Faculty of veterinary medicine

Mansoura university

Name: ‫محمود ياسر بهجت احمد‬

Section:8

Title: general mode of action of toxicant with examples.

Introduction
The mode of toxic action (MOA) is recognized as a key determinant of chemical
toxicity and as an alternative to chemical class-based predictive toxicity modeling.
However, MOA classification has never been standardized in ecotoxicology, and a
comprehensive comparison of classification tools and approaches has never been
reported.
Here we critically evaluate three MOA classification methodologies using an
aquatic toxicity data set of 3448 chemicals, compare the approaches, and assess
utility and limitations in screening and early tier assessments.
The comparisons focused on three commonly used tools: Vehar prediction of
toxicity MOA, the U.S. Environmental Protection Agency (EPA) Assessment Tool
for Evaluating Risk (ASTER) QSAR (quantitative structure activity relationship)
application, and the EPA Mode of Action and Toxicity (MOAtox) database.
Of the 3448 MOAs predicted using the Verhaar scheme, 1165 were classified by
ASTER, and 802 were available in MOAtox. Of the subset of 432 chemicals with
MOA assignments for each of the three schemes, 42% had complete concordance
in MOA classification, and there was no agreement for 7% of the chemicals. The
research shows the potential for large differences in MOA classification between
the five broad groups of the Verhaar scheme and the more mechanism-based
assignments of ASTER and MOAtox.
Harmonization of classification schemes is needed to use MOA classification in
chemical hazard and risk assessment broader.
There has been a shift in the risk assessment paradigm toward a more
mechanistically based understanding of the mechanism and mode of action of
chemicals.
Improved understanding of toxicological mechanisms would reduce reliance on
traditional animal testing, allow chemical grouping to increase assessment
efficiency, and improve toxicity extrapolations. These out- comes would increase
the potential use of animal alternative methods, which is a long-term policy goal of
most chemical management programs.
Various structure-based classification schemes have been developed to categorize
chemicals based on the mode of toxic action (MOA).
Thousands of chemicals are in commerce and most have little available
information other than structure.

Classification:
Understanding how the available MOA schemes were devised, what information
they are based on, and the limitations of each approach are critical. The various
MOA classification schemes provide information on a key determinant of chemical
toxicity and as an alternative to chemical class-based predictive toxicity modeling.
Direct regulatory application of these MOA classifications has been seen most
directly in chemical classification and in the use of Quantitative Structure−Activity
Relationship (QSAR) models in risk assessment.
MOA also plays a role in mixture risk assessment, where both the grouping
of chemicals and the choice of the concentration addition or independent action
hypothesis can be based on knowledge of the MOA.
MOA is an operational term that has been loosely defined in both human health
and ecotoxicology as a functional change at the cellular level, in contrast to the
mechanism of action or molecular initiating event.
The definition of MOA is critical in the context of Adverse Outcome Pathways
(AOPs). Generally, MOA describes key events at various levels of biological
organization, starting with cellular interaction and leading to functional and/or
anatomical changes. MOA is important in classifying chemicals because it
represents an intermediate level of complexity between molecular mechanisms and
physiological or organismal outcomes and provides an organizing scheme for
chemical classification.
MOA has also been used to classify pesticides by the interaction with the receptor
of the target species and to describe the toxicology of human cancer and noncancer
agents by key cytological and biochemical events.
The definitions of MOA used are unique to each of the classification schemes
investigated: Verhaar used five broad categories based on general toxicological
responses, whereas the U.S. Environmental Protection Agency (EPA) Assessment
Tool for Evaluating Risk (ASTER) QSAR (quantitative structure activity
relationship) application and the EPA.
high degree of specificity based on fish behavioral responses or weight of
evidence classification, respectively. It is also important to note that both within
and between classification schemes, different levels of biological organization may
be represented (e.g., at the molecular, cellular, and organism-level).
Research to develop screening and early tier methods to more easily classify or
assess chemicals using approaches such as the ecological threshold of toxicological
concern (eco TTC) and chemical activity is ongoing.
Evaluation of these approaches and determination of their domain of applicability
is partly dependent on the MOA classification used to group chemicals. The
objectives of this study were to critically evaluate available MOA classification
methodologies using a set of unique chemicals from a large aquatic toxicity data
set, compare the various approaches, and evaluate their utility and limitations in
screening early tier assessments.
OVERVIEW OF MOA SCHEMES:
The comparison of classification approaches focused on three currently available
schemes: the Verhaar scheme for prediction of toxicity MOA, the EPA ASTER
QSAR application, and the EPA MOAtox database. The Verhaar scheme classifies
organic compounds into one of four categories: inert chemicals (Class 1), less inert
chemicals (Class 2), reactive chemicals (Class 3), and chemicals acting by a
specific mechanism (Class 4). Chemicals in Class 1 exhibit nonpolar narcosis or
baseline toxicity and can only be predicted if they have log octanol: water
partition coefficient (Know) values between 0 and 6 (e.g., benzenes).
Chemicals in Class 2 are more toxic and cause polar narcosis, and typically possess
hydrogen bond donor acidity (e.g., phenols and anilines). Chemicals in Class 3
demonstrate enhanced toxicity as compared to baseline toxicity and react
nonspecifically with biomolecules (e.g., epoxides) or are metabolized into more
toxic species (e.g., nitriles). Chemicals in Class 4 cause toxicity through a specific
mechanism such as acetylcholinesterase (AChE) inhibition by carbamate
insecticides. The assignment of a chemical to a class is based on a decision tree
that utilizes the presence or absence of certain chemical structures and moieties.
The Verhaar classification scheme has been further modified by changing the order
of some classification rules in the decision tree and by adding a fifth class to which
chemicals are assigned if the chemical could not be placed in Class 1−4.19
Verhaar and modified Verhaar schemes have been implemented into online tools in
the public domain such as the Organization for Economic Cooperation and
Development (OECD) QSAR Toolbox and Toxtree the OECD QSAR Toolbox is a
software application used in the grouping of chemicals and in filling (eco)toxicity
data gaps for assessing the hazards of chemicals.
Chemical profiling is based on the Chemical Abstracts Service numbers (CAS)
and/or SMILES (Simplified Molecular Line Entry System). an open-source
application, groups chemicals into categories using several schemes based on
SMILES notation and decision trees.
A postprocessing filter encoded as a KNIME (Konstanz Information Miner)
workflow adds amendments to the classification rules to improve the Toxtree
classification. The postfilter addresses three types of mis- classification: Class 5 of
some aliphatic alcohols, some halogenated compounds that should both be
assigned to Class 1 (rule 1.5.2 and rule 1.7.1, respectively, of the Verhaar
classification scheme), and non- or weakly acidic phenols (rule 2.1).
Fewer compounds are misclassified as outside of the model domain, further
improving the prediction accuracy of the scheme. ASTER is a rule-based expert
system that screens a chemical structure for structural fragments to determine
MOA.
The tool uses acute toxicity of over 600 chemicals in fathead minnows. Nonpolar
narcosis is the default MOA if no MOA specific structural fragments are identified.
If fragments satisfy requirements for more than one MOA, then the QSAR for the
most potent toxicity prediction is selected. ASTER is based on the MOA categories
and contains information on over 50 000 molecular structures. ASTER was
primarily developed with neutral industrial organic chemicals acting through
narcosis and has a more limited representation of pesticides and other chemicals
with more specific modes of action. The tool is not publicly available.
MOAtox is composed of a database of MOA assignments for 1208 chemicals
including metals, organometallics, pesticides, and other organic compounds. The
categorization scheme was based on earlier work determining chemical modes of
acute toxic action in fish and encompasses six broad and specific MOAs.
The resulting data set used a combination of high confidence MOA assignments,
including biological responses in fish acute toxicity assays pesticide classifications
schemes [e.g., Insecticide Resistance Action Committee (IRAC)], QSAR
predictions (e.g., ASTER), and weight of evidence professional judgment
incorporating an assessment of chemical structure. and available information on
MOA, mechanism of action, and toxicity pathways. Chemicals with an uncertain
MOA assignment and MOAs specific to invertebrates were excluded.
Specific MOAs were developed as subcategories of the broad MOAs based on
either chemical structure or known mechanism of action.
Non-specific acting modes of toxic action result in narcosis; therefore, narcosis is
a mode of toxic action. Narcosis is defined as a generalized depression in
biological activity due to the presence of toxicant molecules in the organism. The
target site and mechanism of toxic action through which narcosis affects organisms
are still unclear, but there are hypotheses that support that it occurs through
alterations in the cell membranes at specific sites of the membranes, such as the
lipid layers or the proteins bound to the membranes. Even though continuous
exposure to a narcotic toxicant can produce death, if the exposure to the toxicant is
stopped, narcosis can be reversible.

Specific Toxicants that at low concentrations modify or inhibit some biological

process by binding at a specific site or molecule have a specific acting mode of
toxic action. However, at high enough concentrations, toxicants with specific
acting modes of toxic actions can produce narcosis that may or may not be
reversible. Nevertheless, the specific action of the toxicant is always shown first
because it requires lower concentrations.
There are several specific acting modes of toxic action:
Uncouplers of oxidative phosphorylation. Involves toxicants that uncouple the two
processes that occur in oxidative phosphorylation: electron transfer and adenosine
triphosphate (ATP) production.
Acetylcholinesterase (AChE) inhibitors. AChE is an enzyme associated with nerve
synapses that it’s designed to regulate nerve impulses by breaking down the
neurotransmitter Acetylcholine (ACh). When toxicants bind to AChE, they inhibit
the breakdown of ACh. This results in continued nerve impulses across the
synapses, which eventually cause nerve system damage. Examples of AChE
inhibitors are organophosphates and carbamates, which are components found in
pesticides (see Acetylcholinesterase inhibitors).
Irritants. These are chemicals that cause an inflammatory effect on living tissue by
chemical action at the site of contact. The resulting effect of irritants is an increase
in the volume of cells due to a change in size (hypertrophy) or an increase in the
number of cells (hyperplasia). Examples of irritants are benzaldehyde, acrolein,
zinc sulphate and chlorine.
Central nervous system (CNS) seizure agents. CNS seizure agents inhibit cellular
signaling by acting as receptor antagonists. They result in the inhibition of
biological responses. Examples of CNS seizure agents are organochlorine
pesticides.
Respiratory blockers. These are toxicants that affect respiration by interfering with
the electron transport chain in the mitochondria. Examples of respiratory blockers
are rotenone and cyanide.
Determination The pioneer work of identifying the major categories of modes of
toxic action (see description above) was conducted by investigators from the U.S.
Environmental Protection Agency (EPA) at the Duluth Laboratory using fish,
reason why they named the categories as Fish Acute Toxicity Syndromes (FATS).
They proposed the FATS by assessing the behavioral and physiological responses
of the fish when subjected to toxicity tests, such as locomotive activities, body
color, ventilation patterns, cough rate, heart rate, and others.
It has been proposed that modes of toxic action could be estimated by developing a
data set of critical body residues (CBR). The CBR is the whole-body concentration
of a chemical that is associated with a given adverse biological response and it is
estimated using a partition coefficient and a bioconcentration factor. The whole-
body residues are reasonable first approximations of the amount of chemical
present at the toxic action site(s). Because different modes of toxic action generally
appear to be associated with different ranges of body residues modes of toxic
action can then be separated into categories. However, it is unlikely that every
chemical has the same mode of toxic action in every organism, so this variability
should be considered.
The effects of mixture toxicity should be considered as well, even though mixture
toxicity it's generally additive chemicals with more than one mode of toxic action
may contribute to toxicity.
Modeling has become a commonly used tool to predict modes of toxic action in the
last decade. The models are based in Quantitative Structure-Activity Relationships
(QSARs), which are mathematical models that relate the biological activity of
molecules to their chemical structures and corresponding chemical and
physicochemical properties. QSARs can then predict modes of toxic action of
unknown compounds by comparing its characteristic toxicity profile and chemical
structure to reference compounds with known toxicity profiles and chemical
structures were one of the first group of researchers being able to classify modes of
toxic action with the use of QSARs; they classified 600 chemicals as narcotics. Even
though QSARs are a useful tool for predicting modes of toxic action, chemicals
having multiple modes of toxic action can obscure QSAR analyses. Therefore, these
models are continuously being developed.
Environmental risk assessment
The objective of environmental risk assessment is to protect the environment from
adverse effects. Researchers are further developing QSAR models with the ultimate
goal providing a clear insight about a mode of toxic action, but also about what the
actual target site is, the concentration of the chemical at this target site, and the
interaction occurring at the target site, as well as to predict the modes of toxic action
in mixtures. Information on the mode of toxic action is crucial not only in
understanding joint toxic effects and potential interactions between chemicals in
mixtures, but also for developing assays for the evaluation of complex mixtures in
the combination of behavioral and physiological responses, CBR estimates, and
chemical fate and bioaccumulation QSAR models can be a powerful regulatory tool
to address pollution and toxicity in areas where effluents are discharged.