molecules

Review
Upregulation of Melanogenesis and Tyrosinase
Activity: Potential Agents for Vitiligo
Chao Niu 1,2 and Haji A. Aisa 1,2, *
1 Key Laboratory of Plant Resources and Chemistry of Arid Zone, Chinese Academy of Sciences,
Urumqi 830011, China; niuchao@ms.xjb.ac.cn
2 State Key Laboratory Basis of Xinjiang Indigenous Medicinal Plants Resource Utilization, Xinjiang Technical
Institute of Physics and Chemistry, Chinese Academy of Sciences, Urumqi 830011, China
* Correspondence: aisa@ms.xjb.ac.cn; Tel.: +86-991-383-5679; Fax: +86-991-383-8957

Academic Editor: Diego Muñoz-Torrero

Received: 11 July 2017; Accepted: 1 August 2017; Published: 4 August 2017

Abstract: Melanin, the compound primarily responsible in humans for hair, eye and skin
pigmentation, is produced by melanocytes through a complicated process called melanogenesis that
is catalyzed by tyrosinase and other tyrosinase-related proteins. The abnormal loss of melanin causes
dermatological problems such as vitiligo. Hence the regulation of melanogenesis and tyrosinase
activity is very important for treating hypopigmentary disorders. Many melanogenesis stimulators
have been discovered during the past decade. This article reviews recent advances in research on
extracts and active ingredients of plants, synthesized compounds with stimulating effect on melanin
synthesis and tyrosinase activity, as well as their influence on the expression of related proteins and
possible signaling pathways for the design and development of novel anti-vitiligo agents.

Keywords: melanogenesis; tyrosinase activity; vitiligo; plant extracts; natural products;

synthesized derivatives; analogues

1. Introduction
Vitiligo is an acquired chronic depigmentation disorder of the skin resulting from selective
destruction of melanocytes. Celsus was the first to use the term vitiligo in his Latin medical classic
De Medicina during the second century B.C [1,2]. It is clinically characterised by the development of
white macules (Figure 1) [3] due to the loss of functioning melanocytes in the skin or hair, or both [4].
The prevalence of the disease is often referred to as 0.05–1% of the world’s population and it is
the most frequent cause of depigmentation worldwide [5]. Although essentially asymptomatic,
the psychosocial impact of vitiligo can be devastating, and affected persons are often desperate
for an effective therapy. Many possible causes of vitiligo, including immunologic, genetic, stress,
neural mechanism, and biochemical factors [6] had been proposed, but the etiopathogenesis of the
disease is still enigmatic. However, it is believed that the disease is mainly a result of destruction of
melanocytes and obstruction of the melanin synthesis pathway [7,8].
Melanin, derived from dopaquinone, serves a number of valuable physiological functions with
the most important being photoprotection of the human skin from ultraviolet (UV) radiation [9].
Melanogenesis takes place in special organelles—the melanosomes in the melanocytes—and can
be triggered by a variety of paracrine cytokines including α-melanocyte-stimulating hormone
(α-MSH) [10], stem cell factor (SCF) [11], endothelin-1 (ET-1) [12], nitric oxide (NO) [13],
adrenocorticotropic hormone (ACTH) [14], prostaglandins [15], thymidine dinucleotide [16] and
histamine [17]. These factors all induce melanogenesis through diverse signaling pathways by
activating the expression and activation of pigment-related proteins such as microphthalmia-associated

Molecules 2017, 22, 1303; doi:10.3390/molecules22081303 www.mdpi.com/journal/molecules

Molecules 2017, 22, 1303 2 of 28

transcription
Molecules 2017, factor
22, 1303(MITF), tyrosinase (TYR), tyrosine-related protein-1 (TRP-1) and tyrosine-related
2 of 27
protein-2 (TRP-2).
Molecules 2017, 22, 1303 2 of 27

Figure 1. Vitiligo with typical lesions of the face and trunk [3].
Figure1.
Figure 1. Vitiligo
Vitiligo with typical
typicallesions
lesionsofofthe
theface
faceand
andtrunk [3].
trunk [3].
Figure 2 [18,19] illustrates the most common signaling pathways involved in the synthesis of
Figure
Figure2
melanin. 2[18,19]
All [18,19]illustrates
illustrates
signaling pathwaysthe
the most
most common
common
involve signaling
MITF,signaling
a master pathways
pathways
regulator involved
involved in in
thethe
of melanogenesis, synthesis
synthesis of of
which
melanin.
melanin. AllAll
upregulates signaling
the pathways
signaling pathways
melanogenesis involve MITF,
involve
enzymes a TRP-1
master
MITF,
TYR, regulator
a and
master of melanogenesis,
regulator
TRP-2 via to thewhich
M-boxupregulates
of melanogenesis,
binding which
motif in
the melanogenesis
upregulates
their promoter enzymes
the regions [20].TYR,
melanogenesis TRP-1 and
enzymes
In addition, TRP-2
TYR,
MITF TRP-1 via
and
regulates binding
TRP-2to viathe
melanocyte M-boxto
binding
function motif in their
the M-box
including promoter
motif in
melanocyte
differentiation,
their
regions promoter pigmentation,
regions
[20]. In addition, [20].
MITF In proliferation
addition,
regulatesMITF andregulates
cell survival
melanocyte [21].
melanocyte
function function
including includingdifferentiation,
melanocyte melanocyte
differentiation,
pigmentation, pigmentation,
proliferation andproliferation
cell survivaland cell survival [21].
[21].

Figure 2. Regulation of melanogenesis through different signaling pathways [18,19].

Among them, TYR, TRP-1 and TRP-2 (otherwise called dopachrome tautomerase or DCT) are
mainly involved in the transformation of tyrosine into melanin pigments [22]. Melanosomes produce
two types Figure
ofFigure
melanin: eumelanin,
2.2.Regulation
Regulation a brown–black
of melanogenesis
of melanogenesis or dark
through
through insoluble
different
different polymer;
signaling and pheomelanin,
pathways
signaling [18,19].
pathways [18,19]. a red-
yellow soluble polymer, formed by the conjugation of cysteine or glutathione [23,24] (Scheme 1).
Among
Although
Among them,
three TYR,
TYR,TRP-1
enzymes
them, (TYR,and
TRP-1 TRP-2
TRP-1
and (otherwise
and (otherwise
TRP-2 calleddopachrome
TRP-2) are called
involved dopachrome tautomerase
in the melanogenesis
tautomerase or or
pathway, DCT)
DCT) areare
only
TYR
mainly is exclusively
involved in necessary
the for melanogenesis.
transformation of tyrosine into melanin pigments [22]. Melanosomes
mainly involved in the transformation tyrosine into melanin pigments [22]. Melanosomes produce produce
two
twotypes
typesofof melanin: eumelanin,
melanin: eumelanin, a brown–black
a brown–black or dark
or dark insoluble
insoluble polymer;
polymer; and pheomelanin,
and pheomelanin, a red-
yellow soluble
a red-yellow polymer,
soluble formed
polymer, by the
formed conjugation
by the of cysteine
conjugation or glutathione
of cysteine [23,24]
or glutathione (Scheme
[23,24] 1). 1).
(Scheme
Althoughthree
Although threeenzymes
enzymes(TYR,
(TYR, TRP-1
TRP-1 and
and TRP-2)
TRP-2)are
areinvolved
involvedininthe
themelanogenesis
melanogenesis pathway,
pathway,only
only
TYR is exclusively necessary for melanogenesis.
TYR is exclusively necessary for melanogenesis.
Molecules 2017, 22, 1303 3 of 28

Molecules 2017, 22, 1303 3 of 27

SchemeScheme 1. Melanogenesis
1. Melanogenesis catalyzed
catalyzed byby tyrosinase(TYR),
tyrosinase (TYR), tyrosinase-related
tyrosinase-related protein
protein1 (TRP-1) andand
1 (TRP-1)
tyrosinase-related protein 2 (TRP-2).
tyrosinase-related protein 2 (TRP-2).

TYR (EC 1.14.18.1) is a multifunctional membrane-bound type-3 copper protein, which is located
TYR (EC 1.14.18.1) is a multifunctional membrane-bound type-3 copper protein, which is located
in the membrane of the melanosome [25]. TYR is produced only by melanocyte cells. Following its
in the membrane of the melanosome [25]. TYR is produced only by melanocyte cells. Following its
synthesis and consequent processing in the endoplasmic reticulum and Golgi, it is trafficked to
synthesis and consequent
melanosomes, wherein processing in the endoplasmic
the melanin pigment is synthesized.reticulum and Golgi,
From the structural pointit of
is view,
trafficked
two to
melanosomes,
copper ions,wherein the melanin
each surrounded pigment
by three is synthesized.
histidines, are responsible forFrom the structural
the catalytic point
activity of TYR of view,
[26].
two copper
Three different states of the active site have been reported in the pigment formation: oxy-, met- and of
ions, each surrounded by three histidines, are responsible for the catalytic activity
TYR [26]. Three different
deoxy-forms. states ofat the
More specifically, activesite,
the active sitecopper
have atoms
been participate
reported in the pigment
directly formation:
in hydroxylation
of monophenols
oxy-, met- to diphenols
and deoxy-forms. More(cresolase
specifically,activity)
at the and in thesite,
active oxidation
copperofatoms
o-diphenols to o-quinones
participate directly in
(catechol of
hydroxylation oxidase activity) that
monophenols enhance melanogenesis
to diphenols [27,28]. and in the oxidation of o-diphenols to
(cresolase activity)
o-quinonesTYR is also catalyzing
(catechol the process
oxidase activity) thatof enhance
neuromelanin production in[27,28].
melanogenesis which the oxidation of dopamine
produces dopaquinones. However, excessive production of dopaquinones results in neuronal damage
TYR is also catalyzing the process of neuromelanin production in which the oxidation of dopamine
and cell death [29]. This suggests that tyrosinase might play a significant role in neuromelanin
produces dopaquinones. However, excessive production of dopaquinones results in neuronal damage
formation in the human brain and responsible for the neurodegeneration associated with Parkinson’s
and cell deathand
disease [29]. This suggests
Huntington’s that
disease tyrosinase
[30,31]. Tyrosinasemight hasplay
alsoabeen
significant
linked to role
theinbrowning
neuromelanin
of
formation
vegetables and fruits during postharvest and handing process, leading to quick degradationParkinson’s
in the human brain and responsible for the neurodegeneration associated with [32,33].
diseaseTheand Huntington’s
application diseasewas
of tyrosinase [30,31].
furtherTyrosinase
extended inhas thealso beenprocess
molting linkedoftoinsects
the browning
[34].The of
vegetables and accumulation
abnormal fruits duringofpostharvest
melanogenesis and handing
products may process, leading
cause cancer to quick degradation
(melanoma), [32,33].
age spots, freckle
and other dermatological
The application of tyrosinase problems
was further [35,36]. The melanogenesis
extended in the molting stimulators
process of as insects
skin-pigmenting
[34].The agents
abnormal
are very important
accumulation to the occurrence
of melanogenesis products of may
vitiligo. Thuscancer
cause developing new melanogenesis
(melanoma), age spots, activators
freckle andwithother
drug-like properties is very much needed. Here, we focus on the recent
dermatological problems [35,36]. The melanogenesis stimulators as skin-pigmenting agents are discovery of melanogenesis
stimulators from all sources, including plant extracts, natural products and laboratory synthetic
very important to the occurrence of vitiligo. Thus developing new melanogenesis activators with
methods. Moreover, we believe that this perspective will comprise a cumulative source for developing
drug-like properties is very much needed. Here, we focus on the recent discovery of melanogenesis
therapeutic agents for inducing repigmentationin vitiligo-affected skin.
stimulators from all sources, including plant extracts, natural products and laboratory synthetic
methods. Moreover, we believe that this perspective will comprise a cumulative source for developing
therapeutic agents for inducing repigmentationin vitiligo-affected skin.
Molecules 2017, 22, 1303 4 of 28

2. Melanogenesis Stimulators from Different Sources

Many efforts have been made in the discovery and development of melanogenesis stimulators in
the last ten years, as it is a very active field of research for academic centres and medical institutions.
Some of these stimulators have synthetic origin, but others have been derived from small molecules
of natural origin. Worth mentioning is the fact that the plant kingdom has been shown recently to
provide a source of chemical structures with promising biological activities. The number of small
molecule melanogenesis stimulators is continuously increasing, with most of them being in the early
discovery phase. The current stimulators from different sources can be classified into the following
two categories:

1. Plant extracts/crude drug extracts (mixtures);

2. Active natural products/synthesized derivatives (single compounds).

2.1. Plant Extracts/Crude Drug Extracts (Mixtures)

Traditional herbs and plants are widely used for the skin hypopigmentation in view of their lesser
side effects and wealth of sources. The search for new pharmaceuticals for the treatment of vitiligo has
increased over the last few years and plant-derived products (e.g., chemical extracts from medicinal
plants, herbs, and spices) are becoming increasingly accepted and adopted by the medical industry for
this purpose (Table 1).

2.1.1. Daphne gnidium

The ethyl-acetate extract of Daphne gnidium was recently proved to significantly stimulate
production of intracellular and extracellular melanin [37]. Likewise, the tyrosinase activity in B16-F0
cells treated with extraction increased in a time-dependent manner. Later, the effect of chloroform
extract of the plant on melanogenesisin murine B16-F0 melanoma was studied as well by the same
group [38], and a concentration-dependent stimulation effect on tyrosinase and melanogenesis
was observed. It was inferred that the chloroform extract was able to induce differentiation of
B16-F0 melanoma cells preventing them from proliferating to the differentiated state. Induction of
melanogenesis is considered as a well-known marker of differentiated melanoma cells.

2.1.2. Moricandia arvensis

Skandrani et al. [39] evaluated the activity of a chloroform extract of Moricandia arvensis on
melanogenesis and tyrosinase activity. The results indicated that the chloroform extract significantly
promoted production of intra- and extracellular melanin when compared to untreated cells, as well as
tyrosinase activity in B16-F0 cells, in a time-dependent manner.

2.1.3. Ecliptae herba, Polygoni multiflori radixpraeparata and Rehmanniae radix praeparata
Polygoni multiflori radix praeparata (PMRP), Ecliptae herba (EH) and Rehmanniae radix praeparata
(RRP) are the most frequently-used herbs by traditional Chinese medicine practitioners for the
treatment of vitiligo. EH aqueous extract was found to exhibit a synergistic effect on melanocytes by
up-regulating tyrosinaseactivity, enhancing melanin synthesis and promoting melanocyte migration
as well as elevating MITF protein expression [40]. RRP showed a significant stimulating effect
on melanogenesis and MITF protein expression, but no stimulatory effect on tyrosinase activity.
In addition, treatment with PMRP and EH promoted the migration of human melanocytes in a type IV
collagen-coated transwell migration assay. Theseresults suggest that EH and RRP contain substances
with direct enhancing effects on melanogenesis and migration, possibly via their effects on MITF
protein expression.
Molecules 2017, 22, 1303 5 of 28

2.1.4. Cassia alata and Cassia occidentalis

Babitha et al. [41] reported the effect of Cassia alata leaf extract on the differentiation, proliferation
and migration of melanoblast cells in melb-a melanoblast cells. Result indicated that melanin content
increased in a dose-dependent manner. In addition, it induced tyrosinase activity and altered melb-a
cell morphology. A transwell migration assay shows that the extract can directly stimulate the
migration of melanoblast cells. Similar to the above findings, pod extracts of Cassia occidentalis, which
is another plant of Cassia L., was found to be effective in inducing differentiation and migration
ofmouse melanoblast cell line by Babitha and co-workers [42]. Methanolic extract redissolved in
DMSO at 12.5 µg/mL was found to cause 3.5 to 3.8-fold melanin induction in melb-a melanoblast
cells. Stimulation of tyrosinase activity, dendritogenesis and migration of treated cells were observed
as well.

2.1.5. Pyrostegia venusta

The flowers of Pyrostegia venusta are used in Brazil in the treatment ofwhite patches on the body
(such as vitiligo) as a popular folk medicine. It wasdemonstrated that both extracts, leaves and flowers
of Pyrostegia venusta increased the melanin content in a concentration dependent manner after 4 days
of incubation on melanoma cells [43]. Leaves extract promoted enhancement of melanogenesis with a
maximum effect of 33.3 ± 3% (3 µg/mL), and the flower extract increased it by 23.4 ± 3% (0.1 µg/mL).
However, neither extract was able to cause any change in the tyrosinase activity.
To investigate the effect of this extract in animal models of vitiligo, the hyperpigmentant activities
of HE in C56BL/6 mice was also studied later by Moreira et al. [44]. The results showed that extract
administered either by gavage (300 mg/kg) ortopically (10%) increased epidermal melanin level
(116.5 ± 13% and 100 ± 16.9%, respectively), diminished dermal depigmentation (36.0 ± 6.7% and
38.2 ± 6.2%, respectively), as well as tissue TNF-α levels (68.2 ± 11.6% and 99.2 ± 12.1%, respectively)
and cell infiltration (basal levels and 94.3 ± 9.17%, respectively). Only topical treatment with extract
altered melanin-specific markers in hair follicles in mice.

2.1.6. Vernonia anthelmintica

The fruit extract of Vernonia anthelmintica is one of the most popular Uyghur medicines used
for vitiligo and initially recorded as Kaliziriin ‘Yao YongZongKu’ around 300 years ago. The extract
was claimed to increase tyrosinase activity and melanin content in a dosage-dependent manner,
and the expression of tyrosinase time-dependently in both B16-F10 cells and normal human primary
melanocytes as well [45]. Besides, it induced MITF protein expression up-regulation and promoted
the level of phosphorylation of p38 mitogen-activated protein kinase (p38-MAPK) markedly at 0–6 h,
which illustrated that the extract exerted its improving effect on melanogenesis mainly by p38-MAPK
activation and MITF induction of tyrosinase.
Tuerxuntayi et al. [46] of our group reported that Kaliziri increased the tyrosinase activity and
melanin content in a dose-dependent manner at concentrations of 5–40 µg/mL, and treatment with
20 µg/mL Kaliziri extract (KZE) enhanced the expression of tyrosinase in B16 melanoma cells in a
time-dependent manner.
Based on the above research, chlorogenic acid (CGA) [47] and ten compounds (including two
novel compounds 1 and 2) [48] were isolated from the Kaliziri by our group as well (Figure 3). It was
speculated that CGA has two side-roles in melanogenesis of B16 melanoma cells as it exhibited
different effects on melanogenesis and tyrosinase as the incubation time was extended. CGA is likely a
substrate of melanin, but its metabolic products may suppress melanogenesis in B16 melanoma cells
by inhibiting tyrosinase activity.
Molecules 2017, 22, 1303 6 of 28

Table 1. Summary of stimulation effect on tyrosinase, melanogenesis and other factors in cultured cells and pigmentation responsein animal of different plants.

Related
Animal
Botanical Name Extraction Parts Solvents Items Up-Regulated Types of Cultured Cells Signaling Ref.
Experiments
Pathway
Daphne gnidium Leaf Ethyl acetate Melanin, Tyrosinase activity B16-F0 Ns a Ns [37]
Daphne gnidium Leaf Chloroform Melanin, Tyrosinase activity B16-F0 Ns Ns [38]
Moricandia arvensis Leaf Chloroform Melanin, Tyrosinase activity B16-F0 Ns Ns [39]
Polygonum multiflorum Root Water MITF, Melanocyte migration Human melanocytes Ns Ns [40]
Melanin, Tyrosinase activity, MITF,
Eclipta Prostrate Whole herb Water Human melanocytes Ns Ns [40]
Melanocyte migration
Rehmannia Glutinosa Root Water Melanin, MITF Human melanocytes Ns Ns [40]
Melanin, Tyrosinase activity,
Cassia alata Leaf Nm b Melb-a melanoblast Ns Ns [41]
Dendritogenesis, Migration
Melanin, Tyrosinase activity,
Cassia occidentalis Leaf Methanol Melb-a melanoblast Ns Ns [42]
Dendritogenesis, Migration
Ethanol:Water
Pyrostegia venusta Leaf, Flower Melanin B16-F10 Ns Ns [43]
(70:30, v/v)
Ethanol:Water
Pyrostegia venusta Leaf Epidermal melanin, Dermal pigmentation - Ns C56BL/6 mice [44]
(70:30, v/v)
Melanin, Tyrosinase activity, Tyrosinase
Ethanol:Water
Vernonia anthelmintica Fruit expression, p38 MAPK phosphorylation, B16-F10, Human melanocytes p38 MAPK Ns [45]
(60:40, v/v)
MITF expression
Ethanol:Water
Vernonia anthelmintica Seed TYR, TRP-1, TRP-2 and MITF expression B16 murinemelanoma Ns Ns [46]
(80:20, v/v)
Melissa officinalis Whole herb Nm Melanin Human keratinocytes Ns Ns [49]
Ethanol:Water
Melia azedarach Nm Melanin, TRP-1 expression B16-F10 Ns NS [50]
(70:30, v/v)
Capparis spinosa Nm Nm Melanin, Tyrosinase expression B16 murinemelanoma Ns Ns [51]
Erica multiflora Nm Nm Melanin, Tyrosinase expression B16 murine melanoma Ns Ns [51]
Citrus paradisi, Citrus grandis,
Fructus aurantii immaturus, Rind Ethyl acetate Melanin, Tyrosinase expression B16 murine melanoma Ns Ns [52]
Fructus aurantii
a Ns means not studied; b Nm means not mentioned.
Molecules 2017, 22,
Molecules 2017, 22, 1303
1303 77 of
of 27
28

Figure
Figure 3.
3. Structures
Structures of
of compounds
compounds isolated
isolated from
from Vernonia anthelmintica [47,48].
Vernonia anthelmintica [47,48].

2.1.7.TwoMelissa compounds, named benzoyl-vernovan (1) and 2-(40 -hydroxyphenyl)-6-methyl-4H-

newofficinalis
pyran-4-one (2), together 0 -bis-(3,4-dihydroxy-phenyl]-7,70 -dihydroxy
Pérez-Sánchez et al.with
[49]eight known
reported compounds
that extract of[2,2
Melissa officinalis may act as a melanogenic
-2,3,2 0 0
,3 -tetrahydro-[3,3 0 0 0 0
activator since it could ]-bichromenyl-4,4
promote endogenous -dionemelanin
(3), 3 ,4 production
,6-trihydroxyaurone (4), butin(5),
in melanogenesis in butein
a human(6),
isocarthamin 0
keratinocytes(7), luteolin
model. (8), isorhamnetin
Thirteen (9) and2,4-trans-7,4
major phenolic compounds (including -dihydroxy-4-methoxyflavan
rosmarinic acid, RA) (10) were
were
isolated from the KZE (Figure 3). Among them, compounds 5 and
identified by HPLC-DAD-ESI-IT-MS/MS. Unfortunately, the effects of these ingredients on9 were proved to increase melanin
content
melanogenesisby 2.2%wereand 30.9% higher than positive control 8-MOP.
not evaluated.
2.1.7. Melissa officinalis
2.1.8. Melia azedarach
Pérez-Sánchez et al. [49] reported that extract of Melissa officinalis may act as a melanogenic
70% ethanol extract of Melia azedarach was demonstrated to rapidly increase melanin content in
activator since it could promote endogenous melanin production in melanogenesis in a human
a concentration-dependent manner within 4 h [50]. Additionally, although the extract did not affect
keratinocytes model. Thirteen major phenolic compounds (including rosmarinic acid, RA)
intracellular tyrosinase activity, protein levels of tyrosinase and TRP-2 at 2 and 4 h after treatment, it
were identified by HPLC-DAD-ESI-IT-MS/MS. Unfortunately, the effects of these ingredients on
could improve TRP-1 protein expression at both time points.
melanogenesis were not evaluated.
2.1.9. Capparis
2.1.8. spinosa and Erica multiflora
Melia azedarach
Matsuyama
70% ethanol and co-workers
extract of Melia [51] also reported
azedarach that extractto
was demonstrated ofrapidly
Capparisincrease
spinosa and Ericacontent
melanin multiflora
in
enhanced the synthesized melanin content in B16 cells without cytotoxity. Western blotting
a concentration-dependent manner within 4 h [50]. Additionally, although the extract did not affect showed
that tyrosinase
intracellular expression
tyrosinase was clearly
activity, proteinincreased
levels of in cells treated
tyrosinase and with
TRP-2the
atextracts
2 and 4 as well.treatment, it
h after
could improve TRP-1 protein expression at both time points.
2.1.10. Citrus paradisi, Citrus grandis, Fructus aurantii immaturus and Fructus aurantii
Cappariscontent
2.1.9.Melanin spinosa and
and Erica multiflora
tyrosinase expression in mouse B16 melanoma cells were assayed after
treatment with four citrus plant extracts and theirthat
Matsuyama and co-workers [51] also reported hydrolysates by Chiang
extract of Capparis et al.
spinosa and[52]. The
Erica results
multiflora
illustratedthe
enhanced that hydrolysismelanin
synthesized increased the in
content naringenin content cytotoxity.
B16 cells without in citrus Western
extracts blotting
and thatshowed
citrus
preparations stimulated cellular melanogenesis and tyrosinase expression.
that tyrosinase expression was clearly increased in cells treated with the extracts as well.

2.1.11. Bee
2.1.10. Venom
Citrus paradisi, Citrus grandis, Fructus aurantii immaturus and Fructus aurantii
Jeon et al.content
Melanin [53] discovered that bee
and tyrosinase venom (BV)
expression increased
in mouse B16 the numbercells
melanoma of human melanocytes
were assayed after
dose and time
treatment withdependently through
four citrus plant PKA, and
extracts ERK,their
and PI3K/Akt activation.
hydrolysates The level
by Chiang et al.of cAMP
[52]. Thewas also
results
increased by
illustrated BVhydrolysis
that treatment. increased
Moreover,theBVnaringenin
induced melanogenesis through
content in citrus increased
extracts tyrosinase
and that citrus
expression and
preparations induced cellular
stimulated melanocyte dendricity and migration
melanogenesis tyrosinase through PLA2 activation.
expression.
Molecules 2017, 22, 1303 8 of 28

2.1.11. Bee Venom

Jeon et al. [53] discovered that bee venom (BV) increased the number of human melanocytes
dose and time dependently through PKA, ERK, and PI3K/Akt activation. The level of cAMP was
also increased by BV treatment. Moreover, BV induced melanogenesis through increased tyrosinase
Molecules 2017,
expression and22, 1303
induced melanocyte dendricity and migration through PLA2 activation. 8 of 27

2.2. Active Natural

2.2. Natural Products/Synthesized
Products/Synthesized Derivatives
Derivatives (Single
(Single Compounds)
Compounds)

2.2.1.
2.2.1. Flavonoids
Flavonoids

Flavanones
Flavanones
In
In 2006,
2006, Ohguchi
Ohguchi et et al.
al. [54]
[54] first
first examined
examined the the effect
effect of
of naringenin
naringenin (a (a naturallyoccurring
naturallyoccurring citrus
citrus
flavanone) on melanogenesis in mouse B16 melanoma cells. The study
flavanone) on melanogenesis in mouse B16 melanoma cells. The study indicated that naringeninindicated that naringenin
induced melanogenesis,and
induced melanogenesis, andthat
thatthe
themajor
major melanogenic
melanogenic signaling
signaling factors,
factors, suchsuch as tyrosinase,
as tyrosinase, Tyrp-1,
Tyrp-1, Dct,
Dct, and MITF, were upregulated by
and MITF, were upregulated by the compound. the compound.
Huang
Huang et et al.
al. [55]
[55] reported
reported that
that exposure
exposure of of melanoma
melanoma cellscells to
to naringenin
naringenin (Figure
(Figure 4)
4) resulted
resulted in
in
morphological
morphological changes accompanied by the induction of melanocyte differentiation-related markers,
changes accompanied by the induction of melanocyte differentiation-related markers,
such
such as
as melanin
melanin synthesis,
synthesis, tyrosinase
tyrosinase activity,
activity, and
and the
the expression
expression of of tyrosinase
tyrosinase and
and MITF.
MITF. They
They also
also
observed
observed an increase in the intracellular accumulation of β-catenin as well as the phosphorylation of
an increase in the intracellular accumulation of β-catenin as well as the phosphorylation of
glycogen
glycogen synthase
synthase kinase-3β
kinase-3β (GSK3β)
(GSK3β) protein
protein after
after treatment
treatment withwith naringenin.
naringenin.

Figure 4.
Figure 4. Structures
Structures of
of flavanones
flavanones isolated
isolated from
from Citrus.
Citrus.

Moreover,the
Moreover, theactivity
activity of
of phosphatidyl-inositol
phosphatidyl-inositol 3-kinase
3-kinase (PI3K)
(PI3K) was
was up-regulated
up-regulated byby naringenin
naringenin
since the
since the phosphorylated
phosphorylated level
level ofof downstream
downstream Akt Akt protein
protein was
was enhanced.
enhanced. It was
was concluded
concluded thatthat
naringenin-induced melanogenesis through the
naringenin-induced the Wnt-β-catenin-signalling
Wnt-β-catenin-signalling pathway based on on these
these
results (Figure
results (Figure 5).
5).
In their further study
In study [56],
[56], ititwas
wasfound
foundthatthatthe
theacid-hydrolyzed
acid-hydrolyzed extracts of of
extracts Citrus
Citrussinensis, C.
sinensis,
reticulata, and C. aurantium enhanced melanin production. Hesperetin,
C. reticulata, and C. aurantium enhanced melanin production. Hesperetin, which was which was the most abundant
abundant
flavonoids in
flavonoids in citrus
citrus hydrolyzed
hydrolyzed extracts
extracts and
and has
has aa similar
similar structure
structure as
as naringenin,
naringenin, exhibited
exhibited the
the best
best
potency on
potency on melanin
melanin synthesis
synthesis and
and induced
induced tyrosinase
tyrosinase andand MITF
MITF expression.
expression. Moreover,
Moreover, itit stimulated
stimulated
the activation
the activation ofof mitogen-activated
mitogen-activated protein
protein kinases
kinases (MAPKs),
(MAPKs), phosphorylation
phosphorylation of of cAMP-responsive
cAMP-responsive
elementbinding
element bindingprotein
protein (CREB)
(CREB) andand glycogen
glycogen synthase
synthase kinase-3β
kinase-3β (GSK3β),(GSK3β), and subsequently
and subsequently induced
induced
the the accumulation
accumulation of β-catenin.
of β-catenin.
Sixteen 3′,4′,7-trihydroxyflavanones were synthesized to improve melanogenesis in B16-F10
mouse melanoma cell line (Scheme 2). In searching for their action target, a novel fluorescent flavanone-
BODIPY probe without affecting its bioactivity was developed after structure-activity relationship (SAR)
analysis of these compounds [57]. Confocal imaging showed that the probe which could increase the
content of B16-F10 cells by selectively binding to its endoplasmic reticulum.
Isosakuranetin (Figure 6), an important component of propolis, Baccharis dracunculifolia,
Molecules 2017, 22, 1303 9 of 27

Molecules 2017, 22, 1303 9 of 28

Molecules 2017, 22, 1303 9 of 27

Figure 5. Model of signalling pathways involved in naringenin-induced melanogenesis [55].

Chalcones
Chalcone, which are one of the major classes of natural products with widespread distribution
in fruits, vegetables, spices, tea and soy-based foodstuffs, display very versatile physiological
activity Figure 5. Model now,
[62–65].
Figure 5. Until
of signalling
most pathways
Model of signalling chalconesinvolved in naringenin-induced
and their
pathways involved
melanogenesis
derivatives have melanogenesis
in naringenin-induced been described[55].
[55].as potent
inhibitors of tyrosinase [66,67].
Chalcones
Chalcone, which are one of the major classes of natural products with widespread distribution
in fruits, vegetables, spices, tea and soy-based foodstuffs, display very versatile physiological
activity [62–65]. Until now, most chalcones and their derivatives have been described as potent
inhibitors of tyrosinase [66,67].

Scheme
Scheme 2. Synthesis of
2. Synthesis of novel
novel fluorescent
fluorescent flavanone-BODIPY
flavanone-BODIPY probe
probe 77 [57].
[57].

Isosakuranetin (Figure 6), an important component of propolis, Baccharis dracunculifolia, Terminalia

fagifolia, Citrus sinensis [58–60], was proved to stimulates melanogenesis in B16 melanoma cells via
up-regulation of MITF. Furthermore, it induced inhibition of ERK1/2 and PI3K/AKT signaling
pathways activate MITF and subsequent expression of TYR, TRP1, and TRP2 [61].
Scheme 2. Synthesis of novel fluorescent flavanone-BODIPY probe 7 [57].
Molecules 2017, 22, 1303 10 of 28
Molecules 2017, 22, 1303 10 of 27
Molecules 2017, 22, 1303 10 of 27

Figure
Figure 6.
6. Structures
Structures of
of isosakuranetin
isosakuranetin and
and sakuranetin.
sakuranetin.
Figure 6. Structures of isosakuranetin and sakuranetin.
Few
Few flavonoids
flavonoids havehave been
beenreported
reported as as activators
activators ofof tyrosinase.
tyrosinase. A A new
new series
series of
of 4-(phenyl-urenyl)
4-(phenyl-urenyl)
Chalcones
chalcones
chalcones 14–23 and 4′-(phenylurenyl/thiourenyl)chalcone derivatives 24–35 were synthesized
14–23 and 4′-(phenylurenyl/thiourenyl)chalcone derivatives 24–35 were synthesized and and
their Chalcone, which are one ofwerethe major classes of natural productsthatwith14–23
widespread distribution
their effects on the tyrosinase were evaluated. The results showed that 14–23 inhibited the
effects on the tyrosinase evaluated. The results showed inhibited the PPO
PPO
in fruits, vegetables,
enzyme spices, tea and soy-based foodstuffs, display very versatile physiological
enzyme activity.
activity. Conversely,
Conversely, 24–35
24–35 exhibited
exhibited activator
activator effect
effect onon tyrosinase
tyrosinase [68].
[68]. Nixha
Nixha et et al.
al. [69]
[69]
activity [62–65].
introduced a Until
series of now, mostchalcone
carbazole chalcones and their
analogues and derivatives
determined have been
their described
activity on as potent
tyrosinase.
introduced a series of carbazole chalcone analogues and determined their activity on tyrosinase.
inhibitors of analogues
From tyrosinase [66,67].could enhance the activity of the enzyme. Unfortunately, these two
From series,
series, analogues 36–39 36–39 could enhance the activity of the enzyme. Unfortunately, these two
series Few
of flavonoids
compounds have
all been reported
showed as activators of tyrosinase. A new series of 4-(phenyl-urenyl)
series of compounds all 0
showed poor
poor activity
activity (Figure
(Figure 7).7).
chalcones
Inspired 14–23 and 4 -(phenylurenyl/thiourenyl)chalcone derivatives 24–35 were synthesized and
Inspired by by above
above results,
results, itit was
was speculated
speculated that that similar
similar groups
groups onon the
the para-position
para-position of of the
the
their effects
chalcone A on may
ring the tyrosinase
cause a were
great evaluated. on
improvement Theactivating
results showedeffect that
on 14–23 inhibited
tyrosinase, but an the PPO
inhibitor
chalcone A ring may cause a great improvement on activating effect on tyrosinase, but an inhibitor
enzyme activity.
effect Conversely, to 24–35 exhibited activator effect on tyrosinaseet[68]. Nixha prepared
et al. [69]
effect when
when itit was
was introduced
introduced to BB ring ring at
at the
the same
same position.
position. Therefore,
Therefore, Niu
Niu et al.
al. [70]
[70] first
first prepared
introduced
sixteen a seriesderivatives
of carbazolecontaining
chalcone analogues and determined their activityand
on tyrosinase. From
sixteen chalcone derivatives containing benzothiazole and amide moieties and evaluated their
chalcone benzothiazole and amide moieties evaluated their
series,
activator analogues 36–39 could enhance the activity of the enzyme. Unfortunately, these two series of
activator effect on tyrosinase. Compared with the reference drug 8-MOP, compounds 40, 41, 42, 43
effect on tyrosinase. Compared with the reference drug 8-MOP, compounds 40, 41, 42, 43
compounds
and all showed pooractivity
activity (Figure 7).
and 44
44 displayed
displayed promising
promising activity on on tyrosinase
tyrosinase with
with EC EC5050 from
from 30.6–9.6
30.6–9.6 μMμM (Figure
(Figure 8). 8).

Structures ofchalcone
Figure 7.Structures
Figure chalcone derivativesand
and analogueswith
with activatoreffect
effect ontyrosinase
tyrosinase [68,69].
Figure 7.
7. Structures of
of chalcone derivatives
derivatives and analogues
analogues with activator
activator effect on
on tyrosinase [68,69].
[68,69].
Inspired by above results, it was speculated that similar groups on the para-position of the chalcone
A ring may cause a great improvement on activating effect on tyrosinase, but an inhibitor effect when
it was introduced to B ring at the same position. Therefore, Niu et al. [70] first prepared sixteen
chalcone derivatives containing benzothiazole and amide moieties and evaluated their activator effect
on tyrosinase. Compared with the reference drug 8-MOP, compounds 40, 41, 42, 43 and 44 displayed
promising activity on tyrosinase with EC50 from 30.6–9.6 µM (Figure 8).
After that, substituted 1,2,3-triazoles were separately introduced into the A (compounds 45–59)
and B (compounds 60–74) rings of the chalcone skeleton using click chemistry by our group [71].
The results showed that most of prepared compounds 45–59 have potent activating effect on tyrosinase,
especially for 47, 52–54 and 58–59. Among them, compounds 54 and 58 demonstrated the best activity
with EC50 =1.71 and 5.60 µM respectively, even better than the positive control (Figure 9).
Figure
Figure 8.
8. Structures
Structures of
of the
the benzothiazole
benzothiazole and
and amide
amide chalcones.
chalcones.
Molecules 2017, 22, 1303 11 of 28
Figure 7. Structures of chalcone derivatives and analogues with activator effect on tyrosinase [68,69].

Molecules 2017, 22, 1303 11 of 27

Molecules 2017, 22, 1303 11 of 27

After that, substituted 1,2,3-triazoles were separately introduced into the A (compounds 45–59)
After that, substituted 1,2,3-triazoles were separately introduced into the A (compounds 45–59)
and B (compounds 60–74) rings of the chalcone skeleton using click chemistry by our group [71]. The
and B (compounds 60–74) rings of the chalcone skeleton using click chemistry by our group [71]. The
results showed that most of prepared compounds 45–59 have potent activating effect on tyrosinase,
results showed that most of prepared compounds 45–59 have potent activating effect on tyrosinase,
especially for 47, 52–54 and 58–59. Among them, compounds 54 and 58 demonstrated the best activity
especially for 47, 52–54Figure
and 58–59. Amongofthem,
8. Structures compoundsand
the benzothiazole 54 and 58chalcones.
amide
amide demonstrated the best activity
chalcones.
with EC50 =1.71 and 5.60 μM respectively, even better than the positive control (Figure 9).
with EC50 =1.71 and 5.60 μM respectively, even better than the positive control (Figure 9).

Figure9.
Figure Structuresof
9.Structures ofthe
thechalcones
chalconesderivatives
derivatives bearing1,2,3-triazole
bearing1,2,3-triazole moieties.
moieties.
Figure 9. Structures of the chalcones derivatives bearing1,2,3-triazole moieties.
On the
On the contrary,
contrary, compounds
compounds 62,
62, 64–65,
64–65, 68–69, and74
68–69,and inducedenzymatic
74induced enzymaticinhibition
inhibitionon ontyrosinase,
tyrosinase,
On the contrary, compounds 62, 64–65, 68–69, and 74 induced enzymatic inhibition on tyrosinase,
which was
which was consistent
consistent with
with our
our previous
previous speculation.
speculation.
which was consistent with our previous speculation.
In 2016, Niu and co-workers [72]
In 2016, Niu and co-workers [72] also also reported
reported the
the design
design and
and synthesis
synthesis of
of novel
novel chalcone
chalcone
In 2016, Niu and co-workers [72] also reported the design and synthesis of novel chalcone
derivatives 75–92 bearing isoxazole moieties as activators on tyrosinase and melanogenesis
derivatives 75–92 bearing isoxazole moieties as activators on tyrosinase and melanogenesis in murine in murine
derivatives 75–92 bearing isoxazole moieties as activators on tyrosinase and melanogenesis in murine
B16 cells.
B16 cells. Among
Among the the synthesized
synthesizedmolecules,
molecules,compounds
compounds76, 76,78, and 83
78,and exhibited the
83 exhibited the most
most potent
potent
B16 cells. Among the synthesized molecules, compounds 76, 78, and 83 exhibited the most potent
activating effect on tyrosinase, with EC = 1.3, 2.5 and 3.0 µM, respectively. In B16 cells, it was
activating effect on tyrosinase, with EC50 = 1.3, 2.5 and 3.0 μM, respectively. In B16 cells, it was interesting
50
activating effect on tyrosinase, with EC50 = 1.3, 2.5 and 3.0 μM, respectively. In B16 cells, it was interesting
interesting that derivatives substituted with halogens were generally more potent.
that derivatives substituted with halogens were generally more potent. Compounds 76 (463%) and Compounds 76
that derivatives substituted with halogens were generally more potent. Compounds 76 (463%) and
(463%)
92 (438%)and 92 (438%)
with with 3potency
3 and 4-fold and 4-fold potencywith
compared compared
8-MOPwith 8-MOP
(Figure (Figure
10), were 10), wereas
recognized recognized
the most
92 (438%) with 3 and 4-fold potency compared with 8-MOP (Figure 10), were recognized as the most
as the most promising candidate hits for further pharmacological study
promising candidate hits for further pharmacological study of anti-vitiligo. of anti-vitiligo.
promising candidate hits for further pharmacological study of anti-vitiligo.

Figure 10. Structures of the chalcones derivatives bearing isoxazole moieties.

To find theFigure 10. Structures of thechalcone

pharmacophore chalcones derivatives bearing isoxazole moieties.
Figure 10. Structures ofof on tyrosinase,
the chalcones derivatives bearingtwenty-one chalcones and nine
isoxazole moieties.
analogues were synthesized in view of three different components of chalcone (A, B ring and
To find the pharmacophore of chalcone on tyrosinase, twenty-one chalcones and nine analogues
To find the pharmacophore of chalcone on tyrosinase, twenty-one chalcones and nine analogues
were synthesized in view of three different components of chalcone (A, B ring and α-,β-unsaturated
were synthesized in view of three different components of chalcone (A, B ring and α-,β-unsaturated
carbonyl) [73]. The biological evaluation revealed that most compounds (except polyhydroxy-
carbonyl) [73]. The biological evaluation revealed that most compounds (except polyhydroxy-
chalcones) possess activator effects on tyrosinase, especially for 93–97 (Figure 11). Finally, compound
chalcones) possess activator effects on tyrosinase, especially for 93–97 (Figure 11). Finally, compound
93 was found to increase melanin contents and tyrosinase activity 1.75- and 1.3-fold in B16 cells,
93 was found to increase melanin contents and tyrosinase activity 1.75- and 1.3-fold in B16 cells,
respectively and the SAR of these tyrosinase activator was summed up for the first time as well.
 Molecules 2017, 22, 1303 12 of 28

α-,β-unsaturated carbonyl) [73]. The biological evaluation revealed that most compounds (except
polyhydroxy-chalcones) possess activator effects on tyrosinase, especially for 93–97 (Figure 11).
Finally, compound 93 was found to increase melanin contents and tyrosinase activity 1.75- and 1.3-fold
in B16 cells, respectively and the SAR of these tyrosinase activator was summed up for the first time
Molecules 2017, 22, 1303 12 of 27
as well.

Molecules 2017, 22, 1303 12 of 27

Figure 11.11.
Figure Structures ofof
Structures thethe
chalcones with
chalcones activator
with effect
activator and
effect SAR.
and SAR.
Figure 11. Structures of the chalcones with activator effect and SAR.
Flavonoid glycosides
Flavonoid Glycosides
Flavonoid glycosides
Two
Two novel
novelquercetin
quercetin glucosides,
glucosides,were
wereisolated
isolated from
from Helminthostachys
Helminthostachys zeylanica root
zeylanica 50%
root 50%ethanol
ethanol
extract. Of Two
the novelquercetin-glucosides
two quercetin glucosides, were isolated
(Figure 12), from Helminthostachys
compound 98 zeylanica
exhibited a root 50%
high ethanol
melanogenic
extract. Of the two quercetin-glucosides (Figure 12), compound 98 exhibited a high melanogenic
extract. Of the two quercetin-glucosides (Figure 12), compound 98 exhibited a high melanogenic
acceleratory
acceleratoryeffect, 2.72.7
effect, times
timeshigher
higherthan control atat
than 10 μM concentration inin
murine B16 melanoma cells,
acceleratory effect, 2.7 times higher thancontrol
control at 10
10 µM concentration
μM concentration murine
in murine B16B16 melanoma
melanoma cells,cells,
with no cytotoxic effect [74].
with no cytotoxic effect [74].
with no cytotoxic effect [74].

Figure 12. Structures

Figure of of
12. Structures thethe
compounds
compoundsisolated Helminthostachys
fromHelminthostachys
isolated from zeylanica.
zeylanica.
Figure 12. Structures of the compounds isolated from Helminthostachys zeylanica.
In 2014, theirtheir
In 2014, group
groupalso
alsoderivatized
derivatized aa series
seriesofofquercetin
quercetin glycosides
glycosides and evaluated
and evaluated them asthem
In melanogenesis
2014, their group
as melanogenesis also compounds
acceleration
acceleration derivatized
compounds a (Figure
[75] series of
13),quercetin
[75] (FigureSAR13),
wasSAR glycosides
carried and evaluated
out carried
was to correlate them as
outthetoimportance
correlate the
melanogenesis
importance acceleration
of manyofsubstituents compounds [75]
with the observed
many substituents (Figure
with theactivity.
observed 13),
From SAR was carried
the series,
activity. From out
compound to correlate
100,compound
the series, the importance
101 and 102100,showed
101 and
of102
many substituents
more
showed potent with the
more intracellular
potent observed activity.
melanogenesis
intracellular From the
acceleration
melanogenesis series, compound
activities
acceleration than 100,
than101
theophyline
activities andas102
used
theophyline showed
positive
used as
more controlintracellular
potent in a dose-dependent manner with
melanogenesis no cytotoxicactivities
acceleration effect. than theophyline used as positive
positive control in a dose-dependent manner with no cytotoxic effect.
control in a Later, Yamauchi and
dose-dependent co-workers
manner with no[76]cytotoxic
also attempted
effect. structure-guided synthesis of quercetin
derivatives as melanogenesis activators based on their previous research. Among the prepared
Later, Yamauchi and co-workers [76] also attempted structure-guided synthesis of quercetin
compounds, 3-O-methylquercetin (103) and 3′,4′,7-O-trimethylquercetin (104) increased melanin
derivatives as melanogenesis activators based on their previous research. Among the prepared
content more potently than the positive control with low cytotoxicity (Figure 14). However, the former
compounds, 3-O-methylquercetin (103) and 3′,4′,7-O-trimethylquercetin (104) increased melanin
increased the expression of tyrosinase and TRP-1 to a greater extent than the latter. Furthermore,
contentcompound
more potently than the positive
104 stimulated controlofwith
the expression low
MITF cytotoxicity
and p-p38 MAPK (Figure 14).while
as well, However, the former
they were not
increased the expression of tyrosinase and TRP-1 to a greater extent than the latter.
increased by 103. These results suggested that 103 may enhance the expression of tyrosinase and TRP- Furthermore,
compound1 by 104 stimulated
regulating the expression
the proteasomal of MITF
degradation of and p-p38 MAPK
melanogenic enzymes asand/or
well, while they were
by activating othernot
increased by 103. These
transcriptional results
factors suggested
regulating thatexpression.
enzyme 103 may enhance the expression of tyrosinase and TRP-
1 by regulating the proteasomal degradation of melanogenic enzymes and/or by activating other
Molecules 2017, 22, 1303 13 of 28
Molecules 2017, 22, 1303 13 of 27

Molecules 2017, 22, 1303 13 of 27

Molecules 2017, 22, 1303 13 of 27

Figure 13. Structures of the synthesized quercetin glycosides.

Figure 13. Structures of the synthesized quercetin glycosides.

Later, Yamauchi and co-workers [76] also attempted structure-guided synthesis of quercetin
derivatives as melanogenesis activators based on their previous research. Among the prepared
compounds, 3-O-methylquercetin (103) and 30 ,40 ,7-O-trimethylquercetin (104) increased melanin
content more potently than the positive control with low cytotoxicity (Figure 14). However, the former
increased the expression of tyrosinase and TRP-1 to a greater extent than the latter. Furthermore,
compound 104 stimulated the expression of MITF and p-p38 MAPK as well, while they were not
increased by 103. These results
Figure suggested
14. Structures thatsynthesized
of the 103 may enhance
quercetin the expression of tyrosinase and
derivatives.
TRP-1 by regulating the proteasomal degradation of melanogenic enzymes and/or by activating other
Ye et al. [77]
transcriptional first screened
factors Figure 13.for
regulating tyrosinase
Structures
enzyme activity
of the enhancers
synthesized
expression. among
quercetin 35 phytocompounds in B16
glycosides.
mouse melanoma cells in 2010. Among them, three compounds (apigenin, icariin, hyperoside) were
more potent than 8-MOPFigure for 13. Structures of
enhancing the synthesized
tyrosinase quercetin
activity andglycosides.
significantly increased cellular
melanin contents without affecting cell proliferation (Figure 15). Western blot analysis demonstrated
that these compounds could differentially increase the expression levels of tyrosinase, and TRP-1 and
TRP-2. Together these data suggest that apigenin and icariin exert potent melanogenic activities through,
at least in part, upregulating the protein expression levels of melanogenic enzymes in B16 cells.

Figure14.
Figure 14.Structures
Structuresof
ofthe
thesynthesized
synthesizedquercetin
quercetinderivatives.
derivatives.
Figure 14. Structures of the synthesized quercetin derivatives.
Yeetetal.
Ye al.[77]
[77]first
firstscreened
screenedfor fortyrosinase
tyrosinaseactivity
activityenhancers
enhancersamong among35 35phytocompounds
phytocompoundsin inB16
B16
mousemelanoma
mouse Ye et al.
melanomacells [77] first
cellsin screened
in2010.
2010.Among for tyrosinase
Amongthem, activity
them,three enhancers
threecompounds among
compounds(apigenin, 35 phytocompounds
(apigenin,icariin,
icariin,hyperoside) in B16
hyperoside)were were
moremouse
more potent
potent melanoma
thanthan 8-MOP cells
8-MOP inenhancing
for 2010.enhancing
for Among them, three
tyrosinase
tyrosinase compounds
activityactivity (apigenin, icariin,
and significantly
and significantly hyperoside)
increased cellularwere
increased cellular
melanin
melanin
contentsmore potentaffecting
contents
without than 8-MOP
without cell for enhancing
affecting
proliferation tyrosinase
cell proliferation
(Figure activity15).
(Figure
15). Western and significantly
Western
blot blot
analysis increased
analysis
demonstrated cellular
demonstrated
that these
melanin contents without affecting cell proliferation (Figure 15). Western blot analysis demonstrated
that these compounds
compounds could differentially
could differentially increase theincrease the expression
expression levels of levels of tyrosinase,
tyrosinase, and TRP-1 andandTRP-1 and
TRP-2.
that these compounds could differentially increase the expression levels of tyrosinase, and TRP-1 and
TRP-2. Together
Together these data these data suggest
suggest that apigenin
that apigenin and icariin
and icariin exert exert
potent potent melanogenic
melanogenic activities
activities through,
through, at
TRP-2. Together these data suggest that apigenin and icariin exert potent melanogenic activities through,
at least
least in in part,
at part,
least upregulating
inupregulating
part, the
the
upregulating protein
protein
the expression
expression
protein levels
expressionlevels ofmelanogenic
levels of
of melanogenic
melanogenic enzymes
enzymes
enzymes in
in B16in B16
B16cells.
cells. cells.
Figure 15. Structures of three compounds with activator effect on tyrosinase.

Flavones
Yoon et al. [78] reported the stimulating effects of polymethoxylated flavones (nobiletin, tangeretin,
sinensetin) on the production of melanin in murine B16-F10 melanoma cells (Figure 16). The results
indicated all the tested compounds significantly increased melanin content, especially nobiletin.
Further studies showed that nobiletin induced the expression of major melanogenic proteins such as
tyrosinase, TRP-1 and that effect was probably related to the ERK–MAPK pathway.

Figure 15. Structures

Figure of of
15. Structures three
threecompounds
compounds with activatoreffect
with activator effectonon tyrosinase.
tyrosinase.
Figure 15. Structures of three compounds with activator effect on tyrosinase.
Flavones
FlavonesYoon et al. [78] reported the stimulating effects of polymethoxylated flavones (nobiletin, tangeretin,
sinensetin)
Yoon on the
et al. [78] production
reported of melanin ineffects
the stimulating murine ofB16-F10 melanoma cells
polymethoxylated (Figure
flavones 16). The results
(nobiletin, tangeretin,
indicated all the tested compounds significantly increased melanin content,
sinensetin) on the production of melanin in murine B16-F10 melanoma cells (Figure 16). especially nobiletin.
The results
Further studies showed that nobiletin induced the expression of major melanogenic proteins such as
Molecules 2017, 22, 1303 14 of 28

Flavones
Yoon et al. [78] reported the stimulating effects of polymethoxylated flavones (nobiletin, tangeretin,
sinensetin) on the production of melanin in murine B16-F10 melanoma cells (Figure 16). The results
indicated
Molecules
Molecules all22,
2017,
2017, 22,the
1303tested compounds significantly increased melanin content, especially nobiletin.
1303 14
14 of
of 27
27
Further studies showed that nobiletin induced the expression of major melanogenic proteins such as
tyrosinase,
MoleculesTRP-1
2017, 22,and
1303 that effect was probably related to the ERK–MAPK pathway. 14 of 27

Figure
Figure 16.
16. Structures
Structures of
of polymethoxylated
polymethoxylated flavones.
flavones.
Figure 16. Structures of polymethoxylated flavones.
Figure 16. Structures of polymethoxylated flavones.
In
In 2013,
2013, itit was
was found
found thatthat incubation
incubation of
of the
the B16-F10
B16-F10 cells
cells with
with 10
10 μM
μM 4′-O-methylated
4′-O-methylated flavonoidsflavonoids
(diosmetin,
(diosmetin, In acacetin,
acacetin,
2013, it waskaempferide)
kaempferide) (Figure
(Figure
found that incubation 17),
17),
of theincreased
increased the
B16-F10 cellsthe melanin
melanin
with 10 μM contents
contents of
of
4′-O-methylated
0 the
the cells
cells 3-
3- to
to 7-fold
flavonoids 7-fold
In 2013, it was found that incubation of the B16-F10 cells with 10 µM 4 -O-methylated flavonoids
higher
higher (diosmetin,
than the acacetin,
control
than acacetin,
the control kaempferide)
[79] and 20 (Figure
μM
[79] and 20(Figure 17),
acacetin
μM acacetin increased
exhibited
exhibited the melanin
the most contents
promising of the cells
activity 3- to 7-fold
with 33-fold
(diosmetin, kaempferide) 17), increased thethe most promising
melanin contents ofactivity
the cellswith3- to33-fold
7-fold
higher
higher higher than than
activity
activity the
thancontrol
the
the [79] and 20
vehicle.
vehicle. On
On μM acacetin
the
the other
other exhibited
hand,
hand, thethecorresponding
the most promising activity
corresponding 4′-OH-type
4′-OH-typewith 33-fold
flavonoids
flavonoids
higher than
higher the control [79]
activity kaempferol) and
than the vehicle. 20 µM acacetin
On theanother exhibited
hand, smaller
the the most promising
corresponding activity
4′-OH-type with 33-fold
(luteolin,
(luteolin,
higher
apigenin,
apigenin,
activity than kaempferol)
thekaempferol)
vehicle. On
possessed
possessed
the otheran
obviously
obviously
hand, smaller
the corresponding
effect.
effect. In
In addition,
addition,
40 -OH-type theflavonoids
the upregulation
upregulation
flavonoids (luteolin,
(luteolin, apigenin, possessed an obviously smaller effect. In addition, the upregulation
of
of tyrosinase
tyrosinase expression,
expression, preceded
preceded by
by activation
activation of
of cAMP
cAMP response
response element
element binding
binding protein
protein (CREB)
(CREB)
apigenin, kaempferol)
of tyrosinase possessed
expression, an obviously
preceded smaller
by activation effect.
of cAMP In addition,
response elementthebinding
upregulation
proteinof tyrosinase
(CREB)
and
and extracellular
extracellular signal-regulated
signal-regulated kinases
kinases types
types 11 and
and 22 (ERK1/2)
(ERK1/2) was
was observed
observed via
via melanogenic
melanogenic
expression, preceded by
and extracellular activation of cAMP
signal-regulated kinasesresponse
types 1 and element binding
2 (ERK1/2) protein
was observed(CREB) and extracellular
via melanogenic
protein
protein evaluation.
evaluation.
protein evaluation.
signal-regulated kinases types 1 and 2 (ERK1/2) was observed via melanogenic protein evaluation.

Figure 17. Structures of the4′-O-methylated and corresponding 4′-OH flavonoids.

Figure
Figure 17.
17. Structures
Structures of the40 -O-methylatedand
of the4′-O-methylated
the4′-O-methylated and corresponding 40 -OHflavonoids.
corresponding 4′-OH
4′-OH flavonoids.
flavonoids.
Isoflavones
Isoflavones
Isoflavones
Park et al. [80] reported that the Pueraria thunbergiana (PT) extract and its major active ingredient
puerarin (Figure 18) stimulated the melanogenesis via cAMP/MITF-M signaling pathway in vitro,
Park
Park
Parket
etal.
et al.[80]
al. [80] reported
reported
[80]the
reported that
that
thatthe
thePueraria
the Puerariathunbergiana
Pueraria thunbergiana
thunbergiana (PT)
(PT) extract
extractand
extract and its
itsmajor
its major active
activeingredient
active ingredient
and prevented follicular depigmentation and vitiligo by(PT)
stimulating and
melanin major
synthesis. ingredient
puerarin
puerarin (Figure
(Figure 18)18) stimulated
stimulated the the melanogenesis
melanogenesis via via cAMP/MITF-M
cAMP/MITF-M signaling
cAMP/MITF-M signalingpathway
signaling pathwayin
pathway invitro,
in vitro,
vitro,
and
and prevented
prevented thethe follicular
follicular depigmentation
depigmentation and and vitiligo
vitiligo by
by stimulating
stimulating melanin
melanin synthesis.
synthesis.

Figure 18. S tructure of puerarin.

2.2.2. Coumarins
Figure
Figure 18.
18. SS tructure
tructure ofof puerarin.
puerarin.
For over a thousand years, theFigure 18. Structure
plant species Ammiof puerarin.
majus L., Psoralen corylifolia L. and Ficus carica
L. have often been used for repigmentation of skin with natural sunlight in India, Egypt and other
2.2.2. Coumarins
2.2.2.oriental
Coumarins
countries [81–83]. Similarly, their extracts were popular Uygur medicines used for vitiligo
alone
For overin
or
For over combination
aa thousand with the
thousand years,
years, XinJiang
plantasspecies
the plant well. Coumarins
species Ammi
Ammi majus are widely
majus L., distributed
L., Psoralen
Psoralen in these
corylifolia
corylifolia L. plants
L. and
and andcarica
Ficus
Ficus carica
L.
L. havehave
have oftenbeen isolated
often been
been used from
used fortheir seeds, leaves
for repigmentation
repigmentation of and fruits
of skin [84–86].
skin with
with natural
natural sunlight
sunlight inin India,
India, Egypt
Egypt and
and other
other
oriental
oriental countries
countries [81–83].
[81–83]. Similarly,
Similarly, their
their extracts
extracts were
were popular
popular Uygur
Uygur medicines
medicines used
used for
for vitiligo
vitiligo
 Molecules 2017, 22, 1303 15 of 28

Molecules 2017, 22, 1303 15 of 27

2.2.2. Coumarins
Continuous studies have proved that these compounds show strong photosensitivity, which
For over a thousand years, the plant species Ammi majus L., Psoralen corylifolia L. and Ficus carica
may be used foroften
L. have the treatment
been used for of repigmentation
vitiligo with subsequent exposure
of skin with natural to long-wave
sunlight ultraviolet
in India, Egypt and other radiation.
Althoughoriental
the therapy
countries was accompanied
[81–83]. withextracts
Similarly, their somewere undesired
popular side
Uygureffects [87,88],
medicines useditforisvitiligo
still the most
alone or
successfulMolecules
one forin combination with XinJiang as well. Coumarins are widely distributed in these
the disease today. Unfortunately, few coumarin derivatives possessing15anti-vitiligo plants and
2017, 22, 1303 of 27
have been isolated from their seeds, leaves and fruits [84–86].
activity were reported.
Continuous studies have proved that these compounds show strong photosensitivity, which may
Early inContinuous
2005, seven studies have proved
ethanolic extractsthatfrom
these Umbelliferae
compounds show strong
crude photosensitivity,
drugs and sixteen
be used for the treatment of vitiligo with subsequent exposure to long-wave ultraviolet radiation.
which
coumarins
may be used for the treatment of vitiligo with subsequent exposure to long-wave ultraviolet radiation.
(105–120)Although
isolatedthe were evaluated
therapy for theirwith
was accompanied effects
some onundesired
melaninsidecontent
effects using
[87,88],murine B16most
it is still the melanoma
Although the therapy was accompanied with some undesired side effects [87,88], it is still the most
cells [89].successful
Amongone them, thedisease
for the extract of Unfortunately,
today. Heracleum lanatumand
few coumarin and four compounds—psoralen
derivatives possessing anti-vitiligo (105),
successful one for the disease today. Unfortunately, few coumarin derivatives possessing anti-vitiligo
activity
xanthotoxin were
(106), reported.
activity were bergapten
reported. (107), isopimpinellin (108) and sphondin (117)—showed a potent
Early in 2005, seven ethanolic extracts from Umbelliferae crude drugs and sixteen coumarins
stimulatory effect
Early in on2005,
melanogenesis.
seven ethanolicMoreover,
extracts from the SAR was crude
Umbelliferae summarized
drugs andaccording to the results
sixteen coumarins
(105–120) isolated were evaluated for their effects on melanin content using murine B16 melanoma
(105–120)
(Figure 19). isolated were evaluated for their effects on melanin content using murine B16 melanoma
cells [89]. Among them, the extract of Heracleum lanatumand and four compounds—psoralen (105),
cells [89]. Among them, the extract of Heracleum lanatumand and four compounds—psoralen (105),
xanthotoxin (106), bergapten (107), isopimpinellin (108) and sphondin (117)—showed a potent
xanthotoxin (106), bergapten (107), isopimpinellin (108) and sphondin (117)—showed a potent
stimulatory effect on melanogenesis. Moreover, the SAR was summarized according to the results
stimulatory effect on melanogenesis. Moreover, the SAR was summarized according to the results
(Figure
(Figure 19).
19).

Figure 19. Structures of coumarins isolated from seven Umbelliferae crude drugs.
Figure 19. Structures of coumarins isolated from seven Umbelliferae crude drugs.
Figure 19. Structures of coumarins isolated from seven Umbelliferae crude drugs.
Chodurek et al. [90] reported a new treament for malignant melanoma consisting of a
Chodurek
combination Chodurek etetal.al.
of valproic [90][90]
acid reported
(VPA)
reported newa treament
aand new treament for malignant
5,7-dimethoxycoumarin
for malignant melanoma
(DMC).
melanoma consisting
In A375
consisting of cells,ofthe
a combination a results
combination
of valproic of valproic acid (VPA) and 5,7-dimethoxycoumarin (DMC). In A375 cells, the results
demonstrated thatacid
both (VPA) and 5,7-dimethoxycoumarin
compunds could enhance the (DMC) (Figure 20).
synthesis In A375 cells, the
of melaninand the results
formation of
demonstrated that
demonstrated that both
both compunds
compunds could
could enhance
enhance the
the synthesis
synthesis ofof melaninand
melaninand thethe formation
formation of of
dendrite dendrite
and star-shaped cells.
and star-shaped
star-shaped cells.
Moreover,
cells. Moreover,
upregulation
Moreover, upregulation
upregulation of
of tyrosinase
of tyrosinase
activity
tyrosinase activity
activity and
and
and gene
gene
gene expression
expression
expression
dendrite and
were observed
were in response
observed in to
response VPA
to treatment.
VPA treatment.
were observed in response to VPA treatment.

Figure 20. Structures of valproic acid and 5,7-dimethoxycoumarin.

FigureFigure 20. Structures
20. Structures of of valproicacid
valproic acid and
and5,7-dimethoxycoumarin.
5,7-dimethoxycoumarin.
Recently, Pang et al. [91] of our group prepared coumarin derivatives bearing isoxazole moieties
Recently,
(121–135) as Pang et al. [91] stimulator
melanogenic of our group fromprepared
suitablecoumarin derivatives bearing isoxazole
5-(bromomethyl)isoxazoles and 4-
Recently,
moietiesPang et al. [91]
(121–135) asAmong
methylumbelliferone. of our
melanogenicgroup prepared
stimulator
the synthesized coumarin
from compounds
molecules, derivatives bearing isoxazole
suitable 5-(bromomethyl)isoxazoles andmoieties
124 and 126 exhibited excellent
(121–135)4-methylumbelliferone.
as on
potency melanogenic Among
melanin synthesis stimulator
with nearly from
the synthesized and suitable
1.6-molecules, 5-(bromomethyl)isoxazoles
compounds
2.6-fold potency 124 and 126
compared exhibited
with 8-MOP (149%)and 4-
excellent
methylumbelliferone.
potency on AmongB16
melanin
respectively, in murine the
synthesis synthesized
cells (Figure 21). molecules, compounds 124 and 126 exhibited excellent
with nearly 1.6- and 2.6-fold potency compared with 8-MOP (149%)
potency on melanin synthesis with nearly21).
respectively,
In in
another murine
study, B16
our cells (Figure
researchers also
1.6- reported the synthesis
and 2.6-fold potencyofcompared
twenty-five with
furocoumarin
8-MOP (149%)
derivatives and evaluated the stimulatory effect of them on melanogenesis in murine B16 cells [92].
respectively, in murine B16 cells (Figure 21).
In this series, twenty-three compounds were more potent than the positive control (8-MOP).
In another
Compounds study, our researchers
137 (350.5%) also based
and 138 (313.1%) reported
on thethe synthesis
scaffold of twenty-five
of 136 were furocoumarin
nearly 3-fold stronger
derivatives
thanand evaluated
8-MOP the stimulatory effect of them on melanogenesis in murine B16 cells [92].
(Figure 22).
In this series, twenty-three compounds were more potent than the positive control (8-MOP).
Compounds 137 (350.5%) and 138 (313.1%) based on the scaffold of 136 were nearly 3-fold stronger
Molecules 2017, 22, 1303 16 of 28
Molecules 2017, 22, 1303 16 of 27

Molecules 2017, 22, 1303 16 of 27

Molecules 2017, 22, 1303 16 of 27

Figure 21. Structures of the coumarin derivatives bearing isoxazole moieties.

Figure 21. Structures of the coumarin derivatives bearing isoxazole moieties.
Figure 21. Structures of the coumarin derivatives bearing isoxazole moieties.
In another study, our researchers also reported the synthesis of twenty-five furocoumarin
derivatives and evaluated the stimulatory effect of them on melanogenesis in murine B16 cells [92]. In
this series, twenty-three compounds were more potent than the positive control (8-MOP). Compounds
137 (350.5%) and 138 (313.1%) based on the scaffold of 136 were nearly 3-fold stronger than 8-MOP
(Figure 22). Figure 21. Structures of the coumarin derivatives bearing isoxazole moieties.

Figure 22. Structures of the synthesized furocoumarin derivatives.

2.2.3. Terpenoids Figure 22. Structures of the synthesized furocoumarin derivatives.

Following the report of stimulation of melanogenesis by glycyrrhizin (GR, Figure 23) via increasing
2.2.3. Terpenoids
tyrosinase expression at mRNA and protein levels, Lee et al. [93] studied the molecular mechanism of the
Following
process. the report
The results of stimulation
indicated of melanogenesis
that GR induces melanogenesisby glycyrrhizin
by elevating(GR, Figure 23)cAMP
intracellular via increasing
level. In
tyrosinase
addition, itexpression at
to mRNA
was able Figureactivate and
bothprotein
AP-1 levels,
and CRELee et al.
pathways[93] studied
by the
increasing
22. Structures of the synthesized furocoumarin derivatives.
molecular mechanism
intracellular cAMP of the
level.
process. The results
Geniposide Figure
(Figure 24)Structures
22.
indicated that GR from
isolated of the
induces thesynthesized
melanogenesis furocoumarin
fruit of Gardenia elevatingderivatives.
byjasminoides intracellular
Ellis was used cAMP as alevel.
ChineseIn
addition,
traditionalit was able
medicine to activate
for both
treatment AP-1
of and
generalizedCRE pathways
vitiligo. In by
2008,increasing
Lan et intracellular
al. [94] studiedcAMP the level.
action
2.2.3. 2.2.3. Terpenoids
Terpenoids
and Geniposide
mechanism (Figure 24) isolated
of geniposide’s from the fruit
enhancement of Gardenia jasminoides
of melanogenesis Ellis was used as a Chinese
in norepinephrine-exposed normal
Following the report of stimulation
traditional
human
Following medicine
epidermal
the report for
oftreatment
melanocytes.
stimulation From ofof melanogenesis
of generalized
the vitiligo.
results,
melanogenesis it wasbyInglycyrrhizin
2008, Lanthat
bysuggested
glycyrrhizin (GR,
et (GR,Figure
al.geniposide
[94]
Figure23) via
studied can
23) increasing
the action
enhance
via increasing
tyrosinase
and
melanogenesisexpression
mechanism of
by at mRNA
geniposide’s
stem cell and protein signalling
enhancement
factor/c-kit levels,
of Lee et al.which
[93] studied
melanogenesis
in in
the the molecular
norepinephrine-exposed
expression of mechanism
c-kit of the
normal
receptor is
tyrosinase expression at mRNA and protein levels, Lee et al. [93] studied the molecular mechanism
process.
human
augmented The results
epidermal
in indicated
melanocytes. that
norepinephrine-exposed GR
From induces
the melanogenesis
results,
normal humanit was by elevating
suggested
epidermal thatintracellular
melanocyte. geniposide cAMP
can level.
enhance In
of theaddition,
process.itThe wasby
results
ablestem
indicated
to activate
that GRand induces
both AP-1 signalling
melanogenesis
CRE pathways
by elevating intracellular cAMP
in whichbythe increasing intracellular
of c-kit cAMP level.
level.melanogenesis
In addition,
Geniposide it was cell factor/c-kit
able to activate
(Figure 24) isolated from both AP-1
the fruit and
of GardeniaCRE expression
pathways
jasminoides by increasing receptor
Ellis was used as a Chinese
is
intracellular
augmented in norepinephrine-exposed normal human epidermal melanocyte.
cAMPtraditional
level. medicine for treatment of generalized vitiligo. In 2008, Lan et al. [94] studied the action
and mechanism of geniposide’s enhancement of melanogenesis in norepinephrine-exposed normal
human epidermal melanocytes. From the results, it was suggested that geniposide can enhance
melanogenesis by stem cell factor/c-kit signalling in which the expression of c-kit receptor is
augmented in norepinephrine-exposed normal human epidermal melanocyte.

Figure 23. Structure of glycyrrhizin isolated from Glycyrrhiza glabra L.

Figure
Figure 23.23. Structureofofglycyrrhizin
Structure glycyrrhizin isolated
isolatedfrom Glycyrrhiza
from glabra
Glycyrrhiza L. L.
glabra

Geniposide (Figure 24) isolated from the fruit of Gardenia jasminoides Ellis was used as a Chinese
traditional medicine for treatment of generalized vitiligo. In 2008, Lan et al. [94] studied the
action and mechanismFigure
of geniposide’s enhancement of melanogenesis in norepinephrine-exposed
23. Structure of glycyrrhizin isolated from Glycyrrhiza glabra L.
normal human epidermal melanocytes. From the results, it was suggested that geniposide can
Molecules 2017, 22, 1303 17 of 28

Molecules 2017, 22, 1303 17 of 27

enhance melanogenesis by stem cell factor/c-kit signalling in which the expression of c-kit receptor is
augmented in norepinephrine-exposed normal human epidermal melanocyte.
Molecules 2017, 22, 1303 17 of 27
Molecules 2017, 22, 1303 17 of 27

Figure 24. Structure of geniposide isolated from Gardenia jasminoides Ellis fruits.
Figure24.
Figure 24.Structure
Structureofofgeniposide
geniposideisolated
isolatedfrom
fromGardenia
Gardeniajasminoides
jasminoidesEllis
Ellisfruits.
fruits.
Villareal andFigure 24. Structure
co-workers [95]ofevaluated
geniposidethe isolated from Gardenia
melanogenesis jasminoides effects
stimulatory Ellis fruits.
of leaf extracts of
Erica multiflora and
Villarealand
Villareal its active
andco-workers component
co-workers[95] [95]evaluated lupenone
evaluatedthe (Figure 25) as
the melanogenesisstimulatory possible
stimulatoryeffects therapeutic
effectsof agents
of leafextracts
extracts of to
Villareal and co-workers [95] evaluated the melanogenesis
melanogenesis stimulatory effectsleaf of
of leaf extracts of
address
Ericahypopigmentation
Erica multiflora and
multiflora and its disorders.
its active
active The results
component
component showed
lupenone
lupenone that
(Figure
(Figure 25)E.
25) asmultiflora
as ethyl-acetate
possible therapeutic
possible therapeutic agentsextract
agents to and
to
Erica multiflora and its active component lupenone (Figure 25) as possible therapeutic agents to address
address
lupenone
address hypopigmentation
enhanced melanogenesis
hypopigmentation disorders. The
byThe
disorders. resultsshowed
increasing
results showed thatE.
E.multiflora
the tyrosinase
that multiflora
enzyme ethyl-acetate
expression
ethyl-acetate extract and
via mitogen-
extract and
hypopigmentation
lupenone
lupenone disorders.
enhanced
enhanced The results
melanogenesis showed
increasingthat
byextracellular the E. multifloraenzyme
tyrosinase ethyl-acetate
enzyme extract
expression via and lupenone
mitogen-
activated protein kinase melanogenesis
phosphorylated by increasing the tyrosinase
signal-regulated expression
kinases 1 and 2via mitogen-
phosphorylation
enhanced melanogenesis
activated
activated proteinkinase
protein kinase by increasing theextracellular
phosphorylated
phosphorylated tyrosinasesignal-regulated
extracellular enzyme expression
signal-regulated via11and
kinases
kinases mitogen-activated
and22phosphorylation
phosphorylation protein
inhibition.
kinase phosphorylated
inhibition.
inhibition. extracellular signal-regulated kinases 1 and 2 phosphorylation inhibition.

Figure
Figure
Figure 25.
25.25.
Figure
25. Structure
Structure ofofof
Structure
Structure of lupenoneisolated
lupenone
lupenone
lupenone isolatedfrom
isolated
isolated fromleaf
from
from leaf
leaf effects
leafeffects extracts
extracts
effects ofofErica
extracts Erica
of multiflora.
multiflora.
Erica multiflora.

Three new triterpene glycosides(lonicerosides

(lonicerosidesK, K,LLLand
Three
Three Three
newnew
new triterpene
triterpene
triterpene glycosides
glycosides
glycosides (lonicerosides
(lonicerosides K, L and
and M)
andM)M)
M) and
andand
and eleven
eleven known
known
eleven
eleven known
knowncompounds
compounds
compounds
compounds werewere
were were
isolated
isolated
isolated from from
from the
thethe aerial parts
aerialparts
aerial of Weigela
parts of Weigela
Weigela subsessilis
subsessilis [96]. Among
[96].[96].
subsessilis Among them, lonicerosides
them, them,
lonicerosides A (140) and
A (140) and
AL L (139)
(139)
isolated from the aerial parts ofof Weigela subsessilis [96]. AmongAmong lonicerosides
them, lonicerosides A (140) (140)
and Land L
(139)
were
were
(139) were proved
proved
proved to
to stimulate
stimulate
to stimulate melanogenesis
melanogenesis
melanogenesis without
without
without cytotoxicity
cytotoxicity in
in
cytotoxicity murine
murine B16-F0
B16-F0 melanoma
melanoma cells
cells
were proved
(Figure26).
toFurthermore,
stimulate
26).Furthermore,
melanogenesis
theexpression
expressionof
without
oftyrosinase
cytotoxicity
tyrosinaseand andMITF
in inmurine
MITFproteins
murineB16-F0
proteinswere
B16-F0melanoma
melanoma cells
wereupregulated
upregulatedbybyboth
both
cells
(Figure the
(Figure
(Figure 26). Furthermore,
26). according
Furthermore, the expression
the expression of tyrosinase and MITF proteins were upregulated
of tyrosinase and MITF proteins were upregulated by both by both of
ofthem
of them accordingto towestern
western blotanalysis.
blot analysis.
them according to western blot analysis.
of them according to western blot analysis.

Figure26.
Figure 26.Structure
Structureofoftriterpene
triterpeneglycosides
glycosidesisolated
isolatedfrom
fromaerial
aerialparts
partsofofWeigela
Weigelasubsessilis.
subsessilis.

Ren etet26.
Ren
Figure al. [97]
al. [97] reported
reported the isolaton
the isolaton and
and structure
structurefrom
identification
identification of two
of two new
new triterpenoids
triterpenoids
Figure 26. Structure
Structure ofof triterpene
triterpene glycosides
glycosides isolated
isolated from aerial
aerial parts
parts of
of Weigela
Weigela subsessilis.
subsessilis.
(hispindicacids
(hispindic acidsAA(141)
(141)and
andBB(142)
(142)andandaanew
newphenolic
phenoliccompound
compoundhispinine
hispinine(145),
(145),along
alongwith
withnine
nine
known compounds,
known compounds, from from the
the fruiting
fruiting bodies
bodies of
of Inonotus
Inonotus hispidus.
hispidus. Isolates
Isolates 141–146
141–146 were
were found
found toto
Ren etetal.al.[97]
[97] reported
reported the the isolaton
isolaton and structure
and structure identification
identification of two newof two new triterpenoids
triterpenoids (hispindic
exhibitedstronger
exhibited strongeractivate
activateabilities
abilitiesofofmelanogenesis
melanogenesisand andtyrosinase
tyrosinasein inB16
B16melanoma
melanomacellscellsthan
thanthose
those
(hispindic acids
acidsofApositive
(141) and A (141)
B (142)andandB (142)
a 50
new and a new phenolic
phenolic compound compound
hispininehispinine (145),with
(145), along along with
nine nine
known
control
of positive control (8-MOP)
(8-MOP) atat50 μmol/L
μmol/L (Figure
(Figure 27).
27).
known compounds,
compounds, from thefrom the fruiting
fruiting bodies ofbodies of Inonotus
Inonotus hispidus. hispidus. Isolates were
Isolates 141–146 141–146 were
found found to
to exhibited
exhibited stronger
stronger activate activate
abilities ofabilities of melanogenesis
melanogenesis and tyrosinase
and tyrosinase in B16 melanoma
in B16 melanoma cells than
cells than those those
of positive
of positive
control control
(8-MOP) at(8-MOP)
50 µmol/L at 50 μmol/L
(Figure 27).(Figure 27).
Molecules 2017, 22, 1303 18 of 28
Molecules 2017, 22, 1303 18 of 27

Molecules 2017, 22, 1303 18 of 27

Molecules 2017, 22, 1303 18 of 27

Figure 27.
Figure 27. Structure
Structureof triterpenoids
of triterpenoidsandand
phenolic compound
phenolic isolated
compound from fruiting
isolated bodies of
from fruiting Inonotus
bodies of
hispidus.
Inonotus hispidus.
Figure 27. Structure of triterpenoids and phenolic compound isolated from fruiting bodies of Inonotus
Figure 27. Structure of triterpenoids and phenolic compound isolated from fruiting bodies of Inonotus
hispidus.
2.2.4. Resveratrols
2.2.4. Resveratrols
hispidus.
2.2.4. Resveratrols
EarlyResveratrols
2.2.4.
Early in 2008,
in 2008,Guan
Guanand andco-workers
co-workers[98] [98]reported
reported2,3,5,4
2,3,5,4′-tetrahydroxystilbene-2-O-β- -glucoside
0 -tetrahydroxystilbene-2-O-β- DD-glucoside
Early
(THSG) (Figure in
(Figure 28) 2008,
28) as Guan and
astyrosinase co-workers
tyrosinaseactivator [98] reported 2,3,5,4′-tetrahydroxystilbene-2-O-β-
activatorandmelanogenesisstimulator
andmelanogenesisstimulator without -glucoside
without cytotoxicity in B16
D
(THSG) Early(Figure
in 2008, Guan and co-workers [98]andmelanogenesisstimulator cytotoxicity
reported 2,3,5,4′-tetrahydroxystilbene-2-O-β- D-glucoside in B16
(THSG)
melanoma cells. In 28) as tyrosinase
further studies, activator
it was found to induce melanin without cytotoxicity
production via in B16 the
increasing
melanoma
(THSG)
melanoma
cells.
(Figure In further
cells.28) as studies,
tyrosinase
In levels
further
ititwas
activator
studies,
found to inducemelanin
melanin
was andmelanogenesisstimulator
found to induce
production via increasing
without cytotoxicity
production via increasingin B16 the
the its
mRNA
mRNA andprotein
and protein levels of of tyrosinase.
tyrosinase. Western Western blotanalysis
blotanalysis revealed revealed
that THSG that
exertsTHSG
its exerts
stimulatory
melanoma
mRNA and cells. In further
protein levelsstudies, it was found
of tyrosinase. to induce
Western melaninrevealed
blotanalysis production
thatvia
THSGincreasing
exerts the
its
stimulatory
mRNA
effect effect
on melanogenesis on melanogenesis
and protein levels
by MAP by MAP
of kinase
tyrosinase. kinaseand
Western
activation activation
blotanalysis andrevealed
MITF induction MITFofinduction
that THSG
tyrosinase of[99].
tyrosinase
exerts its [99].
stimulatory effect on melanogenesis by MAP kinase activation and MITF induction of tyrosinase [99].
stimulatory effect on melanogenesis by MAP kinase activation and MITF induction of tyrosinase [99].

Figure 28. Structure

Structure of of THSGisolated
isolated from
from dried tuber root ofof
Polygonum multiflorum.
Polygonum multiflorum.
Figure
Figure 28.
28. 28.
Figure Structure ofofTHSG
Structure THSG
THSGisolated
isolated from
dried
fromdried tuber
driedtuber
tuber root
root
root of of Polygonum
Polygonum multiflorum.
multiflorum.
Recently,
Recently, OodeOode
et et
al.al.[100]
[100] described
described a a facile synthesis
facile of cellobioside
synthesis (149) and
of cellobioside xylobioside
(149) (150)
and xylobioside
Recently,
based Oode
Recently, et al.
Oode
on naturally [100]
[100]described
etoccurring
al. described aa facile synthesisofagent
facile synthesis
melanogenesis-controlling ofcellobioside
cellobioside (149)
(149) andand
dihydroresveratrol xylobioside
xylobioside
glucoside via (150)
(150)
(150)based
basedononnaturally
naturally occurring
occurring
melanogenesis-controlling
melanogenesis-controlling agent
agent dihydroresveratrol glucoside
basedSchmidt
on naturally occurring
glycosylation (Figure melanogenesis-controlling
29). Both analogues 149 and 150dihydroresveratrol
agent dihydroresveratrol
stimulated glucoside via via
glucoside
melanogenesis with
via Schmidt
Schmidt glycosylation
glycosylation (Figure29).29).Both
Bothanalogues
analogues 149 and 150 stimulated melanogenesiswith with
Schmidt glycosylation
efficacies to(Figure
comparable(Figure that of29). Bothwhich
8-MOP, analogues 149
149and
suggested and150
that 150stimulated
stimulated
diglycosyl melanogenesis
melanogenesis
modification of the 4′-OH with
0 -OH
efficacies comparable
efficacies comparable
on thecomparable
to that
to
dihydroresveratrol thatof 8-MOP,
of 8-MOP, which
which suggested
suggested that
that diglycosyl
diglycosyl modification
modification of of
the the
4′-OH 4
efficacies to thatskeleton
of 8-MOP, leadswhich
to the suggested
activation ofthatmelanogenesis, both with and of
diglycosyl modification without
the 4′-OH
on the
on dihydroresveratrol
the dihydroresveratrol
hydroxymethyl groups in skeleton
skeleton
the sugarleads
leads to
to the
moieties.the activation
activation ofofmelanogenesis,
melanogenesis, both
both withwith
and and without
without
on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without
hydroxymethyl
hydroxymethyl groups
groups in in thethe sugarmoieties.
sugar moieties.
hydroxymethyl groups in the sugar moieties.

Figure 29. Structures of dihydroresveratrol glucosides.

Figure
Figure 29.Structures
29. Structures of
of dihydroresveratrol
dihydroresveratrol glucosides.
glucosides.
2.2.5. Aurones
2.2.5.2.2.5.
Aurones
Aurones Figure 29. Structures of dihydroresveratrol glucosides.
In 2012, Dubois et al. [101] synthesized several aurones 151–155 and discovered that 153 and 154
In 2012, Dubois
etetal.al. [101] synthesized
synthesized several aurones 151–155 and ofdiscovered thataurones
153that
andwith
154
2.2.5.In 2012,
behaved
Aurones Dubois
as hyperbolic [101]
activators of mushroom several aurones
tyrosinase; Later,151–155
aseries and discovered
twenty-four 153 and
behaved
154 behaved as
different as hyperbolic activators
hyperbolic activators
hydroxylation of
patterns onof mushroom
A,mushroom tyrosinase;
B rings were Later,
tyrosinase;
prepared and aseries
Later, of twenty-four
aseries for
evaluated aurones
of twenty-four with
their abilitiesaurones
on
with different
In
tyrosinase
different hydroxylation
2012, Dubois
from mushroom
hydroxylation patterns
et al. [101] on on
A, respectively
tosynthesized
bacterial
patterns B rings
A, were
Bseveral
rings byprepared
aurones
were theprepared
same and
151–155 evaluated
group
andand
[102], thefor
discovered
evaluated their
results abilities
that
showed
for their 153 on
and 154
that
abilities on
tyrosinase
behaved asfrom
156–157, from mushroom
hyperbolic
158–159, activators
160–161 to bacterial respectively
of mushroom bythethe
tyrosinase; same group
Later, [102],
aseries ofthe results showed
twenty-four that
aurones
30). with
tyrosinase mushroom toand 162–163
bacterial can improve
respectively by activity
the same ofgroup
mushroom[102], tyrosinase
the results (Figure
showed that
156–157,
different 158–159, 160–161
hydroxylation patternsand 162–163
on A, can improve
B rings weretheprepared
activity of and
mushroom tyrosinase
evaluated (Figure
for their 30). on
abilities
156–157, 158–159, 160–161 and 162–163 can improve the activity of mushroom tyrosinase (Figure 30).
tyrosinase from mushroom to bacterial respectively by the same group [102], the results showed that
156–157, 158–159, 160–161 and 162–163 can improve the activity of mushroom tyrosinase (Figure 30).
Molecules 2017, 22, 1303 19 of 28
Molecules 2017, 22, 1303 19 of 27
Molecules 2017, 22, 1303 19 of 27

Figure 30. Structure of the synthetic aurones.

Figure 30. Structure of the synthetic aurones.

2.2.6. Polyphenols
2.2.6. Polyphenols
It has been reported that the extracts of Salvia officinalis L. increased the melanin production
It has been reported that the extracts of Salvia officinalis L. increased the melanin production
without necessarily changing the enzymatic activity in B16-F10 cells. Moreover, rosmarinic acid
without necessarily changing the enzymatic
enzymatic activity
activity in
in B16-F10
B16-F10 cells.
cells. Moreover, rosmarinic acid
(Figure 31), the main phenol derivative isolated from the extracts was found to exhibit a dual behavior
(Figure 31), the main phenol derivative
(Figure derivative isolated
isolated from
from the extracts was found to exhibit a dual behavior
on melanogenesis, increasing melanin biosynthesis and tyrosinase activity at low concentrations and
on melanogenesis, increasing melanin biosynthesis and tyrosinase activity at low concentrations and
decreasing them at higher levels [103]. Based on these findings, Lee et al. [104] studied the molecular
decreasing them at higher levels [103]. Based on these findings, Lee et al. [104] studied the molecular
events of pigmenting effect of rosmarinic acid in B16 melanoma cells. After several experiments, it
events of pigmenting effect of rosmarinic acid in B16B16 melanoma
melanoma cells.
cells. After several experiments, it
was believed that the compound induced melanogenesis through the PKA activation signaling.
compound induced
was believed that the compound induced melanogenesis
melanogenesis through
through the
the PKA
PKA activation
activation signaling.
signaling.

Figure
Figure 31.
31. Structure of rosmarinic acid.
Figure 31. Structure of rosmarinic acid.

2.2.7. Other Compounds

2.2.7. Other Compounds
In 2014, Zhu et al. [105] assessed the role of epigallocatechin-3-gallate (EGCG) (Figure 32) in
In 2014, Zhu et al. [105] assessed
assessed the role
role of
of epigallocatechin-3-gallate
epigallocatechin-3-gallate (EGCG) (Figure 32) in
vitiligo induced by monobenzone in mice. It was demonstrated that EGCG could delay the time of
vitiligo induced by monobenzone in mice. It was demonstrated that EGCG could delay the time of
depigmentation, reduce the prevalence of depigmentation and decrease the area of depigmentation
depigmentation, reduce the prevalence of depigmentation and decrease the area of depigmentation
by reducing excessive inflammatory responses, especially infiltration of CD8+ T cells and inhibiting
by reducing excessive inflammatory responses, especially infiltration of CD8+ T cells and inhibiting
the levels of inflammatory mediators. In addition, gene-expression profile of the model in relation to
the levels ofof inflammatory
inflammatorymediators.
mediators.InInaddition,
addition,gene-expression
gene-expression profile ofof
profile the model
the modelin in
relation
relationto
EGCG was studied as well. A total of 1264 down-regulated genes and 1332 up-regulated genes were
EGCG
to EGCG was wasstudied
studiedas well. A total
as well. of 1264
A total down-regulated
of 1264 down-regulated genesgenes
and 1332 up-regulated
and 1332 genesgenes
up-regulated were
recorded in the 5% EGCG group compared with the model group using whole genome oligo
recorded
were in thein 5%
recorded the EGCG
5% EGCG group compared
group compared with thethe
with model
modelgroup
groupusing
usingwhole
whole genome
genome oligo
microarray assay. Thesere sults suggested that EGCG may significantly decrease the risk of vitiligo.
microarray assay.
Molecules 2017,assay. Thesere sults
22, 1303Thesere sults suggested
suggested that
that EGCG
EGCG maymay significantly
significantly decrease
decreasethetherisk
riskof
ofvitiligo.
vitiligo.
20 of 27
The extract of Piper methysticum rhizome (Kava) was identified as the most potent agent on
The extract of Piper methysticum rhizome (Kava) was identified as the most potent agent on
melanogenesis in B16 cells among different parts of five Piper species [106]. Activity-guided
melanogenesis in B16 cells among different parts of five Piper species [106]. Activity-guided
fractionation of Kava extract led to the isolation of six compounds, with two active kavalactones,
fractionation of Kava extract led to the isolation of six compounds, with two active kavalactones,
yangonin (165) and 7,8-epoxyyangonin (168), a glucosyl-steroldaucosterin (169), which all possess a
yangonin (165) and 7,8-epoxyyangonin (168), a glucosyl-steroldaucosterin (169), which all possess a
significant stimulatory effect on melanogenesis (Figure 33).
significant stimulatory effect on melanogenesis (Figure 33).
Hirata and co-workers had discovered that the natural product (−)-cubebin (Figure 34) isolated
Hirata and co-workers had discovered that the natural product (−)-cubebin (Figure 34) isolated
from leaves of Piper nigrum L. wa sproved to have a stimulator effect on melanogenesis and tyrosinase
from leaves of Piper nigrum L. wa sproved to have a stimulator effect on melanogenesis and tyrosinase
activity in murine B16 melanoma cells without any significant effects on cell proliferation [107]. The
activity in murine B16 melanoma cells without any significant effects on cell proliferation [107]. The
expression levels of tyrosinase and tyrosinase
Figure
mRNA
Structure mRNA
were both enhanced after addition of cubebin.
of epigallocatechin-3-gallate.
expression levels of tyrosinase and32.
tyrosinase were both enhanced after addition of cubebin.
Western blot analysis revealed that melanogenesis induced by cubebin was attributable to the
Western blot analysis revealed that melanogenesis induced by cubebin was attributable to the
increase
Theof tyrosinasegene
of Piper expression
methysticum through positive regulator, MITF,initiated
as the by cubebin-induced
increase ofextract
tyrosinasegene expression rhizome
through (Kava)
positive was identified
regulator, MITF,initiated most
by potent agent
cubebin-induced
activation
on of p38-MAPK,
melanogenesis in B16ascells
shown in Figure
among 35. parts of five Piper species [106]. Activity-guided
different
activation of p38-MAPK, as shown in Figure 35.
fractionation of Kava extract led to the isolation of six compounds, with two active kavalactones,
 Molecules 2017, 22, 1303 20 of 27

Molecules 2017, 22,

Molecules 2017, 22, 1303
1303 20
20 of
of 28
27
Molecules 2017, 22, 1303 20 of 27

yangonin (165) and 7,8-epoxyyangonin (168), a glucosyl-steroldaucosterin (169), which all possess a
Figure
significant stimulatory effect on 32. Structure of(Figure
melanogenesis epigallocatechin-3-gallate.
33).

Figure 32. Structure of epigallocatechin-3-gallate.

Figure 32. Structure of epigallocatechin-3-gallate.

Figure 33. Structure of kavalactones and glucosylsterols isolated from the rhizome extract of Piper
Figure 33. Structure of kavalactones and glucosylsterols isolated from the rhizome extract of
methysticum.
Piper methysticum.

Hirata and co-workers had discovered that the natural product (−)-cubebin (Figure 34) isolated
from leaves of Piper nigrum L. wa sproved to have a stimulator effect on melanogenesis and tyrosinase
activity in murine B16 melanoma cells without any significant effects on cell proliferation [107]. The
expression levels of tyrosinase and tyrosinase mRNA were both enhanced after addition of cubebin.
Western blot33.
Figure analysis revealed
Structure that melanogenesis
Figure
of kavalactones and34. induced
Structure by cubebin
of (−)-cubebin.
glucosylsterols isolated from was attributable
the rhizome to of
extract thePiper
increase
Figure 33. Structure of kavalactones and glucosylsterols isolated from the rhizome extract of Piper
of tyrosinasegene
methysticum. expression through positive regulator, MITF, initiated by cubebin-induced activation
methysticum.
of p38-MAPK, as shown in Figure 35.

Figure 34. Structure

Structure of ((−)-cubebin.
−)-cubebin.
Figure 34. Structure of
Figure 34. of (−)-cubebin.

Figure 35. A proposed scheme showing the activation mechanism of cubebin on melanogenesis [107].

In 2008, Faas et al. [108] investigated the ability of PIP and three analogues (THP, CHP and
rCHP, Figure 36) to stimulate pigmentation in a strain of sparsely pigmented mice. The results
showed that treatment with PIP, THP or rCHP and UVR induced a marked pigmentation response
in HRA/Skh-II mice, with clinically better results than UVR alone, which supported the potential use
of these compounds in treating vitiligo.

Figure 35. A proposed scheme showing the activation mechanism of cubebin on melanogenesis [107].
Figure 35. A
A proposed
proposed scheme
scheme showing
showing the
the activation
activation mechanism
mechanism of cubebin
cubebin on
on melanogenesis
melanogenesis [107].

In 2008,
In 2008, Faasetetal.al.[108]
[108] investigated the ability of PIP and three analogues (THP, CHP and
In 2008, Faas
Faas et al. [108]investigated
investigated thethe
ability of PIP
ability andand
of PIP threethree
analogues (THP,(THP,
analogues CHP and
CHPrCHP,
and
rCHP,
Figure Figure
36) to 36) to
stimulate stimulate
pigmentation pigmentation
in a strain in
of a strain
sparsely of sparsely
pigmented pigmented
mice. The mice.
results The results
showed that
rCHP, Figure 36) to stimulate pigmentation in a strain of sparsely pigmented mice. The results
showed that
treatment treatment
with PIP, THP with
or PIP, THP
rCHP and or rCHP
UVR and aUVR
induced induced
marked a markedresponse
pigmentation pigmentation
in response
HRA/Skh-II
showed that treatment with PIP, THP or rCHP and UVR induced a marked pigmentation response
in HRA/Skh-II
mice, mice, with
with clinically betterclinically better
resultsbetter results
than results than UVR
UVR alone, whichalone, which supported
supported the potential
the potential use
use of these
in HRA/Skh-II mice, with clinically than UVR alone, which supported the potential use
of these
compounds compounds in
in treating treating vitiligo.
vitiligo. vitiligo.
of these compounds in treating
Molecules 2017,
Molecules 22, 1303
2017, 22, 1303 21 of
21 of 27
28
Molecules 2017, 22, 1303 21 of 27

Figure 36. Structure of PIP and three analogues.

Figure
Figure 36.
36. Structure
Structure of
of PIP
PIP and
and three
three analogues.
3. Conclusion
3. Conclusion
Conclusions
Melanogenesis stimulators are important as skin-pigmentation agents in several dermatological
Melanogenesis
Melanogenesis
problems, such asstimulators
stimulators
vitiligo. Overaretheimportant
are
last ten as skin-pigmentation
important
years, as skin-pigmentation
a number agents in
of melanogenesis several
agents
stimulators dermatological
haveinbeen several
problems,
dermatological
reported suchfromas vitiligo.
problems,
natural and Over
such the
as last
synthetic ten years,
vitiligo.
sources. Over a the
number
Although last
none of
tenofmelanogenesis
years, a number
them have stimulators
revealed of the have been
melanogenesis
precise
reported
stimulators from
therapeutic havenatural
target
been and
of reportedsynthetic
the vitiligo, many
from sources.
possible
natural Although
andmechanisms
synthetic noneof of them
these
sources. have in
stimulators
Although revealed of the
melanogenesis
none themprecise
have
were
therapeutic proposed
revealed thetarget
precise to help us
oftherapeutic understand
the vitiligo,target this complicated
manyofpossible disease.
mechanisms
the vitiligo, many possible of these stimulators
mechanisms ofin melanogenesis
these stimulators
were Many melanogenic
proposed
in melanogenesis to were
help us stimulating
understand
proposed agents
to help target catalytic
thisuscomplicated
understand activity
disease.
this of TYR. TYR
complicated catalyzes the process
disease.
of neuromelanin production, in which oxidation of dopamine produces dopaquinones.
Many
Many melanogenic
melanogenic stimulating
stimulating agents
agentstarget catalytic
target activity
catalytic of TYR.
activity of TYR TYR However,
TYR.catalyzes the process
catalyzes the
excessive production of dopaquinones results in neuronal damage and cell death [109]. This links
of neuromelanin
process production,
of neuromelanin in which in
production, oxidation of dopamine
which oxidation produces dopaquinones.
of dopamine produces dopaquinones.However,
TYR to several neurodegenerative disorders such as Parkinson’s as mentioned above [110].
excessive
However, production
excessive of dopaquinones
production of results in neuronal
dopaquinones results indamage
neuronal and cell death
damage and [109].
cell This [109].
death links
In addition to that, other approaches to melanogenesis stimulators include an increase and activation
TYR
This ofto
links several
the TYR neurodegenerative
to several
expression disorders
neurodegenerativeproteins
of melanogenesis-related such
disorderssuch as
such Parkinson’s
as Parkinson’s
as MITF, as
TYR, TRP-1asandmentioned
mentioned above [110].
above [110].
TRP-2, resulting
In addition
from thetomodulation
addition that,
that, other
otherofapproaches
approaches to
to melanogenesis
various signaling pathways. stimulators
melanogenesis stimulators include include an an increase
increase and and activation
activation
of the expression
However,of
expression melanogenesis-related
a pity that most of theseproteins
melanogenesis-related
it is proteins such
such as
melanogenesis as MITF,
MITF, TYR,
stimulators TRP-1
are still in theand
drug TRP-2, resulting
discovery
fromphase,
the modulation of various signaling
and few pharmacologic actions and pathways.
adverse reactions in vivo are reported as a result of the
difficulty
However,initbuilding
is a pitythe thatrelated
most of animal
thesemodels. Accordingstimulators
melanogenesis
melanogenesis to the current are research
are still
still in
in theprogress
the drug and
drug discovery
discovery
problems,
phase, and
and few it was more
few pharmacologic efficient
pharmacologic actions to develop
actions and new
and adverse drugs from
adverse reactions traditional
reactions in in vivo Chinese
vivo are medicines,
are reported
reported as as aa resultasof the
such
Uighur medicine. Many drugs for vitiligo have already been to
difficulty available on the market for years.
difficulty in building the related animal models. According According to the the current
current research
research progress
progress and
Qubaibabuqi and Kaliziri [46,111,112], which exhibit attractive therapeutic effects clinically with few
problems,
problems, itit waswas moremoreefficient
efficienttotodevelop
developnew newdrugs
drugsfrom from traditional
traditional Chinese
Chinese medicines,
medicines, suchsuchas
side effects, were popular in Xinjiang and its neighboring central Asian countries. Besides, JAK
Uighur
as Uighur medicine.
medicine. Many Many drugs
drugsfor vitiligo have already been available on the market for years.
inhibitors (Figure 37), often used forfor vitiligo have already
myelodysplastic disorders,been available
were proven on theefficacious
to be market for foryears.
Qubaibabuqi
vitiligo recently and a large number of clinical trials are currently underway [113,114].JAK inhibitors few
and Kaliziri [46,111,112],
[46,111,112], which
which exhibit
exhibit attractive
attractive therapeutic
therapeutic effects
effects clinically
clinically with
with few
side effects,
side are were popular in Xinjiang and its
its neighboring
neighboring central
central Asian
Asian
likely to have broad applicability in dermatology and more lead compounds for vitiligo may be countries.
countries. Besides,
Besides, JAK
JAK
inhibitors
inhibitors
explored (Figure
among37),
(Figure 37), often
often
these used
used in
inhibitors for myelodysplastic
forview
myelodysplastic disorders,
disorders, were
of their clear target. were proven
proven to to be
be efficacious
efficacious for
vitiligo recently and a large number of clinical trials are currently
currently underway
underway [113,114].
[113,114].JAK
JAK inhibitors
inhibitors
are likely to have broad applicability in dermatology
dermatology and and more lead compounds
compounds for vitiligo may be
explored among these inhibitors in view of their clear target.

Figure 37. Structure of JAK inhibitors.

In conclusion, we hope that this perspective will be useful to medicinal chemists working on
melanogenesis and related proteins to identify novel melanogenesis stimulators with drug-like
properties.
Figure 37. Structure of JAK inhibitors.
Figure 37. Structure of JAK inhibitors.

In
In conclusion,
conclusion, we
wehope
hopethat
thatthis
thisperspective
perspectivewill bebe
will useful to to
useful medicinal chemists
medicinal working
chemists on
working
melanogenesis and related
on melanogenesis proteins
and related to identify
proteins novel melanogenesis
to identify stimulators
novel melanogenesis with drug-like
stimulators with
properties.
drug-like properties.
Molecules 2017, 22, 1303 22 of 28

Acknowledgments: This work was supported by the Funds for the Xinjiang Key Research and Development
Program (2016B03038-3); “Personalized Medicines-Molecular Signature-based Drug Discovery and Development”,
Strategic Priority Research Program of the Chinese Academy of Sciences (XDA12050301); West Light Foundation
of the Chinese Academy of Science (No. XBBS201403).
Author Contributions: Haji A. Aisa conceived the papers. Chao Niu wrote the manuscript. Both authors have
read and approved the final manuscript.
Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations
UV Ultra-violet
α-MSH α-Melanocyte-stimulating hormone
SCF Stem cell factor
ET-1 Endothelin-1
NO Nitric oxide
ACTH Adrenocorticotropic hormone
MITF Microphthalmia-associated transcription factor
TYR Tyrosinase
TRP-1 Tyrosinase-related protein 1
TRP-2 Tyrosinase-related protein 2
Dct Dopachrome tautomerase
L-DOPA 3,4-Dihydroxyphenylalanine
DQ Dopaquinone
DHI 5,6-Dihydroxyindole
DHICA 5,6-Dihydroxyindole-2-carboxylic acid
IQ Indole-5,6-quinone
5-S-CD 5-S-cysteinyldopa
2-S-CD 2-S-cysteinyldopa
p38 MAPK p38 Mitogen-activated protein kinase
8-MOP 8-Methoxypsoralen
PMRP Polygoni multiflori radix praeparata
EH Ecliptae herba
RRP Rehmanniae radix praeparata
TNF-α Tumor necrosis factor
CGA Chlorogenic acid
RA Rosmarinic acid
BV Bee venom
GSK3β Glycogen synthase kinase-3β
PI3K Phosphatidylinositol 3-kinase
MAPKs Mitogen-activated protein kinases
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
EDCI
hydrochloride
HOBt 1-Hydroxybenzotriazole
SAR Structure-activity relationship
CREB cAMP response element binding protein
ERK Extracellular signal-regulated kinases
AP-1 Activator protein-1
PKA Protein kinase AA

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