Psychopharmacology (1985) 86:392-399
Psychopharmacology
Comparative pharmacokinetics and pharmacodynamics
of lorazepam, alprazolam and diazepam
Everett H. Ellinwood, Jr., Douglas G. Heatherly, Arlene M. Nikaido,
Thorir D. Bjornsson, and Clinton Kilts
Behavioral Neuropharmacology Section, Department of Psychiatry, and Division of Clinical Pharmacology, Department of Medicine,
Duke University Medical Center, P.O. Box 3870, Durham, NC 27710, USA
Abstract. The contribution of differential absorption-dis-
tribution pharmacokinetics to drug activity can be partially
determined by comparing simultaneous estimates of drug
serum level with pharmacodynamic effects. In the present
paper we have contrasted the effects of clinically equipotent
doses of lorazepam, alprazolam, and diazepam on the
performance of tracking and digit symbol substitution
tasks. Eight young males were tested for 12 h after ingesting
the drug. The three benzodiazepines and placebo were
administered to each subject according to a balanced
double-blind Latin square design. A model is presented
that describes the relationship between drug concentration
and the degree of impairment across time after the final
peak effect. Exponential rate constants were determined
for each drug using a Marquardt nonlinear fit of the pooled
data. Basically, the constants relate offset serum drug
values to the impairment curves at a time when serum-brain
equilibrium is assumed to have occurred. The values
indicate markedly rapid acute tolerance for alprazolam and
diazepam but relatively little acute tolerance for lorazepam.
Whether these constants reflect adaptation or differential
association-dissociation receptor rate constants cannot be
determined, but they do highlight the need to consider
receptor kinetics as an important factor in benzodiazepine
pharmacodynamics.
Key words: Lorazepam - Alprazolam - Diazepam -
Acute tolerance - Pharmacokinetics - Pharmacodynam-
ics
Benzodiazepine pharmacodynamic profiles are not pre-
dicted well by dispositional pharmacokinetics following
acute dosing. The offset of diazepam-induced psychomotor
impairment is much more rapid than the corresponding
decline of concurrent bound and unbound diazepam
concentration (Ellinwood et al. 1983a, b; MacLeod et al.
1977). In contrast, lorazepam has a prolonged period of
impairment relative to diazepam (Seppala et al. 1976;
George and Dundee 1977), which is clearly not a function
of drug elimination rates, since the elimination half-life of
lorazepam is one-third that of diazepam.
Either differential absorption-distribution pharmacoki-
netics (Arendt et al. 1983) or receptor kinetics appear to
be a more likely explanation of the observed differences in
pharmacodynamics between the two drugs than their
elimination kinetics. The contribution of differential
Offprint requests to: E. H. Ellinwood, Jr.
9 Springer-Verlag 1985
absorption-distribution pharmacokinetics to drug activity
can be partially determined by comparing simultaneous
estimates of drug serum level with behavioral effects. In the
present study we have contrasted the effects of clinically
equivalent doses of lorazepam, alprazolam, and diazepam
on the performance of cognitive-neuromotor tasks. We
selected a high dose, i.e., twice the recommended higher
anxiolytic dosage, to obtain values closer to the maximum
effect and to be in the more linear segment of the
dose-response curve. The three benzodiazepines were
chosen on the basis of their diversity in pharmacologic
properties including elimination half-life, lipid solubility,
plasma binding, and high-affinity receptor binding.
Materials and methods
Eight males, who were between the ages of 21 and 26 and
within + 10% of ideal height-weight ratios, participated in
the study. The purpose and procedure were explained to
the subjects and written informed consent was
obtained.
Procedure
Subjects were trained to a plateau level of performance on
the tasks in three 2-h sessions. Four drug sessions were
scheduled at 2-week intervals according tO a Latin square
design balanced for drug order. Lorazepam, alprazolam,
diazepam, and placebo were administered using a dou-
ble-blind procedure at doses of 0.057, 0.028, 0.286, and
0.000 mg/kg body weight, respectively. The powered drug
was given PO in a starch lactose solution.
Each test session started at 7:30 AM with a predrug trial
followed by repeated performance testing on various
cognitive-neuromotor tasks. Two of the most frequently
presented tasks, continuous subcritical tracking (SCT) and
digit symbol substitution (DSS), are discussed in this paper.
Results of the other tests will be described in other papers
(Heatherly et al. in preparation). To assess the time course of
the drug effect, particularly during the initial period, the SCT
task was presented at predrug and at 5, 20, 25, 30, 35, 65,
80, 95, 110, and 155 min and 3.5, 5.5, 7.5, 9.5, 10.5, and
ll.5'h postdrug ingestion. The DSS task was performed
concurrently at the same time intervals as the SCT task,
except at 20, 25, and 30 min; only the SCT task was done at
the latter three time periods.
A 7-ml venous blood sample was drawn before and at
20, 35, 65, 85,100, and 135 min and at 3, 4, 6, 8, 10, 11, 24,
and 48 h after drug ingestion from an obturated indwelling
catheter in the subject's left forearm. Lorazepam, diaze-

pam, and alprazolam serum levels were quantified after
extraction using electron-capture detection gas chromato-
graphy' as described by Greenblatt et al. (1978, 1981) and
Skinner (1979) with modifications. Only very small
amounts of N-desmethyldiazepam were detected on test
sessions following the diazepam treatment and did not
interfere with the assays of the other drugs.
Psychomotor tasks
Apparatus. The experimental chamber was a sound-atten-
"uated room, containing a 98- x 128-cm rearview video
display screen and an adjustable chair facing the screen. A
car steering wheel and nine-digit keypad were attached in
front of and to the right of the chair, respectively. All
subjects were right-handed. The images were controlled by
a Digital Equipment PDP 11/34 computer and an LPS 11
Analog/Digital System interfaced to a custom-designed
Raster Scan unit. The computer system recorded digital
data on-line.
Continuous subcritical tracking. A 3-cm wide illuminated
bar was displayed from top to bottom in the center of the
darkened TV projection screen. A central portion of the
bar, 34 cm in length, moved laterally across the entire
screen. The movement of the tracking bar was controlled by
the subject manipulating the steering wheel and by the
computer. The wheel was connected to a 10-turn poten-
tiometer of 100 g2, which was connected to the analog/di-
gital input of the computer. The position of the poten-
tiometer was controlled by the steering input, added to the
position of the bar by the computer and multiplied by a
constant value, termed the level of difficulty (2). A
simplified equation of this relationship would be: change in
bar position = (bar position + potentiometer position) x 2.
Using this equation a new bar position was calculated 60
times/s. The duration of the tracking task was 3 rain; the 2
was 2 for the first 30 s and increased to 3 during the last
150 s. Subjects were instructed to maintain the moving bar
in the center of the screen. The measure of merit was the
root mean square (RMS) deviation of the bar from the
center.
Digit symbol substitution. This task was an automated
version of the DSS subtest from the Wechsler Adult
Intelligence Scale. A" code table, consisting of one row of
sequential numbers from 1 to 9 and a second row of the nine
DSS symbols, was displayed on the screen. A symbol was
presented below the code table and the subject pressed on
the keypad the number paired with the symbol in the code
table. The number-symbol pairings were randomly rear-
ranged at the start of each battery. There were 12
presentations of each symbol for a total of 108 trials for the
entire task. The primary measure of merit was the power
score which was calculated by dividing the total number of
correct responses by the average reaction time.
Pharmacokinetic analysis
The pharmacokinetic parameters, elimination half-life
(t 89 apparent total clearance (C1), and apparent volume
of distribution (Vdarea), w e r e calculated from the serum
concentration versus time data using conventional expres-
sions (Gibaldi and Perrier 1975). The elimination half-life
was determined from the elimination rate constant (Kel),
393
which was calculated from the terminal linear phase of the
natural log concentration versus time plot. The K d for
diazepam is a rough estimate since the last blood sample
was taken at only 48 h. The total area under the serum
concentration versus time curve (AUC) was computed
using the trapezoidal method for the observed data and
extrapolated to infinity from the last data point using the
Kel determined earlier. Apparent total clearance was
determined from the ratio of dose administered to AUC.
Apparent volume of distribution was computed as the ratio
of C1 to K d. The values reported as the maximum
concentration (C~x) and the time at which Cm~ occurs
(tmax) are the actual observed values.
Statistical treatment of the data
Analysis of the drug effect across all batteries was performed
by using the repeated measures analysis of variance program
of the 1983 version of the Biomedical Statistics Package
(BMDP). Battery was the repeated factor. Subsequent
contrasts of means were conduced with an ct level of 0.001
due to the number of multiple comparisons.
Pharmacodynamic/pharmacokinetic model. Since previous
studies suggest that drug serum concentrations account for
only a small percentage of the variance in. the performance
scores, we defined the following mathematical model to
describe more accurately the relationship between drug
concentration and the degree of performance impairment
across time after the final impairment peak:
E = FF x P + A o x e(--KATc X t) Eq. (1)
where E = performance score, P = serum drug concen-
tration, FF = flee fraction for drug, t = time after drug
ingestion,/(AT c = a hybrid acute tolerance constant, Ao =
coefficient or intercept term for KaTC.
Due to the nature of the impairment time course (fast
rise, double peak, and offset), the model was applied only
to the offset phase from the final impairment peak at 95 min
for lorazepam and alprazolam and at 80 rain for diazepam
to the last measured time point at 690 rain postingestion.
Both parameters, /(ATc and Ao, were determined by
performing nonlinear least-squares regression analysis on
all the known parameters. Since the free fraction (FF)
change over time for each drug was negligible compared to
the change in serum concentration, the FF used in the
equation was a constant determined from values reported
in the literature (Greenblatt et al. 1980, 1983; Johnson et al.
1979). The nonlinear method used was the Marquardt
method of the NLIN procedure in the Statistical Analysis
System (SAS) package. Performance scores were repre-
sented as delta scores (postdrug score minus predrug
score). Regression analyses were performed to determine
how well the model predicted the observed performance
measures.
Results
Pharmacokinetic parameters for the three benzodiazepines
are listed in Tables 1, 2, and 3. In general, they are within
the range of published results for PO dosing; however, tmax
is early, representing primarily the fast absorption of the
suspension formulation. To define more precisely the time
course of the drug effect during the initial period of rapid
394

Table 1, Pharmacokinetic parameters of diazepam after a PO dose of 0.286 mg/kg body weight

Subject Weight Age Dose tm C1 Vd~ea Cm,x t,n~

(ID) (kg) (years) (mg) (h) (ml/min/kg) (l/kg) (ng/ml) (h)

L1 -SK 71.8 25 20.5 19.4 0.41 0.69 740 1.583

L2-DB 84.1 25 24.1 22.7 0.39 0.76 1,143 1.333
L3-GH 61.4 25 17.6 36.8 0.16 0.50 1,039 1.083
L4-PZ 61.4 26 17.6 24.8 0.31 0.66 840 1.833
L5-CD 86.4 24 24.7 22.9 0.36 0.71 1,067 0.583
L6-DT 72.7 26 20.8 85.0 0.15 1.13 1,086 1.083
L8-TS 70.0 26 20.0 27.2 0.31 0.73 1,347 0.583
L9-RC 84.1 22 24.1 27.4 0.35 0.83 891 1.333
J~- 73.9 25 21.1 33.3 0.31 0.75 1,019 1.178
SD 9.9 1 2.9 21.5 0.10 0.18 191 0.442

Table 2. Pharmacoldnetic parameters of alprazolam after a PO dose of 0.028 mg/kg body weight

Subject Weight Age Dose tv2 C1 Vd.... Cmax tmax

(ID) (kg) (years) (rag) (h) (ml/min/kg) (1/kg) (ng/ml) (h)

L1-SK 71.8 25 2.05 13.6 1.03 1.21 26 1,333

L2-DB 84.1 25 2.41 14.7 1.09 1.39 25 2.583
L3-GH 61.4 25 1.76 13.5 0.72 0.85 62 0.333
L4-PZ 61.4 26 1.76 18.6 0.94 1.51 37 0.333
L5-CD 86.4 24 2.47 11.6 0.74 0.74 39 1.583
L6-DT 72.7 26 2.08 22.0 0.40 0.77 58 0.333
L8-TS 70.0 26 2.00 1.92 1.25 1.29 31 1.583
L9-RC 84.1 22 2.41 9.7 0.72 0.61 43 3.500
2( 73.9 25 2.11 14,5 0.86 1.05 40 1.448
SD 9.9 1 0.29 4.0 t3.27 0.34 14 1.15I

Table 3. Pharmacokinetic parameters of lorazepam after a PO dose of 0.057 mg/kg body weight

Subject Weight Age Dose tv2 CI Vd.... Cmax /max

(ID) (kg) (years) (mg) (h) (ml/min/kg) (1/kg) (ng/ml) (h)

L1-SK 71.8 25 4.10 19.9 1.20 2.05 46 1.583

L2-DB 84.1 25 4.80 20.0 1.08 1.87 81 0.583
L3-GH 61.4 25 3.50 19.4 1.08 1.81 78 0.583
L4-PZ 61.4 26 3.50 12.1 1.35 1.41 63 0.333
L5-CD 86.4 24 4.93 17.6 0.84 1.29 77 0.583
L6-DT 72.7 26 4.15 9.1 1.80 1.41 40 0.583
L8-TS 70.0 26 4.00 14.8 1.40 1.80 54 0.583
L9-RC 84.1 22 4.80 12.9 1.63 1.82 52 0.583
2( 73.9 25 4.22 15.7 1.30 1.56 61 0.670
SD 9.9 1 0.57 4.1 0.31 0.40 16 0.380

changes in p e r f o r m a n c e , SCT, the m o t o r coordination task,
was p r e s e n t e d frequently during the first 2 h and is the
principal test described in this report, DSS is also discussed
to d e m o n s t r a t e differential offset effects for a cognitive
task.
T h e tracking profiles are shown in Fig. 1A. The
i m p a i r m e n t profile is quite similar for d i a z e p a m and
alprazolam b u t different for lorazepam. O n e r e m a r k a b l e
feature of the tracking profiles for individual subjects is the
a p p e a r a n c e of two distinct m a j o r p e a k s of i m p a i r m e n t for
d i a z e p a m and alprazolam. H o w e v e r , the difference
b e t w e e n the m e a n of each p e a k and the lowest intervening
m e a n p o i n t b e t w e e n the p e a k s (trough) was not significant
for either drug, p r o b a b l y due to intersubject variability in
the latencies of the p e a k s and the trough. T h e r e was no
consistent relationship across drugs between times of p e a k
drug concentrations and p e a k e f f e c t s .
A f t e r the second p e a k effect all three benzodiazepines
show a r a p i d offset of the SCT effect; d i a z e p a m and
a l p r a z o l a m rapidly return to baseline within 6 0 - 1 0 0 min.
L o r a z e p a m has both an initial r a p i d offset and a substantial
late, shallow offset slope, and returns to baseline only after
500 rain. In general, l o r a z e p a m had m o r e p r o n o u n c e d and
p r o l o n g e d impairing effects, while alprazolam p r o d u c e d a
level of i m p a i r m e n t b e t w e e n that of l o r a z e p a m and
diazepam. T h e time course of i m p a i r m e n t on the DSS task
was very similar for all three drugs to that of tracking.
H o w e v e r , the i m p a i r m e n t falloff for later periods was
slower for DSS than SCT (Fig. 1B).
T h e r e p e a t e d measures analysis indicated signficant
 395
275

drug [F (3,28) = 7.39, P < 0.0009], testing time [F (15,420)

250
= 24.14, P < 0.0001], and drug x testing time [F (45,420) =
4.63, P < 0.0001] effects. Significant differences between
2~'5 drugs on the SCT task are shown for different testing time
points in Fig. 1A. Similarly, the drug IF (3,27) =26.68, P <
T
2O0 0.0001], testing time [F (12,336) = 50.08, P < 0.0001], and
R drug x testing time [F (36,336) = 6.34, P < 0.0001] effects
A
C 175 were significant for the DSS task. As illustrated in Fig. 1B,
K
I differences between drugs for DSS impairment were signifi-
N
G IS0 cant during the first 4 h.
O To compare directly the time course of the drug serum
E
L 125 level to that of the drug effect, the performance of a subject
T
R for the SCT and DSS tasks was represented as a percent of
S 100
the maximum effect shown by that individual for each drug
C
0
condition (Figs. 2 A - C ) . Serum drug levels were similarly
R
E 75
transformed to a percent of the maximum blood levels
S achieved by each subject for the different drugs. In this way
5o
performance and drug measures were normalized for all
subjects within each drug condition allowing the compar-
ison of the time courses of effect and drug concentration for
25 Z
each drug. However, the transformed values do not validly
reflect magnitude of effect or drug levels for comparisons
~ across the three benzodiazepines.
0 }00 200 300 ~00 500 600 700 Over the 11.5-h testing period N-desmethyldiazepam
A M]N POST INGESTION constituted a very small fraction of the total diazepam
concentration. Based on the reported relative potency of
~oi ~ N-desmethyldiazepam (Palva and Linnoila 1978), the
diazepam serum levels in Figs. 2 and 3 reflect diazepam plus
55, half the measured metabolite concentration. To simplify
the data presentation in Fig. 2 and 3, the free fraction of the
5D. three drugs was not used in the calculation of the blood
O
$
values since protein binding would not vary substantially
S ~5. over the testing interval and thus would not alter the
0
V
time-dependent relationship between performance and
E qO-
R
serum levels.
B
L
The curves in Figs. 2A and 2C indicate that the
L 35-
diazepam impairment effect had a leading phase relation-
P
0 ship to serum drug level, whereas the opposite was the case
W 30-
E for lorazepam. As reported previously (Ellinwood et al.
R
1983a, b), acute tolerance is demonstrated for diazepam by
I 25-
M the precipitous drop in tracking impairment effect between
P
R
I 20-
110 and 155 rain, while serum levels fall more slowly.
R Alprazolam also demonstrated a 70% recovery of the peak
N 15-
impairment effect between 110 and 210 min during which
T time mean serum drug concentrations remained constant
S
io T
(Fig. 2B). In fact, serum values did not start to decline
) significantly until after the impairement effect had almost
S S- returned to baseline, suggesting the involvement of other
mechanisms before a decrement in drug level begins to
O2 affect performance.
The DSS task demonstrated a more prolonged effect
-5 for all three drugs, especially in the late phase offset. By
,' . . . .

I00
i , , .... , ....... i ...... , , - . , 20.0rain tracking impairment had returned to 15% of
200 300 qO0 500 600 700

B MIN POST INGESTION

maximum effect, whereas DSS performance improved at a
much slower rate. The varied time courses for the effect
Fig. 1. Impairment on subcritical tracking (A) and digit symbol falloff indicate that different underlying processes may be
substitution (B) tasks for diazepam (curve D), alprazolam (A), involved in the effect of the benzodiazepines on the SCT
lorazepam (L) and placebo (P). Performance is represented as unit and DSS tasks.
changes from the predrug score (delta score). A smooth spline
fitting procedure was used to generate the curves connecting all Similar concentration-response relationships are re-
time points. Symbols indicate significance levels for diazepam flected in the clockwise and counterclockwise hysteresis
versus alprazolam (A), alprazolam versus lorazepam (+), and curves for diazepam and lorazepam, respectively (Figs, 3A
diazepam versus lorazepam (C)) mean comparisons (one symbol and 3C). Only the tracking hysteresis is shown, since there
represents P < 0.05; two symbols P < 0.01; and three symbols P < were no early DSS measures. Compared to diazepam and
0.001) lorazepam, alprazolam had a very distinct hysteresis profile
396
lOO-I I O0-~ P~
r
90L
9o ~ i
oo-] ~1 N ',

7o

T 6D T 60

0
50
B
S !
E ~0 s ~o
R

D 30-

X 20.
1 T "-..~_ . . . . . . . . D. 2~ T ,, ,,,
M ~o- M I0 I'; " " ' " .... ""T-. ~ "I)
"..-.T- ..... "-'D

oZ "" ......... ~ ............. T---....T. __ ..-T O "'T ...... T

~]0 .,% -lO

0 I00 200 300 lO0 500 600 700 0 I00 200 300 ~00 500 600 700
A MIN POSTINOESTION B MIN POSTINGESTION

!/"~?'~$'"'\
I00--

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'lDl~"'~"
: %%

//
R 70 m

EPN I
T 60. i
0 ', ""
F ~\
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B$405" !
ER "T."'.........
O 30- T ""I x\ --B

X 20. i \
I "'" "~
M . "~..
M lo- "'"'-... Fig. 2. Time-dependent relationship between subcritical tracking
""'T'-.-._T.....~ " (curve T), digit symbol substitution (D) and drug serum concen-
tration (B) for diazepam (A), alprazolan (B), and lorazepam (C).
Task performance was measured as unit changes relative to
-I0 'qr predrug level. Task and blood values are expressed as percent of
0 lO0 200 SO0 400 500 600 700 maximum value and the curves were generated using a smooth
C HIN POSTINGESTION spline fitting procedure

(Fig. 3B); the onset and offset of the entire major unbound drug serum concentration alone. Alprazolam
impairment effect occurred within a very narrow average showed the largest increase in r2 from 16% to 52% for SCT
drug serum range, between 2 8 - 3 1 ng/ml. In contrast, there and from 32% to 57% for DSS. There was a more modest
was a much more systematic linear relation between the improvement for lorazepam from 33% to 43% for SCT and
offset time courses of the impairment and serum drug levels from 47% to 53% for DSS. Similarly, use of the KATC
of lorazepam. model changed to a lesser extent the r2 values for diazepam
The exponential constants (KATc) that represent the from 19% to 27% for SCT and from 30% to 34% for
best relationship of the offset of the impairment curves to DSS.
drug serum values were 1.06 h -1 for diazepam, 0.91 h -1 for
alprazolam, and a much slower constant of 0.23 h -1 for
lorazepam. The KATC constants for DSS were all consid-
erably lower, i.e., 0.08 h -1, 0.14 h -1 and 0.09 h -I, respec- Discussion
tively. Regression analyses indicated that for all three drugs The clockwise hysteresis loop observed for diazepam in
the model described by Eq. 1 accounted for more of the Fig. 3A has been designated as pathognomic of tolerance
performance variability during the offset phase than the (Holford and Sheiner 1981). Tolerance is defined as a
 397

reduced drug effect at the same concentration that earlier more robust indicator of acute tolerance is the contrast
produced a greater effect. Clockwise hysteresis loops have between the onset and offset areas of the loop. For
been reported previously for the same tracking task example, diazepam impairment at 25 min is nine times the
following each of three different doses of diazepam level at 210 min, yet serum concentrations for both are
(Ellinwood et al. 1983a), demonstrating that the diazepam around 500 ng/ml. The KATCis an indicator of a late phase
acute tolerance profile is reproducible across dose levels9 of acute tolerance. The KATC values for diazepam and
Several components of the hysteresis loop are important in alprazolam quantitatively represent the faster effect offset
the formulation of the contributions to acute tolerance9 The as compared to the serum level offset. Both drugs thus

160

150 I

140 200.
i 110
130
100,
120 -

LiO - 160,
100 - 80, 15
T
R lqO
90- fl
C
B0 - I
M 120,
r.
70-
0
s
60- L 100, lS
T
Z5
$
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~~ 0
I
E
3O $
/9 ~0.
20'
q0.

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++go%+,~........ +" ""' 20,
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-20
., 9 , .... J . . . . . . . . . + . . . . . . . . . i . . . . . . , 9 + . . . . . . . + . . . . . . . . , . . . . . . . . . ,

IO0 200 300 qO0 500 600 700 800 gO0 10 15 20 2S 30

A SERUM DIflZEPflH CONCENTRRTION CNG/ML) B SERUNALPRAZOLAMCONCENTRATION(NG/ML)

275 -
IP
/
110"
250.

225 9

2~ -
T
R

C 175-
K
!
N
S ISO-
D
s
L }.25
T +
fl
330
+ I00,
C
0
R
SE 75,
Fig. 3. Hysteresis curves of subcritical tracking performance versus
drug serum concentration for diazepam (A), alprazolam (B), and
lorazepam (C). Impairment is expressed as unit changes from the
predrug level (delta score). Mean values of both actual (curve A)
%;'~ and predicted (P) scores are shown. Predicted values are based on
the KATCmodel. Points represent testing time periods, which are
0 S 10 J$ ~0 2$ 30 35 q~ q~ $0 S$ indicated as number of minutes post ingestion. Arrows denote the
C SERUN LOP&ZEPANCONCE~ITP.ATIOtl(HG/NL) sequence of the observations
398
induce a very rapid initial offset of tracking impairment that
is not reflected in the shallower slope of the drug level fall
off. Between the onset and offset arms of the hysteresis
loop there is a plateau period lasting 4 5 - 6 0 min for both
drug concentration level and effect.
The alprazolam hysteresis curve does not fit classical
notions about hysteresis since the delay in the onset drug
effect produces a counterclockwise hysteresis and the fast
offset of effect is consistent with a clockwise hysteresis
indicating acute tolerance. Arendt et al. (1983)found in
dogs a late (28 min) latency time to peak cerebrospinal fluid
(CSF) concentations for alprazolam in contrast to 3 min for
diazepam and 7 min for lorazepam. If confirmed, this lag in
central nervous system (CNS) entry for alprazolam may
prove to be the reason for the abrupt rapid onset of
alprazolam-induced impairment following an initial onset
delay of 1 5 - 2 0 min. The alprazolam offset KATC, calcu-
lated from time periods when the equilibrium between
serum and CSF should be established, is very similar to that
of diazepam and probably reflects receptor kinetics or
adaptation.
The counterclockwise hysteresis loop manifested with
lorazepam indicates either a reverse tolerance (sensitiza-
tion) or the presence of a drug equilibrium delay between
the plasma and CNS receptor sites. However, an equili-
brium delay as the only contributor to the reverse tolerance
is doubtful for this lipid-soluble drug, since Arendt et al.
(1983) have demonstrated peak CSF levels in dogs 7 min
after intravenous dosing. Furthermore, while the onset
hysteresis limb for lorazepam is counterclockwise, the
offset slope demonstrates no evidence of reverse tolerance.
In fact, the small positive KAXCconstant is one-third that of
diazepam or alprazolam. Finally, although the desmethyl
metabolite of diazepam was the only active metabolite
measured, there is no evidence in the literature that other
metabolites of any of the three drugs contribute to the early
acute effects.
The recovery curve for DSS is characterized by a late
substantially slow offset component. The contrast to the
tracking recovery profile was distinct for diazepam and
alprazolam and the KATCconstants for all three drugs were
below 0.14 h -1. The DSS task incorporates components of
learning and memory as well as psychomotor speed
(Heatherly et al. in preparation), whereas tracking reflects
mainly coordination ability. Elsewhere we have reported
that the learning component of the DSS displays a more
prolonged impairment offset than the memory portion
(Heatherly et al. in preparation). The data raise the intrigu-
ing question as to what mechanisms may be responsible for
the differential profiles of the benzodiazepine effect demon-
strated by the different types of tasks.
Since it is difficult to conceive of a differential receptor
or effector mechanism operating for these drugs, the
present data are more consistent with the hypothesis of
differences in receptor association-dissociation rate con-
stants as recently proposed by Jack et al. (1983)~ These
authors argued that the pharmacodynamic half-life of
benzodiazepine may relate to the pharmacokinetic profile
as well as drug differences in association-dissociation
receptor rate constants; they conclude on a basis of
differences in ex vivo binding studies that diazepam has a
dissociation rate approximately ten times faster than
lorazepam. That SCT impairment for diazepam leads and
lorazepam lags serum drug concentration during the onset
phase in our data may indicate that receptor association
rate constants m a y be involved significantly in the early
process. In addition, the finding that for the SCT task the
diazepam KATC was three times that of lorazepam is
consistent with the Jack et al. (1983) hypothesis. However,
there is no direct data to support this hypothesis.
The comparison between lorazepam and alprazolam
indicates that the prolonged effect of the former is not
simply a function of relative binding affinity (Kaplan and
Jack 1983). Both drugs have similar binding affinity
(Baestrup and Nielsen 1983), but our findings demonstrated
a much shorter duration of impairment for alprazolam.
Similarly, ex vivo binding studies have shown that diazepam
has a dissociation rate that is about nine times that of
lorazepam (Jack et al. 1983). The structure of lorazepam, as
well as triazolam and clonazepam, includes a chloride ion on
the ortho position of the ,,C" ring, which has been associated
with the very high affinity of these benzodiazepines and may
account for a specific long-lasting lorazepam effect.
Recently emerging evidence has fostered the hypotheti-
cal position that various areas of the CNS have differential
distributions of at least two benzodiazepine receptors with
differential binding and dissociation as a function of time,
detergent solubilization, temperature, interaction with
triazolopyridazines, G A B A , and the presence of chloride
ion (Dubnick et al. 1983; Lo et al. 1982; Squires et al. 1979).
In contrast, on the basis of their research Gee and
Yamamura (1982) and Chin and Rosenberg (1982) have
suggested an alternative explanation, namely that there is a
single benzodiazepine receptor population with at least two
conformations. Thus, the hypothesis of multiple benzod-
iazepine receptor subtypes or conformations with differ-
ential dissociation rates represent one possible explanation
for the findings of fast and slow offset slopes for the
different tasks.
Squires et al. (1979) have described two components of
3H-diazepam dissociation with half-lives of 2 and 9 min
after addition of either diazepam or flunitrazepam. When
CL-218-872, a 3-triazolopyridazine (a BZ1 type) anxiolytic,
is added, the fast component of the 3H-diazepam disso-
ciation is unaffected while the slow displacement compo-
nent becomes much slower, suggesting to these authors that
CL-218-872 has a lower affinity to "slow receptors" than
"fast receptors." Recently, Reeves and Schweizer (1983)
found evidence that in young rats acute treatment with
diazepam results in a 147% increase in BZ 1 (fast) receptor
density while BZ2 (slow) receptors remain unchanged.
Thus, an acute adaptation of the BZ1 receptor type could be
involved in diazepam's short pharmacodynamic half-life.
Dubnick et al. (1983) has also reported greater distribution
of BZ1 receptors in the cerebellum in contrast to increased
distribution of BZ2 receptors in the hippocampus and
cortex. Thus, differential rate of recovery for learn-
ing-memory versus coordination skills could be related to
anatomical distribution of receptor types. Although these
hypotheses are currently untestable in man, they do
highlight the point that receptor kinetics need to be
considered in the pharmacodynamic profiles of benzodia-
zepines.
Acknowledgements. This research was supported by NIDA grant
DA01883.

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Received August 27, 1984; Final version November 27, 1984