Review

published: 26 November 2019

doi: 10.3389/fphar.2019.01421

Dermaseptins, Multifunctional
Antimicrobial Peptides: A Review
of Their Pharmacology, Effectivity,
Mechanism of Action, and Possible
Future Directions
Emiel Jacob Henri Bartels 1, Douwe Dekker 2* and Mohamed Amiche 3*
1University Medical Center, University Utrecht, Utrecht, Netherlands, 2 Dutch Poisons Information Center, University Medical
Center Utrecht, Utrecht, Netherlands, 3 Mondor Institute of Biomedical Research, INSERM U955 Team 7, School of
Medicine, University Paris Est Créteil, Créteil, France

Dermaseptins are a group of α-helical shaped polycationic peptides isolated from the
Hylid frogs, with antimicrobial effects against bacteria, parasites, protozoa, viruses in vitro.
Besides, anti-tumor effects have been demonstrated. However, few animal experiments
Edited by:
and no clinical trials have been conducted thus far. This review summarizes the current
Banasri Hazra,
Jadavpur University, India knowledge on the pharmacology, ethno pharmacology, effectivity against infectious
Reviewed by: pathogens and tumors cells and the mechanism of action of the Dermaseptins. Future
Sol Cristians, research should focus on further clarification of the mechanisms of action, the effectivity
National Autonomous University of
Mexico, Mexico of Dermaseptins against several cancer cell lines and their applicability in humans.
Ran Wang,
Keywords: dermaseptin, Phyllomedusa bicolor, peptide, amphibian, infectious, antimicrobial, tumor
Tianjin Medical University, China

*Correspondence:
Douwe Dekker
d.dekker@umcutrecht.nl
INTRODUCTION
Mohamed Amiche
mohamed.amiche@u-pec.fr
Dermaseptins (DRSs) are a family of peptides that are part of the skin secretions of several Hylid
frogs, particularly from the Agalychnis and Phyllomedusa family (Nicolas and Amiche, 2006; Amiche
Specialty section:
et al, 2008). In 1991, the first DRS was identified and characterized as a peptide rich in basic amino
This article was submitted to acids with a high propensity to adopt an α-helical structure in a hydrophobic medium (Mor et al,
Ethnopharmacology, 1991). The DRS-S1 was purified by reverse phase high-pressure liquid chromatography (RP-HPLC)
a section of the journal from a skin extract of the Phyllomedusa sauvagii and sequenced by Edman’s method. By now, there
Frontiers in Pharmacology are more than a hundred DRS-like peptides classified in the large family of DRSs that share a strong
Received: 22 April 2019 identity in the cDNA sequences encoding their biosynthetic precursors (Nicolas and El Amri, 2009).
Accepted: 07 November 2019 DRSs are often classified as antimicrobial peptides (AMPs), since they show effectivity in vitro
Published: 26 November 2019 against some gram positive and gram negative bacteria, parasites, yeasts, protozoa, viruses and
Citation: display immune modulatory effects (Mor et al., 1994a; Mor et al., 1994b; Strahilevitz et al., 1994;
Bartels EJH, Dekker D and Amiche M Charpentier et al., 1998; Brand et al., 2002; Navon-Venezia et al., 2002; Brand et al., 2006; Conceicao
(2019) Dermaseptins, Multifunctional et al., 2006; Conlon et al., 2007; Leite et al., 2008; Galanth et al., 2009; Nicolas and El Amri, 2009;
Antimicrobial Peptides: A Review of
Jiang et al., 2014; Zairi et al., 2014; Huang et al., 2017). Beside these antimicrobial properties, DRSs
Their Pharmacology, Effectivity,
Mechanism of Action, and
show activity against several human cancer types (Charpentier et al., 1998; Conlon et al., 2007;
Possible Future Directions. Nicolas and El Amri, 2009; Galanth et al., 2009; Van Zoggel et al., 2012; Shi et al., 2016; Huang
Front. Pharmacol. 10:1421. et al., 2017; Dos Santos et al., 2017; Zhu et al., 2018) and can therefore be classified as an anticancer
doi: 10.3389/fphar.2019.01421 peptide as well.

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Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

In view of the current increasing bacterial resistance to or without experienced guidance. These include a transient
conventional antibiotics (Laxminarayan et al., 2013) the demand syndrome of inappropriate antidiuretic hormone secretion
for novel antibacterial pharmaceuticals is high. Likewise, there (Leban et al., 2016), presumed drug induced liver injury
is a need for novel anti-tumor treatments as cancer is rapidly in a chronic alcoholic (Pogorzelska and Lapinski, 2017),
becoming the leading cause of death in the Western world sudden death upon chronic kambo use in which autopsy
and conventional therapeutics are both cytotoxic and prone to revealed underlying heart disease possibly related to reduced
therapy resistance due to microevolution of the tumor tissue myocardial perfusion (Aquila et al., 2018) and delayed kambo
(Corrie, 2008; Colak and Medema, 2014). related symptoms in a 24-year-old woman 22 hours after the
The aim of this review is to summarize the current knowledge ritual (Li et al., 2018).
on DRSs, to elaborate on the ethnopharmacology, the potential The short-lasting effects of kambo are related to a diversity of
therapeutic values with respect to their anti-microbial and anti- biologically active peptides besides DRS including Phyllokinin
tumor potency, and to suggest future directions for research. We (a bradykinin), Phyllomedusa (a tachykinin), Sauvagine
thereby restricted ourselves to DRSs (sensu stricto) according to (vasodilator), Caerulin (a CCK like peptide) and Deltorphin/
the nomenclature proposed by Amiche et al. (2008). Demorphin (opioid receptor agonists). Studies on these
peptides have contributed greatly to our knowledge concerning
ETHNOPHARMACOLOGY IN PEPTIDE the μ/δ-opioid and serotonin receptor (Erspamer et al., 1993),
DISCOVERY but are beyond the scope of this review. Many of the immediate
effects of Kambo can be explained by peptides in the frog-
Many plants and animal products have yet found their way from secretion. Vasoactive properties of some of these peptides
traditional use to Western medicine leading to the discovery for example might contribute to the rapid absorption of the
of for example morphine, codeine, quinine, aspirin, curare, secretion into the fresh burn. Amongst these peptides, DRS is
pilocarpine and ACE-inhibitors (Bisset, 1991; Alves and Alves, the primary candidate for the anti-microbial effects reported
2011; Dias et al., 2012). Likewise, DRS-B’s from the Phyllomedusa upon the use of Kambo.
bicolor secretions are yet known to be applied for human use in
traditional medicine. This Phyllomedusa bicolor is an Amazonian
amphibian found in the forests of Brazil, the Guianas, Venezuela, STRUCTURAL AND PHARMACOLOGICAL
Colombia, Peru, and Bolivia. The secretions of this frog are ANALYSIS OF THE DRS FAMILY
referred to by ‘Kambo’,’Kampu, or ‘Sapo’, and are used by natives
as a medicine and part of a cleansing ritual. The use of Kambo Even though each DRS has a unique amino acid sequence
by South American Indians was first described by Constantin and selectivity patterns towards microorganisms and tumors,
Tastevin in 1925 in the Kachinaua, Kurina, and the Kanamari there are also a lot of pharmacological similarities. Figure 1
tribes (De Lima and Labate, 2008). Later, the traditional use of shows the amino acid sequence alignment of 57 DRSs
kambo was also documented in the Katukina tribe, the Mayoruna sequences extracted from the antimicrobial peptide databases
tribe, and the Matse tribe (De Lima and Labate, 2008; Gorman, (http://aps.unmc.edu/AP/main.php). There is amino-acid
2015). Other DRS secreting frogs have not been documented to sequence similarity within DRS from the same frog (DRS-B1–
be useful in rituals. B6; 33–62%) (Charpentier et al., 1998), but also between DRS
The Kambo-ritual is characterized by an immediate and from different species (DRS-B1 and DRS-S1; 81%, DRS-B2,
short-lived physical response followed by a longer lasting mental and DRS-D 84.8%) (Mor et al., 1994b; Auvynet et al., 2008).
and minor physical effects. The Indians ‘harvest’ the frog by They are mostly rather short peptides (21–34 residues) with
collecting the secretion from a live frog and transfer its secretions a highly-preserved tryptophan residue on the 3rd position
to a bamboo stick. The medicine may then be used to cure or from the N-terminus (Nicolas and El Amri, 2009) except for
prevent illness, to expel ‘panema’ (bad spirit), or even to induce DRS-S10, DRS-S13, DRS-C3, and DRS-A4. DRSs can be fitted
an abortion (Gorman, 2015). When unlucky in hunting Kambo into an amphipathic α-helix with their hydrophobic residues
reportedly increases stamina, and sharpens senses during long on one face and the polar cationic residues in cluster on the
hunts. Application involves burning dots on the skin, usually on opposite face. They usually do this is an anionic environment,
arms and/or legs, and sticking a small dose (10 mg) ‘dot’ on the or under the influence of certain phospholipids (Hoskin
open wound. The symptoms are severe and immediate; violent and Ramamoorthy, 2008). These charged clusters tend to
nausea, vomiting, diarrhea, edema of the face and headaches. differ among DRS, for example; DRS-B1 has a narrow polar
Symptoms last until the secretion is removed from the wound face of a mean radial angle of 115°, DRS-B2 has a polar face
usually after 15–20 minutes. covering almost half of the helix; 175° and DRS-S1 a polar
Besides its traditional use in the Amazon, Kambo has face of 145° (Mor et al., 1994a). Furthermore, they show
found its way into the Western alternative healing scene as great variation in net charge and density of charge. DRS-S9
well. While the reports on the beneficial effects of this ritual seems to be an exception as it has a highly hydrophobic core
are numerous and range from relieving symptoms of pain flanked by cationic residues (Lequin et al., 2006; Caillon et al.,
syndromes, autoimmune diseases, skin disease, and cancer 2013). Thus far, several DRS have been in vitro tested for their
to substance abuse and depression (Hesselink, 2018), so are activity against various microorganisms (Lorin et al., 2005;
the accounts of adverse effects of participating in a rite, with Savoia et al., 2010; de Moraes et al., 2011; van Zoggel et al.,

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Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

FIGURE 1 | Amino acid sequence alignment of 57 DRS peptides (http://aps.unmc.edu/AP/main.php) using CLUSTAL O (1.2.4) multiple sequence alignment
(https://www.ebi.ac.uk/Tools/msa/clustalo/). The letters represent an amino acid according to conventional nomenclature. The dashes are introduced to optimize
amino-acids alignments. Blue colors indicate a well-preserved amino acid (>55% of identity), green colors indicate higher rates of preservation within this DRS group
(>80% of identity).

2012a; Zairi et al., 2014), DRS-B2 has been tested in mice RESEARCH IN INFECTIOUS DISEASES
and rats in vivo for tumor and antimicrobial activity (Navon-
Venezia et al, 2002; Huang et al., 2017). Though reportedly Antibacterial Activity of Peptides From the
administered to humans in non-experimental settings (DRS- DRS Family
B’s), robust data on pharmacokinetics, efficacy and safety in In 1991, Mor was the first to publish on the anti-microbial
humans are currently lacking. properties of DRS (Mor et al., 1991). DRS-B1 and S1 show in

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Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

vitro activity against gram positive and negative bacteria with Simplex Virus 1 & 2, including the acyclovir resistant strain in
various specificities (Strahilevitz et al., 1994). Derivatives vitro (Belaid et al., 2002; Bergaoui et al., 2013). For the activity
of DRS-S4, DRS-CA1, DRS-DU1 and DRS-PH show in peaks in case of DRS administration prior to incubation with
vitro activity against Staphylococcus aureus (including the virus, interference early in the viral replication cycle is
the methicillin resistant strain), Pseudomonas aeruginosa hypothesized (Belaid et al., 2002; Mechlia et al., 2018). DRS-
and E. coli, even when they are formed in biofilm (Navon- S4 and S9 both demonstrate in vitro activity against HIV-1
Venezia et al., 2002; Zairi et al., 2014; Liu et al., 2017; Zhu virus (Lorin et al., 2005; Wang et al., 2010) by inhibition of
et al., 2018). Though less cytotoxic compared to conventional HIV attachment to endometrial cells, uptake by the dendritic
antibiotics (Zairi et al., 2014), the DRS-S4 derivatives used cells and subsequent transmission to T-cells (Lorin et al.,
demonstrated similar or even higher efficacy in vitro(Porat 2005). Again, interference of DRS-S4 in an early phase of virus
et al., 2006; Rotem et al., 2006; Marynka et al., 2007; Jiang replication is suggested as less reduction of HIV is observed
et al., 2014). These results were confirmed in vitro and in once the T-cells have been infected. Substitution of methionine
vivo with mice infected with P. aeruginosa (Navon-Venezia for lysine on the 4th position DRS-S4 to reduce cytotoxicity to
et al., 2002; Marynka et al., 2007) and in various incubation mammalian cells, did not affect the anti-HIV activity observed
media varying in temperature and acidity (Rydlo et al., 2006). (Lorin et al., 2005). More recently, DRS-S4 has shown effectivity
Table 1 provides an overview of the activity of a few DRSs against Rabies virus in mice (Mechlia et al., 2018).
against selected pathogens frequently related to infections and
associated with anti-biotic resistance. For a complete list of Antifungal Activity of Peptides From the
DRSs and pathogens with corresponding MIC and source, we DRS Family
refer to Supplementary Table 1s. DRSs also show activity against fungi in vitro. So far, DRS-B1-B2,
DRS-S1-S5 DRS-O1, DRS-CA1 DRS-DU1 all demonstrate
Antiviral Activity of Peptides From the cytotoxicity against Candida albicans (Strahilevitz et al., 1994;
DRS Family Mor et al., 1994a; Mor et al., 1994b; Leite et al., 2008; Nicolas
In addition to the antibacterial efficacy, DRS-S1 (and and El Amri, 2009; Shi et al., 2016; Huang et al., 2017; Zhu
derivatives) show activity against pathogens causing genital et al., 2018). In particular, DRS-S3 shows anti-fungal activity
infections such as human papilloma virus (HPV) and herpes by means of triggering apoptosis (Morton et al., 2007). In
simplex virus (HSV) (Savoia et al., 2010). Furthermore, Aspergillus fumigates, cytotoxic activity was demonstrated for
modified DRS-S4 has shown antiviral activity against Herpes DRS-B1-B2 and DRS-S1, but not DRS-S5. Minimal inhibitory

TABLE 1 | Effectivity of Dermaseptins in vitro against various pathogens.

E. coli E. faecalis P. aeruginosa S. aureus C. albicans A. fumigatus Human

erythrocytes

DRS-B1
DRS-B2
DRS-B3
DRS-B4
DRS-S1
DRS-S2
DRS-S3
DRS-S4
DRS-S4 K4-S4(1-16)
DRS-S5
DRS-PH
DRS-H3
DRS-L1
DRS-O1
DRS-DI06
DRS-CA1
DRS-DU1
DRS-PD1
DRS-PD2
DRS-PS4

This table gives an overview of the effectivity of DRS activity against a selected number of pathogens. Cells are colored dark green if: more than one study agrees on high
activity (MIC <10 μM); light green if: 1 study finds high activity (MIC <10 μM); orange if studies do not agree on MIC; red if one or more studies agree on low activity (MIC >
10 μM); white indicates that there was no published data available. In the Human erythrocyte column, the opposite is instinctively true; green color indicates low, and red
indicates high activity (Strahilevitz et al., 1994; Mor et al., 1994a; Mor et al., 1994b; Carpentier et al., 1998; Brand et al., 2002; Navon-Venezia et al., 2002; Conceicao et al.,
2006; Brand et al., 2006; Conlon et al., 2007; Galanth et al., 2009; Zairi et al., 2014; Huang et al., 2017; Nicolas and El Amri, 2009; Leite et al., 2008; Jiang et al., 2014; De
Assis et al., 2016; Shi et al., 2016; Belmadani et al., 2018; Zhu et al., 2018; Shams et al., 2019) for more information, see Supplementary Table S1.

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Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

concentrations (MIC) ranging from 3.1 μM to 30 μM were positive stained endothelial cells. No side-effects or differences in
observed. In addition, others (Belmadani et al., 2018) found total blood count were observed (van Zoggel et al., 2012).
that DRS-S1 inhibits C. albicans in biofilm formation when Furthermore, DRS-PH shows IC50’s of 0.69 µM, 2.01 µM, and
using concentrations of 100 μM. These findings are summarized 2.36 µM, against breast cancer adenoma (MCF-7), non-small cell
in Table 1. lung carcinoma (NSCLC) (H157), and glioblastoma (U251MG)
in vitro respectively (Huang et al., 2017). For DRS-PH less efficacy
was observed against PC3 cell lines, compared to the activity
Antiparasitic Activity of Peptides From the profile of DRS-B2. DRS-PH did also show some cytotoxicity
DRS Family against human dermal endothelium, and mammalian red blood
Finally, DRS-S3 and S4 derivatives appear able to target malarial cells (Huang et al., 2017).
parasites within a host erythrocyte without disrupting the host
(Ghosh et al., 1997; Krugliak et al., 2000; Dagan et al., 2002).
In addition DRS from the Phyllomedusa oreades (DRS-O1) has MECHANISM OF ACTION
shown activity against Schistosoma mansoni (de Moraes et al.,
2011), Trypanosoma cruzi (Brand et al., 2002; Leite et al., 2005) The mode of action by which antimicrobial peptides kill microbes
and Leishmania amazonesis (Brand et al., 2006). is mainly known for α-helix cationic peptides which have been
extensively studied (Amiche and Galanth, 2011; Melo and
Castanho, 2012; Bahar and Ren, 2013). Two models explaining the
RESEARCH IN ONCOLOGY interaction of α-helical cationic AMPs with membranes have been
proposed: the barrel-stave model (Ehrenstein and Lecar, 1977) and
Parallel to the efficacy against microbials, efficacy against tumor the carpet or carpet model (Pouny et al, 1992), both taken over
cells has been studied as well (Hoskin and Ramamoorthy, 2008; by Shai in 1999 (Shai, 1999). The cationic antimicrobial peptides,
Balandin et al., 2016). Table 2 summarizes the data published on destructured in aqueous media, adopt an α-helical structure in
the impact of DRS on selected cancer cell lines. contact with the plasma membranes of the host cell and then interact
DRS-B2 for example, shows a dose dependent growth with the negative charges of the components of the membrane
inhibition of prostatic adenocarcinomas (GI50 = 0.71–2.65µM) surface (Zasloff, 2002). After binding, the peptide will disrupt the
and some pancreatic cancer cell lines. Administration of 1µM permeability of the membrane and either cause the death of the
DRS-B2 was enough for 50% reduction in colony formation in microorganism or enter the cell compartment and interact with
both prostate adenoma and mammary carcinoma cell lines (van intracellular targets. Note that most cationic antimicrobial peptides
Zoggel et al., 2012). No growth inhibition on glioblastoma and have a direct action on the membrane of bacteria, but some such
mammary carcinoma cell lines was observed (van Zoggel et al., as buforine II act intracellularly (Park et al, 1998). In most of the
2012; Dos Santos et al., 2017). At a concentration of 15 µM, no mechanisms described, the binding of antimicrobial peptide to the
activity against stromal prostate fibroblasts and skin fibroblasts membrane is followed by permeabilization of the membrane, which
was observed, indicating low cytotoxicity in surrounding tissue alone can cause cell death, or as a step in more complex processes.
(van Zoggel et al., 2012; Dos Santos et al., 2017). In vivo, DRS did In addition, very few studies on the anti-tumour action
not arrest human PC3 tumor growth in mice, but inhibited growth mechanism(s) of antimicrobial peptides have been conducted.
with more than 50% vs. controls attributed to a 24% reduction of The most important results show that their oncolytic mechanisms
angiogenesis in tumors of treatment groups, quantified by CD34+ include: (i) induction of necrosis via cell membrane lysis, (ii)

TABLE 2 | Overview of the activity DRSs against various cancer cell types.

HEPG2 MCF-7 U251MG H157 MDA-MB-435S PC-3 Human

erythrocytes

DRS-B2
DRS-B3
DRS-B4
DRS-PH
DRS-L1
DRS-CA1
DRS-DU1
DRS-PD1
DRS-PD2
DRS-PS4

This table gives an overview of the DRS that have been tested for activity against some human cancer cell lines in vitro. Cells are colored in green if: high activity
(< 10 μM to reach EC50) was found, orange if: two studies did not agree on high activity, red if they agreed on low activity (> 10 μM to reach EC50), white if: no
published data was available. In the Human erythrocyte column, the opposite is instinctively true; green color indicates low, and red indicates high activity (van Zoggel
et al., 2012; Dos Santos et al., 2017; Mor et al., 1994b; Nicolas and El Amri, 2009; Charpentier et al., 1998; Conlon et al., 2007; Galanth et al., 2009; Shi et al., 2016;
Huang et al., 2017; Zhu et al., 2018)

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initiation of apoptosis via mitochondrial membrane rupture and binding and other communications between cancer cell and
(iii) non-membranolytic modes of action (Ellerby et al., 1999; environment (Hoskin and Ramamoorthy, 2008). This may
Chen et al., 2001; Deslouches and Di, 2017). account for DRSs being more effective against one cancer
Below, we will elaborate on how DRSs disrupt the lipid bilayer type (PC3) than another (Leuschner and Hansel, 2004; van
of microbes and cancer cells, and which features are possibly Zoggel et al., 2012a). It has been observed that membrane
responsible for DRSs affinity to microbes and cancer cells. environment with a strong positive curvature strain influences
Additionally, we will discuss how DRSs modulate host immune the DRS-B2 into a state that facilitates insertion into the
systems and pathogen’s gene expression. Last, we summarize the membrane (Galanth et al., 2009).
evidence of DRS acting as a receptor (ant)agonist.
Modulation of the Host Immune System
Disruption of the Lipid Bilayer Besides the membrane disrupting activity, DRS likely modulates
One common feature DRSs often demonstrate is their disruption the host defense system as well. By introducing DRS-S1,
of the lipid bilayer of a target cell. Early evidence for this is the neutrophils of rat and humans stimulate their microbicidal
depolarization of bacterial membranes in vitro (Fleury and activities such as their production of reactive oxygen species
Longeron, 1998). To do this, DRSs likely form tetramers in their (Ammar et al., 1998). Moreover, DRS-S9 is chemotactic for
quaternary structure that form toroidal pores. Using the planar human leukocytes (Auvynet et al., 2008). On the other hand,
lipid bilayer technique, it was shown that the DRS peptides DRS-S4 has been observed to bind Lipopolysaccharides (LPS),
accumulate in a carpet like manner on the outside of a lipid which would rather suppress activation of macrophages and
bilayer until a threshold concentration is reached, causing them decrease the production of inflammatory cytokines (Navon-
to form pores in which the peptides are intercalated with the Venezia et al., 2002). Furthermore, DRS also shows angiostatic
phospholipid headgroups of the membrane (Duclohier, 2006). activities, which may influence tumor growth (van Zoggel et al.,
The selectivity of these pores appears to be determined by the 2010; van Zoggel et al., 2012).
phospholipid headgroups of the plasma membrane. The carpet
of DRSs on the outside of the lipid bilayer as well as tetramer Modulation of the Pathogen’s Gene
channel with intercalated phospholipid headgroups causes the Expression
membrane to lose its integrity (Yeaman and Yount, 2003). The We previously mentioned that DRSs can permeate membranes of
reason why DRS favor binding to some pathogens and tumors is different types of cells and cell nuclei, however it is also possible
still under investigation and several hypotheses exist, discussed that DRS changes gene expression in a pathogen cell as well.
here are the possible influence of; membrane charge, membrane DRS-S1 Modulates the expression of C. albicans genes, such as
sulfatation and membrane fluidity. the Hyphal wall protein 1 (HWP1) gene (Belmadani et al., 2018).
First DRSs are generally rather cationic peptides and thus In vitro, its expression was downregulated which likely accounts
more prone to bind negatively charged membranes. Bacteria for the modified cell morphology that was found. Furthermore,
and cancer cell membranes typically have a net negative charge, changes in aspartic protease genes were found.
the former due to negatively charged phospholipids on inner
(gram negative) and single (gram positive) membranes (Shai,
2002), the latter due to expression of phosphatidylserine and
Interaction With Cell Membrane Receptors
Lastly, it seems unlikely that there is a partner protein on the
negatively charged mucin proteins (Utsugi et al., 1991; Hoskin
surface or cytoplasm of the tumor cells that can account for the
and Ramamoorthy, 2008). Interestingly, erythrocytes have net
effects of DRS. This is illustrated by the example in which DRS-
negatively charged membranes as well, but are generally less
B2’s activity on PC3 cell lines is almost identical when composed
affected by DRS (Caillon et al., 2013).
of only amino acids in D configuration, (Dos Santos et al., 2017).
Second the sulfatation of glycosaminoglycans (GAG)’s on the
Since receptors are usually stereo-selective, it is unlikely DRS-B2
membrane surface seems to be of high importance for DRS-B2’s
works through a receptor. However, there is evidence that some
effectivity against prostate cancer cell lines (PC3) (Dos Santos
DRS can inhibit adenosine triphosphate (ATP) production
et al., 2017). Low concentrations of Chondroitin Sulfate C (CS-
through receptor binding (Laughlin and Ahmad, 2010).
C), which is a sulfated GAG, contribute to the α-helical shape of
DRS-B2, which is its biological active form. Interestingly, sulfated
GAG’s seem to be essential to the effectivity of cell penetrating FUTURE PERSPECTIVES
peptides (CPPs) as well (Yang et al., 2014). Furthermore, when
Zhu et al. (2018) investigated the activity of DRS-DP1 and 2, As DRSs act against a wide variety of pathogens and tumor cell
they found that by introducing a TAT (GRKKRRQRRR) peptide lines, it is tempting to speculate on their therapeutic potential.
at the N-terminal, affinity to the cell membrane and interaction Nevertheless, current knowledge on DRS is fragmented with
with GAG’s increased. respect to mechanism of action and diverse with respect to
Third, cancer cells often have increased membrane pathogens and cancer cell lines studied. Consecutively, we
fluidity and irregularities of cell surface which contributes will discuss remaining questions on the mechanism of action,
to membrane destabilization and could affect receptor potential clinical applications and safety.

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Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

Future Research on Available DRSs chemotherapies. This idea finds its roots in a hypothesis
For research on DRSs to move forward on the already investigated regarding the evolutionary origins of DRSs in frogs. The idea is
DRSs described in this paper, it is essential to understand the that DRSs function as an accessory protein that lyses cells and
mechanism of action of a specific DRS against a pathogen to fine- penetrates tissue to allow effectiveness of other neuromodulators
tune this DRS into a molecule that can target a specific clinical and enzymes in frog secretions (Konig et al., 2015). This
problem. These potential clinical therapies include treatment of contrasts with the view that DRSs are part of an innate immune
infections caused by specific micro-organisms, the treatment system. Indeed, there is striking similarity in the amino acid
of infections caused by multiple micro-organisms such as skin sequences of preproDRS and precursors of demorphin and
infections, and as a component of cancer treatment. Furthermore, deltorphin (Amiche et al., 1994) (which are powerful opioid
we discuss the current use of DRS in biotechnology. agonists), suggesting these peptides work together to achieve
It seems likely that the mode of action of DRSs relies, a common goal. This idea might extend to clinical practice as
similarly to many AMPs, in part on membrane disruption well. Potentially DRS could be part of a drug delivery system
of target cells. Important factors in this include the cationic and play similar role to other cell penetrating peptides in cancer
nature of the peptide, and its ability to retain an α-helical shape. research (Bolhassani, 2011; Bolhassani et al., 2011; Yang et al.,
The presence of certain phospholipids, GAGs, and proteins 2014). DRS would penetrate and disintegrate cell membranes of
contribute to this. However, the means to identify more of the specific cancer cells and (attached) conventional cytostatic agents
cell membrane factors, including the shape and fluidity of the may then – more effectively – affect the tumor tissue. Some
cell membrane are not readily available, and would require support for this idea comes from a study using Cecropin A and
innovative tools and techniques at the interface of chemistry conventional chemotherapy on leukemic patients, Cecropin A
biology and biophysics. increased the effect of the conventional therapy (Hui et al., 2002).
DRSs can be tested for specific clinical problems in vivo. Possibly the same idea can be used when targeting microbial
For example, some DRSs act against specific, anti-biotic pathogens (Balaban et al., 2004). Potentially, red blood cells or
resistant pathogens. From Table 1 we can infer that DRS- chitosan nanoparticles could function as a potential carrier for
B4, DRS-O1, DRS-DI06, DRS-CA1, DRS-DU1, DRS-PD2 DRS (Feder et al., 2001; Medeiros et al., 2014).
and DRS-PS4 are all promising candidates for specifically Meanwhile the field of applied biotechnology has moved
targeting P. aeruginosa, a pan-resistant bacterium notorious forward on these developments and researchers have
for causing severe infections in hospital settings. Another incorporated DRS in the genetic material of potato and citrus
example is DRS-O1 which has thus far been the only DRS plants. The plants can express these peptides and protect the
to show activity against the S. mansoni, a neglected tropical crops from disease (Rivero et al., 2012; Furman et al., 2013).
disease in need of treatment possibilities. More recently, DRS-B1 was modified (N-terminally modified
On the other hand, there are some DRSs that can act against and recombined respectively) to protect poplar plants and
a range of pathogens. This makes some DRSs ideal for clinical tobacco plants from infections by inserting the DRS protein in
applications in which several pathogens need to be targeted. the genome of the host (Yevtushenko and Misra, 2019; Shams
Skin infections, for example in diabetic foot ulcers or catheter et al., 2019). On the one hand pesticides can now be avoided on
infections are caused by organisms such as gram-positive these crops, on the other hand studies already report resistance
bacteria as well as some fungi (Schittek et al., 2008). From of bacteria to anti-microbial peptides by producing positively
Table 1 we can conclude that DRS-S3, DRS-S5, DRS-CA1, charged molecules on the membrane and pumping AMP’s
DRS-DU1, DRS-PS4, DRS-PD2 and DRS-O1 all show activity out of the cells (Joo, Fu & Otto, 2016; Andersson Hughes &
against the S. aureus (some including the methicillin resistant Kubicek Sutherland, 2016). Genetically altering plants on a
variant), as well as the C. albicans fungus, without damaging large scale could potentially endanger DRS’s use as an antibiotic
erythrocytes. Gomes and colleagues (2015) have already in humans.
positively assessed DRS’s incorporation in cotton gauzes and Lastly, the safety of DRS in humans remains to be investigated.
show potential in fast-release medical applications. Therefore Even though several DRS have been administered to mice
they are potential starting points for therapeutic applications and rats, to our knowledge, no phase-1 clinical trials have
in skin infections. been conducted on the safety of any DRS in humans thus far.
Some even speculate that there is potential for DRS as a Encouragingly, many of the documented short-term adverse
contraceptive, as DRS-S4 is spermicidal (Zairi et al., 2005). effects of the Kambo ritual such as nausea and tachycardia can
DRS-S4 shows activity against HIV, and several other genital be ascribed to other molecules in the cocktail (Erspamer et al.,
pathogens (Lorin et al., 2005; Savoia et al., 2010). However, it 1993), nevertheless the safety of the DRS remain to be shown.
appears the recent literature prefers antimicrobials that are within Researchers in many fields could benefit from the knowledge of
(or close to) the human genome for the use as contraceptives a safe DRS peptide.
(Tanphaichitr et al., 2016). Moreover, the clinical relevance of
such a contraceptive as well as the high degree of effectivity that Future Research on Novel DRSs
would be demanded are questionable. The investigation of other Anuran species is likely to yield
There is the potential of DRS as an aiding therapeutic in anti-microbial or DRS like peptides that can contribute to
cancer treatment by working in tandem with conventional illuminating the mechanism of action of these peptides and

Frontiers in Pharmacology | www.frontiersin.org 7 November 2019 | Volume 10 | Article 1421

Bartels et al. Dermaseptins, Multifunctional Antimicrobial Peptides

provide starting points for therapeutical treatments. According DRS are a complex family of bioactive peptides. Accumulating
to some estimations, skin compounds have been detected and evidence suggests their efficacy in a wide variety of medical
isolated from 400 anuran species, which means that more than applications. Despite the still puzzling mechanisms of action,
90% of all documented frog species still await screening (Konig DRSs are extremely suitable for specific medical problems.
et al., 2015). The isolated peptides from different species
thus far are very often unique and sometimes useful which
makes it very likely that new, useful and novel biomolecules AUTHOR CONTRIBUTIONS
await discovery.
To ensure these new molecules contribute to scientific EB is first author and writer of this manuscript. DD supervised
literature in a less fragmented way, it is important to identify the initial draft of this manuscript, helped identify the aims and
the questions that remain on the mechanism of action of research question and came up with the initial structure. MA
DRS. Whenever a new DRS or a modification is tested, ideally critically reviewed the drafts several times, elaborated on the
these mechanisms should be evaluated: 1) disruption of mechanisms of action, revised structure and added figures.
plasma/mitochondrial membranes; 2) necrosis; 3) apoptosis;
4) mechanisms of mediated immunity; 5) membrane receptor
involvement; 6) inhibition of DNA synthesis; 7) anti-angiogenic SUPPLEMENTARY MATERIAL
effects (Gaspar et al., 2013). Additionally, their activity against
human erythrocytes and epithelial cells should be evaluated to The Supplementary Material for this article can be found online at:
assess clinical relevance and help understand how DRS recognize https://www.frontiersin.org/articles/10.3389/fphar.2019.01421/
target cells. full#supplementary-material

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dermaseptin family, Biochim. Biophys. Acta 1388, 279–283. doi: 10.1016/ absence of any commercial or financial relationships that could be construed as a
s0167-4838(98)00202-7 potential conflict of interest.
Yang, J., Tsutsumi, H., Furuta, T., Sakurai, M., and Mihara, H. (2014). Interaction
of amphiphilic alpha-helical cell-penetrating peptides with heparan sulfate. Copyright © 2019 Bartels, Dekker and Amiche. This is an open-access article
Org. Biomol. Chem. 12 (26), 4673–4681. doi: 10.1039/c4ob00673a distributed under the terms of the Creative Commons Attribution License (CC
Yeaman, M. R., and Yount, N. Y. (2003). Mechanisms of antimicrobial peptide BY). The use, distribution or reproduction in other forums is permitted, provided
action and resistance. Pharmacol. Rev. 55 (1), 27–55. doi: 10.1124/pr.55.1.2 the original author(s) and the copyright owner(s) are credited and that the
Yevtushenko, D. P., and Misra, S. (2019). Enhancing disease resistance in poplar original publication in this journal is cited, in accordance with accepted academic
through modification of its natural defense pathway. Plant Mol. Biol. 100, 481. practice. No use, distribution or reproduction is permitted which does not comply
doi: 10.1007/s11103-019-00874-2 with these terms.

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