Pharmacology 27; 223-236 (1983)

The Effects of SKF 525-A on the Analgesic and

Barbiturate-Potentiating Activity of A9-Tetrahydrocannabinol
in M ice and Rats
R.D. Sofia, II. Barry, III
Department of Pharmacology. University of Pittsburgh School of Pharmacy. Pittsburgh, Pa., USA

Key Words. A9-Tetrahydrocannabinol • SKF 525-A A9-Tctrahydrocannabinol/SKF

525-A interaction • Analgesia • Sleeping time

Abstract. A9-Tetrahydrocannabinol (THC) markedly potentiated barbital Na sleeping time

in mice and rats. The magnitude and duration o f this effect were markedly enhanced when the
animals were pretreated with SKF 525-A, a nonspecific inhibitor o f liver microsomal
enzymes responsible for drug metabolism. Moreover, depending on the method and species of
animal used, THC was found to be one half (mouse hot plate), one third (mouse tail flick), and
one eighth (rat tail flick) as potent an analgesic as morphine S 0 4. However, pretreatment with
SKF 525-A significantly potentiated the analgesic activity in mice. These data suggest that
intact THC and not necessarily a metabolite(s) is principally responsible for the CNS depres­
sant and analgesic effects observed.

Introduction cal activity o f systemically administered

According to several reports [Isbell et al.,
1967; Gershon. 1970; Mechoulam, 1970;
Hollister, 1971] A9-tetrahydrocannabinol
(THC) appears to be the major psychoactive
ingredient of marihuana. Metabolic studies
have indicated that THC is rapidly hydroxy-
lated in the body via liver microsomal en­
zymes to form the metabolite ll-hydroxy-
THC [Nilsson ct al., 1970; Agurell et al.,
1970]. This biotransformation has been
shown to be mediated by nonspecific hepatic
microsomal oxidases [Truitt, 1971]. Several
THC [Mechoulam, 1970; Foltz et al., 1970;
Christensen et al., 1971; Martin et al.. 1978],
If this hypothesis is valid, inhibition o f the
metabolism o f THC should attenuate its ef­
fects. However, if the parent compound is the
principal source o f the activity, an inhibitor
o f drug metabolism should enhance and pro­
long the effects o f THC. In a preliminary-
study Sofia and Barn> [1970] have shown
that prolongation o f barbital sleeping time in
mice by THC was enhanced in animals that
had been pretreated with SKF 525-A, a com­
pound known to nonspecifically inhibit liver
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investigators believe that the 11-hydroxy me­ microsomal enzyme systems responsible for
tabolite is responsible for the pharmacologi­ drug metabolism [Anders, 1971], It has been
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224
demonstrated both in vitro [Burstein and
Kupfer, 1971; Dingell et al., 1973] and in
vivo [Gill and Jones, 1972; Estevez et al.,
1974] that SKF 525-A does indeed inhibit
THC metabolism, which supports the earlier
findings o f Sofia and Barry [1970] that the
barbital sleeping potentiating activity of
THC resides primarily in the unmetabolized
molecule. Subsequent to this it was observed
by Adams and Sofia [1973] that chloram­
phenicol, another inhibitor o f liver micro­
somal enzymes responsible for drug metabo­
lism. also further prolonged barbital sleeping
time in rats treated with THC.
Methods
These experiments were conducted on male, al­
bino mice (Swiss-Webster) and rats (Sprague-Dawley)
weighing 20-28 and 140-190 g, respectively, at the
time they were tested, i.e.. at least 5 days after arriving
from the supplier (Hilltop Laboratories. Inc., Scott-
dale. Pa.. USA). Upon receipt from the supplier all
animals were randomly assigned and housed in groups
of 6 (rats) or 8 (mice) for use in the following experi­
ments.
THC was kept in a stock solution of 20 mg/ml of
undiluted propylene glycol at -4 °C. The vehicle for
THC in rats was undiluted propylene glycol while a
10% propylene glycol-1% Tween 80-saline solution
was used for mice [Sofia et al., 1971. and 1974]. Bar­
bital Na, morphine SOj and SKF 525-A were dis­
solved in saline. All compounds were prepared for
injection in such a way as to permit a volume of 10
and I ml/kg body weight to be administered to mice
and rats, respectively. Each drug and its vehicle were
administered intraperitoneally except for morphine
SO4, which was given subcutaneously.
Sleeping Time Studies
Sleeping time following barbital Na administra­
tion was measured by the duration in minutes when
the righting reflex was completely absent during a 5-
second observation period while the animal lay on its
back. Whenever one of the animals was tested for res­
toration of the righting reflex, a similar test was made
Sofia/ Barry
on all the other sleeping mice to equalize the source of
stimulation. In all cases the onset (in minutes) to loss
of righting (induction time) was also measured. Mean
induction and sleeping times were calculated and sta­
tistically evaluated using the Student’s t test for paired
comparisons.
Hot Plate Method
The hot plate method of Eddy and Leimbach
[1953] based on the reaction time in seconds measured
from the onset of the painful stimulus to the response
of lickingof the forelimbs and jumping was used. Mice
were placed individually in a 600-ml Pyrex beaker on
a hot plate (U.L., Type 1900, Model HP-A1915B,
Thermodyne Corp.. Dubuque. Iowa) maintained at a
temperature of 50-60 °C. A control reaction time to
the painful stimulus was obtained for all mice 24 h
prior to the test for drug effects. The drug or vehicle
treatment was given only to those animals with a con­
trol reaction time of less than 10 s. On the next day, all
drugs were given 30 min prior to the second exposure
to the hot plate. A maximum reaction time of 30 s was
scored when a mouse made no response within that
time span after exposure. Mean drug reaction times
were compared with pre-drug control reaction times
and statistically analyzed by the Student's t test for
paired comparisons.
Tail Flick Method
The tail flick method used for both mice and rats is
essentially the procedure described by D'Amour and
Smith [1941] except that the heat source was electrical
rather than a high intensity of light. The apparatus
consisted of a grooved asbestos board which was con­
structed in such a way that only the tail of the animal
was exposed to a high resistence Nichrome wire. The
intensity of the electrical current passing through the
heating element was controlled by a rheostat (Type
500B, Staco Inc.) set at the arbitrary value of nine
which delivered 10 V to the system. A single manual
switch controlled the entire apparatus including an
electric timer. The animal’s tail was placed in the
groove, the switch turned on to activate both the heat
source and timer, and the test terminated when a sud­
den twitch of the animal's tail indicated the response
to the painful stimulus. Control reaction times were
measured 24 h prior to administration of the drugs or
their vehicles. Animals with a control reaction time
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greater than 7.0 s in this preliminary test were not
used.
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Interaction of SKF 525-A and THC 225

Results and II were used. In schedule III, THC was

Sleeping Time Studies
In the initial experiment, three dosing
schedules (I, II or III) were used to determine
the optimal time interval between dosing and
test for subsequent studies. A 25-mg/kg dose
o f SKF 525-A was given either 30 min before
(I), 15 min after (II) or 60 min after (III) a
300-mg/kg dose o f barbital Na. A 20-mg/kg
dose o f THC was administered to these same
groups 25 min after the SKF 525-A injection
(5 min before, 40 or 85 min after the barbitu­
rate, respectively). Changes in the induction
time to barbital Na-induced anesthesia fol­
lowing these different dosing schedules are
shown in table 1. In each dosing schedule,
comparison o f drug treatment group I (con­
trol) with group 2 shows that pretreatment
with SKF 525-A only did not significantly
alter the induction time to barbital Na-in­
duced loss o f righting. However, pretreat­
ment with THC alone (group 3) produced a
significant reduction in the onset to barbital
anesthesia but only when dosing schedules I
given after the animals had already lost their
righting reflex. Group 4 shows that SKF 525-
A pretreatment did not further lower, to a
significant degree, the reduction in induction
time produced by THC in dosing schedules II
and III.
Table II shows the effects o f the three dos­
ing schedules on barbital Na-induced sleep­
ing time. Comparison o f the control group ( 1)
with each o f the other three groups revealed
highly significant increases in the duration of
sleeping for each dosing schedule. A combi­
nation of SKF 525-A and THC in all dosing
regimens resulted in a larger prolongation of
sleeping time than either drug when given
alone. The most profound effect o f THC
alone was observed when it w'as given 40 min
after administration o f barbital Na (schedule
II). Moreover, this activity w'as significantly
greater than that noted for either dosing
schedules I and III. In addition, the combina­
tion o f SKF 525-A and THC prolonged
sleeping time equally in dosing schedules I
and II, whereas III produced a lesser effect.

Table I. The effect of various dosing time schedules on induction of barbital Na (300 mg/kg) sleeping time in
mice administered THC (20 mg/kg) and SKF 525-A (25 mg/kg) alone or in combination

Drug treatment group Induction time at various dosing schedules, min

1 11 III
-5 1 40 85

1 Vehicle + vehicle + barbital Na 71.38 ± 7.81 61.38 ± 2.87 53.63 ± 1.83

2 SKF 525-A + vehicle + barbital Na 61.88 ± 0.93 57.63 ± 4.74 57.50 ± 1.84
3 Vehicle + THC + barbital Na 51.25 ± 0.84* 50.88 ± 0.89** 53.13 ± 2.27
4 SKF 525-A + THC + barbital Na 47.00 ± 1.55** 48.00 ± 1.54** 50.50 ± 1.30

Each value represents the mean ± the standard error for each group (n = 8). * p < 0.05: ** p < 0.001.
1 Interval from barbital to THC or its vehicle (in min). SKF 525-A or its vehicle was injected 25 min prior to
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THC or its vehicle.

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226 Sofia/Barry

Using only dosing schedule I and replacing
the barbital Na injection with an equal vol­
ume o f saline, THC (20 mg/kg) and SKF 525-
A (25 mg/kg) alone or in combination did not
cause any mice to sleep.
In the next study the effect o f various
combinations o f doses o f THC and SKF 525-
A on barbital sleeping time were tested in
mice and rats. Using dosing schedule II, all
animals were injected intraperitoneally with
barbital sodium (300 mg/kg in mice and 240
mg/kg in rats), followed 15 min later by SKF
525-A (12.5 or 25.0 mg/kg) or saline, and 40
min after barbital Na by THC (10 or 20
mg/kg) or its vehicle. The experiments were
replicated and the data pooled for each ex­
perimental condition (n = 16 for mice and n =
12 for rats).
Figure 1 shows the mean sleeping time
and standard error o f the mean for the 16
mice in each o f the treatment groups. The
first three bars show minimal effects o f SKF
525-A with a significant difference only be­
tween zero and 12.5 mg/kg (p < 0.05). THC
alone reliably prolonged barbital sleeping
time at both 10 mg/kg (p < 0.001) and 20
mg/kg (p < 0.001). In addition, a dose-
related response to THC is indicated by a sig­
nificantly longer sleeping time with 20 than
10 mg/kg (p < 0.01). Figure 1 also shows that
the effect o f THC was greatly enhanced by
pretreatment with SKF 525-A. With 10
mg/kg of THC plus 12.5 mg/kg o f SKF 525-
A sleeping time was significantly longer than
after 10 mg/kg o f THC alone (p < 0.001);
likewise, 20 mg/kg o f THC plus the lower
dose o f SKF 525-A prolonged sleeping time
over 20 mg/kg o f THC alone (p < 0.01). The
higher dose of SKF 525-A (25.0 mg/kg) also
enhanced the effect o f both doses o f THC,
but there was no significant difference be­
tween the two doses o f SKF 525-A in con­
junction with any o f the three dosage condi­
tions for THC (0, 10 and 20 mg/kg). Poten­
tiating effects o f THC and SKF 525-A when
given in combination, compared with the
effect of either drug alone, were shown by the
fact that the lower doses o f both compounds

Table II. The effect of various dosing lime schedules on the duration of barbital Na (300 mg/kg) sleeping time
in mice administered THC (20 mg/kg) and SKF 525-A (25 mg/kg) alone or in combination

Drug treatment group Sleeping time of various dosing schedules. min

I 11 iu
-5> 40 85

1 Vehicle + vehicle + barbital Na 42.50 ± 6.29 31.88 ± 1.26 38.63 ± 1.43

2 SKF 525-A + vehicle + barbital Na 68.25 ± 3.66* 46.75 ± 3.46* 52.13 ± 5.39*
3 Vehicle + THC + barbital Na 80.50 ± 10.13*** 134.00 ± 15.98*** 53.38 ± 3.12**
4 SKF 525-A + THC + barbital Na 244.75 ± 19.95*** 223.63 ± 10.46*** 140.88 ± 28.09***

Each value represents the mean + the standard error for each group (n = 8). * p < 0.05: ** p < 0.005:
*** p < 0.001.
1 Interval from barbital to THC or its vehicle (min). SKF 525-A or its vehicle was injected 25 min prior to
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THC or its vehicle.

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Interaction of SKF 525-A and THC 227

resulted in reliably longer mean sleeping

times than twice the dose (25.0 mg/kg) of
SKF 525-A alone (p < 0.001) or twice the
dose (20 mg/kg) o f THC alone (p < 0.05).
The mean interval from barbital administra­
tion to the loss of righting reflex ranged from
49 to 55 min among the nine treatment
groups with no statistically reliable difference
among them. Since this was almost the same
as the 40-min interval from barbital to THC
administration, table III shows that the aver­
age waking time was approximately 2-3 h
after administration o f THC alone and more
than 3 h following THC preceded by SKF
525-A.
Figure 2 graphically represents the mean
sleeping time and standard error o f the mean
for the 12 rats in each o f the treatment condi­
tions. The results are closely similar to those
obtained with mice. The principal difference
between the two species was that neither dose
Fig. 1. Duration of sleeping time (mean ± SE)
o f SKF 525-A when given alone significantly
after administration of barbital (300 mg/kg) in mice
altered barbital sleeping time in rats. Both treated with THC alone or in combination with SKF
doses o f THC when given alone significantly 525-A.

Table III. The effect of treatment with SKF 525-A (25 mg/kg) and THC (20 mg/kg). alone or in combination,
on sleeping time induced by various doses of barbital Na in mice

Drug treatment group Sleeping time with barbital Na. min

100 mg/kg 200 mg/kg 300 mg/kg

Vehicle + vehicle _ 34.14 ± 5.05 31.88 ± 1.26

SKF 525-A + vehicle - 52.13 ± 7.90 46.75 ± 3.46*
Vehicle + THC - 66.88 ± 10.83* 134.00 ± 15.98***
SKF 525-A + THC “ 116.88 ± 20.33** 223.68 ± 10.46***

Each value represents the mean ± the standard error for each group (n = 8). * p < 0.05 for difference from
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vehicle + vehicle treatment. ** p < 0.005 for difference from vehicle + vehicle treatment. *** p < 0.001 for
difference from vehicle + vehicle treatment.
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228
750 - SKF 525-A, 250 mg /kg
THC, mg/kg
Fig. 2. Duration of sleeping time (mean ± SE)
after administration of barbital (240 mg/kg) in rats
treated with THC alone or in combination with SKF
525-A.
enhanced barbital sleeping time (p < 0.001)
in a dose-related manner (10 mg/kg < 20
mg/kg; p < 0.05). The combination o f THC
with 12.5 mg/kg o f SKF 525-A resulted in a
significantly longer (p < 0.005) duration o f
barbital sleeping time than after the same
dose o f THC alone (p < 0.005 for 10 mg/kg:
p < 0.001 for 20 mg/kg). Potentiation was
evidenced by the fact that the effect on sleep­
ing time o f the lower doses of both drugs was
significantly greater than twice the dose (25
mg/kg) o f SKF 525-A alone (p < 0.001) or
twice the dose (20 mg/kg) o f THC alone (p <
0.05).
In the next experiment various doses o f
barbital were used to determine whether
lower doses of the barbiturate might also be
potentiated by THC when administered
alone or in combination with SKF 525-A. As
in the previous study dosing schedule II was
Sofia/Barry
followed. Mice were administered 100, 200
and 300 mg/kg o f barbital Na intraperitone-
ally, followed 15 min later by 25 mg/kg of
KF 525-A or saline, and 40 min after the bar­
biturate by 20 mg/kg o f THC or its vehicle.
Table III shows that 100 mg/kg of barbital
alone or in combination with THC and SKF
525-A failed to induce anesthesia in mice.
However, ‘preanesthetic excitation' was ob­
served after this dose o f barbital and was con­
spicuously enhanced in mice given the barbi­
turate in combination with THC alone or
THC plus SKF 525-A. Sleeping time for mice
given 200 or 300 mg/kg o f barbital only did
not result in a dose-related response. How­
ever, when pretreated with THC alone or
THC plus SKF 525-A, barbital Na-induced
anesthesia was markedly potentiated and in a
dose-related manner.
The duration o f effect for THC and the
combination o f THC with SKF 525-A on
barbital sleeping time were studied (table IV).
Groups o f 8 mice each were given 25 mg/kg
o f SKF 525-A or its vehicle followed 25 min
later by 20 mg/kg o f THC or its vehicle, alone
or in combination with SKF 525-A. Barbital
(300 mg/kg) was then administered at var­
ious time intervals following THC or its ve­
hicle from 5 min to 48 h and 5 min. Mean
control (vehicle + vehicle) sleeping time was
quite consistent except for a significantly
lower value at 4 h and 5 min preceding bar­
bital administration. A significant elevation
o f barbital sleeping time was seen after ad­
ministration o f SKF 525-A alone, compared
with the longer interval, only when given 30
min before the barbiturate (5 min time inter­
val for the THC shown in table IV). Mice
treated with THC slept significantly longer
than control animals through 4 h and 5 min
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prior to barbital injection with the peak effect
occurring at the 1-hour and 5-min interval.
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Interaction ofSK F S25-A and THC 229

Table IV. The effect ofSKF 525-A (25 mg/kg) and THC (20 mg'kg) alone or in combination on barbital Na
(300 mg/kg) sleeping time in mice at various time intervals following barbiturate administration

Time Sleeping time, min

interval1
vehicle + vehicle SKF 525-A + vehicle vehicle + THC SKF 525-A + THC

5 min 52.50 ± 6.29 68.25 ± 3.66* 80.50 ± 10.13** 244.75 ± 19.95**

1 h. 5 min 49.38 ± 2.95 50.13 ± 3.78 109.75 ± 6.84** 285.13 ± 14.18**
2 h, 5 min 45.00 ± 4.94 39.50 ± 4.73 76.63 ± 7.07** 204.75 ± 18.26**
4 h, 5 min 33.38 ± 1.75* 44.00 ± 4.52 65.88 ± 6.06* 172.50 ± 12.77**
24 h. 5 min 47.13 ± 2.32 49.25 ± 2.60 54.00 ± 2.83 72.38 ± 6.14**
48 h. 5 min 47.13 ± 2.20 40.00 ± 2.88 55.13 ± 1.53 56.75 ± 2.06

Each value represents the mean + the standard error for each group (n = 8). * p < 0.01 for difference from
48 h. 5 min. ** p < 0.001 for difference from 48 h. 5 min.
1 For THC administration before barbital Na.

Table V. Analgesic activity of THC and morphine S 0 4 in mice using the hot plate method

Test drug and dose Reaction time, s Drugxontrol

mg/kg response ratio
control1 drug treated

Vehicle 2.61 ± 0.28 3.33 + 0.22 1.28

THC
5.0 2.45 + 0.28 4.28 ± 0.33* 1.75
10.0 3.04 + 0.19 6.95 + 0.63* 2.29
20.0 2.78 ± 0.19 11.16 + 1.37* 4.02
40.0 2.38 ± 0.16 11.08 ± 1.54* 4.65

Saline 4.35 ± 0.31 4.88 ± 0.17 1.12

Morphine S 04 administered subcutaneously

1.25 4.20 + 0.19 5.43 ± 0.34 1.29
2.5 4.55 ± 0.26 7.70 + 0.48* 1.69
5.0 4.11 ± 0 . 1 5 9.47 ± 0.63* 2.30
10.0 3.97 ± 0.20 17.53 + 1.54* 4.42

Each value represents the mean + the standard error for each group (n = 8). * p < 0.001.
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1 Control reaction limes were obtained 24 h prior to drug treatment.

2 Administered subcutaneously.
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230
Fig. 3. Dose response curves for the analgesic ac­
tivity of THC and morphine SO.i in mice using the hot
plate test.
Fig. 4. The analgesic activity of THC alone or in
combination with SKF 525-A using the hot plate
method in mice.
Pretreatment with SKF 525-A plus THC
caused a significant prolongation of barbital
sleeping time when given 24 h and 5 min
before the barbiturate. Only 48 h and 5 min
after pretreatment was there no enhancement
o f the THC effect by SKF 525-A as indicated
by sleeping time returning to control values.
These data suggest a relatively long duration
o f effect for this combination of drugs. Simi­
larly, the peak effect also occurred after the
1-hour and 5-min pretreatment time inter­
val.
Hot Plate Studies
Table V shows that all doses of THC and
all but the lowest dose of morphine (1.25
mg/kg) displayed highly significant analgesic
activity. The vehicles for both drugs were
without significant analgesic effect. In addi­
Sofia/Barry
tion, a dose-related response for the analgesic
activity of both THC and morphine S 0 4 is
indicated by an elevation in reaction time
with increasing doses. However, for THC
similar elevations were obtained after both
the 20- and 40-mg/kg doses, indicating that a
ceiling effect had occurred.
The ratios between drug and predrug reac­
tion times, as shown in figure 3, indicate that
the analgesic potency of THC is approxi­
mately half that of morphine, since the dose
necessary to give a ratio of 2.30 is 5.0 mg/kg
for morphine S 0 4 and approximately 10.0
mg/kg for THC.
The effect o f SKF 525-A pretreatment on
THC-induced analgesia in the hot plate test is
shown in figure 4. On the drug day, 25 mg/kg
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of SKF 525-A or its vehicle (saline) was given
intraperitoneally 30 min prior to the admin-
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interaction of SKF 525-A and THC 231

Table VI. Duration of action for the analgesic effect produced by THC (20 mg/ kg) alone or in combination
with SKF 525-A (25 mg/kg)

Treatment and Reaction time, s Testtcontrol

time interval, h
control1 test2

Saline + THC
0.25 4.98 ± 0.33 8.75 ± 0.42* 1.76
0.5 5.01 ± 0.15 8.27 ± 0.32* 1.65
1 4.46 ± 0.26 7.81 ± 0.33* 1.75
2 4.81 ± 0.32 7.43 ± 0.48* 1.54
4 5.03 ± 0.19 6.07 ± 0.30** 1.21
8 5.16 ± 0.22 5.03 ± 0.21 0.98
24 4.90 ± 0.31 5.40 ± 0.24 1.10

SKF 525-A + THC

0.25 5.11 ± 0.30 9.05 ± 0.47* 1.77
0.5 4.86 ± 0.31 9.45 ± 0.65* 1.94
1 4.52 ± 0.30 9.52 ± 0.50* 2.11
2 5.02 ± 0.25 8.15 ± 0.59* 1.62
4 4.95 ± 0.33 7.68 ± 0.59* 1.55
8 4.90 ± 0.21 7.08 ± 0.30* 1.44
24 4.53 ± 0.27 5.26 ± 0.26*** 1.16

Each value represents the mean ± the standard error for each group (n = 8). * p < 0.001; ** p < 0.025;
*** p < 0.05.
1 Tested 24 h prior to treatment.
2 Tested 30 min after treatment.

istration o f various doses o f THC (5, 10 and
20 mg/kg). Drug reaction times were mea­
sured 30 min following the injection o f THC.
SKF 525-A alone did not elicit an analgesic
response. All doses o f THC in combination
with SKF 525-A produced greater analgesic
effects than the same doses o f THC adminis­
tered alone. The potency o f THC following
pretreatment with SKF 525-A was approxi­
mately three times greater than THC alone
shown by the same ratio o f 3.50 with 15.0
mg/kg o f THC alone and only 5.0 mg/kg of
seen because at the high dose o f 20 mg/kg the
combination with SKF 525-A increased the
ratio only slightly (from 4.80 to 5.30).
In the next experiment the duration o f the
analgesic effect o f THC in combination with
SKF 525-A or saline was studied. 24 h prior
to drug administration, control reaction
times were obtained for all animals. On the
test day, half the groups were pretreated in-
traperitoneally with 25 mg/kg ofSK F 525-A
and the remaining groups received saline, fol­
lowed 30 min later by THC (20 mg/kg) or its
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THC when combined with SKF 525-A. A vehicle. At time intervals o f 0.25, 0.5, 1,2, 4,
ceiling effect for THC plus SKF 525-A can be 8 and 24 h after THC administration, the
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232
Fig. 5. Dose response curves for the analgesic ac­
tivity of THC and morphine S 0 4 in mice using the tail
(lick method.
mice were subjected to the painful stimulus
o f the hot plate and drug reaction times
recorded. Table VI reveals that the onset o f
analgesic activity following administration of
THC alone was quite rapid, i.e.. a highly sig­
nificant response at 0.25 h which persisted
for up to 4 h. In mice pretreated with SKF
525-A. the onset was equally rapid but the
duration o f action for the analgesic effect o f
THC was statistically elevated even 24 h fol­
lowing its injection.
Tail Flick Studies
Figure 5 depicts the ratio o f drug to con­
trol reaction times as a function o f dose for
each drug in the mouse tail flick test. Accord­
ing to these data, THC is one third as potent
as morphine S 0 4 since a ratio o f 2.00 was
obtained with 7.5 mg/kg o f morphine S 0 4
Sofia/Barr\
Fig. 6. Dose response curves for the analgesic ac­
tivity of THC and morphine S 0 4 in rats using the tail
flick method.
and 21.5 for THC. Figure 6 indicates that in
rats THC has only one eighth the analgesic
potency o f morphine, with a drug to control
ratio of 2.00 being obtained with approxi­
mately 4.0 mg/kg o f morphine compared to
33.5 mg/kg o f THC. Neither saline nor the
vehicle for THC displayed any significant
analgesic effect in mice or rats.
The effect o f SKF 525-A pretreatment (25
mg/kg i.p.) 30 min before injection o f THC is
shown in figure 7. It can be observed that in
mice all doses o f THC in combination with
SKF 525-A produced a greater analgesic ef­
fect than the same doses o f THC alone. The
difference was statistically reliable (p < 0.05)
for all doses except the lowest (2.5 mg/kg). A
threefold increase in the potency o f THC
130.237.165.40 - 1/19/2019 5:13:34 AM
after pretreatment with SKF 525-A was indi­
cated by the drug to control response ratio of
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Interaction of SKF 525-A and THC
THC, mg/kg
Fig. 7. The analgesic activity of THC alone or in
combination with SKF 525-A in mice using the tail
flick method.
2.00 obtained with 7.0 mg/kg with THC
alone and 21.5 mg/kg with both drugs. A ceil­
ing effect, at a ratio between 2.50 and 3.00,
was indicated by the failure o f 20 mg/kg o f
THC after SKF 525-A pretreatment to in­
crease the ratio above this level found with
the lower dose o f 10 mg/kg after SKF 525-A
and with 40 mg/kg o f THC given alone. Fig­
ure 8 shows that in rats SKF 525-A also
increased the analgesic response o f THC, but
at all doses tested this effect was only margin­
ally reliably significant.
Discussion
In the present study THC markedly pro­
longed barbital Na-induced sleeping time in
mice and rats, which is in good agreement
233
THC. mg/kg
Fig. 8. The analgesic activity of THC alone or in
combination with SKF 525-A in rats using the tail
flick method.
with the data presented by Kubena and Barry
[1970]. Since barbital Na is virtually unme­
tabolized and excreted as the intact drug
[Burns ct al., 1957], a central, synergistic de­
pressant activity o f THC can be suggested.
However, this explanation for the potentiat­
ing effect o f marihuana extract and THC on
pentobarbital and hexobarbital sleeping
times [Bcycet al., 1963: Garriott et al., 1967]
cannot be made with complete certainty
since both barbiturates are metabolized by
the liver microsomal enzymes. Enhancement
o f their depressant action could be due to
inhibition by THC o f liver microsomal en­
zyme systems responsible for their degrada­
tion.
The analgesic action o f THC [Bicher and
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Mechoulam, 1968; Dewey et al., 1970; Bux-
baum, 1972; Sofia et al., 1973, 1975; Chesher
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234
et al., 1973; Kaymakcalan et al., 1974] was
confirmed in the present study using the hot
plate and tail flick methods on mice and rats.
THC, following intraperitoneal administra­
tion to mice, was approximately half as po­
tent an analgesic as morphine SO4 when the
latter was given subcutaneously and evalu­
ated in the hot plate test. This is precisely the
potency observed in the mouse hot plate test
as reported by Bicher and Mechoulam
[1968]. However, the analgesic potency o f
THC compared to morphine was only 1 to 3
in mice when measured by the tail flick
response. This same degree o f analgesic po­
tency for THC was reported by Dewey et al.
[1970] when comparing percent analgesic ef­
fect in mice using the above methods. In the
rat tail flick bioassay, THC was only one
eighth as potent as morphine. Moreover, a
definite species difference exists for the
analgesic response to THC. Using the tail
flick method, the latency to the pain response
was 50% higher than control in mice with an
intraperitoneal dose o f 11.0 mg/kg, while in
rats it took approximately 19.0 mg/kg for the
same drug effect. These data suggest that
mice are nearly twice as sensitive as rats to
the analgesic actions o f THC. In addition, the
hot plate method appears to be a more reli­
able procedure than the tail flick method for
determining the analgesic effect o f THC in
mice.
Several investigators [Mechoulam, 1970;
Truitt, 1971; Foltz et al., 1970; Christensen et
al.. 1971; Martin et al.. 1978] have suggested
that a metabolite o f THC, i.e., 11-hydroxy-
THC, is a more potent psychoactive agent
than the parent compound and is largely
responsible for the effects observed following
THC administration. Results o f the present
investigation clearly show that pretreatment
o f mice and rats with SKF 525-A, a nonspe­
Sofia/Barry
cific inhibitor o f liver microsomal enzymes
responsible for drug metabolism [Anders,
1971], enhanced and prolonged the analgesic
and barbital sleeping time potentiating ac­
tions o f THC. These data along with our ear­
lier observations [Sofia and Barry, 1970] sug­
gest that THC itself is primarily responsible
for the depressant and analgesic effects ob­
served after its administration. Moreover,
Sofia [1974] has clearly demonstrated that
the lethal effect o f THC is also markedly
enhanced by pretreatment with either SKF
525-A or chloramphenicol.
Cook et al. [1954] have reported that the
duration o f action for the inhibiting effect of
SKF 525-A on liver microsomal enzymes is
at least 15 h. The enhancement o f the action
o f THC on barbital sleeping time and analge­
sia following pretreatment with SKF 525-A
was greater than 24 h. Further evidence sup­
porting a potentiating rather than an additive
effect of SKF 525-A on THC behavioral
effects was provided by the fact that SKF
525-A alone had minimal to no effects on
barbital sleeping time or analgesia. Only
when the rat tail flick procedure was used, a
method which was less sensitive to the
analgesic actions o f THC, were the pain-
inhibiting actions o f the drug not signifi­
cantly potentiated by SKF 525-A. Based on
these data one cannot say that the 11 -hy­
droxy metabolite, or any other metabolite,
lacks pharmacological actions; however, it is
likely that THC is at least equally or even the
more potent o f the two compounds in poten­
tiating barbital sleeping time and producing
analgesia. This speculation is in good agree­
ment with findings o f Scheckel et al. [1968]
and Mclssac et al. [1971] who have demon­
strated more pronounced depressant effects
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when concentrations o f unmetabolized THC
in body tissues were the highest.
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Interaction of SKF S25-A and THC
Acknowledgements
THC was received from the National Institute of
Mental Health through the courtesy of Monique
Braude, PhD, Executive Secretary of the FDA-NIMH
Psychotomimetic Agents Advisory Committee. SKF
525-A was generously supplied by Smith, Kline &
French, Inc.. Philadelphia. Pa. This report was sup­
ported by Prcdoctoral Training grant GM-1217 from
the National Institute of General Medical Sciences
and Research grant M H-13595 to Herbert Barry, III,
from the National Institute of Mental Health.
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Received: December 18. 1982
Accepted: January 8, 1983
R.D. Sofia.Vice President.
Preclinical Research,
Wallace Laboratories.
Half Acre Road.
Cranbury. NJ 08512 (USA)
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