Y. GONG(1), R. ROBERTS(2), I.
GADAWSKI(1), D. CHEUNG(1), T. TAM(1), AND S.L. SACKS(1).
(1) Viridae Clinical Sciences, Inc. and The University of British Columbia,
Vancouver, B.C.
(2) Bryant Nutrients, 4133 Greenwell St., Baton Rouge, LA 70805, USA
The antiviral effect of Virucept Red Marine Algae
(Virucept) against HSV was evaluated in cultured Vero cells using plague
reduction assay. The crude extracts containing polysaccharide were
prepared by ethanol precipitation. Pre-treatment of cells with the
Virucept extracts showed a strong inhibitory effect against both HSV type 1 and
3 with the 90% effective concentrations (EC90) ~0.1% for HSV-1 and 0.01% for
HSV-2, respectively. Strong inhibition was also observed even after
removing the extracts by washing with PBS on pre-treated cells. Similar
activities were examined on HSV Acyclovir-or Forscanet-resistant strains.
Approximately 66% inhibition was observed on cells that were already infected
with HSV-1. Approximately 20-36% inhibition at a concentration of 0.5% was
seen on HSV-2 and Acyclovir-or Forscarnet-resistant HSV infected cells.
Our data indicate that the extracts from Virucept Red Marine Algae can be
further developed as a vaginal microbicide for preventing HSV infection.
Inhibition of Herpesvirus Replication by Marine Algae Extracts
E. FRANK DEIG, DOUGLAS W. EHRESMANN, MELVIN T. HATCH, AND
DELIMAR J. RIEDLINGER
Naval Biomedical Research Laboratory, School of Public Health,
University of California, Berkeley, California 94720
Received for publication 8 August 1974.
Extracts from two species of marine red algae, Cryptosyphonia
woodii and Farlowia mollis, specifically inhibited herpes simplex
virus replicaton in vitro.
American Society for Microbiology, Vol. 6, No. 4, Copyright 1974
Antiviral Substances from California Marine Algae
D. W. Ehresmann, E. F. Deig, M. T. Hatch, L. H. DiSalvo and N. A. Vedros
Naval Biosicences Laboratory, School of Public Health, University of
California, Berkeley, California 94720
Extracts of 28 species of marine macroscopic algae collected from
various coastal habitats of northern California were examined for
antiviral activity against a broad spectrum of mammalian viruses.
Ten members of Rhodophyta contained substance(s) which caused
greater than a 2 log reduction in the infectivity of herpes simplex
virus types 1 and 2. In addition, anti-Coxsackie B5 virus activity
was detected in extracts of constantinea simplex Setchell. The
physical and chemical properties of the substance in extracts of
Farlowia mollis (Harvey and Bailey), Farlow and Setchell and C.
simplex indicated the active agent was a structural polysaccharide.
Journal of Phycology, Vol. 13, 37-40, Copyright 1977
Antiviral Activity of Extracts from Marine Algae
JAMES T. RICHARDS, EARL R. KERN, LOWELL A. GLASGOW, JAMES C.
OVERALL, JR., E. FRANK DEIGN, AND MELVIN T. HATCH
Department of Pediatrics, University of Utah College of Medicine, Salt
Lake City, Utah 84132, and Naval Biosciences Laboratory, School of
Public Health, University of California, Berkeley, California 94720
Received for publication 2 March 1978
Extracts of two species of marine algae, Constantinea simplex and
Farlowia mollis, were tested for antiviral activity in tissue culture and
in experimental infections of mice. Treatment of confluent mouse embryo
fibroblast cell monolayers with either compound before viral inoculation
was effective in inhibiting the replication of herpes simplex virus type 1
and type 2, vaccinia virus, and vesicular stomatitis virus, but not
encephalomyocarditis virus, Semiliki Forest virus, or murine
cytomegalovirus. Prophylactic administration of these extracts was
effective in reducing final mortality or prolonging the mean day of death
of animals inoculated by the intraperitoneal, intracerebral, or intranasal
routes with herpes simplex virus type 2. When therapy was initiated
after viral inoculation or at a site other than that of viral inoculation, no
significant effect on mortality or on mean day of death was observed.
Neither preparation was effective in mice inoculated intraperitoneally
with encepthalomyocarditis virus, Semliki Forest virus, or murine
cytomegalovirus or in animals infected intravaginally with herpes
simplex virus type 2. The prophylactic but not therapeutic antiviral
activity of these preparations seriously limits their potential use in
human herpes simplex virus infections.
American Society for Microbiology, Vol. 14, No. 4, Copyright 1978
Further Studies on the Chemical Composition and an Initial In Vivo
Evaluation of Antiviral Material in Extracts of Macroscopic Marine Algae.
M. T. Hatch, D. W. Ehresmann, E. F. Deig, and N. A. Vedros
Naval Biosciences Laboratory, School of Public Health, University of
California, Berkeley, California 94720
Previous studies have demonstrated that the antiviral material in extracts
of 8 related species of Rhodophyta is a structural polysaccharide. The
present study extends our knowledge of chemical and biological properties
of the active material. The observation that concanavalin A (Con A)
formed an insoluable complex with antiviral molecules resulted in
development of a rapid in vitro titration procedure. Although purified
active fractions had polysaccharides containing 20% sulfate in ester form,
the material differed from other algal sulfated polysaccarides in that it
was specific for herpesvirus whereas carrageenins had a wide antiviral
spectrum. The reaction between Con A and the anti-herpesvirus
polysaccharide indicated the presence of alpha-D-glycosyl and sterically
related residues. Purified fractions were also tested for activity in cell
cultures and in mice subsequently infected with herpesvirus type 1 and 2.
Significant protection occurred when the material was administered to
mice prior to, or at the same time as virus.
Anti-Viral Properties of Algal Polysaccharides and Related Compounds
D. W. Ehresmann, E. F. Deig, M. T. Hatch
Naval Biosciences Laboratory, School of Public Health, University of
California, Berkeley, California 94720
Received for publication 8 August 1974
Research directed toward the discovery and development of drugs which
inhibit the replication of disease-causing mammalian viruses has only
recently yielded agents with clinical promise. The success of vaccines
in controlling many important viral diseases (e.g., poliomyelitis and
small pox), coupled with the toxicity of many anti-viral compounds,
lead to the concept that development of effective anti-viral drugs both
fruitless and unnecessary. However, the more recent recognition of a
cellular anti-viral system (i.e., interferon) and the discovery of
differences between the molecular biology of viral replication and
normal cellular metabolism suggested that viral pathogenesis could be
inhibited without concomitant disruption of normal cell function.
Additionally, the realization that some viral diseases may be
uncontrollable by immunization (e.g., diseases caused by the
herpesviruses and rhinoviruses) has renewed interest in anti-viral
chemotherapy.
Chemical Characterization and Therapeutic Evaluation of Anti-Herpesvirus
Polysaccharides from Species of Dumontiaceae
M. T. Hatch, D. W. Ehresmann, and E. F. Deig
Naval Biosciences Laboratory, School of Public Health, University of
California, Berkeley, California 94720
Recently we found an antiviral compound in marine red algae that may
have some usefulness in the control of herpes simplex virus (HSV)
infections. The HSV problem is of considerable health importance,
particularly in view of the fact that type 1 viruses cause acute
respiratory illness, recurrent oral lesions, herpes keratitis (an eye
infection responsibile for up to 18,000 cases of blindness in the USA
every year) and meningitis; whereas viruses comprising type 2
initiate veneral disease and are under suspicion as one cause of
cervical cancer. Although several antiviral agents appear to have a
potential application in the treatment of herpes virus infections of man,
prevention of recurrenes of herpetic keratitis, oral lesions and venereal
infectgions attributable to HSV is still a critical unsolved problem.
Moreover, increasing numbers of individuals are currently suffering
from exaggerated herpetic diseases that can be life threatening or can
maim severely. Continued exploration is therefore necessary to
discover new and clinically useful drugs active against HSV. However,
it is noteworthy that the selection of a candidate antiviral for clinical
testing in man is made only after numerous preclinical evaluations
including: (a) identification in terms of accepted chemical nomenclature
of active chemical moieties, (b) examination for possible unique physico-
chemical characteristics of the viral inhibitor, (c) demonstration of
selective antiviral activity in at least one in vivo and one in in vitro
model system and (d) determinaton of drug-host interactions,
particularly with respect to viral replication, cell metabolism and
toxicity of the active component or its degradation products at
potentially effective dosage levels.
In our earlier studies, we presented evidence that the selective
anti-herpesvirus material extracted from red algae consisted of large
molecules which were excluded from Sephadex G-50 and G-200 columns.
The active substance was stable to heat and wide range of pH. It was not
partitioned into common lipid solvents but could be precipitated in the
presence of methanol, ethanol or acetone. Spectroscopic observations
of active crude extracts from one source, Constantinea simplex Setchell,
produced a broad peak at wavelengths from 260 to 280nm. The
cummulative evidence suggested that the active agent consisted of a
major polysaccharide fraction of unknown homogeneity bound in some
way to a minor protein or nucleic acid fraction. This paper is
concerned with further chemical characterization of the antiviral
substance as well as results of our initial experiments to determine
the in vivo efficacy of crude and partially purified algal extracts in
a herpesvirus induced mouse encephalitis model.
ANIMAL AND HUMAN STUDIES. Gerber (1958) reported that the
intransal administration of agar polysaccharides prevented death in
70% of mice receiving 100 LD50 units of murine pneumonia virus
(PVM) provided the polysaccharide was given prior to virus inoculation.
Lehel and Hadhazy (1966) found that heparin injected into rabbit skin
prior to or at the same time as infection with a herpes virus resulted
in a reduction in the severity of virus-induced lesions. Museteanu (1975)
reported that sulfated glycosaminoglycans decreased the death rate in
mice infected with yellow fever virus whens the inhibitor was
administered with virus. More recently, Deig, (unpublished data) found
that aqueous extracts of four members of family Dumontiaceae protected
infant mice against herpes simplex induced encephalitis. The reduction in
mortality afforded by the extracts was greater than 2 Log10, when
extracts were administered two hours prior to, or simultaneous with virus.
Additional studies with the Dumontiaceae inhibitor in other herpes simplex
disease models are currently being conducted in our laboratory and by
investigators of the National Institute of Health's Anti-viral Substances
Program.
The dependence of in vivo effectiveness on early administration of the
inhibitor is consistent with the concept that sulfated polyanions block
virus adsorption and suggests that the clinical value of these agents
may be limited to a prophylactic regimen. However, with many disease-
causing viruses, the spread of virions from infected cells to adjacent non-
infected cells occurs primarily extracellularly. Even with the
herpesviruses, a cell-associated group, significant extracellular spread
occurs (Watson, 1973). The anti-viral polyanions therefore may have
therapeutic potential in treating certain established viral disorders,
particularly limited diseases such as herpes simplex dermatitis, by
reducing the severity of infection. Alternatively, combination of relatively
non-toxic sulfated polyanions with the toxic intracellular anti-viral drugs
in a therapeutic preparation, may allow a reduction in the level of the toxic
agent. These possibilities need exploration.
The only sulfated polyanion which has received clinical examinations in
humans is the sulfated glycosaminoglycan L5. Voos, (1977) reported that
when L5 was evaluated against zoster infections (herpesvirus) by direct
application to the lesions, healing of vesicles was facilitated and the course
of the disease was shortened. This investigation encourages continued
evaluation of sulfated glycosaminoglycan and other sulfated polyanions
as potential human anti-viral therapeutics.
Development of Dermal Lesions in Adult Mice Infected with Herpes Simplex
Virus: Application of the Model in the Evaluation of Antiherpesvirus
Substances from Marine Algae
E. F. Deig, M. T. Hatch, A. M. Nonomura
Naval Biomedical Research Laboratory, School of Public Health, University
of California, Berkeley, California 94720
A procedure was developed for producing dermal lesions in a high
proportion of adult mice infected with Herpes Simplex viruses 1
and type 2. Preliminary data indicated that this model system was
adequate for evaluating antiherpesvirus substances from marine red algae.
Evaluation of Extracts of Marine Algae for Antiviral Activity in
Experimental Herpes Simplex Infections of Infant Mice
E. FRANK DEIG, DOUGLAS W. EHRESMANN, MELVIN T. HATCH, AND
DELIMAR J. RIEDLINGER
Naval Biomedical Research Laboratory, School of Public Health, University
of California, Berkeley, California 94720
Primary and secondary extracts from species of marine algae previously
shown to have antiviral toward herpes simples virus (HSV) in cell culture
were evaluated for their ability to protect infant mice from lethal
infections initiated by this virus. Significant protection was acheived
from extracts active in cell culture if they were adminstered to mice
prior to or at the same time as virus. No protection occurred when
extracts inactive in cell culture were used. The route of administration
of HSV and algal material exerted a significant influence on mouse mortality.
Both type 1 and type 2 strains of HSV were sensitive to the antiviral
material in vivo. The conditions used indicated that the agent probably
acts by blocking an early step in the adsorption of virus to otherwise
susceptible cells.
Antiviral carbohydrates from marine red algae
Michael Neushul
Department of Biological Sciences, University of California, Santa Barbara,
93106, USA and Neushul Mariculature Incorporated, 475 Kellogg Way,
Goleta, CA, 93117, USA
It is possible that heparin-like sulfated polysaccharides from red algae,
or fractions thereof, might be found to be low-cost, broad-spectrum
antiviral agents. The prevailing view among virologists has been that
sulfated polysaccharides inhibit viral action by acting only at the sufraces
of cells. This perception now is changing with the finding that both the
herpes virus (containing DNA) and human immunodeficiency virus
(containing RNA) are inhibited by sulfated polysaccharides that act
within the cell as well as external to it. Aqueous extracts of many
red algae are active against retroviruses. Carrageenan, a common cell
wall polysaccharide from red algae, is co-internalized into infected cells
with the Herpes simplex virus (HSV), inhibiting the virus. Carrageenan
also interferes with fusion (syncytium formation) between cells infected
with the human immunodeficiency virus (HIV) and inhibits the specific
retroviral enzyme reverse transcriptase.
Antiretroviral activity in a marine red alga: reverse transcriptase
inhibition by an aqueous extract of Schizymenia pacifica
H. Nakashima 1, Y. Kido2, N. Kobayashi1, Y. Motoki2, M. Neushul3, and
N. Yamamoto1
1 Department of Virology and Parasitology, Yamaguchi University School
of Medicine, 1144 Kogushi, Ube, Yamaguchi 755, Japan
2 Ube Research Laboratory, Fujirebio Inc. Konan-ku, Ube, Yamaguchi
759-02, Japan
3 Department of Biological Sciences, University of California, Santa
Barbara, Santa Barbara, CA 93106, USA and Neushul Mariculature
Incorporated, 475 Kellogg Way, Goleta, CA 93107, USA
An aqueous extract from the marine red alga, Schizymenia pacifica has
been tested in a cell free system for its effect on reverse transcriptase
from avian retrovirus (avian myeloblastosis virus), and mammalian
retrovirus (Rauscher murine leukemia virus). The extract inhibited
reverse transcriptase from both these retroviruses but showed almost
no effect, if any, on the activity of cellular DNA polymerase alpha and
RNA polymerase II in vitro. Consequently it is unlikely to have an
adverse effect on the growth of cultured cell. The inhibitory activity
of the extract was stable over a relatively wide pH range (pH 1-11)
and was not lost after pronase digestion. Inhibitory activity of the
extract was lost after boiling at 100 degree C in 0.67 N HCL, and after
treatment with 100 mM NaIO4. The active principle in the extract has
an apparent molecular weight in excess of 100,000 daltons. This new
reverse transcriptase inhibitor is probably a polysaccharide.
Polysaccharides as Antiviral Agents: Antiviral Activity of Carrageenan
M. EUGENIA GONZALEZ, BALBINO ALARCON, AND LUIS CARRASCO
Departemento de Microbiologia, Centro de Biologia Molecular, Universidad
Autonoma, Canto Blanco, 28049 Madrid, Spain
Received 26 February 1987/Accepted 8 June 1987
A number of polysaccharides showed good antiviral activity against
several animal viruses. At 5 µg/ml, carrageenan prevented the
cell monolayer from destruction by herpes simplex virus type I (HSV-I)
growth. At 10 µg/ml, carraggeenan reduced the formation of new
infectious HSV-I by almost five logs. No cytotoxic effects were detected
with concentrations of carrageenan up to 200 µg/ml. When 10
micro g of carrageenan per ml was added at the beginning of HSV-I
infection of HeLa cells, there was potent inhibition of viral protein
synthesis, and the cells continued synthesizing cellular proteins. This did
not occur if carrageenan was added 1 h after virions into cells indicated
that carrageenan had no effect on virus attachment or virus entry.
Moreover, carrageenan did not block the early permeabilization of cells
to the toxic protein alpha-sarcin. These results suggest that this
sulfated polysaccharide inhibits a step in virus replication subsequent to
viral internalization but prior to the onset of late viral protein synthesis.
Composition of Matter from Cryptosiphonia woodii Useful for theTreatment of Herpes Simplex Virus Arthur M. Nonomura & Raphael Pappo U.S. Patent No. 4,522,814 A water extract of homogenized Cryptosiphonia woodii, in the family of Dumontiaceae, is useful for the treatment of herpes simplex viral infections. The active agent in the extract is a polysaccharide containing glucose and galactose. Herpes infections may be treated in subjects, e.g. human patients, by administering to the subject an effective amount of an aqueous extract of C. woodii, particularly a water extract. This treatment method is effective for treating subjects both prior to and subsequent to infection. It may involve topical application to alleviate symptoms associated with herpes infections or desirably may be systemic, e.g. by oral administration, to eradicate the virus and thereby prevent symptom recurrence.
Further information about Red Marine Algae can be found at the following:
Red Marine Algae and the immune system Red Marine Algae as a super food Red Marine Algae as an alternative medicinal treatment Red Marine Algae general information
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