peptides as antifungal, antibacterial and immunomodulatory agents

17 Pages·2016·0.61 MB·English

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Yadav et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 6.041
Volume 5, Issue 9, 591-607 Review Article ISSN 2278 – 4357

PEPTIDES AS ANTIFUNGAL, ANTIBACTERIAL AND
IMMUNOMODULATORY AGENTS

Ruchi Omar, Sweta Sharma and Arpita Yadav*

Department of Chemistry University Institute of Engineering and Technology
Chhatrapati Shahuji Maharaj University Kanpur 208024, India.

ABSTRACT
Article Received on
02 August 2016, The review article discusses the importance of developing non

Revised on -----------------,
conventional antibacterial or antimicrobial agents. The article describes
Accepted on 06 August 2016

DOI: 10.20959/wjpps20169-7561 the sources and important properties of some naturally occurring
peptides derived from flora and fauna. It also provides a glimpse of the
volume of structural information available on these peptides that can
*Corresponding Author
Dr. Arpita Yadav be exploited for molecular modeling and docking studies to reveal their
Department of Chemistry mechanistic aspects. The increased usage of antibiotics in modern
University Institute of
society and spread of immunosuppressive diseases like AIDS are day-
Engineering and
by-day increasing the demand for safe, natural antibacterials/
Technology Chhatrapati
antifungals. It is important to understand why these peptides could not
Shahuji Maharaj
University Kanpur emerge as standalone therapeutic agents. Apart from medicinal use
208024, India. these peptides can effectively contribute to agriculture and food

industry.

KEYWORDS: Antimicrobial peptides, antifungal, arginine rich, anti HIV, natural, cysteine-

rich.

INTRODUCTION

Serious concerns, side effects associated with the use of conventional azole antifungal agents,
increasing cases of acquired drug resistance and growing needs for safe antifungals have
induced the search of natural, safe antifungals and antimicrobial agents. A number of
antimicrobial peptides (AMPs) derived from natural sources, flora and fauna both, thus came
into existence with the expectations of being safe therapeutic agents. In this review article we
discuss the sources, properties and challenges related to therapeutic use of these peptides.

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Why are non conventional therapeutic agents needed?
Human suffer from different mycosis like athletes foot, ringworm, candidiasis, severe
systemic infections etc. [1] To treat these different types of mycosis antifungal agents come in
different preparations like creams, sprays, oral tablets etc. containing azole compounds like
ketoconazole, clotrimazole. Few azoles and other compounds like terbinafine, fluconazole
may be taken orally to treat severe internal infections[2,3]. Amphotericin, flucytosine,
itraconazole, micafungin (c.f. fig.1) are administered intravenously[4,5] Antifungals are
required not just for those suffering from any type of mycosis but also for people with poor
immune system, for example, those undergoing chemotherapy for AIDS are at a greater risk
of developing internal fungal infection and are prescribed regular antifungal medications[6]
Regular antifungal medication if not safe may lead to severe health issues. Many are under
the impression that antibiotics may be used to treat fungal infections. However, antibiotics
increase the risk of developing fungal infections by depleting harmless bacteria from our skin
surface.[7,8] Topical preparations of antifungal agents usually cause little side effects but oral.

1b. Intravenously administered antifungal drugs
Fig. 1 Structures of some conventional antifungal drugs

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Antifungals frequently cause nausea, headache, diarrhea etc.[9] The intravenously
administered antifungals are given only when the need outweighs the risk involved.[10]

Sources of non conventional antifungals
A number of AMPs have been isolated from natural flora and fauna, out of which some have
been isolated, characterized and synthesized for therapeutic use.[11-15] Only some AMPs have
been designated as antifungals.

Chitinases are compounds derived from microbes, insects, animals or plants with varying
antifungal spectrum. A chitinase derived from chinese Baozhu (Pyrus ussuriensis Maxim.)
recently[16] exhibited significant antifungal activity. Myticin C and related peptides derived
from Mussels (Mytiles spp.)[17] have been suggested for use in cosmetics and food products
to kill fungal pathogens. Anabaenolysins are lipopeptides derived from cyanobacteria
possessing antifungal activity. When studied by Shishido et al[18] these lipopeptides
demonstrated weak activity due to their hydrophobicity but on mixing with hydrophilic
cyclodextrins their solubility and efficacy were significantly enhanced. An antifungal peptide
has also been derived from gastropod Patella rustica found on seashore in Ghana.[19]

Scolopendin is a microbial derived antifungal that has been considered safe for human use.[20]
Some strains of lactic acid bacteria exhibit antifungal activity.[21,22] Such natural
Lactobacillus strains can be used in food preservation. Small peptides derived from
ambhibian skin[23] and spider venom[24] have been shown to possess antibacterial and
immunomodulatory properties. These studies indicate the important role of AMPs in human
health.

Structural and mechanistic aspects of antimicrobial/ antifungal peptides
As metioned in previous section, several AMPs have been isolated and characterized. Model
structures for some of these peptides have been proposed based on solution NMR data. The
databank, ID’s, sources of peptides have been collected in table 1[25-71] for easy reference.
Only the latest structure from each family has been mentioned here. Established sequences
for some of these AMPs are given in table 2. It is easy to note that the sequences largely
resemble cell permeable peptides being highly cationic or cysteine-rich or tryptophan-rich. It
is also obvious from table 1 that so far there is no standard nomenclature or naming
procedure for these antimicrobial peptides. Most of the names have however been kept taking
initials from their scientific names and indicating the numeral of peptide isolated from the
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species. However, the proposed model structures indicate significant variety as can be seen
from figs. 2 and 3.

The cysteine-rich peptides tend to be cyclic due to the presence of disulphide linkages. The
fact that they all act as antifungal/ antibacterial suggests the possibility of more than one
active mechanisms of action. Despite the advantages of these peptides their mechanistic
aspects have not been explored due to the difficulties in handling them experimentally as well
as theoretically, that is with reference to their proteolytic suspectibility and conformational
flexibility. The conventional antifungals, on the other hand, have been predicted to act by
interfering ergosterol biosynthesis.[72,73] Reduced ergosterol levels weaken fungal cell wall [74]
resulting in eventual oozing out of vital contents of fungal cell. This mechanistic aspect is
quite understandable for azole compounds as ergosterol biosynthesis is catalyzed by 14α-
demethylase enzyme, cocatalyzed by heme protein which is known to possess strong affinity
for imidazole type compounds [75]. The mode of action of antifungal peptides still remains in
its infancy. However, it is almost certain that after proper mechanistic elucidation at least
some may be tuned for the benefit of mankind. A recent study has reported and compared
several characteristics of these peptides like cytotoxicity, protease activity etc.[76]

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Fig. 2 Structures of some cationic AMPs

Fig. 3 Structures of some cysteine-rich AMPs
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Table 1. Structural information on some AMPs
No. of amino Reference
PDB ID Content Source of peptide
acids in peptide no.
Flora derived AMPs
Amaranthus caudatus
1MMC 1H NMR study of the solution structure of Ac-AMP2 30 25
(Love-lies-bleeding)
2ERI Solution structure of Circulin B Chassalia parviflora 31 26
Helical hairpin structure of a novel antimicrobial
Echinochloa crus-galli
2L2R peptide EcAMP1 from seeds of barnyard grass 37 27
(Water grass)
(Echinochloa crus-galli)
Solution structure of the antifungal defensin DEF4
Medicago truncatula
2LR3 (MTR-89070770) from Medicago truncatula (barrel 47 28
(Barrel medick)
clover)
2MN1 Solution structure of Kalata B1[W23WW] Oldenlandia affinis 30 29
Solution structure of PAFP-S, an antifungal peptide Phytolacca americana
1DKC 38 30
from the seeds of Phytolacca americana (Pokeweed)
NMR Solution structure of Pisum sativum defensin-1
1JKZ Pisum sativum (Pea) 46 31
(Psd1)
Hevein-type antifungal peptide with a unique 10-
2LB7 Triticum kiharae (Wheat) 44 32
cysteine motif
Fauna derived AMPs
Solution structure of the antimicrobial 18-residue
1KFP Acanthoscurria gomesiana(Spider) 18 33
gomensin
Solution structure of Alo-3: a new knottin-type
Acrocinus longimanus
1Q3J antifungal peptide from the insect Acrocinus 36 34
(Harlequin beetle)
longimanus
Androctonus australis
1CZ6 Solution structure of Androctonin 25 35
(Desert Scorpian)
NMR structure of antifungal defensin ARD1 from Archaeoprepona demophon
1OZZ 44 36
Archaeoprepona demophon (Banded king showmaker)
2KET Solution Structure of BMAP-27 Bos Taurus (Cattle) 18 37
1LFC NMR structure of Bovine Lactoferricin (LfcinB) Bos taurus (Cattle) 25 38
The solution NMR structure of Maximin-4 in SDS Bombina maxima
2MHW 27 39
micelles (Yunnan firebelly toad)
Structure of an indolicidin peptide derivative with
1QXQ Bos taurus (Cattle) 13 40
higher charge
Solution structure of an potent antifungal peptide Cm- Cenchritis muricatus
2MP9 12 41
p5 derived from C. muricatus. (Sea snail)
The solution structure of DEFA-1, a highly potent
2MXQ Equus caballus (Horse) 34 42
antimicrobial peptide from the horse
Solution structure of the Tachykinin peptide
1N6T Gallus gallus (Red junglefowl) 10 43
Neurokinin A
Gastrophysa atrocyanea
2E2F Solution structure of DSP 40 44
(Leaf beetle)
2OYV Neurotensin in DPC micelles Homo sapiens (Human) 13 45
1MA2 Tachyplesin-1 wildtype peptide NMR Structure (Horseshoe crab) 17 46
Three-Dimensional structure of antimicrobial peptide
1CIX (Horseshoe crab) 44 47
tachystatin A isolated from horseshoe crab
NMR Solution structure & model membrane
interaction studies of the peptide Hylin A1 from the Hypsiboas albopunctatus (Spotted
2N0O 18 48
Arboreal South American Frog Hypsiboas tree frog)
albopunctatus
1M4E Solution Structure of Hepcidin-20 Homo sapiens (Human) 20 49
SP-B C-Terminal(residues 59-80) peptide in DPC
2M1T Homo sapiens (Human) 22 50
micelles
Solution structure of the antimicrobial peptide Human
2LXZ Homo sapiens (Human) 32 51
Defensin-5
Litopenaeus vannamei
2N30 Structure of Ace-PvHct-NH2 23 52
(Whiteleg shrimp)
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Limulus polyphemus
1RKK Polyphemusin-1 NMR Solution structure 18 53
(Atlantic horseshoe crab)
NMR Structure of a novel antimicrobial peptide,
2G9P Lachesana tarabaevi (Ant spider) 26 54
latarcin-2a, from spider (Lachesana tarabaevi) venom
2MBD Lasiocepsin Lasioglossum laticeps (Bees) 27 55
2LLD The insect defensin Lucifensin from Lucilia sericata Lucilia sericata (Green bottle fly) 40 56
Solution structure & activity of the four disulphide Mytilus galloprovincialis
1FJN 39 57
bond Mediterranean mussel defensin MGD-1 (Mediterranean mussel)
1E4R Solution structure of the Mouse Defensin mBD-8 Mus musculus(House mouse) 35 58
Solution structure and cell selectivity of Piscidin-1
2OJM Morone saxatilis (Rock fish) 22 59
and its analogues
Oryctes rhinoceros
1IYC Solution Structure of antifungal peptide, scarabaecin 36 60
(Asiatic rhinoceros beetle)
Solution Structure of the antimicrobial peptide (Aba
2M2X Papio Anubis (Olive baboon) 18 61
3,5,7,12,14,16) BTD-2
Phyllomedusa tarsius
2JQ1 Phylloseptin-3 19 62
(Tarsier leaf frog)
Solution structure of termicin, an antimicrobial
1MM0 Pseudacanthotermes spiniger 36 63
peptide from the termite Pseudacanthotermes spiniger
1ZRV Solution structure of spinigerin in H20/TFE 50% Pseudacanthotermes spiniger 25 64
NMR Solution Structure of the Dermaseptin Phyllomedusa sauvagii
2DCX 13 65
antimicrobial peptide analog NC-12-K4S4(1-13)a (Waxy monkey tree frog)
Solution structure of papiliocin isolated from the
2LA2 Papilio xuthus (Asian swallowtail ) 37 66
swallowtail butterfly, Papilo xuthus
1G92 Solution structure of Poneratoxin Paraponera clavata (Bullet ant) 25 67
High resolution NMR Solution structure of the theta- Rhesus macaque
2LYF 18 68
defensin RTD-1 (Old world monkeys)
2I1D DPC micelle-bound NMR Structure of Tritrp-1 Sus scrofa (Wild boar) 13 69
High–resolution NMR Structure of the protegrin-2
2MUH Sus scrofa (Wild boar) 16 70
docked to DPC micelles
2MAG NMR Structure of Magainin-2 in DPC Micelles Xenopus laevis (African clawed frog) 23 71

Table 2. Sequences for some AMPs
Peptide Sequence
Ac-AMP2 VGECVRGRCPSGMCCSQFGYCGKGPKYCGR
Circulin B CGESCVFIPCISTLLGCSCKNKVCYRNGVIP
EcAMP1 GSGRGSCRSQCMRRHEDEPWRVQECVSQCRRRR
Defensin DEF4 RTCESQSHKFKGPCASDHNCASVCQTERFSGGRCRGFRRRCFCTTHC
Kalata B1 GLPVCGETCVGGTCNTPGCTCSWWPVCTRN
PAFP-S AGCIKNGGRCNASAGPPYCCSSYCFQIAGQSYGVCKNR
Pisum sativum defensin-1 (Psd1) KTCEHLADTYRGVCFTNASCDDHCKNKAHLISGTCHNWKCFCTQNC
Hevein-type Antifungal peptide AQRCGDQARGAKCPNCLCCGKYGFCGSGDAYCGAGSCQSQCRGC
Gomensin ECRRLCYKQRCVTYCRGR
Alo-3 CIKNGNGCQPNGSQGNCCSGYCHKQPGWVAGYCRRK
Androctonin RSVCRQIKICRRRGGCYYKCTNRPY
ARD1 DKLIGSCVWGAVNYTSNCNAECKRRGYKGGHCGSFANVNCWCET
BMAP27 GRFKRFRKKFKKLFKKLS
Bovine Lactoferricin (LfcinB) FKCRRWQWRMKKLGAPSITCVRRAF
Maximin-4 GIGGVLLSAGKAALKGLAKVLAEKYAN
Indolicidin ILKKWPWWPWRRK
Cm-p5 SRSELIVHQRLF
DEFA1 SCTCRRAWICRWGERHSGKCIDQKGSTYRLCCRR
Tachykinin peptide Neurokinin A HKTDSFVGLM
DSP AVRIGPCDQVCPRIVPERHECCRAHGRSGYAYCSGGGMYCN
Neurotensin ELYENKPRRPYIL
Tachyplesin 1 KWCFRVCYRGICYRRCR
Tachystatin A YSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY
Hylin A1 IFGAILPLALGALKNLIK
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Hepcidin ICIFCCGCCHRSKCGMCCKT
SP-B DTLLGRILPQLVCRLVLRCSID
Human defensin 5 ATCYCRTGRCATRESLSGVCEISGRLYRLCCR
Ace-PvHCt-NH2 FEDLPNFGHIQVKVFNHGEHIHH
Polyphemusin 1 RRWCFRVCYRGFCYRKCR
Latarcin 2a GLFGKLIKKFGRKAISYAVKKARGKH
Lasiocepsin GLPRKILCAIAKKKGKCKGPLKLVCKC
Lucifensin ATCDLLSGTGVKHSACAAHCLLRGNRGGYCNGRAICVCRN
MGD-1 GFGCPNNYQCHRHCKSIPGRCGGYCGGWHRLRCTCYRCG
mBD-8 NEPVSCIRNGGICQYRCIGLRHKIGTCGSPFKCCK
Piscidin 1 FFHHIFRGIVHVGKTIHRLVTG
Scarabaecin ELPKLPDDKVLIRSRSNCPKGKVWNGFDCKSPFAFS
BTD-2 GVARAVARRGVARAVARR
Phylloseptin-3 FLSLIPHAINAVSALANHG
Termicin ACNFQSCWATCQAQHSIYFRRAFCDRSQCKCVFVRG
Spinigerin HVDKKVADKVLLLKQLRIMRLLTRL
Dermaseptin ALWKTLLKKVLKA
Papiliocin RWKIFKKIEKVGRNVRDGIIKAGPAVAVVGQAATVVK
Poneratoxin FLPLLILGSLLMTPPVIQAIHDAQR
RTD-1 GFCRCLCRRGVCRCICTR
Tritrpticin VRRFPWWWPFLRR
Protegrin-2 RGGRLCYCRRRFCVCV
Magainin 2 GIGKFLHSAKKFGKAFVGEIMNS

Antiviral and anti-HIV activity of natural AMPs
Some recent studies have shown inhibition of enzymes involved in replication of HIV virus
and immunomodulatory properties of small AMPs[77,78]. Topical preparation of AMPs
derived from amphibian skin were shown to inhibit transmission of HIV infections of T cells
and vaginal epithelial cell lines[23]. Latarcins [24] have been tested for their anti HIV activity.
They may also be co-administered with commercial antibiotics like kanamycin. These recent
findings indicate that these small AMPs have immunomodulatory properties.

Above literature indicates the importance of AMPs in different fields as compared to
conventional drugs. At the same time it is also evident that little computational work has been
done due to their conformational flexibility which increases the complexity of modeling
studies multifold.

Antifungals/ antibacterials in agriculture, food industry and animal husbandry
While discussing antifungal or antibacterial properties of any compound it is imperative to
discuss its utility in agriculture and animal husbandry, as both industries consume large
amounts of antifungals. Use of antifungals in these industries is also indirectly linked to
human health. Day by day increased usage of fungicides to curb new resistant strains of fungi
crosses the limits of safe levels increasing human indirect exposure. Similarly, the routine
usage of coccidiostats in animal feed to avoid coccidiosis in animals has raised concerns over
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exacerbation of coronary diseases in human. Commonly used coccidiostats like monensin,
salinomycin, lasalocid are known to be toxic to human.[79]

Spoilage of food products is of great concern in food industry.[80] Some strains of
Lactobacillus used in fermented food products themselves act as antifungal agents.[21,22] Use
of such auto protective cultures is becoming latest trend in food preservation .[81,82] In a recent
study high throughput screening assay were designed to rapidly test thousands of
Lactobacillus isolates for their antifungal activities.[83]

Recent research attempts have thus been motivated towards enhanced utilization of naturally
occurring AMPs as antifungal agents as opposed to synthetic antifungals. Some natural
AMPs were however found to be cytotoxic against mammalian cells [84] causing hemolysis of
human erythrocytes. Cosmetic industry is also witnessing a drift towards enhanced usage of
natural products as it requires significant amount of antifungals.

Future directives
This review article emphasizes the need to study mechanistic aspects of AMPs as antifungal
agents which would perhaps be more feasible utilizing molecular modeling and docking
studies. Mechanistic elucidation will help us understand how to reduce the cytotoxicity
problems. The conformational flexibility and the size of peptides involved makes the
exploration highly challenging but with advances in modeling techniques and computational
facilities it may be achievable. Day by day increasing cases of antibiotic resistance and HIV
infections instigate research in this direction and may help curb some pressing medical
demands by synergistic approaches. Molecular modeling and docking work in this direction
is underway in our laboratory.

ACKNOWLEDGMENTS
A. Yadav and S. Sharma gratefully acknowledge financial support from Science and
Engineering Research Board (SERB) (SR/S1/OC-82/2012), New Delhi. Infrastructural
support from Chhatrapati ShahuJi Maharaj University, Kanpur is also gratefully
acknowledged.

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