Scientists Design Potent Anthrax Toxin Inhibitor
Technique May Also be Applicable to Other Disease Toxins
Scientists funded by the National Institute of Allergy and Infectious Diseases
(NIAID), part of the National Institutes of Health (NIH), have engineered a powerful
inhibitor of anthrax toxin that worked well in small-scale animal tests.
“This novel approach to the design of anthrax antitoxin is an important advance,
not only for the value it may have in anthrax treatment, but also because this
technique could be used to design better therapies for cholera and other diseases,” says
NIH Director Elias A. Zerhouni, M.D.
The research appears in the April 23 online edition of the journal Nature
Biotechnology.
Led by NIAID grantees Ravi S. Kane, Ph.D., of Rensselaer Polytechnic Institute,
in Troy, NY, and Jeremy Mogridge, Ph.D., of the University of Toronto, the investigators
built a fatty bubble studded with small proteins that can cling tightly to the
cell membrane receptor-binding protein used by anthrax toxin to gain entry into
a host cell.
The protein-spiked fatty bubble, or “functionalized liposome,” hampers a critical
early step in the assembly process that anthrax toxin must undergo to become
fully active. In test-tube experiments, the inhibitor, which is covered with
multiple short proteins (peptides), was 10,000 times more potent than unattached
peptides.
“If the effectiveness of anthrax inhibitors designed in this fashion is confirmed
by additional testing, they could one day be important adjuncts to standard antibiotic
therapy for the treatment of inhalation anthrax,” says NIAID Director Anthony
S. Fauci, M.D.
The spore-forming bacterium Bacillus anthracis produces a toxin that
causes anthrax symptoms. Antibiotics are used to treat anthrax, but even with
such therapy, inhalation anthrax, the most severe form of the disease, has a
fatality rate of 75 percent.
“There would be real value to having an additional form of therapy available
to physicians confronting a case of inhalation anthrax,” notes Phillip J. Baker,
Ph.D., anthrax program officer at NIAID.
Anthrax toxin has three parts: protective antigen (PA), a protein that binds
to a receptor on the target cell surface; and two enzymes that must be transported
into the cell to cause damage. The enzymatic portions of the toxin enter the
cell through a pore created for them by PA after it binds to the cell’s outer
surface. PA can be seen as a bundle of seven cigar-shaped parts, a molecular
arrangement referred to as “polyvalent,” meaning it displays multiple binding
sites.
The inhibitor designed by Dr. Kane and his colleagues is also polyvalent. Just
as a glove matches the shape of a hand more closely than a mitten, and so fits
more snugly, the polyvalent inhibitor binds the toxin at multiple sites and is
orders of magnitude more potent than an inhibitor that binds at a single site.
The multiple peptides on the functionalized liposome are arranged with the same
average spacing as the binding sites of the PA molecule, which permits a firmer
bond between the two, explains Dr. Kane. When the inhibitor is bound tightly
to PA, the subsequent steps of enzyme entry cannot occur and the toxin is effectively
neutralized.
The investigators tested the anthrax inhibitor in rats. When given in relatively
small doses, injection of the inhibitor at the same time as anthrax toxin prevented
five out of nine rats from becoming ill. Slightly higher doses of the inhibitor
prevented eight out of nine rats from being sickened by anthrax toxin. Nine additional
rats were injected with anthrax toxin only. Of these, eight became gravely ill.
This experiment was the first to show the efficacy of a liposome-based polyvalent
inhibitor in animals, says Dr. Kane.
Dr. Kane says the recent experiments demonstrate a proof of principle and suggest
that polyvalent inhibitors could be used along with antibiotics in a clinical
setting. Aside from its inherent toxicity, anthrax toxin also accelerates the
disease process. Thus, combining antibiotics with a toxin inhibitor might act
synergistically to lessen or halt anthrax symptoms, notes Dr. Kane.
Using the same technique of placing multiple peptides on a liposome, the researchers
also created a polyvalent inhibitor of cholera toxin that functioned well in
test-tube experiments.
In the next phase of their research, Drs. Kane and Mogridge and their colleagues
plan to test the action of their inhibitor in animals after infecting them with B.
anthracis and allowing the disease process to begin. They also will evaluate
the inhibitor with and without adjunct antibiotic therapy.
News releases, fact sheets and other NIAID-related materials are available
on the NIAID Web site at http://www.niaid.nih.gov.
NIAID is a component of the National Institutes of Health. NIAID supports
basic and applied research to prevent, diagnose and treat infectious diseases
such as HIV/AIDS and other sexually transmitted infections, influenza, tuberculosis,
malaria and illness from potential agents of bioterrorism. NIAID also supports
research on basic immunology, transplantation and immune-related disorders,
including autoimmune diseases, asthma and allergies.
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Agency — includes 27 Institutes and Centers and is a component of
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agency for conducting and supporting basic, clinical and translational medical
research, and it investigates the causes, treatments, and cures for both common
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