Fig. 3. (continued)
7
P.K. Mukherjee et al.
4.5. Glycyrrhiza glabra L
Liquorice or the root of G. glabra (Family: Fabaceae) is traditionally
and commercially used for skin whitening formulations. In a clinical
trial, 2.5% g of G. glabra cream was applied to 100 females for four
weeks. The study concluded that treatment with G. glabra cream sig-
nificantly improved symptoms of melasma compared to the placebo
group without any side effects (Badria, 2015). A major pyranoisoflavan,
Glabridin (20) one of the main ingredients of liquorice has shown
promising anti-melanogenesis activity on B16 murine melanoma cells.
It was observed that glabridin inhibited tyrosinase activity of melanoma
cells at concentrations 1.0 μg/mL without affecting DNA synthesis. It
was also shown that glabridin inhibited UVB induced pigmentation and
inflammation on guinea-pig skins at 0.5% w/v concentration. Yokota
and his colleagues concluded that two hydroxyl groups at 2 and 4 po-
sition of the glabridin are essential for tyrosinase inhibitory activity
(Yokota et al., 1998). The principle disadvantages of glabridin are its
insufficient skin-entering capacity and its instability in a formulation.
For that reason, ester or ether semi-synthetic derivatives were synthe-
sized with the 2′- and 4′- position hydroxy functional group of glab-
ridin. Interestingly, 70% of cosmetic patents are related to the glabridin
molecule (Simmler et al., 2013). In another study, it was found that
isoliquiritigenin (21) and glabrene (22) from G. glabra competitively
inhibited mono and diphenolase activity of mushroom tyrosinase
(Nerya et al., 2003). On the contrary, liquiritigenin activated the
monophenolase activity of tyrosinase and acted as a cofactor. Ad-
ditionally, it was observed glabridin exhibited a noncompetitive type of
inhibition on mushroom tyrosinase. 3-(2,4-dihydroxyphenyl propionic
acid) (23) isolated from G. glabra leaves showed strong inhibition of
mushroom tyrosinase at one μg/mL concentration (Nerya et al., 2006).
Licuraside (24), isoliquiritin (25) and licochalcone A (26) isolated from
the licorice root have also shown competitive inhibition on mono-
phenolase activity of mushroom tyrosinase, whereas liquiritin (27) did
not have any effect on tyrosinase. A recent study however, indicated
that liquiritin isolated from the leaves of G. glabra had potent tyrosinase
inhibition activity. Its depigmentation activity was clinically in-
vestigated. It was found that a 20% liquiritin containing cream applied
for four weeks was therapeutically useful in melasma (Dong et al.,
2014). Glycyrrhizin (GR) or glycyrrhetinic acid (28) is present in large
quantities in the roots of licorice. Lee et al. (2005) reported that the GR
stimulated melanogenesis in B16 melanoma cells. The underlying me-
chanism of melanogenesis activation by GR may be through increasing
cAMP signalling in the melanoma cells.
4.6. Glycine max (L.) Merr
G. max (family- fabaceae) popularly known as soybean is a nu-
tritious plant product primarily used as a dietary supplement. It is a
source of the antioxidant components such as γ-tocopherol and iso-
flavones, flavonols, flavan-3-ols, proanthocyanidins and anthocyanins
(Correa et al., 2010). Therefore, it has been explored for its tyrosinase
inhibitory potential. Soybean seed extracts contain two proteinase in-
hibitors namely Kunitz-type trypsin inhibitor (soybean trypsin in-
hibitor) and Bowman-Birk protease inhibitor. These inhibitors have
been explored as depigmenting agents. Both have been shown to reduce
pigmentation by attenuating phagocytosis of keratinocyte and melanin
distribution in dark-skinned Yucatan swine (Paine et al., 2001). Gly-
ceollins (a mixture of glyceollin I, II, and III) (29a, b, c) are one of a
group of phytoalexins produced in soybeans under stress conditions.
Depigmentation activity of glyceollins is exerted through the blockade
of a downstream signalling pathway. Experimental evidence suggests
that glyceollins inhibit expression of tyrosinase related proteins (Tyr-
osinase and Tyrp-1) and diminish intracellular cAMP levels of B16
melanoma cells, which are stimulated by α-MSH. Pretreatment of
B16F10 cells with glyceollins dose-dependently inhibited SCF-induced
c-kit, Akt phosphorylation and downregulated cAMP levels. It also
8
Journal of Herbal Medicine 14 (2018) 1–16
significantly impaired the expression and activity of MITF (Shin and
Lee, 2013). Glycinin and β-conglycinin, the two major storage proteins
from soy represent 85% of the total seed protein content. β-conglycinin
has been found to be an active mushroom tyrosinase inhibitor. Its in-
hibition potential may arise due to certain amino acid residues namely
arginine and/or phenylalanine (Schurink et al., 2007). Black soybean is
the black seed of G. max (L.) merr, also known as the black bean. Its
water extracts showed a dose-dependent tyrosinase inhibition capacity
up to 98%. In the human patch tests, the black bean extract did not
produce any signs of allergic reactions (Lai et al., 2012). In another
study, two potent o-dihydroxyisoflavone, 7, 8, 4′-trihydroxyisoflavone
(30) and 7, 3′,4′-trihydroxyisoflavone (31) were isolated from Korean
fermented soybean paste (Doenjang). Both significantly reduced tyr-
osinase activity and melanin formation in melan-A cells. Their inhibi-
tion potential was found to be greater compared to other known iso-
flavones. Compound 30 and 31 inhibited 50% tyrosinase activity at a
concentration of 11.21 μM and 5.23 μM (IC50), respectively (Park et al.,
2010). Moreover, two known isoflavones genistein (32) and daidzein
(33) abundant in soya food also possess tyrosinase inhibition. Genistein
suppressed the growth of B16-BL6 mouse melanoma cells. The mole-
cular mechanisms may involve suppression of the cytoskeleton-asso-
ciated protein tyrosine phosphorylation, which leads to changes in cy-
toskeletal network, cell morphology and expressions of tumor-related
genes. Soya food contains one flavanone naringenin (34) in large
quantities; which has also exhibited tyrosinase inhibition (Yan et al.,
1999; Oskoueian et al., 2011).
4.7. Hemidesmus indicus
The woody rootstocks of an Indian medicinal herb Hemidesmus in-
dicus (Family: Asclepiadaceae) locally known as “Anantmul” possesses a
strong and persistent fragrance (Jain, 1994). The active principle of this
fragrance is 2-hydroxy-4-methoxybenzaldehyde (MBALD) which is
found in its root extracts (Sircar et al., 2007). The extracts of the woody
root of H. indicus are traditionally used in the southern part of India as a
natural fragrance for aromatherapy (Kundu and Mitra, 2014). Root
extracts of H. indicus has a fragrant phenolic compound 2-hydroxy-4-
methoxybenzaldehyde (MBALD) that has shown inhibitory potential
against the biphenolase activity of tyrosinase. MBALD was shown to be
more potent than vanillin in inhibiting monophenolase activity. In-
hibition kinetic analysis confirmed the same observation, with a higher
Km value (0.395 mM) of monophenolase reaction in presence of
MBALD (Das and Bisht, 2013; Kundu and Mitra, 2014).
4.8. Mulberry
Morus comprises 10–16 species of deciduous trees commonly known
as “Mulberries”. The closely related Broussonetia papyrifera is also
known as mulberry, notably the Paper Mulberry, which grows naturally
in Asia and Pacific countries. Different parts of this plant are found to
possess anti-melanogenic property. Zheng and colleagues isolated
eleven compounds through bioassay-guided fractionation from the
chloroform extract of B. papyrifera. Among them, 3,5,7,40-tetra-
hydroxy-30-(2-hydroxy-3-methylbut-3-enyl) flavones, uralenol (35),
quercetin (36) and broussoflavonol F (37) showed mushroom tyr-
osinase inhibitory activity better than arbutin. In contrast to the pre-
vious study, isoliquiritigenin (22) isolated from B. papyrifera possessed
much weaker tyrosinase inhibitor activity compared to arbutin and
kojic acid (Zheng et al., 2008). The anti-tyrosinase activity of black
mulberry species (Morus nigra Linn) was investigated by Chun et al.
(2011) which showed mixed type mushroom tyrosinase inhibition.
Ethanol extract of Morus alba L twig and root bark contains high
amounts of polyphenols. On Chromatographic investigation high
amounts of polyphenols namely maclurin (38), rutin (39), isoquercitrin
(40), resveratrol (41), and morin (42) were observed which exhibited
dose-dependent tyrosinase inhibitory effects (Chang et al., 2011).
P.K. Mukherjee et al.
Mulberroside-A (43) and oxyresveratrol (44) obtained from the root of
M. alba strongly inhibited monophenolase as well as diphenolase ac-
tivity of mushroom tyrosinase. Oxyresveratrol exhibited potent diphe-
nolase inhibition compared to mulberroside-A and both the compounds
showed a mixed type of inhibition of monophenolase activity. However,
Mulberroside-A was a competitive inhibitor of the diphenolase activity
of mushroom tyrosinase (Wang et al., 2014a,b). Several phenolic and
flavonoid compounds have been isolated from its leaves such as gallic
acid (45) and quercetin. Among them, mulberroside F (46) showed
promising inhibitory effects on tyrosinase activity and melanin forma-
tion in melan-A cells. This finding is consistent with another study, in
which Jeong and Associates isolated twenty compounds from M. alba
leaves and evaluated their anti-melanogenesis effects on mushroom
tyrosinase as well as B16F10 melanoma cells. This research suggested
that kaempferide 3-O-β-D-glucoside (47) showed the most potent tyr-
osinase inhibition potential followed by compounds 2-(3,5-dihydrox-
yphenyl)-5,6-dihydroxybenzofuran, wittifuran E, moracin N, moruni-
grol C, kuwanon C and quercetin-3-O-β-D-glucopyranoside. Structure
activity relationship of the compounds showed that the tyrosinase in-
hibitory potential of the compounds was enhanced by the increased
number of hydroxyl and phenyl groups in the compound. Except for
compounds wittifuran E and morachalcone A, all phenolic compounds
decreased the melanin content of murine melanoma cells at 10 μM
concentration (Jeong et al., 2015).
4.9. Nelumbo nucifera Gaertn
Nelumbo nucifera Gaertn. (Family: Nelumbonaceae), the national
flower of India, is an important plant mentioned in Ayurveda having
various medicinal properties (Paudel and Panth, 2015). Phytochemical
and pharmacological investigations carried out on this plant clarified
the multifaceted role of this medicinal herb (Mehta et al., 2013). Water
extract from Nelumbo nucifera showed DOPA-oxidase inhibition effect
(whitening effect) was 59%, 57% and 50% from the leaf, seed and
flower extract respectively. Thus, it is evident that Nelumbo nucifera
leaf, flower and seed extracts have the potential to be used for skin
whitening and as anti-wrinkle functional cosmetic agents (Kim et al.,
2011). The constituents isolated from this plant including nuciferine, N-
methylasimilobine, (-)-lirinidine, and 2-hydroxy-1-methoxy-6a,7-dehy-
droaporphine showed potent inhibition of melanogenesis. The com-
pounds nuciferine and N-methylasimilobine inhibited the expression of
tyrosinase mRNA, N-methylasimilobine inhibited the expression of
TRP-1 mRNA and nuciferine inhibited the expression of TRP-2 mRNA.
Tyrosinase and tyrosinase-related protein 1 (TRP-1) and TRP-2 are
known to catalyze the major steps in melanin synthesis (Nakamura
et al., 2013a,b).
4.10. Vitis Vinifera L
Vitis vinifera (Family- vitaceae) is also known as “wine grape”,
European grape and grapevine. Phenolic compounds are most abundant
in grape's skin, pulp, and seeds. Among all plant parts, grape seeds
contain a higher number of phenols. In an investigation Hsu et al.
(2012) showed grape seed extracts possessed significantly higher tyr-
osinase inhibition activity than the grape peel extracts. These authors
additionally performed a kinetic analysis of the extracts and demon-
strated its mixed type inhibition. It is reported that more than 500
cosmetic formulations contain grape seed and leaf extracts. Further-
more, the polyphenols (proanthocyanidins) that enrich grape seed ex-
tracts possess free radical scavenging activity. In a clinical trial on fe-
male melasma patients, it was found that oral administration of a
proanthocyanidin rich extract from grape seeds reduced the hy-
perpigmentation symptoms in female volunteers after a year's treatment
(Ribeiro et al., 2015). Resveratrol (41) obtained from V. vinifera, was
oxidized by mushroom tyrosinase and irreversibly inhibited the en-
zyme’s activity. It was shown to be an extremely potent inhibitor of
9
Journal of Herbal Medicine 14 (2018) 1–16
human tyrosinase as compared to arbutin and p-coumaric acid. As a
result, it attenuates melanin synthesis in human melanoma cell culture
without producing any cytotoxicity (Park and Boo, 2013). This finding
is consistent with previous studies showing depigmentation effects of
resveratrol in human melanocytes. Notably, resveratrol inhibits mes-
senger ribonucleic acid (mRNA) expression of tyrosinase, Tyrp-1, and
MITF in human melanocytes. In the most recent research, topical ap-
plication of resveratrol was demonstrated to decrease hyperpigmenta-
tion significantly on ultraviolet-B stimulated guinea-pig skin that was
also supported by the histological data. It has been observed that re-
sveratrol inhibits melanin synthesis via a reduction of melanogenic
protein (Tyrp-2) (Lee et al., 2015).
4.11. Miscellaneous
Apart from the mainstream popular tyrosinase inhibitory botanicals,
some important medicinal plants such as Indian gooseberry, citrus, and
ginseng are also widely used in cosmetic skin whitening formulations.
Phyllanthus emblica L. (Family-Phyllanthaceae) is known as Indian
Gooseberry or Amla. It is used as an antioxidant and heals many health
ailments. Gallic acid (45) and vanillic acid (48) are the major phenolic
compounds in Phyllanthus emblica extracts. Methanol extracts of amla
fruit showed significant mushroom tyrosinase inhibition, which may be
due to its high phenolic content. Whereas ethanol extracts of the fruits
significantly inhibited the mRNA expressions of tyrosinase and tyr-
osinase-related proteins (Tyrp-1 and Tyrp-2) in B16 murine melanoma
cells, proving its efficacy as a skin whitening agent (Sripanidkulchai
and Junlatat, 2014).
Citrus fruits are cultivated widely and are the most abundant or-
ganic products on the planet. Citrus fruits belong to the rutaceae family.
C. limon Burmann forma Eureka (Eureka lemon), C. keraji Hort. ex.
Tanaka (Keraji), C. limon Burmann forma Lisbon (Lisbon lemon) and
Cistrus sp (Kiyookadaidai) are some of the essential Chinese lemons used
in traditional medicine. Volatile oils such as citral (49) and myrcene
(50) are abundant in these fruits. The tyrosinase inhibition kinetics
analyzed by a Lineweaver-Burk plot indicated that citral is a non-
competitive inhibitor and myrcene is a competitive inhibitor. In a re-
cent study, it was revealed that citrus hydrosols (stem distillation co-
product) were mixed type tyrosinase inhibitors. The GC analysis in-
dicated that some of the essential oils such as myrcene, sabinene, ger-
aniol and citral were present in hydrosols (Lante and Tinello, 2015).
Nobiletin (51) and hesperidin (52) are citrus flavonoids which are
obtained from citrus peel, as a by-product of the citrus juice industry.
Nobiletin exhibited a competitive type whereas hesperidin showed a
noncompetitive type mushroom tyrosinase inhibitory mechanism
(Zhang et al., 2007).
Proanthocyanidins present in the fruit pericarp of Clausena lansium
(Lour.) Skeels, were shown to have highly potent, reversible and mixed
competitive inhibitors of tyrosinase according to the results from en-
zyme experiments (Chai et al., 2017).
Ginsenosides or panaxosides are a class of natural steroid glycosides
and triterpene saponins that are widely distributed in white ginseng and
red ginseng. Ginsenoside Rh4 (53) isolated from Korean red ginseng has
exhibited depigmentation activity by regulating protein kinase A (PKA)
pathway and cAMP level in B16 melanoma cells resulting in the in-
hibition of melanin synthesis. It also attenuated melanin production
and tyrosinase activity in α-MSH and forskolin-stimulated B16 mela-
noma cells. Additionally, ginsenoside Rb1 (54) inhibited melanogenesis
through the inhibition of tyrosinase activity in α-MSH-stimulated B16
cells in a dose-dependent manner. p-coumaric acid (55) one of the
useful compounds obtained from fresh leaves of P. ginseng showed in-
hibitory activity on mushroom tyrosinase. Interestingly, it showed po-
tent inhibition of human or murine tyrosinase rather than the mush-
room tyrosinase. Its inhibition potential for human tyrosinase was
significantly higher compared to kojic acid and arbutin. Enzyme ki-
netic’s analysis indicates that p-coumaric acid is a mixed type (for L-
P.K. Mukherjee et al. Journal of Herbal Medicine 14 (2018) 1–16

Table 1
Important tyrosinase inhibitors from medicinal plants.
Plant name Phyto-constituent Plant part Type of Percentage of inhibition Mechanism of References
(Family) used tyrosinase Inhibition

Alhagi camelorum Fisch. Calycosin, kaempferol, isorhamnetin, Aerial parts M.T-DP 63% at 1.14 g/L−1 Competitive (Gholamhoseinian and
(Fabaceae) and kaempferol-3-galactorhamnoside Razmi, 2012)
Berberis aristata DC. Berberine Steam and M.T-MP 50% at 45.45 and Mixed (Biswas et al., 2016a)
(Berberidaceae) Bark M.T-DP 431.11 μg/mL
Betula pendula L. Chlorogenic and p-coumaric acid, Leaves M.T-DP 50% at Non- (Germanò et al., 2012)
(Betulaceae) quercetin and quercetin 119.08 ± 2.04 μg/mL competitive
3-O-galactoside, myricetin and
kaempferol glycosides
Bletilla striata (Thunb.) Polyphenols Fibrous roots M.T-DP 50% at 4.3 mg/L Dose dependent (Jiang et al., 2013)
Reichb.f. (Orchidaceae) part inhibition
Carthamus tinctorius L. Carthamus yellow: Plant M.T-MP 50% at Competitive (Chen et al., 2013)
(Compositae) Safflomin A, and Pigment B16F10 1.01 ± 0.03 mg/mL Decreased
Safflomin B melanoma 82.3 ± 0.4% melanin
cells at 4 mg/mL production
Casta neahenryi (Skan) Lignan glycosides Exocarp of M.T-DP 50% in between 15–21 μM Dose dependent (Wu et al., 2012a,b)
Rehd. et Wils fruit inhibition
(Fagaceae)
Citrus mitis Blanco 3’,5’-di-C-β-glucopyranosylphloretin Peels M.T-MP 0.87 mg/mL Competitive (Lou et al., 2012)
(Rutaceae) (Calamondin)
Naringin and Hesperidin
Crataegus pinnatifida Bunge 7R,8S-dihydrodehydrodiconiferyl Seeds M.T-MP 50–60% at 500 μg/mL Dose dependent (Huang et al., 2014)
(Rosaceae) alcohol-9-O-β-D-glucoside inhibition
and 7R,8S-dihydrodehydrodiconiferyl
alcohol-9′-O-β-D-glucoside
Cudrania tricuspidata Trans-dihydromorin, Oxyresveratrol, Twig M.T-MP 21.54 μM N.D (Zheng et al., 2013)
(Carrière) Bureau Steppogenin 2.85 μM
(Moraceae) 2.52 μM
Eugenia dysenterica DC. Gallicacid and catechins Leaves M.T-MP 90% at N.D (Souza et al., 2012)
(Myrtaceae) 1mg/mL
Eupatorium triplinerveVahl 7-methoxycoumarin Leaves M.T-MP 50% at 2360 μM Dose dependent (Arung et al., 2012)
(Asteraceae) M.T-DP 2840 μM inhibition
Mouse 1780 μM
melanoma,
B16
Ficus virens L. Condensed Tannins Leaves, fruit, M.T-MP 50% in between Mixed (Chen et al., 2014)
(Moraceae) and stem M.T-DP 99–131 mg/mL
bark 43-128mg/mL
Filipendula ulmaria (L.) Rutin, chlorogenic acid, quercetin3-β-D- Whole plant M.T-DP 98.30 ± 3.91% Dose dependent (Neagu et al., 2015)
Maxim. glucoside at 3 mg/mL inhibition
(Rosaceae)
Flemingia philippinensis Polyphenols: Roots M.T-MP 50% at Competitive (Wang et al., 2014a,b)
(Merr. et Rolfe) Li Fleminchalcone C M.T-DP 1.28 μM
(Fabaceae) Flavone: M.T-MP 5.22 μM
Flemichin D M.T-DP 1.79 μM
7.48 μM
Gnetum gnemon L. Gnetin C, Seeds Murine B16 50% in between 7–7.2 μM Decreased (Yanagihara et al.,
(Gnetaceae) Resveratrol cells melanin content 2012)
Hemidesmus indicus L. R.Br. 2-hydroxy-4-methoxybenzaldehyde Roots M.T-MP 50% at 0.03 mM Mixed (Kundu and Mitra,
(Asclepiadaceae) (MBALD) 2014)
Hypericum perforatum L. Hypericin and derivatives Aerial parts M.T-DP 19.21 ± 1.44% Dose dependent (Altun et al., 2013)
(Hypericaceae) at 250 μg/mL inhibition
Lawsonia inermis L. Lawsone (2-hydroxy-1,4 napthaquinone) Leaves M.T-DP 65% at 1.14 g/L−1 Mixed (Gholamhoseinian and
(Lythraceae) Razmi, 2012)
Madhuca latifolia (J.Konig) Ursolic acid, p-hydroxy-acetophenone, Fruit pulp M.T-DP 50% in between Dose dependent (Khan et al., 2015)
J.F.Macbr. hydroquinone,taxifolin, madhushazone, and seeds 5–18 μM inhibition
(Sapotaceae) madhusalmone, madhucic acid
Morusa ustralis Poir. Oxyresveratrol, Moracenin D, Roots M.T-MP 2.85 μM Dose dependent (Zheng et al., 2012)
(Moraceae) Sanggenon T, 4.61 μM inhibition
Kuwanon-O 1.20 μM
1.81 μM
Murraya koenigii L. Sprengel Koenimbine, Leaves B16 50% in between 1.2-2.9 Decreased (Nakamura et al.,
(Rutaceae) Mahanimbine, Mahanimbicine, melanoma μM melanin content 2013a,b)
Murrayamine-E 4A5 cells 16.1 and 20.2% at 100 μM Dose dependent
Eustifoline-C and Murrayamine-E M.T-DP inhibition
Piper nigrum L. Piperonylic acid Fruits M.T-DP 50% at Mixed (Si et al., 2013)
(Piperaceae) 0.36 ± 0.02 mM
Pouteria torta Radlk. Myricitrin Leaves M.T-MP 100% at N.D (Souza et al., 2012)
(Sapotaceae) 1 mg/mL
Protea madiensis Oliv., Terpenoids and steroids Barks Human cell > 75.0 μg/mL Decreased (Kamagaju et al., 2013)
(Proteaceae) lines (30.2%) melanin content
LOCE-MM001 > 75.0 μg/mL Dose dependent
LOCE-MM028 (36.3%) inhibition
M.T-DP 31 ± 4 μg/mL
(continued on next page)

10
P.K. Mukherjee et al. Journal of Herbal Medicine 14 (2018) 1–16

Table 1 (continued)

Plant name Phyto-constituent Plant part Type of Percentage of inhibition Mechanism of References
(Family) used tyrosinase Inhibition

Quercus infectoria Oliv. Gallic and ellagic acids Galls M.T-DP 50% at 1.14 g/L−1 Mixed (Gholamhoseinian and
(Fagaceae) Razmi, 2012)
Rabdosiaserra (maxim.) Methyl rosmarinate, – M.T-DP 21.5 μM Non- (Lin et al., 2011)
HARA RosmarinicacidPedalitin, 16.8 μM competitive and
(Labiatae) 0.28 mM Mixed
Rhizophorastylosa Griff. Polyphenols like tannin Barks M.T-DP 89.7% N.D (Suh et al., 2014)
(Rhizophoraceae)
Rubia Cordifolia L. Purpurin Roots M.T-MP 50% at Competitive (Biswas et al., 2015)
(Rubiaceae) M.T-DP 0.11 ± 0.02 mg/mL
0.29 ± 0.09 mg/mL
Sapium sebiferum(L.)Roxb. Kaempferol, kaempferol-3-O-beta-D- Leaves M.T-DP 39.10 ± 3.88% Dose dependent (Fu et al., 2014)
(Euphorbiaceae) glucopyranoside, and quercetin at 2 mg/mL inhibition
Sesamum angolense Welw. Terpenoids and steroids Leaves LOCE-MM001 > 7.0 μg/mL N.D (Kamagaju et al., 2013)
(Pedaliaceae) LOCE-MM028 (20.3%) Dose dependent
M.T-DP > 7.0 μg/mL inhibition
(15.6%)
> 22 μg/mL
Smilax china L. Dioscin Rhizomes M.T-MP 50% at > 100 μg/mL Dose dependent (Liang et al., 2012)
(Smilacaceae) M.T-DP 7.8 μg/mL inhibition
M.T-MP 10.9 μg/mL
M.T-DP
Sonneratia alba Sm. Polyphenols like tannin Barks M.T-DP 82.4% Dose dependent (Suh et al., 2014)
(Lythraceae) inhibition
Stewartia pseudocamellia 4-Hydroxybenzoic acid, Leaves M.T-DP 18% at 500 μg/mL Dose dependent (Roh et al., 2015)
Maxim. (Theaceae) Gallic acid, B16F1 34.0% at 500 μg/mL inhibition
Quercetin melanoma
cells
Vaccinium arctostaphylos L. N.D Fruits M.T-DP 60% at 1.14 g/L−1 Competitive (Gholamhoseinian and
(Ericaceae) Razmi, 2012)
Vaccinium bracteatum p-coumaric acid Whole plant H.T 0.5 μg/mL Mixed (Kim et al., 2012)
Thunberg
Xanthoceras Sorbifolia Saponins Nutshell M.T-DP 52% at 0.72 mg/mL Mixed (Zhang and Zhou,
Bunge 2013)
(Sapindaceae)

M.T-MP- Mushroom tyrosinasemonophenolase, M.T-DP- Mushroom tyrosinasediphenolase, H.T-Human tyrosinase, N.D – Not determined.

tyrosine) or competitive inhibitor (for L-dopa) of human tyrosinase. It
has also shown anti-melanogenesis activity on human epidermal mel-
anocytes. Cinnamic acid is found in the root and seed of P. ginseng.
Cinnamic acid significantly diminishes melanin content and tyrosinase
activity in the melan-A cell, besides that it has also shown depigmenting
activity on the UVB-tanned skin of brown guinea pigs (Kim, 2015).
Considering the facts as outlined above, several medicinal plants pos-
sessing tyrosinase inhibition activity have been summarized in Table 1.
5. Role of phyto-chemicals in tyrosinase inhibition
A vast number of phyto-molecules are reported as tyrosinase in-
hibitors. Most recent reviews are focussed on different phyto-molecules.
In this regard, some major groups of phytochemicals that have been
shown to have tyrosinase inhibitory activities are grouped below.
5.1. Flavonoids
Flavonoid is a group of compounds (including flavanols, flavones,
flavonols, flavanones, isoflavones and anthocyanidins) isolated from
plants and explored for natural tyrosinase inhibitors (Chang, 2009).
Flavonols exert tyrosinase inhibition activity in a competitive manner
by oxidizing L-dopa as a substrate. The 3-hydroxy-4-keto moiety of the
flavonol structure is responsible for the copper chelation present in the
active site of the enzyme (Kubo and Kinst-Hori, 1999). The flavonol
compounds that have been identified as tyrosinase inhibitors are
quercetin, myricetin, kaempferol, galangin, morin, buddlenoid A,
buddlenoid B. (Xie et al., 2003). The other major flavonoids identified
as tyrosinase inhibitors are nobiletin (5,6,7,8,3′,4′-hexamethoxy-
flavone), naringin (5,7,4′-trihydroxyflavanone), and neohesperidin
(5,7,3′-trihydroxy-4′-methoxyflavone) (Itoh et al., 2009). Additionally,
norartocarpetin (5,7,2′,4′-tetrahydroxyflavone), streppogenin
(5,7,2′,4′-tetrahydroxy-flavavone), dihydromorin (5,7,2′,4′-tetra-
hydroxyflavanol), artocarpetin (5,2′,4′-trihydroxy-7-methoxyflavone)
have been identified as tyrosinase inhibitor (Chang, 2009). The tetra-
hydroxyflavanol compound, taxifolin isolated from Polygonum hydro-
piper, exhibited the promising inhibitory activity of mushroom tyr-
osinase (Miyazawa and Tamura, 2007). One flavone-flavanone dimmer
(isolated from Garcinia subelliptica) was shown to be more active than
kojic acid in respect to monophenolase activity of mushroom tyrosinase
(Masuda et al., 2005). Some isoflavonoid compounds like glabridine,
glabridin, glabrene, Glyasperin C obtained from Glycyrrhiza species
have also been identified as potent tyrosinase inhibitors (Kim et al.,
2005). The other isoflavonoid compounds like Haginin-A (2′,3′-di-
methoxy-7,4′-dihdroxyisoflav-3-ene), Dalbergioidin (5,7,2′,4′-tetra-
hyroxyisoflavan), calycosin (4′-methoxy-7,4′-dihydroxyisoflavone) also
have a significant tyrosinase inhibiting property (Chang, 2009). The
hydroquinone class of compounds (mainly catechin, rhamnetin, ar-
butin, deoxy-arbutin) has been found to competitively inhibit mush-
room tyrosinase and is also active therapeutically against the treatment
of hyperpigmentation (Lee et al., 2015).
5.2. Stilbenes
Stilbene derivatives are distributed in the plants belonging to
Dipterocarpaceae, Vitaceae, Gnetaceae, Leguminosae, and Cyperaceae
families (Ohguchi et al., 2003). Stilbenes (diarylethenes) are produced
through the mixed shikimate-acetate pathway. There are numerous
naturally occurring stilbenes (with polyoxygenation) possessing tyr-
osinase inhibitory activity (Lee et al., 2015). The presence of trans-
olefin structure in the parent stilbene skeleton has been shown to be
essential for its inhibitory activity. The inhibitory potency of these

11
P.K. Mukherjee et al.
compounds depends on the position of free hydroxyl groups and also
the presence of a double bond present in the stilbene skeleton (Ohguchi
et al., 2003). The stilbene derivatives differ in their activity mainly due
to oxygenation in the parent structure at the 2, 4, 3′ and 5′ positions
which decreases their binding affinity caused by the bulkiness of the
side chain (Likhitwitayawuid, 2008). The major bioactive compound
belonging to the stilbene group is resveratrol that has been found
abundantly from plant sources such as Artocarpus lakoocha, A. gome-
zianus, Morus alba, and Veratrum album. The result of recent studies
suggests that oxyresveratrol appears to be a potent inhibitor when
compared to Kojic acid. It is considered to be a very promising lead, as
kojic acid has recently been banned in some countries due to its mu-
tagenic potential (Ohguchi et al., 2003; Shin et al., 1998). The other
oligomeric stilbene compounds such as Chlorophorin (Artocarpus in-
cises), Artocarbene (from A. incises), Rhapontigenin (R. undalatum),
Gnetol (Gnetum gnemon), Artogomezianol (Artocarpus gomezianus), An-
dalasin A (A. gomezianus), (–)-ε-Viniferin (V. rassak), Vaticanols A–C
and G (V. rassak), (+)-α-Viniferin (Shorea hemsleyana), (–)-Hope-
aphenol (Hopea utilis) have shown potential tyrosinase inhibition
(Likhitwitayawuid, 2008). It has been found that the monomeric stil-
benes are more potent tyrosinase inhibitors in comparison with the
oligomeric stilbenes, in both monophenolase and diphenolase inhibi-
tion model.
5.3. Other classes of phyto-chemicals
A taraxastene-type triterpene; 3,16-di-hydroxy-21-hydroperoxy-
20(30′)-taraxastene isolated from Arnica montana L. (Family:
Compositae) flowers was found to act as a melanogenesis suppressor
(Maeda et al., 2007). It completely suppressed melanin production at a
dose of 0.53 M in melanoma cultured B16 cells acting through down-
regulation of MITF-M (isoform of MITF) and reduction of tyrosinase
related genes and proteins. Andrographolide, a labdane diterpenoid,
suppressed melanin content and tyrosinase activity in B16F10 and HEM
cells. It also exhibited similar activity in UVB-induced pigmentation of
brown guinea pig skin. Andrographolide exerted anti-melanogenesis
activity via decreasing phosphorylation of glycogen synthase kinase-3β
(GSK-3β) that leads to promote degradation of β-catenin. The resulting
mechanism decreased the expression of MITF as well as tyrosinase re-
lated proteins (Zhu et al., 2015). Betulinic acid, a pentacyclic tri-
terpenoid, isolated from Rhododendron collettianum and Thai Mulberry
plant extracts exhibited a significant in-vitro tyrosinase inhibitory ef-
fect. It is now evident that betulinic acid significantly decreases melanin
production in B16F10 cell line by inhibiting tyrosinase, Tyrp-1, and
Tyrp-2 (Jin et al., 2014). Triterpenoid glycosides from shea (Vitellaria
paradoxa; Sapotaceae) showed promising inhibition of melanogenesis
in B16 melanoma cells induced by α-melanocyte-stimulating hormone
(α-MSH). Further, it was elucidated that paradoxoside E found in shea
downregulates the expression of MITF, tyrosinase, Tyrp-1 and Tyrp-2 to
inhibit melanogenesis (Zhang et al., 2014). Very few alkaloids are re-
ported as tyrosinase inhibitors. Investigation of the melanogenesis in-
hibitory effect of barnyard millet grain extracts led to the isolation of an
n-p-coumaroyl serotonin (Serotonin alkaloid) active against mushroom
tyrosinase and B16 melanoma cells. It exhibited potent non-competitive
tyrosinase inhibition compared with that of kojic acid. More im-
portantly, it decreased melanin content significantly by lowering the
expression of tyrosinase, Tyrp-1 and Tyrp-2 in B16 melanocytes without
cytotoxicity (Seo et al., 2012).
Polysachharides are the most widely found groups of biopolymers
having diverse activities. Polysaccharides may be an important source
of tyrosinase inhibitory agents based on the evidence that a poly-
saccharide fraction isolated from Punica granatum rind exhibited non-
competitive inhibition of tyrosinase showing an inhibitory activity of
43% at 10 μg/mL concentration (Rout and Banerjee, 2007). In another
study polysaccharides from longan fruit pericarp showed potent tyr-
osinase inhibitory activity (Yang et al., 2008). The association of tea
12
Journal of Herbal Medicine 14 (2018) 1–16
polysaccharides with tea polyphenols could prove to be an effective
means to combat skin hyperpigmentation as suggested by their tyr-
osinase inhibitory activity (Wei et al., 2009).
Essential oils are complex compounds volatile in nature having a
sweet smell and are produced in plants as secondary metabolites.
Cinnamaldehyde obtained from the stem bark of Cinnamomum cassia by
steam distillation inhibited tyrosinase in a mixed manner (Chang et al.,
2013). In a study reported by Suganya and associates, essential oil from
leaves of Pogostemon plectranthoides from three different geographical
regions were evaluated for their tyrosinase activity. The result showed
that all the three varieties were capable of inhibiting the tyrosinase
enzyme (Suganya et al., 2015). Sarikurkcu et al. (2015) reported the
tyrosinase inhibition potential of Origanum vulgare (subsp. hirtum) es-
sential oil obtained from the aerial parts of the plant (Sarikurkcu et al.,
2015). Thymol a naturally occurring monoterpenoid has been found to
possess a novel tyrosinase inhibiting property which inhibited the
conversion of leukodopachrome to dopachrome formation (Satooka and
Kubo, 2011). In another study, 19 essential oils from endemic plants
were screened for tyrosinase inhibition activity. Among them, the es-
sential oil extracted from Cinnamonum zeylanicum, Citrus grandis, and
Citrus hystrix showed significant inhibition activity compared to kojic
acid. Kinetic analysis revealed C. grandis extracts showed a competitive
type of inhibition (Aumeeruddy-Elalfi et al., 2016).
6. Traditional Skin Care Formulations with anti-tyrosinase
potential
6.1. Ubtan
Ubtan is a traditional Unani formulation with cosmetic use. This
herbal preparation is widely used in India and its surrounding sub-
continent for the rejuvenation of dermal health. ‘Ubtan’ is a semisolid
powdered preparation used to remove the dirt particles from the skin
and enhance the lustre of the body. Ubtan is prepared from three pri-
mary ingredients, i.e. the powdered rhizomes of Curcuma longa (tur-
meric), seeds of Cicer arietinum (chickpea), and heartwood of Santalum
album (sandalwood) mixed in equal ratio of 1:1: 1 w/w/w (Biswas et al.,
2016a,b; Jamal et al., 2005). There is ample evidence to suggest that
Curcuma longa one of the ingredients of Ubtan improves the complexion
of skin by a skin lightening effect (Dixit and Goyal, 2011). The ritual
‘Solah Shringar’ encompasses sixteen ways of the beautifying of a wo-
man’s body during the marriage ceremony of the Hindu as well as in
Muslim communities in India where Ubtan paste is one of the vital
components of Solah Shringar (Miczak, 2001). It is also used in Ayur-
vedic body massage known as ‘Ubvartan’ which makes skin smooth and
soft (Sachs, 2002). Women in South Asia believe that it lightens the
complexion of the skin making the baby more beautiful (Reissland and
Burghart, 1987) and in Pakistan it is used by the local communities as a
skin tonic and to cleanse and refresh the skin. (Mushtaq et al., 2012). A
tyrosinase inhibition study with the prepared formulation indicated
both diphenolase and monophenolase inhibitory activity (Biswas et al.,
2016a,b).
6.2. Kumkumadi Tailam
According to Ayurveda, healthy skin is a result of the overall health
condition of the individual and prescribes numerous skin care treat-
ments that need to be pursued at every stage of life. Several herbs have
been mentioned in Ayurveda which can be used to obtain healthy skin
and a glowing complexion. Kumkumadi Tailam is an Ayurvedic herbal
oil used for face massage. It is advantageous to improve skin texture,
complexion and also to relieve skin problems such as acne and scars.
Kumkuma means saffron (Crocus sativus), which is the main ingredient
of this topical formulation. Kumkumadi Tailam improves skin com-
plexion and texture, relieves blemishes, acne, acne scars, white and
black heads, dark circles, sun tan, and wrinkles (Kumar et al., 2013). It
P.K. Mukherjee et al.
has both a cleansing and nourishing effect on the skin plus anti aging
qualities as it adds radiance to the skin. There are usually three essential
components in the manufacture of Kumkumadi Tailam; Drava (any li-
quid medium as prescribed in the composition), Kalka (fine paste of the
specified drug), Snehadravya (fatty component in the form of goat milk
or sesame oil) and occasionally, Gandhadravya (perfuming agents);
(Anonymous, 2008). The formulation is mentioned in Bhaisajya Rat-
navali which is a unique classical textbook of Ayurveda composed by
Shri Govind Das during 19th century. The process is known as “Kshir-
pakvidhi” where “Kshir” means milk in Sanskrit and involves boiling of
medicinal herbs in milk. The medicinal herbs used for preparing
Kumkumadi Tailam are collectively called ‘Kwathadravya’” in Ayur-
vedic terminology. This is because these medicinal herbs are extracted
by decocotion termed “Kwatha”in Ayurveda by solvent (water in this
case) termed “dravya” in Ayurveda. These plants include Keshar
(Crocus sativus), Chandan (Santalum album), Laksha (Laccifera lacca),
Manjistha (Rubia cordifolia), Yashtimadhu (Glycerrhiza glabra),
Raktchandan (Pterocarpus santalinus), Usher (Vetiveria zinzanioides),
Padmakasht (Prunus cerasoides), Kamal (Nymphaea stellata), Vatankur
(Ficus benghalensis), Pushkarmool (Inula recemosa), Kamal kesar (Ne-
lumbo nucifera) and Dashmool (Ayurvedic herbal combination)
(Shashtri, 2005). Crocus sativus, the main ingredient of this formulation
has been found to have potent antityrosinase effect (Sariri et al., 2011)
which has been discussed earlier.
7. Discussion
Melasma (from the Greek word, ‘melas’ meaning black) is a
common, acquired, confined hyperpigmentation of skin exposed to
extreme sunlight. It is common among people of light brown skins such
as people of East Asian, Southeast Asian, and Hispanic origin living in
the tropical areas. It is the most common pigmentary disorder among
Asian Indians. Hydroquinone (HQ), is the main treatment for melasma
which acts by inhibiting the tyrosinase enzyme (Bandyopadhyay,
2009). Triple-combination cream (TCC), which contains 4% HQ, 0.05%
tretinoin, and 0.01% fluocinolone acetonide is approved by the United
States Food and Drug Administration for the treatment of melasma.
Apart from HQ, tretinoin present in the formulation show depigmentory
effect by tyrosinase inhibition (Shin and Park, 2014). Disseminated
melanoma is a highly metastatic malignancy with high mortality rate.
The inhibition of melanogenesis by blocking tyrosinase activity has
been found to make melanoma cells prone towards cytotoxic action of
cyclophosphamide, and amplified immunotoxic activities of IL-2 acti-
vated lymphocytes. It has been observed that resistance to a che-
motherapeutic agent or immunotoxic activity of lymphocytes could be
returned by the action of tyrosinase inhibitors. Thus, tyrosinase in-
hibition may prove to be a useful therapeutic approach for the man-
agement of advanced melanomas (Slominski et al., 2009).
The study of Tyrosinase inhibition is an active field of research in
dermatological, biomedical, food and agricultural science and the do-
main of insect physiology. The present review covers commercially
important medicinal plants and their derived phyto-molecules used in
skin whitening cosmetic formulations. These plants are also useful in
the treatment of hyperpigmentation. However, some limitations are yet
to be overcome despite their successful uses as tyrosinase inhibitors.
Natural Hydroquinone (HQ) or benzene-1,4-diol is found in food
and beverages such as tea, coffee and berries that are popularly used in
cosmetic formulations for their effect on tyrosinase inhibition. Natural
Hydroquinone (HQ) acts as an alternative substrate to tyrosinase which
is transformed to highly reactive species like hydroxyhydroquinone
(HHQ) and benzoquinone (BQ) (Garcia-Molina et al., 2014). These
catalyzed products can cause permanent damage to melanosomes and
melanocytes, the site for tyrosinase activity. As a result, long-term uses
of HQ may cause permanent depigmentation, exogenous ochronosis,
and skin allergies. However, more severe side effects may occur due to
their mutagenic and carcinogenic potential (Lee et al., 2015).
13
Journal of Herbal Medicine 14 (2018) 1–16
A vast number of phenols are reported as tyrosinase inhibitors. A
great number of phenols are oxidized to form ortho-quinones through
tyrosinase like catechols. Ortho-quinones act as an endogenous nu-
cleophile. They can attack Michael acceptor thiol group containing
enzymes and lead to the production of reactive oxygen species. They
may also produce an allergic reaction by binding with melanosomal
proteins to generate neo-antigens, thus triggering immunological re-
sponses. A phenolic compound rhododendrol (RH) or 4-(4-hydro-
xyphenyl)-2-butanol was isolated from Acer maximowiczianum Miq.
(Family- Sapindaceae). In Japan, it was used as an active ingredient in
skin whitening cream. It was found to show a similar pattern of toxicity
like ortho-quinone. A large number of consumers developed leuko-
derma on their face, neck, and hands due to RH quinones toxicity.
These RH quinones are bound with cellular glutathione and con-
secutively attack melanocyte’s proteins. RH based skin whitening
creams were withdrawn from the market due to its melanocyte toxicity
via a tyrosinase-dependent mechanism (Ito and Wakamatsu, 2015).
Collectively, it is evident that phenolic compounds may act as an al-
ternate substrate for tyrosinase and produce the highly toxic quinone
moiety. For this reason, preliminary screening is required to dis-
tinguished substrates and inhibitors.
Apart from this, some mutagenic and carcinogenic potential of
plants should be considered before choosing a plant-based active in-
gredient for skin whitening formulation. In Pakistan, five commonly
used skin whitening creams were evaluated for their mutagenic and
oncogenesis effects by Allium test. Surprisingly, results showed all
tested creams possessed muto-depressive action (Udengwu and
Chukwujekwu, 2008). Furthermore, some organic and inorganic mer-
cury salts are widely used in skin whitening products due to its melanin
inhibitor property. These are absorbed through the skin and produce
harmful side-effects in the body, e.g. kidney damage, skin discoloration,
allergic reaction and scarring, and the skin loses its resistance to bac-
terial and fungal infections (Al-Saleh et al., 2004). In India, 61% of the
dermatology market consists of skin lightening or whitening products.
Several tests have been carried out to analyze the heavy metals such as
As, Hg, Cd, Pb and trace elements in herbal cosmetics preparations sold
in the Indian market (Ladizinski et al., 2011). Some of them contain
heavy metals above the safety limits. Therefore, quality related safety
issues should be studied properly for all the medicinal plant based
formulations (Mukherjee, 2015).
Most of the tyrosinase inhibition studies have been performed with
the commercially available tyrosinase from the mushroom Agaricus
bisporus. The 3D crystallographic structure of tropolone that inhibited
tyrosinase from A. bisporus was experimentally developed and de-
posited in the Protein Data Bank (PDB) with access code 2Y9X (Ismaya
et al., 2011). This X-ray crystallographic structure of mushroom tyr-
osinase is extensively used in In-silico modeling for screening potential
lead molecules from synthetic as well as those of natural origin. How-
ever, the 3D atomic structure of the human tyrosinase has not been
determined. The computational drug design has been performed by
preparing human homology modeling. In a study, molecular docking
and dynamic technique were used together to analyze 61,000 com-
pounds from the traditional Chinese medicines (TCM) database. Two
alkaloids, merresectine C, and bufotenine were found to be the most
potent human tyrosinase inhibitors (Tang and Chen, 2015). The po-
pularity of mushroom tyrosinase over human tyrosinase may be due to
their structural connection. Human tyrosinase contains an epidermal
growth factor (EGF)-like domain, which has structural similarity with
mushroom tyrosinase. Furthermore, as with mushroom, human tyr-
osinase’s two copper ions were coordinated by six histidine residues at
the catalytic site. In spite of that resemblance, the amino acid sequence
similarity between tetrameric mushroom tyrosinase and monomeric
human tyrosinase is just 23% (Chang, 2009; Ismaya et al., 2011). Fifty
endemic plants in the Republic of Korea were extracted with ethanol
and subjected to the primary screening assay using cell-free human
tyrosinase from HEK293-TYR cells. Results of this test showed that
P.K. Mukherjee et al.
extracts of Vaccinium bracteatum Thunb compared to mushroom tyr-
osinase possessed potent human tyrosinase inhibition. In contrast,
Morus bombycis Koidz was showed higher mushroom tyrosinase in-
hibition potential contrast to human tyrosinase (Kim et al., 2012).
Moreover, in another study, the isoform of α, β, and γ- thujaplicins
(natural monoterpenoid) were investigated against human and mush-
room tyrosinase. It has been observed that compared to mushroom
tyrosinase human tyrosinase was inhibited at higher thujaplicins con-
centration. Molecular docking analysis of a human homology model
revealed amino acid residues, i.e. histidine-367, Isoleucine -368, and
valine-377 were played a significant role in inhibitor binding
(Yoshimori et al., 2014). Ample evidence suggestes that the differences
in inhibition potential of a compound on mushroom and human tyr-
osinase occurs due to their dissimilar active site oxidative states, sub-
strate specificity, and amino acid sequence. Therefore, human tyr-
osinase should be used in screening assays for exploring new skin
hypopigmentation agents.
A specific objective of the research for new tyrosinase inhibitors is
to translate preclinical information into the clinical application. Due to
the toxicity issue with hydroquinone, there is an urgent need for al-
ternate depigmentation drugs that alone or in combination are more
effective than hydroquinone. In this regard, medicinal plants and their
phytochemicals can play a crucial role. Phytomolecules like, n-butyl
resorcinol, ellagic acid, arbutin, and azelaic acid were clinically in-
vestigated for the topical treatment of hyperpigmentation. However,
only a few medicinal plants were found to be clinically useful, i.e. G.
glabra, G. max, and Mulberry extract (Sardana and Ghunawat, 2015).
Tea and coffeeberry have not been clinically validated for skin lightning
activity. They have powerful antioxidant and UV protective activity.
Therefore, these plants are used in many skin whitening formulations
(Leyden et al., 2011).
8. Conclusions
The medicinal plants from traditional medicine systems and other
sources need to be evaluated to find productive leads that may be useful
for the treatment of hyperpigmentation as well as equally functional as
anti-browning agents. The scientific validation of these herbs as tyr-
osinase inhibitors should be explored further based on in-vitro human
tyrosinase inhibition study, in-vivo animal models along with clinical
studies.
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgements
The authors are thankful to the Department of Biotechnology,
Government of India, New Delhi, for financial support for this work
through DBT - Tata Innovation fellowship vide sanction no BT/HRD/
35/01/04/2014. We would like to thank the Emami Limited, Kolkata
for providing fellowship to Ms Akanksha Sharma for her work.
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