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

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.

A.S. Kane

Lets 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, lets 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.

A.S. Kane

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.

A.S. Kane

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 Id 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%).

A.S. Kane

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.

A.S. Kane

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.

Lets 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.

A.S. Kane

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 wed 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.

A.S. Kane

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.

A.S. Kane

Lets 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, lets 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.

A.S. Kane

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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).

A.S. Kane

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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. CIs 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 CIs 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.

A.S. Kane

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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 (19091994)

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?

A.S. Kane

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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.

A.S. Kane

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