Applied Toxicology

NURS 735 - Module 1

DOSE RESPONSE CONCEPTS

Effect

Dose

The underlying principles of toxicology rely on an understanding of the causal

relationships between exposure and effect. In order to better comprehend how
exposure-related effects can be explained, the concept of dose-response is
important. Although this is covered in your textbook as well as the on-line
materials, it may be helpful, particularly for non-science majors, to spend
some time looking at how dose response relationships are graphed and
interpreted. In this PowerPoint presentation, we will focus on the concept
of dose response.

A.S. Kane 1
 “All substances are poisons: there is none
which is not a poison. The right dose
differentiates a poison and a remedy.”
Paracelsus (1493-1541)

Effect

Dose

And as a review of some of the “History of Toxicology” handout, Paracelus is

noted for stating the concept that “THE DOSE MAKES THE POISON.”
At extremely low doses, a given substance may be non-toxic and even beneficial
(a concept known as hormesis), while at intermediate doses, it may be toxic.
At high doses, it may be lethal. This again underscores the importance of
understanding dose response relationships.

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 Let’s start by looking at some dose response data. To do this, we can begin by
graphing some datapoints. Experimental data is typically plotted with the x-axis
representing the different doses or exposure concentrations, often on a log scale.
The units may be mg/kg or ppm, for example. The concentrations go from low
to high (left to right). The y-axis shows the response data for each
experimentally-derived dose exposure. The units for the y-axis in
this graph is percent cumulative response over a given time period. For now,
let’s assume these these data represent mouse mortality responses taken from
preliminary trials with a new drug. Mortality data are often used to determine
what is known as the LD50 for a drug or compound. Determining the LD50
of a drug, compound or toxicant is an important first step in discerning its
relative toxicity.

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 Here is the experimental data from 10 mice exposed to each of eight
drug doses (80 mice total in the experiment). Each datapoint represents
the percent mortality at each dose. In the first, low dose we see a 0%
response (in other words 0 out of 10 mice succumbed due to the
exposure). At higher and higher doses, there was an increase in the %
response.

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 If we fit a line to these datapoints we see that there is a sigmoid shape to the data.
This is typical for dose response graphs. Note that the curve is a “best fit” line;
it does not necessarily go through each and every datapoint. What I’d like to
point out is that the mid-range experimental dose groups responded in
a linear fashion with respect to the dose (I am referring to the responses
between 16 and 84%).

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 NOEL

The lowest experimental dose where there is no measurable effect is known

as the No Observable Effect Level (or NOEL). Your textbook refers to this
at the No Observable Adverse Effect Level (same thing). In our example, this is the
concentration that the experimenters failed to see any mortality. But it could
have just as well been an experiment looking at the highest dose that failed to
cause enzyme inhibition, or skin pathology or whatever the measured endpoint
of the study was. As we will learn later on, the NOEL is a useful measurement
for extrapolating risk and safe exposure concentrations. Keep in mind that
If the lowest dose tested elicits a response, it is possible to have a dataset
for a drug or toxicant where there no NOEL derived.

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 Dose-response curves for 4 different chemicals

Potency = range of doses over which a chemical

produces increasing responses. A > B; C > D.

Maximal efficacy = limit of dose-response

relationship. A=B; C<D.

Let’s look at a comparison between the dose-response curves for four different
compounds and bring up the notions of POTENCY and MAXIMAL
EFFACACY. In these two graphs, the sigmoid shape of the dose-response curves
Is no longer evident because the response (y) axis is now on a probit scale. Plotting
concentration on a log scale and response on a probit scale mathematically
transforms the data to appear linear; at times this facilitates making
comparisons between toxicants, and generating LC50s (which we will get
to shortly). Looking at the left panel, we see the dose-response
lines for compounds A and B. Note that the slopes of the two response
lines are similar, but the line for compound A is to the left of the line for
compound B. In other words, it takes less of compound A to illicit a similar
response with compound B. Therefore, we can say that compound A is more
POTENT than compound B. And, since the definition of POTENCY is the range
of concentrations over which a chemical produces increasing responses, we
can similarly say that compound C is more POTENT than compound D.

If we now focus on the right panel, we can address the concept of MAXIMAL
EFFICACY. This concept reflects the limit of the dose-response relationship
on the response axis to a certain chemical. The MAXIMAL EFFICACY of
compound C is less than that of compound D. The MAX EFFICACY of
compounds A and B are equal.

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 So, here is how transformed data on a log-probit scale can yield
What is known as an LD50. The LD50, or statistically-derived dose
that is lethal to 50% of a modeled population, is a helpful tool to
compare the toxicity of different compounds, or between different
population models. Since the data is linear here, we can extrapolate
a line across from the 50% response on the y-axis, to the
corresponding concentration on the X-axis. In this example, the LD50
is approximately 100 mg/kg.

Note that LD50s are used to compare responses between drugs or

animal species. However, if we look at the toxicity of a compound
as it is exposed to fish in water, for example, we would not be looking
at an administered dose, but at the concentration that the animals
were exposed to. In this case we’d be deriving the LC50 (lethal
concentration). At the same time, we might be interested in
experimentally deriving an effective dose for a compound, and we
would similarly calculate the EC50, or the effective concentration.

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 Comparison of effective, toxic and lethal dosages

Here is an example of data illustrating the EFFECTIVE DOSE RESPONSE,

the TOXIC DOSE RESPONSE, and the LETHAL DOSE RESPONSE for the
same drug in the same animal model. The order of these response lines
from left to right should be intuitive. Think of an example of a drug and
see if this makes sense to you.

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 Let’s go back to our sigmoid-shaped dose response curve and take a closer
look at interpreting the concept of variability. Here we are looking at
percent response at each concentration or dose. The sigmoid curve between
16 and 84 percent is relatively linear. These limits on the response axis
represent +/- 1 standard deviation of the mean in a population with a normal
distribution.

To better understand the concept of SD and its importance in describing

responses, let’s look at the same data plotted such that the total number
of animals used at ALL doses or concentrations are shown. This is
known as a cumulative frequency response.

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 This frequency response graph works well for QUANTAL data
(i.e., all or none responses, such as mortality, tumor production,
etc.). We can see that the data takes the shape of a BELL CURVE,
with the majority of the responders mostly in the middle. It
appears from this dataset that the mean (mathematical average)
response is approx 110 mg/kg. There are fewer responders at
the ends of the bell curve, and this expected in most data that is
normally distributed. The reason for this distribution is that there
are differences in inter-individual responses. This is known as
BIOLOGICICAL VARIABILITY. Responders at the far left of the mean
are typically HYPERSUSCEPTIBLE, whereas the those at the far right
are RESISTANT. In a normally distributed population, 1 SD of mean
represents 68% of the population; 2 SD of the mean represents
95% of the population; 3 SD represents 99% of the popopulation.
So, the SD, derived from some statistical mathematics, helps us
to identify the portion of the test population that tends to deviate
most from those who respond with a tendency around the mean.
From this estimate of deviation, we can estimate a % of the
population that would tend to not response similarly to the “average”
individual (i.e., an individual that has a response tendency around the mean).

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 Comparison of dose-response relationship for two
different chemicals plotted on a log dose-probit scale

As we described, log-dose data provides a linear plot, easier for interpretation

and extrapolation. This graphs shows two curves on either side of the response
line or compound A. These curves are CONFIDENCE INTERVALS for compound A
data. CI’s are related to standard deviation and tell us how confident we can be
that the true population will respond similarly to the experimental (model) population,
based on the experimental variability encountered. Note that CI’s are tightest
closest to the data line nearest the 50% probit response, closest to the mean.
This makes sense since we know that response variability increases as we move
away from the mean response.

The dose-response line for Chemical B is steeper > Chemical A. Take a second look
at the data, and make a judgment call as to which chemical is safer to use?
Please answer this question and state your reasoning on the Discussion Board.

The next two slides are additions to this ppt presentation, and were not included
in the audio version.

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 ?

“The problem with toxicology is not

the practicing toxicologists, but
chemists who can detect, precisely,
toxicologically insignificant amounts
of chemicals”

Rene Truhaut, University of Paris (1909-

1994)

Here is a final concept worth considering. Current technology allows us

to accurately measure trace quantities of chemicals. Yet, the biological
significance of the presence of these extremely small amounts of these
chemicals is sometimes not clear (e.g., carcinogens). Just because a
drug or environmental contaminant is measurable, does that mean
that it poses a threat?

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 This slide is food for thought…

Many environmental contaminants (and even foodstuffs) are

Considered differently by different scientific groups and
agencies with regard to their safety. It is, therefore, up
to the “consumer” in many cases to make a judgment call
as to how much exposure is reasonable, based on available
information. This concept is related to RISK ASSESSMENT.

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