You are on page 1of 15

C H A P T E R

17
Development of New Antiherpetic Drugs
Based on Plant Compounds
Adil M. Allahverdiyev1, Melahat Bagirova1, Serkan Yaman1,
Rabia Cakir Koc2, Emrah Sefik Abamor1, Sezen Canim Ates1,
Serap Yesilkir Baydar1, Serhat Elcicek3, Olga Nehir Oztel1
1
Department of Bioengineering, Yildiz Technical University, Istanbul, Turkey,
2
Department of Biomedical Engineering, Yeni Yuzil University, Istanbul, Turkey and
3
Department of Bioengineering, Firat University, Elazig, Turkey

INTRODUCTION are frequently seen in skin, mouth, oral cavity,


genital region, and central nerve system. HSV-
Herpes simplex viruses (HSVs) are double- 1 and HSV-2 have been characterized as the
stranded, enveloped DNA viruses of the Her- mostly common virus species, responsible for
pesviridae family. HSVs are characterized by a variety of mild to severe diseases such as
their icosahedral symmetry and possess 162 herpes corneae, herpes labialis, herpes genitalis,
capsomers. These viruses consist of four basic herpetic whitlow, gingivostomatitis, Kaposi var-
structures: an envelope, a nucleocapsid, icelliform eruption, and neonatal herpes.
a DNA containing core, and a tegument. The Among these diseases, herpes labialis, gingivos-
virion contains 11 envelope glycoproteins, tomatitis, and herpetic whitlow, which are seen
which are found as bulges on the outer surface. in patients infected by HSV-1, causes painful
Four glycoproteins (gB, gD, gH, and gL) play cold sores and blisters at facial sites such as
important roles as receptors for determining the mouth, tongue, and skin. On the other
virus infectivity. Approximately 100 species of hand, HSV-2 infects genital sites of the body
herpes virus have been identified as pathogens and induces ulcers in the genital mucosa.
both for humans and animals, and 25 of these Primary HSV infections are caused by pene-
have been shown to cause diseases affecting tration of the virus into the skin and mucosa of
humans [1]. HSV infections usually give rise to susceptible individuals from scratches and
painful lesion in tissues originating from the wounds on the skin or mucosal surface. Gener-
embryonic ectoderm [2]. Hence, these infections ally, intact skin or mucosa is resistant to HSV

Fighting Multidrug Resistance with Herbal Extracts,


Essential Oils and Their Components
Copyright 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/B978-0-12-398539-2.00017-3 245
246 17. ANTIHERPETIC PLANT COMPOUNDS

infections. Broadly, HSV-1 infections are trans- chemotherapy, or have had bone-marrow trans-
mitted into individuals via aerosols or saliva, plants. Treatment failure can also be caused by
while HSV-2 infections are frequently trans- the recurrence of latent viruses. Considering
mitted sexually. HSV diseases occur by one of the constant increase on HSV infections world-
two mechanisms: a new infection or reactivation wide and recently developed resistance to
of latent viruses. HSV is known to reside in nerve current therapies, it is clear that there is an
ganglia after initial infection and can survive at urgent need to identify new, more effective anti-
these sites as a latent virus for many years. Viral viral agents to combat HSV infections [12].
reactivation inside nerves can be induced by Medicinal plants have been used for the treat-
fever, ultraviolet rays, stress, fatigue, menstrua- ment of human diseases since ancient times and
tion, sexual intercourse, or immune suppres- their use offers an alternative to traditional anti-
sion; after reactivation, herpetic blisters can be biotics as antimicrobial agents. In contrast to
observed at different sites of the human body [3]. synthetic drugs that may contain several harm-
According to the World Health Organization, ful components, medicinal plants have the
an estimated 536 million people all over the advantage of being 100% natural and biologi-
world were infected with HSV-2 in 2003 [4]. In cally sourced. Recently, the acceptance of tradi-
addition, more than 50 million people are tional medicine as an alternative form of
currently infected with HSV-2 in the USA and health care and the development of microbial
an estimated 0.5 million new cases occur each resistance to the current antibiotics have encour-
year [5]. It is known that HSV-1 infections are aged researchers to investigate the antimicrobial
more commonly seen in humans than are activities of medicinal plants. In several studies,
HSV-2 infections. For example, 57% of the pop- medicinal plant products, such as extracts and
ulation of the United States is reported to be essential oils (EOs), have been proven to possess
seropositive for HSV-1, while 20% are seroposi- significant antimicrobial properties [13].
tive for HSV-2 infection [6]. Worse news is that Additionally, plant products have been
the prevalence of both types of the disease has recently marketed as pharmaceuticals in devel-
been gradually increasing and has reached oped and developing countries, since they
life-threatening levels, especially for immune- include biologically active substances and have
compromised patients. On this subject, it was been indicated to be safe and without side
reported that the prevalence of HSV-2 has been effects. According to the literature, many plant
increasing at a high rate within HIV-positive products obtained from traditional medicinal
patients, indicating that the herpetic infection plants also have strong antiviral activity, and
is a major cause of morbidity, especially in some have already been used in the treatment
immune-suppressed patients. Acyclovir of viral infections in humans and animals. One
(Zovirax) and other nucleoside derivatives, type of viral infection for which researchers
including famciclovir, ganciclovir, penciclovir, are investigating plant products is herpes
and valacyclovir, have been approved for the simplex infection. There are severally reports
treatment of HSV (both HSV-1 and HSV-2) infec- of studies on antiherpetic plants. Therefore, we
tions worldwide [7e11]. The basic efficacy will review the information on various herbal
mechanisms of these drugs involve inhibition compounds available in the current literature,
of thymidine kinase and DNA polymerase compare the antiherpetic efficacy of these
enzymes specific to HSV. However, it was compounds, and determine the potential of
recently reported that resistance can develop these plants for developing new antiherpetic
against acyclovir, especially in individuals that therapies. In this chapter, the antiherpetic
are immune compromised, undergoing cancer activity of herbal extracts and EOs will be
PLANT EXTRACTS WITH ANTIHERPETIC ACTIVITY 247
presented. Such plant extracts and EOs could be sage (23 mg/g) were used, and the healing time
important for the discovery of new and effective of treated patients was 6e7 days for sage cream,
antiviral drugs. 6e7 days for rhubarb-sage cream, and 5e6 days
for acyclovir cream. The results of this study
showed that sage was less effective than
PLANT EXTRACTS WITH rhubarb against HSV, while a combined topical
ANTIHERPETIC ACTIVITY preparation of these plant extracts was as effec-
tive as topical acyclovir [18].
Carissa edulis (Forssk.) Vahl
(Apocynaceae) Hypericum connatum (St. Johns Wort)
Carissa edulis is a medicinal plant that grows The Guttiferae family produces several anti-
particularly in Kenya. C. edulis has been used viral compounds, and species of the Hypericum
as a traditional medicine in Kenyan for the treat- genus have long been used in traditional medi-
ment of various ailments, without any reported cine. Extracts and isolated compounds from
side effects [14]. The roots of this plant have plants of the Hypericum genus are commonly
been also used for antiherpetic purposes. used for the treatment of viral infections. For
Briefly, roots are washed, dried, boiled, and example, H. connatum is utilized for the treat-
then lyophilized for the preparation of extracts. ment of mouth lesions in southern Brazil. In
The antiherpetic effects of C. edulis were investi- a study, ethanol extracts of H. connatum
gated in Vero E6 cells infected with 100 plaque compounds were tested for antiherpetic effects
forming units of wild-type or resistant strains in vitro using the Vero cell line infected with
of HSV; under these conditions, 50 g/mL of HSV-1. This study demonstrated different rates
the plant extract showed antiviral activity. In of antiviral activity against HSV for the crude
vivo animal tests with Balb/c mice showed methanol extract, fractions, and isolated
that the extract at an oral dose of 250 mg/kg compounds. Therefore, compounds obtained
significantly delayed the onset of HSV infections from H. connatum have inhibitory effects against
by > 50% and increased the survival time of HSV. The mechanisms of the antiherpetic effects
treated animals. C. edulis toxicity was low [50% of these compounds may involve a direct viru-
cytotoxic concentration (CC50) value of 480 g/ cidal effect; interference with virus entry across
mL] and it did not cause acute toxicity in the cell membrane; or interference with viral
BALB/c mice at the oral therapeutic dose of genome replication or other early steps in intra-
250 mg/kg [15]. cellular viral replication [19].

Rhubarb and Sage Clinacanthus nutans (Snake Plant)


The antiviral activity of rhubarb root extract Clinacanthus nutans extract is traditionally
has been observed and confirmed by several in used in Thailand for the topical treatment of
vitro studies [16,17]. The antiviral activity of HSV and varicella-zoster virus infection [20].
sage leaf extract was also confirmed, although The active ingredient was purified from the
is showed a low efficiency. In a screening study, heartwood of Artocarpus lakoocha Roxb. (Mora-
the antiherpetic effect of rhubarb-sage cream, ceae), is extracted in methanol and then purified
sage cream, and acyclovir cream were compared by vacuum liquid chromatography. The purified
in an intention to treat analysis. Aqueous- compound was identified as 2,30 , 4,50 -
ethanol extracts of rhubarb (23 mg/g) and dried tetrahydroxy-trans-stilbene (oxyresveratrol) by
248 17. ANTIHERPETIC PLANT COMPOUNDS

mass spectroscopy, and high performance liquid HSV in animal experiments. Cutaneous applica-
chromatography analysis confirmed it to be tion of oxyresveratrol was found to be more
> 99% pure. Antiviral activities of oxyresveratrol effective than oral administration. Therefore,
were demonstrated against HSV-1, HSV-2, oxyresveratrol may be an appropriate topical
measles virus (Tanabe strain), and poliovirus treatment for HSV-1 [21e23].
type 1 (Sabin strain). There were no significant In conclusion, oxyresveratrol has been shown
differences among the IC50 values (P > 0.05) to be effective against both wild-type and
for various strains of HSV-1. Acyclovir-resistant acyclovir-resistant HSV strains. These results
HSV-1 strains were also susceptible to oxyresver- provided evidence that topical rather than oral
atrol, suggesting that the mode of anti-HSV applications of oxyresveratrol provide a greater
activity of oxyresveratrol differs from that of therapeutic efficacy against cutaneous HSV
acyclovir. Concentrations of 25e100 mg/mL had infection in mice.
no effect on viral infectivity. However, treatment
of mice with three applications a day showed
Coptidis rhizoma (Coptis Root or
a significant therapeutic efficacy (P 0.004),
although two applications a day did not show
Goldenthread)
significant efficacy (P 0.10), compared to the Three extracts from the Coptidis rhizoma plant
control group. An ointment containing 30% oxy- are berberine, C. rhizome extract, and Ching Wei
resveratrol showed no significant difference in San. Berberine is an antimicrobial herbal extract
efficacy (P 0.62) compared to 5% acyclovir with activity against a variety of organisms,
cream [21]. Therefore, topical administration of including human immunodeficiency virus [24],
oxyresveratrol is suitable as an anti-HSV thera- human cytomegalovirus (HCMV) [25], and hepa-
peutic route for cutaneous HSV infections in titis B virus [26]. This compound intercalates
mice. Oxyresveratrol also demonstrated efficacy DNA, thus inhibiting DNA synthesis and reverse
on acyclovir-resistant mutant strains, and may transcription [24,27e29]. Berberine does not
therefore be effective against acyclovir-resistant affect membrane permeability, but does affect
virus strains. Oxyresveratrol showed synergy protein biosynthesis [30]. A study compared the
with the anti-HSV-1 activity of acyclovir. The antiviral activity of all three C. rhizome extracts
IC50 for combined acyclovir and oxyresveratrol against HSV and demonstrated that all exhibited
was reduced by two- to four-fold, compared to antiherpetic effects, with IC50 values of 8.2, 2.4,
the IC50 values when these drugs were used and 24.3 mg/mL for berberine, C. rhizome extract,
alone. This provides further supporting and Ching Wei San, respectively. Levels of cyto-
evidence that the mode of oxyresveratrol anti- toxicity induced by these extracts were evaluated
HSV-1 activity differs from that of acyclovir: in Vero cells; the toxicity of berberine was found
acyclovir is a nucleoside analog that exhibits to be lower than those of the other two extracts.
antiherpetic activity after its phosphorylation Thus, the selectivity of berberine appears to be
by viral thymidine kinase; in contrast, oxyresver- 1.2e1.5-fold higher than that of the C. rhizome
atrol affects viral replication by inhibiting late extract and Ching Wei San.
viral protein synthesis [21].
According to the results of topical treatment
Ilex paraguariensis (Yerba Mate)
experiments, ointment containing 30% oxyres-
veratrol applied 4e5 times a day is most effec- Ilex paraguariensis (yerba mate) is an ever-
tive in delaying the skin lesion development green tree from the holly family that grows in
and protecting against death. Further, topical the subtropical forests of Paraguay, Uruguay,
treatment of oxyresveratrol is reported to inhibit and the Parana state of Brazil. In a study, an
PLANT EXTRACTS WITH ANTIHERPETIC ACTIVITY 249
aqueous extract of I. paraguariensis leaves of R. imperialis leaves showed antiviral activity
showed antiviral activity against HSV-1 in vitro against HSV-1 in vitro [31]. The EC50 of
[31]. Caffeoylquinic acid derivatives and triter- R. imperialis extracts was 70 mg/mL, while the
penoid saponins are widely distributed among CC50 was 1390 mg/mL (SI value of 19.8). As
plants, including yerba mate, which shows an a result, the presence of tannins and flavonoids
antiviral effect that may be linked to the pres- may be associated with the high antiviral
ence of these compounds. The 50% effective activity of this extract against HSV-1 [31].
concentrations (EC50) of caffeoylquinic acid
derivatives and triterpenoid saponins derived
from I. paraguariensis were 80 mg/mL and the Slonea guianensis
CC50 was 1260 mg/mL [selectivity index (SI In a study, the methanol extract of Slonea guia-
CC50/EC50) value of 15.8] [31]. Therefore, these nensis leaves showed antiviral activity against
compounds may play important roles in the HSV-1 in vitro. The EC50 of the S. guianensis
drug discovery process in the future. methanol extract was 140 mg/mL and the CC50
was 1400 mg/mL [31].
Lafoensia pacari
Lafoensia pacari is a plant of the Lythraceae Pongamia pinnata (Indian Beech)
family that grows in Brazil and Paraguay. In Pongamia pinnata belongs to the Papiliona-
a study, a methanol extract of L. pacari leaves ceae family. Seeds were collected from dried
showed antiviral activity against HSV-1 in vitro. fruits and, in an in vitro study, a seed extract
A high SI value (19.0) was obtained for tannins completely inhibited HSV-1 and HSV-1 multi-
isolated from L. pacari, indicating that these plication at concentrations of 1 mg/mL (w/v)
compounds are very effective against the HSV- and 20 mg/mL (w/v), respectively [33].
1 virus and that extracts containing tannins
may play an important role against HSV [31].
Other Important Plant Extracts
Passiflora edulis (Passion Flower) In contrast to the other studies, a study per-
formed in Thailand identified specific activity
Passiflora edulis is a vine species of passion against HSV in vitro in extracts of Aglaia edulis,
flower that grows in Brazil, Northern Argentina, Centella asiatica, Glyptopetalum sclerocarpum,
and Paraguay. In a study, an aqueous extract of Maclura cochinchinensis, and Mangifera indica
P. edulis roots showed antiviral activity against [34]. Additionally, extracts of Azadirachta indica,
HSV-1 in vitro [31]. The EC50 of P. edulis extracts Rinorea anguifera, and Protium serratum were also
(containing saponins and flavonoids) was demonstrated to be effective against HSV-1 and
89.9 mg/mL and the CC50 was 1600 mg/mL. poliovirus in vitro [34]. Moreover, extracts of
Therefore, saponins and flavonoids may exhibit Hura crepitans, Moringa oleifera, Schima wallichii,
antiherpetic activity, consistent with previous and Willughbeia edulis specifically inhibited pla-
reports [32]. que formation by HSV-1; in addition, none of
these plant extracts were cytotoxic [34]. The
Moringa oleifera extract was more effective
Rubus imperialis
against a phosphono acetate-resistant (APr)
Rubus imperialis grows abundantly in strain and Aglaia odorata was less effective
southern Brazil. In a study, a methanol extract against a thymidine kinase-deficient strain
250 17. ANTIHERPETIC PLANT COMPOUNDS

compared to their activities against the wild- apigenin, kaempferol, quercetin, and luteolin;
type strain (7401H HSV-1) [34]. In vivo studies phenolic acids and tannins; rosmarinic acid
revealed that A. odorata, Cerbera odollam, M. olei- and glycosidically bound caffeic and chloro-
fera, Ventilago denticulata, and W. edulis either genic acids; and the triterpenes, ursolic and ole-
inhibit plaque formation or exhibit therapeutic anolic acids [36e38]. This plant has been used in
levels of antiviral efficacy against HSV-1 a variety of practical applications in medical
following oral application [34]. The anti-HSV science. M. officinalis extract has been reported
activity of M. oleifera in mice does not involve to inhibit protein synthesis and to exhibit anti-
a direct antiviral effect, because this extract can viral activity against HSV-1 [39e41].
also be used as an immune modulator in mice. Allahverdiyev et al. showed the antiviral
The three plant extracts containing active effect of volatile oils of M. officinalis against
compounds (A. odorata, M. oleifera, and V. dentic- HSV-1 and HSV-2 viruses. They showed that
ulata) were also nontoxic. Therefore, all may be concentrations up to 100 mg/mL are not toxic
used as prophylactic or therapeutic anti-HSV to cells, although concentrations > 100 mg/mL
medicines for the treatment of HSV-1 infections. showed toxic effects on HEp-2 cells. Volatile oil
The M. oleifera extract is particularly promising concentrations up to 100 mg/mL exhibited anti-
because of its immunomodulatory and anti- viral activity against HSV-2. Since the maximal
HSV features. nontoxic concentration was determined to be
Combinations of acyclovir and herbal extracts 100 mg/mL, they used this concentration in all
have also been used to improve the treatment of experiments to test the effects of volatile oils on
HSV-1 infection in humans. Some studies have virus replication. Their in vivo findings indicate
shown that Geum japonicum, Rhus javanica, Syzy- that M. officinalis L. extract inhibit the HSV-2
gium aromaticum, and Terminalia chebula can replication at nontoxic doses [42].
augment the anti-HSV-1 activity of acyclovir in This research indicates that M. officinalis L.
the brain, rather than the skin [35]. Controver- volatile oils combined with synthetic materials
sially, the anti-HSV-1 activity of acyclovir may serve as an effective alternative to the
appeared to be weaker in the brain than in the currently available antiviral drugs by producing
skin. The researchers also suggested that the novel, more effective drugs or using the volatile
combined use of acyclovir with herbal extracts oils to increase the efficacy of available drugs.
at various concentrations showed strong syner-
gistic anti-HSV-1 activity, without a toxic effect. Illicium verum (Star Anise)
Additionally, combined treatment with acyclovir
and T. chebula was more efficient in the brain than Illicium verum is also known as star anise, star
the use of T. chebula alone [35]. Thus, these aniseed, or Chinese star anise, and is a member
extracts may be beneficial for prophylactic use of Schisandraceae family. I. verum is a small
against central nervous system complications. native evergreen tree of northeast Vietnam and
southwest China. In a study, EOs obtained
from I. verum were investigated for their antiher-
petic effect on HSV-2 [43]. The pharmaceutical
ANTIHERPETIC ACTIVITY quality and identity of the EOs and their chem-
OF PLANT ESSENTIAL OILS ical composition was confirmed by quantitative
and qualitative analysis using gas chromatog-
Melissa officinalis (Lemon Balm) raphy (GC) and gas chromatography-mass
Melissa officinalis L. contains the flavonoids, spectrometry (GC-MS) [43]. In this study, EO
quercitrin and rhamnocitrin; the 7-glucosides, derived from star anise showed antiviral
ANTIHERPETIC ACTIVITY OF PLANT ESSENTIAL OILS 251
activity prior to viral adsorption. The antiviral 0.7 ng/mL, and the CC50 was 7 ng/mL, making
activity of star anise EO against HSV-2 is shown the SI of thyme oil 10 [43].
in Table 17-1 [43]. EOs derived from I. verum
seed showed a much higher SI value than those
Zingiber officinale (Ginger)
of other extracts and EOs examined. The 50%
EC50 was 1 mg/mL and the CC50 was 160 mg/ Culinary ginger is the root of the Zingiber offi-
mL (SI value of 160). They concluded that the cinale plant. Ginger is a member of Zingibera-
star anise EO interferes with the virion envelope ceae family and is generally cultivated in
structure or may mask viral compounds that are southern Asia and East Africa. The EO of ginger
necessary for viral adsorption and thus inhibits is very effective against HSV-2, but is relatively
the virulence of the virus. In another study, the cytotoxic (CC50 0.004%), compared to the
inhibitory effect of star anise oil against HSV-1 other oils tested [43]. However, the IC50 of
infection was compared with the antiviral ginger EO (containing zingiberene, limonene/
potential of selected phenylpropanoids and cineol, b-sesquiphellandrene, camphene, and
sesquiterpenes, important constituents of EOs, pinocamphene) was found to be 0.0001% and
and acyclovir. Star anise EO and most isolated the SI value was 40 [43].
compounds exhibited high levels of antiviral
activity against HSV-1 in viral suspension Matricaria chamomilla (German
tests [44].
Chamomile)
Hyssopus officinalis (Hyssop) Matricaria chamomilla, also known as German
chamomile, is a member of a very wide range
Hyssopus officinalis is a species native to of different species belonging to Asteraceae
southern Europe, the Middle East, and the family. Chamomile originated from Europe
region surrounding the Caspian Sea. An EO and western Asia, and is also found in
extract of H. officinalis showed antiviral activity Australia and North America. The toxic
against HSV-2 in vitro [43]. In this study, concentration of chamomile in RC-37 cells
the EC50 was 6 ng/mL, the CC50 was 7.5 ng/ was found to be very low compared to those
mL, and the SI value of this oil was calculated of other oils. In addition, the SI of this plant
as 13. is 20, which indicates an average antiviral effect
compared to controls [43]. Pretreatment of the
virus with this oil showed a relatively strong
Thymus vulgaris (Thyme)
effective compared to acyclovir. The IC50 was
Thymus vulgaris is a gramineous plant that 0.15 ng/mL and the oils exhibited low toxicity
grows widely in southern Europe. This plant is (EC50 0.003 mg/mL) [43].
generally known as thyme. The characterization
of thyme oil by GC and GC-MS methods indi-
Geum japonicum
cated that thymol (40.5%), p-cymene (23.6%),
carvacrol (3.2%), linalool (5.4%), b-caryphyllene Geum japonicum is a yellow flowered peren-
(2.6%), and terpinen-4-ol (0.7%) are present in nial native to North America and eastern Asia.
thyme EO. Investigations into the effects of G. japonicum has been investigated for its anti-
thyme EO on HSV-2 indicated that pretreatment herpetic effects and was reported as being
of cells with the EO has a stronger effect on HSV potentially galenic in vivo. Statistically
virulence than does EO pretreatment of the significant symptom alleviation by G. japonicum
virus. In this study, the EC50 of T. vulgaris was has been reported in HSV-infected
252
TABLE 17-1 Antiviral Activities of Plant Essential Oils and Purified Compounds Against Herpes Simplex Virus

Extract or EC50
Plant part In vivo/ purified Type of CC50 (mg/
Plant used Active compound In vitro compounds herpes (mg/mL) mL) SI Reference(s)
a a a
Melissa officinalis Leaves Volatile oils In vitro Extracted by vapor HSV-2 [42]
distillation method

Illicium verum Seed Sesquiterpene In vitro Ethanol HSV-2 160  30.7 1  0.1 160 [44]
Hyssopus Flowering tops Iso-pinocamphene, In vitro Ethanol HSV-2 0.0075 0.0006 13 [43]
officinalis and leaves pinocamphene,
spathulenol, linalool,
sandalwood oil,
a-santalol, trans-a-

17.
santalol, cis-lanceol,
a-santalene

ANTIHERPETIC PLANT COMPOUNDS


Thymus Flowering tops Thymol, p-cymene, In vitro Ethanol HSV-2 0.007 0.0007 10
vulgaris and leaves carvacrol, linalool,
b-caryphyllene,
terpinen-4-ol

Zingiber Root Zingiberene, In vitro Ethanol HSV-2 0.004 0.0001 40


officinale limonene/cineol,
b-sesquiphellandrene,
camphene,
pinocamphene
Matricaria Flowers bisabolol, In vitro Ethanol HSV-2 0.003 0.00015 20
chamomilla b-farnesene,
bisabolol oxide A,
bisabolol oxide B
Geum Whole plant e In vivo Dry-boil filtered HSV-1 d d d
[45]
japonicum aqueous extract
Rhus javanica Gall e In vivo Dry-boil filtered HSV-1 d d d

aqueous extract
Syzygium Flower buds e In vivo Dry-boil filtered HSV-1 d d d

aromaticum aqueous extract


Terminalia Fruit e In vivo Dry-boil filtered HSV-1 d d d

chebula aqueous extract


Santalum album Commercially e e e HSV-1 e
25 e
[46]
obtained HSV-2
Minthostachys Leaves and e In vitro e HSV-1 613 20.25 30.3 [47]
verticillata Stems
Aloysia Fruits and leaves e In vitro HSV-1 150 65 2.31 [48]
gratissima
Artemisia Leaves e In vitro HSV-1 313 83 3,77
douglasiana
Eupatorium Flowers, fruits, e In vitro HSV-1 294 125 2,35
patens and leaves

ANTIHERPETIC ACTIVITY OF PLANT ESSENTIAL OILS


Tessaria Leaves e In vitro HSV-1 263 105 2,50
absinthioides

Melaleuca Commercially e b b b b
[49]
alternifolia obtained

Menthae piperitae Commercially Menthol, In vitro Ethanol HSV-1, ~0.003, ~0.014 ~0.215, [50]
aetheroleum obtained menthone, HSV-2 ~0.0009 ~0,06
isomenthone,
menthyl acetate,
cineole

Rosmarinus Leaves (in spice e In vitro Aqueous HSV-1 1800 e e [51]


officinalis L. form)

Artemisia Whole plant e In vitro Ethanol, HSV-1, 2.4, 4.1 e e [52]


arborescens aqueous HSV-2
Santolina Full blossom e In vitro e HSV-1 112 e 127 [53]
insularis
Santolina Full blossom e In vitro e HSV-2 112 e 160
insularis
a
Evaluated tissue culture infective dose decreasing method at three different concentrations.
b
Statistically significant difference compared to placebo.
d
Statistically evaluated (analysis of variance, Fishers exact test, or Chi-square test and Students t-test).
e
Inhibition of virus replication was measured by the plaque reduction assay.
CC50, 50% cytotoxic concentration; EC50, 50% effective concentration; HSV, herpes simplex virus; SI, selectivity index.
SI CC50/EC50.
c
Determined as the viral titer reduction factor (log10), compared with untreated controls.

253
254 17. ANTIHERPETIC PLANT COMPOUNDS

mice [45]. However, other parameters were not Asia, from India and Nepal to Southwestern
investigated. China. In an antiherpetic treatment study, T.
chebula was examined with acyclovir; in two of
the six mice treated, disease recurrence was
Rhus javanica (Sumac) observed. However, T. chebula was able to
Rhus javanica is a member of the Anacardia- induce a statistically significant improvement
ceae family, which contains a large number of in acyclovir efficiency in vivo [45].
species in the Rhus genus. Rhus spp. are
commonly known as sumac and grow in
temperate and subtropical regions throughout
Santalum album L. (Sandalwood)
the world. R. javanica is native to Asia. For inves- Santalum album L. is a small tropical tree
tigating the potential antiherpetic activity of R. known as sandalwood, which grows abun-
javanica, researchers have studied compounds dantly in China, India, Indonesia, Malaysia,
present in the whole or separate parts of the northwestern Australia, the Philippines, and
plant. In an in vivo animal study, R. javanica Sri Lanka,. In a study, sandalwood EO showed
promoted significant healing when applied in antiviral activity against HSV-1 and HSV-2 in
combination with acyclovir onto the pinna and vitro. However, this effect was not due to a viru-
ganglia of HSV-1-infected mice [45]. cidal activity of the EO, but rather to inhibition
of herpes virus replication [46].

Syzygium aromaticum (Cloves)


S. aromaticum is commonly known as cloves
Minthostachys verticillata (Peperina)
and has a wide range of uses in both medicine Minthostachys verticillata, known as peperina,
and cooking. Cloves are the flower buds of the is the only species of the genus Minthostachys
S. aromaticum tree, a member of the Myrtaceae that grows in Argentina. Its EO has pharmaco-
family. In general, cloves are harvested in India, logic importance. In a study, pulegone, a compo-
Indonesia, Madagascar, Pakistan, and Sri Lanka nent of this EO, showed antiviral activity
and are used as a spice all over the world. The against HSV-1 in vitro. In this study, the EC50
antiherpetic activity of different EOs of clove of M. verticillata was 20.25 mg/mL and the
was investigated and satisfactory results were CC50 was 613 mg/mL (Table 17-2). The SI value
obtained. The recurrence of HSV infection was of M. verticillata EO was calculated as 30.3, while
investigated in mice infected with HSV-1. Ultra- the SI values of its components were: pulegone,
violet irradiation of skin was also applied during 17.6; menthone and limonene were both < 1
the prophylactic treatment with clove EO. Of the [47]. Thus, the antiviral activity of pulegone
seven mice investigated, no HSV DNA was and the EO may play a future role in the drug
detected in samples taken from the pinna of discovery process.
one HSV-infected and clove oil-treated mouse,
indicating that S. aromaticum may be capable of
preventing recurrence of the disease [45]. Melaleuca alternifolia (Tea Tree)
In a clinical trial, Melaleuca alternifolia oil gel
(6%) was used for the treatment of recurrent
Terminalia chebula (Black Myrobalan)
herpes labialis. The oil gel was applied to
The common name for Terminalia chebula is patients five times a day, and assessed for anti-
black myrobalan and it is native to southern herpetic (evaluated by HSV DNA analysis)
TABLE 17-2 Antiviral Activity of Plant Crude Extracts Against Herpes Simplex Virus

Plant part In vivo/ Extract/purified Herpes CC50 EC50


Plant used Active compound In vitro compounds subtype (mg/mL) (mg/mL) SI Reference(s)

Ilex paraguariensis Leaves Caffeoylquinic In vitro Aqueous HSV-1 1260 80.0 15.8 [31]
acid, triterpenoid
Lafoensia pacari Leaves Tannin In vitro Methanol HSV-1 1140 60.0 19.0
Passiflora edulis Roots Saponin, flavonoid In vitro Aqueous HSV-1 1600 89.9 17.8
Rubus imperialis Leaves Tannin, flavonoid In vitro Methanol HSV-1 1390 70.0 19.8
Sloanea guianensis Leaves e In vitro Methanol HSV-1 1400 140.0 10.0

ANTIHERPETIC ACTIVITY OF PLANT ESSENTIAL OILS


Clinacanthus nutans Heartwood Oxyresveratrol In vitro Commercially HSV-1 237,6 24.0 9.9 [21]
produced
Artocarpus lakoocha Heartwood Oxyresveratrol In vitro Methanol HSV-1, 237.5 18,70 12.7
HSV-2
Coptidis rhizoma Whole Berberine, Coptidis In vitro Dried plant HSV-1, 13.2, 8.2, 2.4, 147, [54]
plant rhizoma extract HSV-2 321, 2500 24.3 111, 99
Ching Wei San
extract
Pongamia pinnata Seed e In vitro Ethanol-aqueous HSV-1, 1000 e e [33]
HSV-2 20000

Combination of Leaf, Root Tannin, In vivo Aqueous extract, HSV-1 e e e [55]


Salvia officinalis and anthraquinones aqueous-Ethanol
Rheum palmatum extract
Carissa edulis Root Unknown In vivo Boiled-lyophilized HSV-1, 480 15.1  0.57, 31.79, [15]
In vitro aqueous HSV-2 6.9  1.27 69.57
Hypericum Aerial Hexane extract, In vitro Ethanol HSV-1 e e e [19]
connatum parts flavonoid
Aglaia odorata Leaf e In vivo and Ethanol HSV-1 312 9.5 32.9 [34]
In vitro
Moringa oleifera Leaf e In vivo and Ethanol HSV-1 875 100.0 8.8 [34]
In vitro
Ventilago denticulata Leaf e In vitro Ethanol HSV-1 838 46.3 18.1

255
CC50, 50% cytotoxic concentration; EC50, 50% effective concentration; HSV, herpes simplex virus; SI, selectivity index.
SI CC50/EC50.
256 17. ANTIHERPETIC PLANT COMPOUNDS

and adverse effects. Their findings indicated significant protection in a mouse model of intra-
some benefit from tea tree oil treatment; vaginal HSV-2 [56].
however, none of the differences between
groups reached statistical significance. The SI
of this compound was determined to be 0.215 HIGHLY EFFECTIVE PLANT
against HSV-1 and approximately 0.06 against EXTRACTS AND ESSENTIAL OILS
HSV-2 (Table 17-1) [49].
The most promising herbs, such as A. odorata
(SI value of 32.9) [34], C. edulis (SI value of 69.57)
Menthae piperitae aetheroleum
[15], I. verum (SI value of 160), M. verticillata (SI
(Peppermint Oil) value of 30.3) [47], Santolina insularis (SI value
Menthae piperitae aetheroleum (peppermint oil) of 127e160), and Z. officinale (SI value of 40)
is an EO derived from leaves of M. piperitae [53] are shown as very active in Tables 17-1
aetheroleum L. Peppermint oil is used as a tradi- and 17-2. In addition, most of the other prom-
tional treatment for different human diseases ising herbals showed SI values of > 2. In conclu-
and various pain conditions, such as headaches sion, natural treatments comprising the
[54], postherpetic neuralgia [55], or mild bacte- different herbal compounds described in this
rial or fungal infections of the skin. Recently, chapter can be effective, safe, and without
the antiviral properties of peppermint oil unpleasant side effects. However, information
against HSV-1 and HSV-2 were reported. Viral on the antiviral activities and identities of the
inhibitory activity was tested in vitro on RC-37 active substances present in these extracts and
cells using a plaque reduction assay, and the EOs is not complete. The information provided
findings showed that peppermint oil exhibits in this chapter should enable further studies
high levels of virucidal activity against both should be conducted to enable the development
HSV-1 and HSV-2 in viral suspension tests at of effective antiherpetic drugs based on herbal
noncytotoxic concentrations of the oil. The lipo- compounds. In addition, the literature review
philic nature of this oil provides penetration into indicates that herbal extracts are more effective
the skin and it may therefore have topical thera- when combined with acyclovir. Hence, it is
peutic use as a virucidal agent against recurrent possible that through its combination with
herpes infections [50]. herbs, the increasing resistance to acyclovir
may be prevented and therapeutic dose of
acyclovir may be reduced.
Other Important Plant Essential Oils
Other plant extracts, chemical compounds,
and products have been studied for activity REVIEW CRITERIA
against HSV-2 both in vitro and in vivo. The
extracts contained a very wide range of chemi- The Science Citation Index of the Web of
cal compounds (including borneol, caffeic acid, Science and the PubMed database were
carrageenan polysaccharide, cineole, cinnamon searched using various combinations of the
oil, citral, curcumin) and the effects of these terms Herpes simplex, HSV-1, HSV-2, plant
against HSV-2 were investigated. Following in compounds (and derivative words), natural prod-
vitro analysis, the compounds with the highest ucts, antiherpetic drugs, clinical trial, in vivo,
antiviral activities were tested in mice. Four mouse, in vitro, and therapeutic index to identify
compounds, carrageenan lambda type IV, relevant articles published between 1954 and
cineole, curcumin, and eugenol, provided 2010. Full-length articles published in English
REFERENCES 257
were selected. For in vitro and in vivo studies, the detail the data obtained from several antiher-
reference lists of recent research papers and petic studies in the tables. Herbal extracts and
reviews were used to identify further articles. EOs were evaluated for their antiherpetic effects
according to different parameters. Therefore, we
reviewed the data according to their SI values.
CONCLUSIONS AND FUTURE Generally, for most of these in vitro studies, the
PROSPECTS SI value was calculated to determine whether
the extracts contained sufficient antiviral
Several drugs such as acyclovir are available activity. The SI measures the antiviral efficiency
for the treatment of HSV infections. Acyclovir is and should exceed the level of toxicity. Herbal
a nucleoside analog and selective antiherpetic components with an SI of  2 are considered
agent. It inhibits viral DNA replication and active and those with an SI of  10 are consid-
DNA synthesis by targeting viral thymidine ered very active.
kinase. However, acyclovir resistance is being Consequently, this chapter reveals that plant
increasingly described, caused by mutations in extracts and EOs show great potential for use
either the thymidine kinase or the DNA poly- in the treatment of HSV infections. However,
merase genes [59]. Acyclovir-resistant herpes we found that many studies are restricted to in
viruses have been isolated from the patients vitro investigations and that there are insuffi-
with AIDS or cancer, as well as recipients of cient numbers of in vivo studies and clinical
bone marrow or organ transplants [60,61]. trials. However, considering recent develop-
Therefore new anti-HSV drugs are urgently ments in this field, we believe that the applica-
needed, and many studies have focused on tions of plant compounds for treating HSV
herbals. Medicinal and aromatic plants are infections in humans will begin shortly and
widely used today in modern phytotherapy that the promising results described in this
[62]. Recently, anti-HSV activity has been chapter will guide these applications.
reported in some plant extracts and EOs. Several
herbal components with activity against HSV-1
and HSV-2 have been proposed as promising References
alternative therapeutic tools [44]. These herbals [1] Snoeck R. Antiviral therapy of herpes simplex. Int J
are often also effective against acyclovir-resis- Antimicrob Agents 2000;16:157e9.
tant strains. Many herbal extracts and oils [2] Ablin J, Symon Z, Mevorach D. Sacral herpes-zoster
have been investigated against HSV; however, infection presenting as sciatic pain. J Clin Rheumatol
1996;2:167e9.
there was no comprehensive review of the [3] Aurelian L, Specter S, Hodinka R, Young S,
knowledge gleaned from these studies. There- Wiedbrauk D. Herpes simplex viruses. Clin Virol
fore, for the first time, we have tried to evaluate Manual 2009:424e53.
and review the anti-HSV effects of different [4] Looker KJ, Garnett GP, Schmid GP. An estimate of the
herbal extracts and oils. Our aim was to bring global prevalence and incidence of herpes simplex
virus type 2 infection. Bull WHO 2008;86:805e12.
together the information about various herbal [5] Fatahzadeh M, Schwartz RA. Human herpes simplex
compounds available in the literature, investi- virus infections: epidemiology, pathogenesis, symp-
gate the antiherpetic effectiveness of these tomatology, diagnosis, and management. J Am Acad
compounds, and identify the potential of these Dermatol 2007;57:737e63. quiz 764e36.
plants for developing novel herpetic treatments. [6] Whitley RJ, Kimberlin DW, Roizman B. Herpes
simplex viruses. Clin Infectious Diseases
In this chapter, plants investigated for their 1998;26:541e53.
antiherpetic effectiveness are presented as [7] Brady RC, Bernstein DI. Treatment of herpes simplex
herbal extracts and EOs. We have described in virus infections. Antivir Res. 2004;61:73e81.
258 17. ANTIHERPETIC PLANT COMPOUNDS

[8] De Clercq E. Discovery and development of BVDU herpes simplex virus replication. Antivir Res
(brivudin) as a therapeutic for the treatment of herpes 1999;43:145e55.
zoster. Biochem Pharmacol 2004;68:2301e15. [23] Docherty JJ, Smith JS, Fu MM, Stoner T, Booth T.
[9] De Clercq E. Antiviral drugs in current clinical use. J Effect of topically applied resveratrol on cutaneous
Clin Virol 2004;30:115e33. herpes simplex virus infections in hairless mice.
[10] De Clercq E, Andrei G, Snoeck R, De Bolle L, Antivir Res 2004;61:19e26.
Naesens L, Degreve B, et al. Acyclic/carbocyclic [24] Gudima S, Memelova L, Borodulin V, Pokholok D,
guanosine analogues as anti-herpesvirus agents. Mednikov B, Tolkachev O, et al. [Kinetic analysis of
Nucleosides Nucleotides Nucleic Acids interaction of human immunodeficiency virus reverse
2001;20:271e85. transcriptase with alkaloids]. Molekuliarnaia Bio-
[11] Leung DT, Sacks SL. Current recommendations for the logiia 1994;28:1308.
treatment of genital herpes. Drugs 2000;60:1329e52. [25] Hayashi K, Minoda K, Nagaoka Y, Hayashi T,
[12] Collins P, Darby G. Laboratory studies of herpes Uesato S. Antiviral activity of berberine and related
simplex virus strains resistant to acyclovir. Rev Med compounds against human cytomegalovirus. Bio-
Virol 1991;1:19e28. organic & medicinal chem let 2007;17:1562e4.
[13] Khan MTH, Ather A, Thompson KD, Gambari R. [26] Li HL, Han T, Liu RH, Zhang C, Chen HS, Zhang WD.
Extracts and molecules from medicinal plants against Alkaloids from Corydalis saxicola and their anti-
herpes simplex viruses. Antivir Res 2005;67:107e19. hepatitis B virus activity. Chem & Biodiversity
[14] Gachathi FN. Kikuyu botanical dictionary of plant 2008;5:777e83.
names and uses. Nairobi, Kenya: AMREF; 1989. [27] Schmeller T, Latz-Bruning B, Wink M. Biochemical
[15] Tolo FM, Rukunga GM, Muli FW, Njagi ENM, activities of berberine, palmatine and sanguinarine
Njue W, Kumon K, et al. Anti-viral activity of the mediating chemical defence against microorganisms
extracts of a Kenyan medicinal plant Carissa edulis and herbivores. Phytochemistry 1997;44:257e66.
against herpes simplex virus. J Ethnopharmacol [28] Sethi ML. Enzyme inhibition VI: inhibition of reverse
2006;104:92e9. transcriptase activity by protoberberine alkaloids and
[16] Taylor A, Mc KG, Burlage HM, Stokes DM. Plant structureeactivity relationships. J Pharm Sci
extracts tested against egg cultivated viruses. Tex Rep 1983;72:538e41.
Biol Med 1954;12:551e7. [29] Sethi MI. Comparison of inhibition of reverse tran-
[17] Sydiskis R, Owen D, Lohr J, Rosler K, Blomster R. scriptase and antileukemic activities exhibited by
Inactivation of enveloped viruses by anthraquinones protoberberine and benzophenanthridine alkaloids
extracted from plants. Antimicrob Agents Chemother and structure-activity relationships. Phytochemistry
1991;35:2463. 1985;24:447e54.
[18] Saller R, Buechi S, Meyrat R, Schmidhauser C. [30] Roizman B, Sears A. Herpes simplex viruses and their
Combined herbal preparation for topical treatment of replication. In: The human herpesviruses. New York,
Herpes labialis. Forsch Komplementarmed Klass NY: Raven Press; 1993. p. 11e68.
Naturheilkd. 2001;8:373e82. [31] Muller V, Chavez JH, Reginatto FH, Zucolotto SM,
[19] Fritz D, Venturi CR, Cargnin S, Schripsema J, Niero R, Navarro D, et al. Evaluation of antiviral
Roehe PM, Montanha JA, et al. Herpes virus inhibi- activity of South American plant extracts against
tory substances from Hypericum connatum Lam., herpes simplex virus type 1 and rabies virus. Phyt-
a plant used in southern Brazil to treat oral lesions. J other Res 2007;21:970e4.
Ethnopharmacol 2007;113:517e20. [32] Goncalves JL, Leitao SG, Monache FD, Miranda MM,
[20] Sangkitporn S, Chaiwat S, Balachandra K, Na- Santos MG, Romanos MT, et al. In vitro antiviral effect
Ayudhaya TD, Bunjob M, Jayavasu C. Treatment of of flavonoid-rich extracts of Vitex polygama (Verbe-
herpes zoster with Clinacanthus nutans (bi phaya naceae) against acyclovir-resistant herpes simplex
yaw) extract. J Med Assoc Thai 1995;78:624e7. virus type 1. Phytomedicine 2001;8:477e80.
[21] Chuanasa T, Phromjai J, Lipipun V, [33] Elanchezhiyan M, Rajarajan S, Rajendran P,
Likhitwitayawuid K, Suzuki M, Pramyothin P, et al. Subramanian S, Thyagarajan S. Antiviral properties of
Anti-herpes simplex virus (HSV-1) activity of oxy- the seed extract of an Indian medicinal plant, Pon-
resveratrol derived from Thai medicinal plant: mech- gamia pinnata, Linn., against herpes simplex viruses:
anism of action and therapeutic efficacy on cutaneous in vitro studies on Vero cells. J Med Microbiol
HSV-1 infection in mice. Antivira Res 2008;80:62e70. 1993;38:262e4.
[22] Docherty JJ, Fu MM, Stiffler BS, Limperos RJ, [34] Lipipun V, Kurokawa M, Suttisri R, Taweechotipatr P,
Pokabla CM, DeLucia AL. Resveratrol inhibition of Pramyothin P, Hattori M, et al. Efficacy of Thai medicinal
REFERENCES 259
plant extracts against herpes simplex virus type 1 infec- [49] Carson C, Ashton L, Dry L, Smith D, Riley T. Mela-
tion in vitro and in vivo. Antivir Res 2003;60:175e80. leuca alternifolia (tea tree) oil gel (6%) for the treat-
[35] Kurokawa M, Nagasaka K, Hirabayashi T, Uyama S, ment of recurrent herpes labialis. J Antimicrob
Sato H, Kageyama T, et al. Efficacy of traditional Chemoth 2001;48:450.
herbal medicines in combination with acyclovir [50] Schuhmacher A, Reichling J, Schnitzler P. Virucidal
against herpes simplex virus type 1 infection in vitro effect of peppermint oil on the enveloped viruses
and in vivo. Antivir Res 1995;27:19e37. herpes simplex virus type 1 and type 2 in vitro.
[36] Bruneton J, Pharmacognosy P. Medicinal Plants. Phytomedicine 2003;10:504e10.
Intercept, Hampshire 1995. [51] Mancini DAP, Torres RP, Pinto JR, Mancini-Filho J.
[37] Khan IA, Abourashed EA. Leungs Encyclopedia of Inhibition of DNA virus: Herpes-1 (HSV-1) in cellular
Common Natural Ingredients: Used in Food, Drugs culture replication, through an antioxidant treatment
and Cosmetics. NY, USA: John Wiley & Sons; 2010. extracted from rosemary spice. Braz J Pharm Sci
[38] Bisset NG, Czygan FC. Herbal drugs and phyto- 2009;45:127e33.
pharmaceuticals: a handbook for practice on a scien- [52] Saddi M, Sanna A, Cottiglia F, Chisu L, Casu L,
tific basis; [with reference to German Commission E Bonsignore L, et al. Antiherpevirus activity of Arte-
monographs]. Germany: Medpharm GmbH; 2001. misia arborescens essential oil and inhibition of lateral
[39] Cohen RA, Kucera LS, Herrmann Jr EC. Antiviral diffusion in Vero cells. Ann Clin Microbiol Anti-
activity of Melissa officinalis (lemon balm) extract. microb 2007;6:10.
Exp Biol Med 1964;117:431. [53] De Logu A, Loy G, Pellerano ML, Bonsignore L,
[40] Kucera LS, Herrmann Jr EC. Antiviral substances in Schivo ML. Inactivation of HSV-1 and HSV-2 and
plants of the mint family (Labiatae). I. Tannin of prevention of cell-to-cell virus spread by Santolina
Melissa officinalis. Exp Biol Med 1967;124:865. insularis essential oil. Antivir Res 2000;48:177e85.
[41] May G, Willuhn G. Antiviral effect of aqueous plant [54] Gobel H, Schmidt G, Dworschak M, Stolze H,
extracts in tissue culture]. Arzneimittel-Forschung Heuss D. Essential plant oils and headache mecha-
1978;28:1. nisms. Phytomedicine 1995;2:93e102.
[42] Allahverdiyev A, Duran N, Ozguven M, Koltas S. [55] Davies SJ, Harding LM, Baranowski AP. A novel
Antiviral activity of the volatile oils of Melissa offici- treatment of postherpetic neuralgia using peppermint
nalis L. against Herpes simplex virus type-2. Phyto- oil. Clin. J Pain 2002;18:200.
medicine 2004;11:657e61. [56] Bourne KZ, Bourne N, Reising SF, Stanberry LR. Plant
[43] Koch C, Reichling J, Schneele J, Schnitzler P. Inhibi- products as topical microbicide candidates: assess-
tory effect of essential oils against herpes simplex ment of in vitro and in vivo activity against herpes
virus type 2. Phytomedicine 2008;15:71e8. simplex virus type 2. Antivir Res 1999;42:219e26.
[44] Astani A, Reichling J, Schnitzler P. Screening for [57] Chin LW, Cheng YW, Lin SS, Lai YY, Lin LY, Chou MY,
antiviral activities of isolated compounds from et al. Anti-herpes simplex virus effects of berberine
essential oils. Evidence-based Complement. Alternat from Coptidis rhizoma, a major component of
Med 2011;2011:253643. a Chinese herbal medicine, Ching-Wei-San. Arch Virol
[45] Kurokawa M, Nakano M, Ohyama H, Hozumi T, 2010;155:1933e41.
Kageyama S, Namba T, et al. Prophylactic efficacy of [58] Reuter J, Wolfle U, Weckesser S, Schempp C. Which
traditional herbal medicines against recurrent herpes plant for which skin disease? Part 1: Atopic derma-
simplex virus type 1 infection from latently infected titis, psoriasis, acne, condyloma and herpes simplex. J
ganglia in mice. J Dermatol Sci 1997;14:76e84. Dtsch Dermatol Ges 2010;8:788e96.
[46] Benencia F, Courreges MC. Antiviral activity of [59] Pottage Jr J, Kessler H. Herpes simplex virus resis-
sandalwood oil against herpes simplex viruses-1 and -2. tance to acyclovir: clinical relevance. Infect Agent Dis
Phytomedicine 1999;6:119e23. 1995;4:115.
[47] Mara V, Vogt SBSFME, Sabini Mara C, Cariddi Laura [60] Bacon TH, Levin MJ, Leary JJ, Sarisky RT, Sutton D.
N, Torres Cristina V, Zanon Silvia M, et al. Minthos- Herpes simplex virus resistance to acyclovir and
tachys verticillata essentials oil and its major compo- penciclovir after two decades of antiviral therapy.
nents: antiherpetic selective action in HEp-2 cells. Mol Clin Microbiol Rev 2003;16:114.
Med Chem 2010;21:117e20. [61] Morfin F, Thouvenot D. Herpes simplex virus resis-
[48] Garcia C, Talarico L, Almeida N, Colombres S, tance to antiviral drugs. J Clin Virol 2003;26:29e37.
Duschatzky C, Damonte E. Virucidal activity of [62] Demirci B, Demirci F. Essential oil of Betula pendula
essential oils from aromatic plants of San Luis, 17. Roth. Buds. Evidence-based complement. Alternat
Argentina: Phytother. Res 2003; 1073e1075. Med 2004;1:301e3.