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Antiviral activity and mode of action of Dianthus caryophyllus L. and Lupinus termes L. Seed extracts against in vitro herpes simplex and hepatitis A viruses infection

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Crude extracts of sixteen seeds belonging to different plant species were tested for their antiviral activity against herpes simplex virus-1 (HSV-1) and hepatitis A virus-27 (HAV-27). Non-toxic concentration (20 µg/ml) of Dianthus caryophyllus and Lupinus termes seed extracts to both Vero and HepG2 cells showed potent antiviral activity against HSV-1 and HAV-27 using plaque infectivity count assay. The mechanism of action D. caryophyllus revealed its virucidal activity against HSV-1 and HAV-27 as 92.3 and 92.6%, respectively, while, the virucidal activity of L. termes was observed only against HAV-27 giving 93.7% of inhibition. No effect was detected for both extracts on adsorption or on the stages of virus replication. A comparison has been done between the antiviral activity of two therapeutic drugs (Acyclovir and Amentadine used as controls for HSV-1 and HAV-MBB, respectively) and the two tested seed extracts. The results revealed that these seed extracts were more efficient in their inhibitory activity than synthetic chemical drugs against the same viruses. This may open the way to give more attention to use the natural botanical origin in treatmenting viral infection with or without therapeutic agents to obtain better recovery with least side effects.
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Journal of Microbiology and Antimicrobials Vol. 2(3), pp. 23-29, May 2010
Available online http://www.academicjournals.org/JMA
ISSN 1996-0875 © 2010 Academic Journals
Full Length Research Paper
Antiviral activity and mode of action of Dianthus
caryophyllus L. and Lupinus termes L. seed extracts
against in vitro herpes simplex and hepatitis A viruses
infection
Ahmed B. Barakat1*, Sahar A. Shoman1, N. Dina2 and Omar R. Alfarouk1
1Department of Microbiology, Faculty of Science, Ain Shams University, Cairo, Egypt.
2Department of water pollution, National Research Center, Dokki, Egypt.
Accepted 28 January, 2010
Crude extracts of sixteen seeds belonging to different plant species were tested for their antiviral
activity against herpes simplex virus-1 (HSV-1) and hepatitis A virus-27 (HAV-27). Non-toxic
concentration (20 µg/ml) of Dianthus caryophyllus and Lupinus termes seed extracts to both Vero and
HepG2 cells showed potent antiviral activity against HSV-1 and HAV-27 using plaque infectivity count
assay. The mechanism of action D. caryophyllus revealed its virucidal activity against HSV-1 and HAV-
27 as 92.3 and 92.6%, respectively, while, the virucidal activity of L. termes was observed only against
HAV-27 giving 93.7% of inhibition. No effect was detected for both extracts on adsorption or on the
stages of virus replication. A comparison has been done between the antiviral activity of two
therapeutic drugs (Acyclovir and Amentadine used as controls for HSV-1 and HAV-MBB, respectively)
and the two tested seed extracts. The results revealed that these seed extracts were more efficient in
their inhibitory activity than synthetic chemical drugs against the same viruses. This may open the way
to give more attention to use the natural botanical origin in treatmenting viral infection with or without
therapeutic agents to obtain better recovery with least side effects.
Key words: Antiviral seed extract, herpes virus infection, hepatitis virus infection, amantadine, acyclovir.
INTRODUCTION
There is currently a large and ever-expanding global
population base that prefers the use of natural products
in treating and preventing medical problems. This has
influenced many pharmaceutical companies to produce
new antimicrobial formulations extracted from plants or
herbs. At present, plant and herb resources are unlimited,
have provided mankind remedies for many infectious
diseases and continue to play a major role in primary
health care as therapeutic remedies in developing
countries (Sokmen et al., 1999). The search for biological
active extracts based traditionally used plants is still
relevant due to induction of resistance of pathogens to
chemical drugs and the prevalence of the fatal different
infections (Rabindran et al., 2003). Human herpes
viruses are found worldwide and are among the most
*Corresponding author. E-mail: karimbahaa2004@yahoo.com.
frequent causes of viral infections in immunocompetent
as well as in immunocompromised patients. During the
past two decade a better understanding of the replication
and disease causing state of herpes simplex virus type 1
and 2 (HSV-1 and HSV-2), has been achieved due the
development of potent antiviral compounds that target
these viruses. While some of the antiviral therapies are
considered safe and efficacious (acyclovir, pencyclovir),
others have toxicities associated with them (gancyclovir
and foscarnet). In addition, the increased and prolonged
use of these compounds in clinical setting, especially for
the treatment of immunocompromised patients, has led to
the emergence of viral resistance against most of these
drugs (Villarreal, 2001).
Fulminant hepatitis is a severe complication of hepatitis
A virus infection (HAV). Its mechanism is unknown but
spontaneous recovery is frequent. There are no data on
the level of viral replication according to the clinical form
of HAV (Rezende et al., 2003). A high fatality rate among
24 J. Microbiol. Antimicrob.
chronic hepatitis B or C patients with HAV super-infection
was observed (Lee, 2003). Although there are no
commercial antiviral drugs specifically licensed for
treating HAV infection, ribavirin, amentadine, and 2-
deoxy-D-glucose are among several antiviral substances
known to interfere with HAV replication (Hollinger and
Emerson, 2001).
Although a significant number of studies have used
known purified plant chemicals as antiviral drugs (Binnus
et al., 2002; Guarino and Sciarrillo, 2003; Jassim and
Naji, 2003), very few screening programmes have been
initiated on crude plant materials. Crude extracts of plant
seeds are also a promising source of systemic broad-
spectrum antiviral that may cause less damage to host
cells than do pharmaceuticals. Topical antiviral
substances are also important areas of study for the
treatment of viral lesions such as in HSV, and plant-
based substances offer promise as virucidal alternates
(Hudson, 1990). The seed extracts of Phyllanthus
amarus (Euphorbiaceae) species are known to reduce or
eliminate detectable hepatitis B virus surface antigen in
humans and show in vitro inhibition of viral DNA
polymerase (DNAp) (Unander and Blumberg, 1991). The
repeated oral administration of extracts of Strychnos
potatrum seeds appreciably suppressed the development
of skin lesions induced by HSV-1 in mice (Hattori et al.,
1995). The Pinus nigra seed cones extract has anti-HIV
activity (Eberhardt and Young, 1996). Incubation of
acyclovir-resistant HSV-1 (ACVr-HSV-1), during infection
of the HEp-2 cell culture, with an extract prepared from
the seeds of Licania tomentosa species impaired the
productive replication of this virus in a concentration-
dependent manner. The extract was able to inhibit
extracellular virus (virucidal effect) and also interfered
with a very early event of cell infection at a non-cytotoxic
concentration (Miranda et al., 2002).
The current investigation was undertaken to test the
extracts of 16 plant seeds for their antiviral activity
against herpes simples virus -1 (HSV-1, a DNA virus) and
hepatitis A virus (HAV, a RNA virus). The mode of action
of the most promising extracts was also studied.
MATERIALS AND METHODS
Plant material
Sixteen species of seeds belonging to different families were
collected from seed bank of botanical garden, Ain Shams
University; Ministry of Agriculture and land reclamation, Giza,
Egypt; Orman botanical garden, Giza, Egypt and Flora and
phytotaxonomy research department, Agriculture museum, Giza,
Egypt.
Preparation of seed extracts for bioassay
10 mg of each crushed seed was resuspended in 1 ml solvent (10%
Dimethyl sulfoxide (DMSO) in deionized water). Decontamination
was carried out by adding 1% antibiotic-antimycotic mixture (10,000
IU Penicillin G sodium, 10,000 µg Streptomycin sulfate and
250 µg Amphotericin B) and the extracts were incubated at 37°C for
30 min then stored at -20°C. Sterility test was performed to ensure
the sterility of the prepared extracts.
Cells
Both Vero and HepG2 cells were propagated in minimum essential
medium (MEM) and RPMI 1640 medium respectively. They were
supplemented with 10% foetal bovine serum, 1% antibiotic-
antimycotic mixture. The cell culture was kindly provided by faculty
of medicine, El-azhar University as confluent monolayer in 25 cm2
tissues culture flasks.
Cytotoxicity assays
The cell culture safety doses of the dissolved seed extracts were
performed by cell morphology technique (Aquino et al., 1989). Seed
extracts were inoculated (100 µL each) into both cell lines with
concentration, 5, 10, 20, 30, 40, 50 µg /100 µL and observed
microscopically for any morphological changes after 24 h incubation
at 37o C in a humidified incubator with 5% CO2 .
Viruses
Egyptian isolate of Herpes simplex virus type 1, was provided by
Virology Lab., Department of water pollution, NRC. Hepatitis A
virus-MBB strain was kindly provided by Prof. Dr. Verena gauss-
Muller, Molecular virology Institute, Luebeck University for
Medicine, Germany.
Antiviral bioassay
Plaque infectivity count assay is the most widely accepted method
for determining the % inhibition of virus as a result of being
subjected to a given material (Tebas et al., 1995). A 6 well plate
was cultivated with the specific cell type (105cell/mL) and incubated
for 1 - 2 days at 37°C. Virus was diluted to final concentration of 107
PFU/mL and mixed with the safe concentrations of each seed
extract as mentioned previously and incubated for 1 h at 37°C.
Growth medium was removed from the multi-well plate and virus-
extract mixture was inoculated (100 µl/ well). After 1 h contact time
for virus adsorption, the inoculum was aspirated and 3 ml of cell-
specific 2× medium 2% agarose was overlaid the cell sheet. The
plates were left to solidify and incubated at 37°C until the
development of the viral plaques. Formalin was added for two hours
then plates were stained with crystal violet staining solution. Control
virus and cells were treated identically without seed extract. Viral
plaques were counted and the percentage of virus reduction was
calculated.
Mechanism of virus inhibition
Virus inhibition mechanism for the most potent crude seed extracts
was studied in three categories:
A) Virucidal; tested by subjecting virus to extract directly
(Schuhmacher et al., 2003).
B) Viral Adsorption; tested by subjecting cells to extract for 2 h
before virus inoculation (Zhang et al., 1995).
C) Viral replication; tested by post inoculation of extract after virus
application to cells (Amoros et al., 1994).
Barakat et al. 25
Table 1. Cytotoxicity of the sixteen seed extracts on Vero and HepG2 cells.
Seed extracts / Cell culture
Conc. of extracts (µ
µµ
µg)
5 10 20 30 40 50
Vero/HepG2
A. precatorius
L. +/+ +/+ +/+ +/+ +/+ +/+
A. cepa L. -/- -/- -/- -/- -/- -/-
A. nobilis L. -/- -/- -/- -/- -/- -/-
C. frutescens L. -/- -/- -/- -/- -/- -/-
D. caryophyllus L. -/- -/- -/- +/- +/+ +/+
E. sativa Mill. -/- -/- -/- +/+ +/+ +/+
G. hispida L. -/- -/- -/- +/+ +/+ +/+
L. usitatissimum L. -/- -/- -/- +/+ +/+ +/+
L. termes L. -/- -/- -/- -/- +/+ +/+
N. sativa L. -/- -/- -/- +/+ +/+ +/+
P. harmala L. -/- -/- -/- +/+ +/+ +/+
P. vulgaris L. -/- -/- -/- +/+ +/+ +/+
P. sativum L. -/- -/- -/- -/+ +/+ +/+
P. armeniaca Marshall -/- -/- -/- +/- +/+ +/+
S. alba L. -/- -/- -/- +/+ +/+ +/+
T. f. graecum L. -/- -/- -/- -/- -/- -/-
(-): means safe to cells / (+): means toxic to cells.
RESULTS AND DISCUSSION
Cytotoxicity of tested seed extracts on VERO and
HepG2 cells
Results as shown in Table (1) indicate that the accepted
safe concentrations on both Vero and HepG2 cells were
less than 30 µg/100 µl. The results also showed that the
rate of cell death increased with increasing the
concentration of the tested seed extract. However, 4 out
of 16 tested seed extracts (Allium cepa, Capsicum
frutescens, Anthemis nobilis and Trigonella foenum
graceum) had no toxic effect on both Vero and HepG2
cells even when applied at high concentrations. On the
other hand, Abrus precatorius showed high toxicity on
both Vero and HepG2 cells even when applied at low
concentrations.
The antiviral activity of seed extracts against HSV-1
and HAV-27
To evaluate the antiviral activities of 16 plant seeds, the
inhibitory effects on the plaque formation were examined.
The results in Table 2 show that out of sixteen seed
extracts, D. caryophyllus has strong inhibitory activity
against both HSV-1 and HAV-27 giving 92.3 and 92.6%
inhibition at 20 µg, respectively. L. termes extract showed
strong inhibitory activity against HAV-27 only giving
93.7% inhibition at 20 µg (Table 2). Except for these two
seed species, all other tested seed extracts showed
moderate or negligible inhibitory activity, giving us a
green light to put both seeds under focus as promising
natural extracts to be used for therapeutic purposes.
The effectiveness of seed extracts inhibiting several
human viruses has been demonstrated. For examples,
the hot water extract from seeds of Arachis hypogaea
blocked HSV infection while, the hot water extract from
seeds of Pisum sativum blocked adenoviruses (ADV)
infection (Chiang et al., 2003). The hot-water extract of
black soyabean showed significant antiviral activity
against human adenovirus type 1 and coxsackievirus B1
(Yamai et al., 2003). The crude seed extract of Quercus
lusitanica plant also has a good inhibitory effect on the
replication of dengue virus type 2 (Muliawan et al., 2006).
The purified Egyptian pea (Pisum sativum) lectin which
was isolated from its seed showed a high inhibitory effect
on HCV replication (Al-Sohaimy et al., 2007). In addition,
the black cumin seed (Nigella sativa) exhibited antiviral
activity against infectious Laryngotracheitis virus (Zaher
et al., 2008).
Inhibitory action of D. caryophyllus L. and L. termes L.
extracts comparing with Antiviral drugs (Acyclovir and
Amentadine) against HSV-1 and HAV-MBB viruses.
Effectness of D. caryophyllus extract and anti HSV-1
therapeutic agent (Acyclovir) was compared. Using
similar concentrations of acyclovir and extract starting
from 10 to 50 µg /100 µL were tested against the same
virus (HSV-1). In similar manner, L. termes extract and
anti HAV-MBB control (Amentadine) was also compared.
Similar concentrations of amentadine and each extract
(10 to 50 µg /100 µl) were individually tested against the
26 J. Microbiol. Antimicrob.
Table 2. Inhibitory activity of seed extracts (n = 16) using plaque reduction assay against HSV-1 and HAV-27.
Antiviral effect
Conc.
(µg)
Seed extract
HAV- 27 HSV- 1
% of
virucidal
effect
Viral count
(PFU/ml) × 107
Initial viral
count ×107
% of
virucidal
effect
Viral count
(PFU/ml) × 107
Initial viral
count ×107
0 High toxicity 3.5 0 High toxicity 2.6 10
A. precatorius L. 0 1.6 3.5 0 2 2.6 20
49.7 1.76 3.5 0 2.6 2.6 10
A. cepa L. 54.3 1.6 3.5 0 2.8 2.6 20
20 2.8 3.5 0 3 2.6 10
A. nobilis L. 25.7 2.6 3.5 2.3 2.54 2.6 20
20 2.8
2
3.5 0 2.8
2.88
2.6 10
C. frutescens L. 42.9 3.5 0 2.6 20
88 0.42 3.5 88.5 0.3 2.6 10
D. caryophyllus L. 92.6 0.26 3.5 92.3 0.2 2.6 20
5.7 3.3 3.5 7.7 2.4 2.6 10
E. sativa Mill.
20 2.8 3.5 24.6 1.96 2.6 20
25.7 2.6 3.5 0 2.7 2.6 10
G. hispida L. 42.9 2 3.5 0 2.65 2.6 20
51.4 1.7 3.5 0 3.2 2.6 10
L. usitatissimum L. 65.7 1.2 3.5 34.6 1.7 2.6 20
92 0.28 3.5 0 2.6 2.6 10
L. termes L. 93.7 0.22 3.5 0 2.64 2.6 20
31.43 2.4 3.5 0 2.6 2.6 10
N. sativa L. 42.6 2 3.5 30.77 1.8 2.6 20
45.7 1.9 3.5 3.9 2.5 2.6 10
P.
harmala L. 65.7 1.2 3.5 7.7 2.4 2.6 20
65.7 1.2 3.5 0 3.2 2.6 10
P. vulgaris L. 71.4 1 3.5 3.1 2.52 2.6 20
48.6 1.8 3.5 30.77 1.8 2.6 10
P. sativum L. 51.4 1.7 3.5 46.15 1.4 2.6 20
42.6 2 3.5 0 2.7 2.6 10
P. armenia Marshell 48.6 1.8 3.5 0 2.8 2.6 20
31.4 2.4 3.5 11.5 2.3 2.6 10
Sinapis alba L. 54.3 1.6 3.5 23 2 2.6 20
0 3.6 3.5 0 2.8 2.6 10
Trigonella f. graecum 25.7 2.6 3.5 11.5 2.3 2.6 20
Barakat et al. 27
same virus (HAV-27). The results in Figures 1 and 2
show that the inhibitory activity of all applied
concentrations of the natural seed extracts were higher
than that shown by Acyclovir and Amentadine at the
same concentrations. Treatment of viral infection either
using herbal extracts or combination between natural
extracts and therapeutic agents has been reported in
many investigations. Soybean oil showed significantly
higher activity in vitro against both Herpes simplex virus
and Para-influenza–3 virus as compared to acyclovir and
Oseltamivir (Orhan et al., 2007). The synergistic effect of
betulin, a pentacyclic triterpenoid, isolated from the bark
of Betula papyrifera with acyclovir against herpes simplex
viruses (Yunhao et al., 2004). A combined application of
flowers of Verbascum thapsiforme and three amentadine
derivatives resulted in a marked enhancement of the
inhibitory effect of the natural extract on the reproduction
of influenza virus (Serkedjieva, 2000).
Corina et al. (1999) examined the effect of extracts of
Romanian medicinal plants in combination with acyclovir
in the treatment of 52 patients suffering herpetic keratitis.
Better results and faster healing of ulceration were
obtained using Actium lappa, Calendula officinalis and
Geranium robertianum extracts than with the usual
acyclovir treatment only. These herbal extracts may have
different mechanisms of anti-HSV-1 action from Acyclovir
thus the combination of Acyclovir with herbal extracts
might have worked synergistically. It is also observed that
patients in the Far East are incorporating orthodox
medical drugs into herbal medicinal preparations for
alleviating their illnesses (Chan and Cheung, 2000). The
rationale for doing so is to reduce the side effects of
orthodox medical drugs, and to produce synergistic
effects for better treatment outcome.
Mechanism of action of D. caryophyllus and L.
termes extracts against HSV-1 and HAV-MBB
The results obtained by plaque infectivity count assay
when the seed extracts A and B (D. caryophyllus) and C
(L. termes) (Figure 3) were applied with pre and post viral
treatment revealed that both seed extracts have strong
virucidal activity (97.1, 88.7 and 96.9% at 60 g) either
by their effect on the virus or forming a complex with the
virus preventing it from being adsorbed to its binding sites
on Vero or HepG2 cells. However, no effects were shown
either on the early adsorption or on the replication of
HSV-1 and HAV-27. These results agreed with the study
on Peppermint oil which has antiviral activity against an
acyclovir resistant strain of HSV-1 (HSV-1-Acv). This
essential oil is capable to exert a direct virucidal effect on
HSV (Schuhmacher et al., 2003). While, the mannose-
specific plant lectins showed strong antiviral activity in
vitro against the two corona viruses severe acute
respiratory syndrome (SARS) and the feline infectious
peritonitis virus (FIPV) at 50 - 100 µg/mL by interfering
with two targets in the viral replication cycle. The first
0
10
20
30
40
50
60
70
80
90
100
10
20
30
40
50
Extract conc. in microgram
% of Inhibition
Dianthus
caryophyllus
Acyclovir
Figure 1. Comparison between Dianthus caryophyllus seed
extract and Acyclovir for HSV-1 inhibition.
0
10
20
30
40
50
60
70
80
90
100
10
20
30
40
50
Extract conc. in microgram
% of Inhibition
Dianthus
caryophyllus
Amentadine
Lupinus termes
Figure 2. Comparison between Dianthus caryophyllu,
Lupinus termes seed extracts and Amentadine for
HAV-27 inhibition.
target is located early in the replication cycle, most
probably viral attachment, and the second target is
located at the end of the infectious virus cycle (Keyaerts
et al., 2007).Generally, there are many antiviral
compounds can be found in botanical sources which
have the ability to inhibit human DNA and RNA viruses
which causing serious diseases to humans without
damaging or affecting the host cells. From this
investigation, we hope to open the way for several
studies in this field on these promising effectiveness
natural seed extracts to be used with or without
commercial therapeutic agents against human viral
infections.
28 J. Microbiol. Antimicrob
A B C
Virucidal effect of Dianthus
caryophyllus extract on HSV-1
(60 µg gives 97.1 % viral
inhibition)
Virucidal effect of Dianthus
caryophyllus extract on HAV-27
(60 µg gives 88.7 % viral
inhibition)
Virucidal effect of Lupinus
termes extract on HAV-27
(60 µg gives 96.9 % viral
inhibition)
Effect of pre-treatment of Vero
cells with Dianthus
caryophyllus extract before its
inoculation with HSV-1
(60 µg gives 2.9 % viral
inhibition)
Effect of pre-treatment of HepG2
cells with Dianthus caryophyllus
extract before its inoculation with
HAV-27
(60 µg gives 6 % viral inhibition)
Effect of pre-treatment of
HepG2 cells with Lupinus
termes extract before its
inoculation with HAV-27
(60 µg gives 36.8 % viral
inhibition)
Effect of post-treatment of
Vero cells with Dianthus
caryophyllus extract after its
inoculation with HSV-1
(60 µg gives 47.4 % viral
inhibition)
Effect of post-treatment of
HepG2 cells with Dianthus
caryophyllus extract after its
inoculation with HAV-27
(60 µg gives 65 % viral inhibition)
Effect of post-treatment of
HepG2 cells with Lupinus
termes extract after its
inoculation with HAV-27
(60 µg gives 25 % viral
inhibition)
Figure 3. Studying the mechanism of action of Dianthus caryophyllus L. and Lupinus termes L.
seed extracts against human viruses (HSV-1 and HAV-27).
C: Cell control; V: Virus control; 30, 40, 50, and 60: concentration / µg of seed extract used in
treating each well. Color wells: no viral growth; dotted wells: obvious virus growth.
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... Curcuma longa L. (Zingiberaceae) is enriched with curcumenone, bisacumol, bisacurone, curcumenol, curcumadiol, and demethoxycurcumin. Curcumin inhibits SARS-CoV-2 replication in human cells, as previously reported for HIV-AIDS [44,45], chikungunya virus, Zika virus, and herpes simplex virus (HSV). ...
... The root and flower of clove pink, also known as Dianthus caryophyllus L., has been reported to contain dianthin30, dianthin32, dianthramides, flavonoids against SARS, herpes simplex virus-I (HSV-I), hepatitis-A Virus-27 (HSV-27) [45]. The antiviral activity against herpes simplex virus-I and hepatitis A virus-27 of crude seed extract are found in this plant. ...
... The antiviral activity against herpes simplex virus-I and hepatitis A virus-27 of crude seed extract are found in this plant. At a nontoxic concentration (20 µg/mL), the extract applied to both Vero and HepG-2 cells led to potent antiviral effects on HSV-I and HAV-27, as determined using a plaque infectivity count assay [45]. Dianthus caryophyllus L. as reported to exhibit antiviral activity against HSV-I and HAV-27, so this plant could also be a potential source for antiviral activity against SARS-CoV-2. ...
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Several human diseases are caused by viruses, including cancer, Type I diabetes, Alzheimer’s disease, and hepatocellular carcinoma. In the past, people have suffered greatly from viral diseases such as polio, mumps, measles, dengue fever, SARS, MERS, AIDS, chikungunya fever, encephalitis, and influenza. Recently, COVID-19 has become a pandemic in most parts of the world. Although vaccines are available to fight the infection, their safety and clinical trial data are still questionable. Social distancing, isolation, the use of sanitizer, and personal productive strategies have been implemented to prevent the spread of the virus. Moreover, the search for a potential therapeutic molecule is ongoing. Based on experiences with outbreaks of SARS and MERS, many research studies reveal the potential of medicinal herbs/plants or chemical compounds extracted from them to counteract the effects of these viral diseases. COVID-19′s current status includes a decrease in infection rates as a result of large-scale vaccination program implementation by several countries. But it is still very close and needs to boost people’s natural immunity in a cost-effective way through phytomedicines because many underdeveloped countries do not have their own vaccination facilities. In this article, phytomedicines as plant parts or plant-derived metabolites that can affect the entry of a virus or its infectiousness inside hosts are described. Finally, it is concluded that the therapeutic potential of medicinal plants must be analyzed and evaluated entirely in the control of COVID-19 in cases of uncontrollable SARS infection.
... Aplidium albicans (48) Gastrointistis corona virus (216) SARS Cov2 (194) SARS-CoV-2 B.1.1.7 (154) Cov 19 (144) Targeting the eukaryotic translation elongation factor 1A (eEF1A) a cellular factor required for the enzymatic delivery of aminoacyl tRNAs to the ribosome (216) (159) Cox A21 OSW-1 binds to one of the two established OSBP ligand binding sites and induces prophylactic antiviral activity (100) In vitro unclear 10 Saikosaponin C Isolated substance B Bulperum (100) In vitro Prevent HBe Ag expression and the virus nuclear acid multyplication 11 Not identified Alcoholic extracts B Polygonum cuspidatum (100) In vitro Unclear 12 Methyl ester dehydrochebulic acid, methyl brevifolincarboxylat e Isolated substances B Phyllanthus urinaria (200) In vitro Unclear 13 Betulinic acid B Pulsatilla chinensis (201) Host cellular factor targeting 14 PRP-Et B Liriope platyphylla L (61) Host cellular factor targeting 15 unidentified aqueous extract A Spirulina platensis (106) In vitro Unclear 16 unidentified Seed Extract A Dianthus caryophyllus (13) In vitro Adsorption or in virus replication 17 Anthocyanidine glycoside Seeds extract A Vitis vinifera L (173) In vitro Attachment, adsorption stage 18 Silymarin compounds Standard extracts C Silybum marianum (123) In vitro promotes the JAKATSTAT pathway associated with IFN 19 Flavo lignans C Silybum marianum (100) In vitro Anti oxidant 20 Curcumin Isolated substance C Curcuma Longa (100) In vitro Inhibitor of virus replication by suppressing the pathway AKT-SREBP-1 21 Epigallocathine3gallate C Green tea (100) In vitro Block the entry to the target cell (9) In vitro Inhibition of viral protease 20 Benzoquinon fruit extract Metanol C Embelia schimperi (9) In vitro Protease inhibition 21 unidentified and chloroform Metanol seed extract C Solanum nigrum (9) In vitro Protease inhibition 22 unidentified Seeds C Daucus maritmus (120) In vitro Unclear 23 anthocyanidine Solanum genus (55) (115) In vitro unclear 2 Biflavons Ethyl acetate Rhus succedanea (100) Garcinia multiflora In vitro unclear 3 Calcium spirolan Spirulina platensis (100) In vitro unclear 4 unidentified Snake toxin Crotolus durissus terrificus (100) In vitro Inhibit virus entry to the target 5 Unidentified Plant extract Zanthoxylum chalybeum (100) Warburgia ugandensis ...
... In vitro unclear 6 unidentified Olinia rochetiana (100) In vitro Stops viral infection 7 unidentified Stem, root extract Cajanus cajan (100) In vitro unclear 8 Chebulagic acid , punicalagin Isolated substances Terminalia chebula Retz (100) In vitro Interfere with infusion and virus transmission 9 Podophyllotoxin aqueous extract Podophyllum peltateum (100) In vitro Unclear (175) In vitro unclear 11 Essential oil HSV 1 Eucalyptus globulus (175) In vitro unclear 12 eugenol Essential oil HSV1 Eugenia caryophyllata (175) In vitro unclear 13 Citral , citronellal Essential oil HSV1, 2 Melissa officinale (175) In vitro unclear 14 citral Essential oil HSV1 HSV2 ...
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Viral infections are vital issue in pathology. Recent growing pandemics due to globalization and the easiness of travel confirmed the necessity of preventing these infections as a major public health risk. In spite of significant progress in drug development and disease immunization, there is a lack in preventive vaccines and effective antivirus, because of viruses tendency to genetic mutations in when they intrude on the living cell, and generating new virual patterns iffer from the original d. The research into the molecular virology has opened up new approaches to understand the properties of viruses and their mandatory intrusione nature, in addition to their pathogenesis. Unfortunately, the required antiviral drugs must possess selective antiviral efficacy without interfering with the biological pathways of the host cells. As Viruses are the main cause of death worldwide, in addition to the side effects of their chemical inhibitors, Natural products representing in medicinal plants emerge as a new alternative source of antiviral agents which can fulfill the renew need to such safe effective antiviral medications. Search goal: This review aims to study and analyze puplished data about plants containing antiviral chemical compounds with antiviral efficacy from 1995 to 2021, It discusses their role in curbing the spread of viruses and treating their infections. Inclusion and exclusion criteria: Clinical trials, in vitro and in vivo trials are included. In addition, reference articles, only English articles were accepted, during the period (1995-2021). Articles that need further analysis were excluded. Quality assessment: to assess the quality of each article, different parts such as title, abstract, introduction, methods, results, and conclusion were examined. Ⅰ. INTRODUCTION Current therapeutic routs for viral disease and its limitation: The antiviral medication could be classified into viral adsorption and entry inhibitors, uncoating inhibitors, viral nucleic acid synthesis inhibitors, integration inhibitors, protease inhibitors, release inhibitors (5,147). The interaction between the virus and the host cell membrane (receptors) is the first stage of the viral life circle, which is called entry and infusion, the integration inhibitors which prevent the virus in this stage can be used successfully, and that is where the anti-acquired immune deficiency virus (161,178,43) and respiratory syncytial virus prevention (61,192) were used. Chemokine receptors targeting (84,99) and the glycoprotein receptors interactions supposed to have a crucial role in the entry stage. A new fatty peptide was discovered, it was named Myrcludex B, it blocks Na + taurocholate co-transporting (NTCP) which is the main receptor for entering hepatic B virus, and hepatic disease virus into hepatic cells (132,7). Moreover, the low acidity (PH) for the endosome activated the proton channels after the virus entry to increase the internal viral acidity and weaken the electrostatic interaction; helping to detach the viral coat with the host cell. (145) More of coating inhibitors were used against
... The in silico study conducted so far has confirmed that several plant (NS) compounds have the potential to target the SARS-CoV-2 replication and host cell attachment. Isolated compound thymoquinone and thymohydroquinone have the potential to target main protease, heat shock protein A5, endoribonuclease, RNA-dependent RNA polymerase, and angiotensinconverting enzyme 2, of SARS-CoV-2 with moderate binding affinity (Barakat et al., 2010;Onifade et al., 2013a;Onifade et al., 2013b;Mani et al., 2020). Another study showed that nigellidine and α-hederin ( Figure No. 2) had a significant binding affinity to the protease and peptidase of the virus comparative to the control (Ulasli et al., 2014;Maiti et al., 2020). ...
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Currently, the whole world is facing a life-threatening novel coronavirus 2019 (COVID-19) pandemic. Natural products are well-known for their potential role against viral disease, and some anti-viral agents have been developed to combat these diseases. Herein, the authors investigated the possible effects of this Holy plant Nigella sativa L. (NS), against coronavirus, using evidence-based and mechanistic approaches to conclude the immune-boosting and alleviation of respiratory system effects of NS. The pharmacological studies established a prominent role in treating various respiratory, immune systems, cardiovascular, skin, and gastrointestinal disorders. Literature supported the significant anti-viral role and showed an inhibitory role for NS against MHV-A59 CoV (mouse-hepatitis virus-A59) infected Hela, i.e., HeLaCEACAM1a (HeLa-epithelial carcinoembryonic antigen-related cell adhesion molecule 1a) cell. NS is a safe herbal product or dietary supplement and could be an effective and affordable community adjuvant treatment for coronavirus in the current scenario. Resumen: Actualmente, el mundo entero se enfrenta a una pandemia del nuevo coronavirus 2019 (COVID-19) que amenaza la vida. Los productos naturales son bien conocidos por su papel potencial contra las enfermedades virales, y se han desarrollado algunos agentes antivirales para combatir estas enfermedades. En este documento, los autores investigaron los posibles efectos de esta planta sagrada Nigella sativa L. (NS), contra el coronavirus, utilizando enfoques mecanicistas y basados en la evidencia para concluir el refuerzo inmunológico y el alivio de los efectos del SN en el sistema respiratorio. Los estudios farmacológicos establecieron un papel destacado en el tratamiento de diversos trastornos respiratorios, del sistema inmunológico, cardiovasculares, cutáneos y gastrointestinales. La literatura apoyó el importante papel antivírico y mostró un papel inhibidor de NS contra células Hela infectadas con MHV-A59 CoV (virus de la hepatitis de ratón-A59), es decir, HeLaCEACAM1a (molécula de adhesión celular 1a relacionada con el antígeno carcinoembrionario epitelial de HeLa). NS es un producto a base de hierbas o un suplemento dietético seguro y podría ser un tratamiento adyuvante comunitario eficaz y asequible para el coronavirus en el escenario actual.
... According to the literature, nigellidine, and α-hederin were isolated from N. sativa, and had the best potential to act as SARS-CoV-2 treatment (Ulasli et al., 2014). Some of N. sativa compounds, including as nigellidine (Banerjee et al., 2021;Maiti et al., 2022), α-hederin (Ulasli et al., 2014;Dorra et al., 2019;Mir et al., 2022), rutin and nigellamine A2 (Baig and Srinivasan, 2022), hederagenin (Barakat et al., 2013;Oyero et al., 2016), thymohydroquinone (Romano and Tatonetti, 2019;Esharkawy et al., 2022;Mani et al., 2022), thymoquinone (Barakat et al., 2010;Onifade et al., 2013a,b;Xu et al., 2021), caryophyllene oxide, β-bisabolene (Duru et al., 2021), dithymoquinone (Ahmad et al., 2021;Pandey et al., 2021;Rizvi et al., 2021) were shown to have diverse mechanisms of action against SARS-CoV-2 using in vitro, in vivo, and in silico investigations (Koshak and Koshak, 2020). Therefore, it was considered that these phytoconstituents could be potential inhibitors of SARS-CoV-2. ...
Article
In Wuhan, China, severe acute respiratory syndrome caused by coronavirus-2 (SARS-CoV-2) has emerged, causing serious symptoms in patients such as fever, dry cough, and exhaustion. This fatal pandemic spreads over the globe, causing significant infections in humans, mainly in the respiratory tract. COVID-19 is still without a treatment. Until date, researchers have paid close attention to new therapeutic methods, particularly promising antiviral medicines and vaccines. Especially, existing synthetic antivirals have been used against viruses that prevent replication, entry into the cell, and transmission of the virus. These antiviral agents have been the subject the basis of drug discovery studies that directly affect COVID 19. Therefore, we investigated on the potential herbal medicines against various RNA, and DNA viruses, including SARS-CoV-2. Finally, we recommend that natural substances and plants be investigated further, including clinical trials, for the treatment of COVID-19.
... The energy complex score of nigelledine, an alkaloid in Nigella sativa, docked with 6LU7 active sites was similar to chloroquine and better than hydroxychloroquine and favipiravir. Hederin, a saponin found in N. sativa, docked with 2GTB active sites and outperformed chloroquine, hydroxychloroquine, and favipiravir in terms of energy [12,16,22,23]. Thymoquinone, the primary essential oil ingredient of N sativa, displayed a lower affinity for the active sites of 6LU7, ACE2, and heat shock protein A5 than hydroxychloroquine did in 6LU7 and ACE2. ...
Article
The severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) causes Coronavirus Disease-19 (COVID-19), which began in Wuhan, Hubei Province, China and has since spread around the world. There are currently no effective antiviral medicines authorised for these coronavirus strains. Chloroquine, hydroxychloroquine, lopinavir/ritonavir, ribavirin, remdesivir, favipiravir, umifenovir, interferon-α, interferon-β, and other drugs are used to treat COVID19. There is no specific, effective, or proven conventional treatment to treat COVID-19 patients. As a result, herbal medicine can be used as an option to treat COVID-19 patients, as many traditional herbs include antiviral and other medical characteristics. Natural products contain a large number of physiologically active compounds with antiviral activity, suggesting that they could be useful as antiviral therapeutics for coronavirus infections. Nigella sativa has shown antiviral effects in numerous of these items. Other pharmacological characteristics of N sativa include anti-inflammatory, antibacterial, and immunostimulatory capabilities. The antiviral properties of N sativa have been demonstrated in the literature for a variety of viruses. Natural chemicals identified in N. sativa, such as nigellidine, α-hederin, hederagenin, thymohydroquinone, and thymoquinone, were revealed to be potentially active inhibitors of coronavirus. To establish the activity of N sativa against coronavirus, preclinical proof is required. If preclinical studies show that N sativa has clinical activity, a clinical Phase I trial in patients with COVID19 is recommended to further investigate its clinical potential.
... The in silico study conducted so far has confirmed that several plant (NS) compounds have the potential to target the SARS-CoV-2 replication and host cell attachment. Isolated compound thymoquinone and thymohydroquinone have the potential to target main protease, heat shock protein A5, endoribonuclease, RNA-dependent RNA polymerase, and angiotensinconverting enzyme 2, of SARS-CoV-2 with moderate binding affinity (Barakat et al., 2010;Onifade et al., 2013a;Onifade et al., 2013b;Mani et al., 2020). Another study showed that nigellidine and α-hederin ( Figure No. 2) had a significant binding affinity to the protease and peptidase of the virus comparative to the control (Ulasli et al., 2014;Maiti et al., 2020). ...
Article
Full-text available
Currently, the whole world is facing a life-threatening novel coronavirus 2019 (COVID-19) pandemic. Natural products are well-known for their potential role against viral disease, and some anti-viral agents have been developed to combat these diseases. Herein, the authors investigated the possible effects of this Holy plant Nigella sativaL. (NS), against coronavirus, using evidence-based and mechanistic approaches to conclude the immune-boosting and alleviation of respiratory systemeffects of NS. The pharmacological studies established a prominent role in treating various respiratory, immune systems, cardiovascular, skin, and gastrointestinal disorders. Literature supported the significant anti-viral role and showed an inhibitory role for NS against MHV-A59 CoV (mouse-hepatitis virus–A59) infected Hela, i.e., HeLaCEACAM1a (HeLa-epithelial carcinoembryonic antigen-related cell adhesion molecule 1a) cell. NS is a safe herbal product or dietary supplement and could be an effective and affordable community adjuvant treatment for coronavirus in the current scenario.
... Herpes simplex and hepatitis A virus infections were virucidal when N. sativa was used. With induced interleukin 8 secretion and downregulation of transient receptor potential (TRP) genes such as TRPM6, TRPA1, TRPC4, and TRPM7, N. sativa lowered the coronavirus load in infected HeLa cells [43,45]. Koshak & Koshak (2020) [43] reported a case study where a six-month therapy with N. sativa resulted in prolonged seroreversion in a 46-year-old HIV patient, and six additional HIV cases were identified. ...
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The world is now facing a public health emergency caused by the coronavirus pandemic (COVID-19). The World Health Organization (WHO) has called it an emerging pandemic due to its sudden appearance and distribution. Coronavirus disease 2019 (COVID-19) is a pandemic caused by the Coronavirus2 of the Extreme Acute Respiratory Syndrome (SARS-CoV2). Nigella sativa (N. sativa) (Family Ranunculaceae) is a globally used medicinal herb. Unani and Tibb, Ayurveda, and Siddha are only a few of the western medical systems that use it. N. sativa, also known as black seed, is an essential medicinal plant that has long been used as a multipurpose medicinal agent in various countries. Immune deficiency, autophagy deficiency, oxidative stress, pathological inflammation, diabetes, cardiovascular diseases, and bacterial and viral infections are all treated effectively with the essential oil and other preparations of the N. sativa crop. It comprises of many essential groups of bioactive compounds, one of which, thymoquinone, has piqued the scientific community's interest due to its active function in treating a wide variety of diseases. The therapeutic effectiveness of N. sativa, as well as recent computational results, clearly suggest that it may be used to tackle the COVID-19 pandemic that has recently arisen. The aim of this review is to highlight the therapeutic importance of N. sativa in conventional medicine, as well as the potential for its use as an antiviral agent against the SARS-CoV2 virus and for further preclinical research.
Chapter
Surprisingly little overlap has been found in the studies of the several hundred plant and herb species with promise as new antiviral medicines. Given the rise in migration, international travel, and urbanization today, viral infections are one of the leading causes of disease around the world. Researchers are looking to the plant world as a source of potential novel antiviral treatments due to the numerous adverse effects of synthetic medicines and the growing problem of resistance. For their various therapeutic purposes, a large number of biologically active compounds have been discovered. Over the past few decades, thorough research into phytochemicals’ antiviral properties has gained relevance. This chapter demonstrates the abundance of potentially beneficial medicinal plants and herbs that are only waiting to be assessed and used for therapeutic applications against genetically and functionally varied plant and viral families.
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The chemical composition of the secondary metabolites is of great significance to the quality control of agricultural products. The genus Dianthus is famous for its beautiful flowers in the cut flower trade and also used in the traditional Chinese medicinal system and food market. However, the chemical composition in Dianthus is still unknown. The current study examined the levels of different metabolites of the flowers in Dianthus caryophyllus, Dianthus chinensis and Dianthus superbus via the use of the widely targeted metabolomic strategy. We obtained the structure and content of 423 metabolites in Dianthus, which included the primary and secondary metabolites. The principal component analysis was able to clearly seperate Dianthus caryophyllus, Dianthus chinensis and Dianthus superbus based on the flower metabolites. The differential metabolites can be categorised into 11 different classes, the majority were flavonoids, amino acids and their derivatives, phenolic acids and lipids. The results of this study provide important information for the effective use of Dianthus flowers in edible, medicinal and therapeutic aspects.
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We used reversed-phase high performance liquid chromatography (HPLC) to analyze the amounts of daidzein and genistein, well-known isoflavonoid aglycones, in methanolic extracts (80%) prepared from soy sprouts and also two types of soybeans (Soja hispida L.), one purchased at the local market and the other one cultivated in Turkey. Some commercially sold preparations containing either soy extract or soy isoflavones were also analyzed by HPLC for their daidzein and genistein content. Three oils obtained from the same soybean and soy sprout samples were analyzed for fatty acids by capillary gas chromatography-mass spectrometry (GC-MS). Several in vitro biological activities of the soybean oils were examined, including anticholinesterase, antioxidant, antibacterial, antifungal and antiviral activity. The soy sprouts were much richer in genistein (232.1 μg/g) and daidzein (177.0 μg/g) than the soybean samples. The cultivated soybean sample also showed higher genistein (3.771 μg/g) and daidzein (3.366 μg/g) levels than the soybean sample of market origin (2.971 μg/g and 2.579 μg/g. respectively). The soybean oils were found to be quite rich in essential fatty acids, and the soy sprout oil also contained essential fatty acids in appreciable amounts. The soybean oil of market origin had a notable antiviral effect against Herpes simplex as well as antifungal activity against Candida albicans at 8 μg/ml.
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The biological effect of Black Cumin seed (Nigella sativa; Ns) and Green tea (Camellia sinensis; GT) has been tested using antiviral, anti-cancer, anti-angiogenic and antioxidant assays. Results showed that Ns and GT have antiviral activity against Infectious Laryngotrachietis Virus (ILTV) at concentration of 35 and 4.22 µM, respectively. Also, both plants protected rats from Diethylnitrosamine (DEN) induced hepatocellular adenoma. Moreover, Ns showed inhibition rate of 80 and 65% at concentration of 80µM, while GT showed respective values of 75 and 45% and 90 µM on Hella and Vero cells. Ns and GT showed marked anti-angiogenic activity on endothelial cells of rat's aorta as well as antioxidant activity by Diphenyl Picryl Hydrazyl (DPPH) radical scavenging activity, Nitric Oxide (NO) radical inhibition assay and lipid peroxidation assay. Current data augmented the efficacy of Nigilla sativa and green tea as remedies for viral diseases, cancer, angiogenetic disorders and oxidative stress.
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Extracts of 40 Ayurvedic medicines and 39 Panamanian medicinal plants were screened for their inhibitory activity on the plaque formation of herpes simplex virus type 1 (HSV-1) in cultured cells. The extracts of 11 plant species showed potent inhibitory activity at a concentration of 100 μg/mL, while those of ten species showed moderate activities. Repeated oral administration of each extract of Rhus acuminata (galls), Saraca indica (bark), Strychnos potatrum (seeds) appreciably suppressed the development of typical skin lesions induced by infection of HSV-1 in BALB/c mice. The extract of S. potatrum prolonged both development of skin lesion and mean survival time. This indicates that S. potatrum is a possible candidate for therapeutic application for HSV-1 infection.
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Extracts of Phyllanthus amarus (Euphorbiaceae) are known to reduce or eliminate detectable hepatitis B virus surface antigen in humans or surface antigen of woodchuck hepatitis virus in woodchucks. The objectives of the experiments in this paper were to evaluate the in vitro inhibición of viral DNA polymerase (DNAp) among (1) different species of Phyllanthus; (2) within the most promising species; and (3) to evaluate differences in activity that might be correlated with environmental effects. The ultimate goal of our studies is to develop a system for producing high quality plants for large scale extraction. Inhibitory activity varied significantly among species. InP. urinaria there were also significant differences among intraspecific accessions. Accessions ofP. amarus from various locations did not differ significantly. Experiments with one seedlot showed highly significant plant-to-plant differences in anti-viral activity, but clear-cut heritable differences were not established among progeny of the plants at extremes in the distribution. Differences in general soil fertility, soil moisture, pH or Ca generally did not significantly affect inhibición of viral DNAp but temperature differences did. Except for temperature, the detectable environmental effects were generally less than the size of the genetic differences among species.P. amarus was successfully grown as a row crop with inputs of fertilizer and irrigation and showed no loss of activity when compared to samples from the wild. Thus, once an accession of Phyllanthus with potent inhibición of viral DNAp is identified, it can be grown under cultivated conditions for extraction without a loss of activity.
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A total of 76 extracts from 35 plants available in the Turkish flora were assayed for their in vitro antibacterial activities against five pathogenic bacteria and a yeast. Sixteen crude extracts from eight plant species were found to possess an activity against at least one or more test microorganisms. Bioassay-guided fractionation of the most active crude extracts was also carried out with the most active extracts. Activity against Staphylococcus aureus, Bacillus cereus, Branhamella catarrhalis, Escherichia coli, Clostridium perfringens and Candida albicans (yeast) is discussed.
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A reinvestigation of the bark of Uncaria tomentosa afforded, in addition to the major quinovic acid glycosides 1-3, three further glycosides 4-6. The structures were elucidated by spectral and chemical studies. Furthermore, a series of antiviral tests were performed on all these glycosides and on the related glycosides 7-9, previously isolated from Guettarda platypoda.
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Despite increasing concern about drug-resistant herpes simplex virus (HSV), antiviral susceptibility testing is not routinely performed by most clinical virology laboratories. This omission is in large part because the most widely accepted method, the plaque reduction assay (PRA), is cumbersome to perform and results are rarely available in time to influence treatment. We report here the development of a sensitivity test for HSV which utilizes a cell line (VeroICP6LacZ#7) that expresses beta-galactosidase activity after infection with HSV such that infected cells can be detected by histochemical staining. We designed an assay in which 10-fold dilutions of virus stocks with undetermined titers were inoculated onto VeroICP6LacZ#7 cells in a 24-well tissue culture dish. Forty-eight hours after infection, the cell monolayers were histochemically stained. Plaques appear blue against a clear background and are thus easily visualized at 48 h. As with the standard PRA, the 50% inhibitory concentration (IC50) was reported as the concentration of an antiviral drug that reduces the number of plaques by 50%. Evaluation of 10 well-characterized laboratory strains and 12 clinical HSV isolates showed that the IC50 determined by this method correlated in all instances with the IC50 determined by the PRA. This method is easy to use and eliminates the need to determine the titer of the virus, and results are available within 48 h of the detection of the virus. VeroICP6Lac#7 cells are a useful tool for performing HSV antiviral susceptibility testing and could be used in a number of different formats to facilitate the identification of drug-resistant isolates of HSV.
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The in vitro activity against herpes simplex virus type 1 of 3-methyl-but-2-enyl caffeate isolated from poplar buds or prepared by synthesis was investigated. Under conditions of one or multiple multiplication cycles, this compound, which is a minor constituent of propolis, was found to reduce the viral titer by 3 log10, and viral DNA synthesis by 32-fold.