PHARMACOKETICS AND ADME

Workshop

Introduction
The following pharmacokinetic and metabolism data is derived from data in
the literature on a candidate new chemical entity, amosulalol.

H2N

O
S

O
H
C

H3C

CH2NHCH2CH2

OH
H3CO

Amosulalol

RMM 379

Pharmacokinetic investigations
1. Single dose intravenous bolus study
Experiments were conducted in rats, dogs and monkeys. A solution of amosulalol
hydrochloride in physiological saline was administered at a dose of 1mg/kg body
weight. Blood samples were then taken at set time intervals and analysed for parent
compound. Urine was collected for 48 hr after dosing and analysed for parent
compound and in certain instances for total metabolites.
2. Single dose oral study
Experiments were conducted in rats, dogs and monkeys. Animals were fasted
overnight and given an aqueous solution of amosulalol hydrochloride in physiological
saline at a dose of 10mg/kg body weight by gastric intubation. Parent compound was
determined in plasma at set time intervals and in urine samples. Urine samples were
also investigated for metabolites.
Urine samples were collected from three human volunteers (60-65 kg body weight)
who took 50 mg amosulalol hydrochloride in the form of a tablet.
3. Repeat oral dosing study
Dogs were allowed normal access to food and were given amosulalol hydrochloride,
10 mg/kg body weight, in a gelatine capsule daily for 15 days. Timed blood samples
were taken throughout the day on various days throughout the study for the analysis
of parent compound.

RESULTS
1. Single dose intravenous bolus study

Time (hr)
Rat
0.5
1.0
2.0
3.0
5.0
7.0
8.0

60.9
53.1
40.2
30.4
17.5
10.1
-

Plasma concentration of amosulalol (ng/ml)

Dog
Monkey
127.0
108.0
77.5
55.7
28.8
10.7

396.0
327.0
222.0
151.0
70.0
22.0

PK exercise 1
1. Plot the data on 2 cycle semi-log graph paper.
What compartmental model is the data consistent with?
2. For each species, determine:
(a) Biological half life (t1/2)
(b) Elimination rate constant (Ke)
(c) Apparent volume of distribution (VD)
(d) Area under the plasma concentration-time curve (AUCiv)
Hint: AUCiv = Co
Ke
(e) Plasma clearance (Cl), what relationship have you chosen to
use?

2. Single dose oral study

Time (hr)

Plasma concentration of amosulalol (ng/ml)

Rat
Dog
Monkey

0.2
0.4
0.6
0.8
1.0
2.0
3.0
5.0
6.0
8.0

40.2
69.4
90.1
104.0
113.0
118.0
97.2
56.3
41.9
23.0

317.0
509.0
618.0
673.0
693.0
591.0
437.0
227.0
164.0
84.5

610.0
1080.0
1420.0
1670.0
1840.0
1980.0
1620.0
826.0
560.0
243.0

1. Plot the data on 3 cycle semi-log graph paper.

2. For each species determine
(a) Tmax
(b) Cmax
(c) Biological half life (t1/2)
(d) Elimination rate constant (Ke)
(e) Absorption rate constant (Kabs)
(f) Area under the plasma concentration-time curve (AUCpo)
Hint: AUCpo = Z - Z
Ke Kabs
3. Using both the IV and oral data determine the oral bioavailability for each
species.
4. Comment on the Cmax and AUCpo values.

Excretion of unchanged amosulalol and its metabolites in urine

The amounts excreted in urine over 48 hr after oral dosing.

Species

Urinary excretion (% of dose)

Amosulalol

Total metabolites

IV

Oral

IV

Oral

Rat

17.1

5.6

28.2

20.6

Dog

13.4

6.4

34.7

38.8

Monkey

14.8

10.3

35.8

Man

30.1

12.8

1. Explain the proposition that the ratio:

Excretion of parent compound in urine after oral dose
Excretion of parent compound in urine after IV dose
Is a measure of the oral bioavailability of a drug.
2. Determine the oral bioavailability for each species using this approach and
compare the values from the plasma data.
3. Comment on the urinary excretion data in light of the biological half life of
amosulalol in the species studied.
3. Repeat oral dosing study in dogs
Day

Tmax
(hr)

Cmax
(g/ml)

AUC
(g hr ml-1)

t1/2
(hr)

1.0
2.0
3.0
8.0
15.0

1.4
1.2
2.0
1.8
1.9

0.66
1.14
0.65
1.11
1.15

3.2
3.4
2.5*
2.8
3.4

2.5
2.4
2.2
2.1
2.2

1. What is the purpose of conducting a study like this?

2. Compare the results of this study with the single dose oral study in dogs.
What conclusions would you draw?

3. These results are the mean levels of 4 animals and statistical analysis of
the results showed that the only value that differed significantly from the
other values in the same column was indicated by * for which the level of
significance was P< 0.05. What interpretation would you offer for this
observation?
Dr Glyn B Steventon, ADMET Solutions Ltd, UK

The pathways shown are the major pathways in each species. In monkeys, the
metabolites found were for both man and dog with the former being the more
important.
1. What tentative conclusions can be drawn about the functionalization (phase I)
metabolism of amosulalol?
2. What significance, if any, does this knowledge of metabolism have in the
interpretation of the pharmacokinetic data?
3. Does the metabolic information have any importance in the interpretation of
animal toxicity studies?