Pharmacology Biochemistry & Behavior, Vol. 7, pp. 357--364. Copyright 1977 by ANKHO International Inc.

All rights of reproduction in any form reserved. Printed in the U.S.A.

Evaluation of a Suspension System

for Intravenous Self-Administration
Studies of Water-Insoluble Compounds
in the Rhesus Monkey
J O H N M. C A R N E Y /

I B R A H I M M. U W A Y D A H 3 A N D R O B E R T L. B A L S T E R

Department o f Pharmacology, Medical College o f Virginia, Virginia Commonwealth University

Richmond, VA 23298
(Recieved 8 July 1977)
CARNEY, J. M., I. M. UWAYDAH AND R. L. BALSTER. Evaluation of a suspension system for intravenous
self-administration studies of water-insoluble compounds in the rhesus monkey. PHARMAC. BIOCHEM. BEHAV. 7(4)
357-364, 1977. - A suspension system for water-insoluble drugs was evaluated using the mouse taft-flick test for analgesia
and schedule-controlled responding in rhesus monkeys. One group of monkeys was trained to respond for food
reinforcement under a chain fixed-interval 9 min fixed-ratio 10 schedule of reinforcement (chain FIFR). Another group of
monkeys was trained to respond for intravenous cocaine injections (100t~g/kg/inj) under an F R - 1 0 schedule.
Water-insoluble compounds were suspended in emulphor:ethanol:saline. The bioavailabftity of these drug suspensions was
demonstrated to be equivalent to water-soluble compounds by comparing the activity of the water-soluble and
water-insoluble salts of (+)-propoxyphene in the mouse taft-flick test. The behavioral effects of these drug suspensions were
also determined in the group of monkeys which was trained to respond for food under the chain F I - F R schedule.
Dose-related decreases in responding were produced by (+)-propoxyphene HC1 (water-soluble), (+)-propoxyphene
napsylate (water-insoluble), and a cannabinoid, (-+)-9-nor-9/3-OH-hexahydroeannabinol (/3-HHC) (water insoluble).
Doses of (+)-propoxyphene HC1 (in saline) and (+)-propoxyphene napsylate (in the emulphor suspension system) were
equi-effective in disrupting food-reinforced responding. In addition, these two salts of propoxyphene were equi-effective as
reinforcers of self-administration behavior in rhesus monkeys. /~-HHC was more potent than (+)-propoxyphene in
producing decreases in food-reinforced responding but failed to maintain self-administration behavior in the monkey.
Operant behavior
Self-administration
(+)-Propoxyphene
A 9 -THC
Chain schedule
Rhesus monkey

Suspension vehicle

Cannabinols

saline. S e l f - a d m i n i s t r a t i o n studies generally e m p l o y IV

delivery o f t h e test drug. Because o f t h i s r o u t e o f
a d m i n i s t r a t i o n it is i m p o r t a n t t o c o n s i d e r t h e possible
h e m a t o l o g i c a n d o t h e r effects of a n y vehicle s y s t e m for t h e
delivery of w a t e r - i n s o l u b l e drugs ( h e m o l y s i s , s e r u m sickness, etc.). A n u m b e r of vehicles have b e e n e m p l o y e d t o
deliver w a t e r - i n s o l u b l e c o m p o u n d s for single dose a n d
c h r o n i c i n t o x i c a t i o n studies [5, 13, 3 0 ] . U n f o r t u n a t e l y ,
m a n y o f t h e s e vehicles are n o t well suited f o r use in IV
s e l f - a d m i n i s t r a t i o n studies in m o n k e y s . P o l y v i n y l p y r r o l i d o n e (PVP) has b e e n used in c a n n a b i n o i d studies [13, 21,
2 6 ] . H o w e v e r , t h e 4 0 , 0 0 0 average m o l e c u l a r w e i g h t P V P
used in t h e s e studies has t h e u n d e s i r a b l e e f f e c t o f n o t b e i n g
m e t a b o l i z e d b y t h e a n i m a l , a n d studies have s h o w n t h a t

N A R C O T I C analgesics a n d a wide variety o f o t h e r p s y c h o active c o m p o u n d s have b e e n d e m o n s t r a t e d to f u n c t i o n as

r e i n f o r c i n g stimuli in l a b o r a t o r y a n i m a l s [ 2 9 ] . W i t h t h e
e x c e p t i o n of p r o c a i n e a n d s o m e p s y c h o t o m i m e t i c s (e.g.,
LSD a n d mescaline), t h e r e is g o o d c o r r e l a t i o n b e t w e e n t h e
ability of a drug t o r e i n f o r c e s e l f - a d m i n i s t r a t i o n b e h a v i o r in
l a b o r a t o r y a n i m a l s a n d its l i k e l i h o o d o f h u m a n abuse. F o r
this reason, it has b e e n p r o p o s e d t h a t l a b o r a t o r y investigat i o n s o f t h e a b u s e p o t e n t i a l o f n e w a n d p o t e n t i a l l y useful
p s y c h o a c t i v e drugs s h o u l d i n c l u d e a n e v a l u a t i o n of t h e i r
ability t o m a i n t a i n s e l f - a d m i n i s t r a t i o n b e h a v i o r in laborat o r y a n i m a l s [29 ].
O n e o b s t a c l e in evaluating drugs for s e l f - a d m i n i s t r a t i o n
is t h e degree to w h i c h t h e c o m p o u n d is soluble in w a t e r or

Supported in part by a contract from NAS-NRC Committee on Problems of Drug Dependence and NIDA grant DA-00490. Send reprint
requests to Robert L. Balster, Box 726, Medical College of Virginia, Richmond, VA 23298.
2NIDA postdoctoral fellow, supported by NIDA grant DA-05017. Present address: Department of Pharmacology, School of Medicine,
University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190.
3 NIDA postdoctoral fellow, supported by NIDA training grant DA-07027.
357

358

CARNEY, UWAYDAH AND BALSTER

PVP is deposited in the reticuloendothelial cells of the liver

and other organs of the body [27]. PVP is excreted over a
period of weeks or months and this long-term retention of
the polymer could result in toxic accumulations. The use of
albumin as a vehicle for cannabinoids and other waterinsoluble compounds is also unsafe in monkeys because of
the development of antibodies against the foreign albumin.
Rhesus monkey albumin could be used, but the procedure
would be complicated and awkward for routine use.
Propylene glycol has been used as a solubilizing agent for
sedative-hypnotics and minor tranquilizers. However, these
glycols are alcohols and may themselves function as
reinforcers at the concentrations necessary to keep the
drugs in solution since ethanol has been demonstrated to
function as a reinforcer of self-administration behavior in
monkeys [7,35 ].
The present study was designed to evaluate the use of a
lipoid suspension for the IV delivery of water-insoluble
compounds. The vehicle consists of a polyoxyethylated
vegetable oil (emulphor, E L - 6 2 0 ) , ethanol and saline, the
properties of which have been described by Craddock et al.
[8]. The emulphor vehicle system was selected because it
yields a very stable suspension and it appeared to have little
if any toxic effects when injected intravenously to monkeys
[28]. At the dilute concentrations of the vehicle which
would be used in self-administration studies the amount of
ethanol in the vehicle is too low to function as a reinforcer.

(1:1) and then rapidly diluted to the final concentration

with sterile saline. The weighed sample of the waterinsoluble compound was placed in a vial and a volume of
the emulphor:ethanol ( 1 : 1 ) j u s t sufficient to solubize the
material was added. For example, for ~ 9 - T H C this results
in a stock solution of t00 mg/cc. The vial was then placed
in an ultrasonic cleaner (Cole-Parmer, model 8 8 4 8 - 4 ) and
sonicated for 15 rain. Care was taken to maintain the
temperature of the sample at or below room temperature.
These stock solutions are stable and can be stored.
9 - T H C , for example, can be stored for periods of months
with less than 1% loss of activity. Prior to testing, saline was
rapidly added to an aliquot of the stock and shaken. This
diluted solution is a clear liquid which will remain as a
stable suspension for several weeks. In the case of
zx9 - T H C , concentrations in the range of 20 mg/ml and
lower can be easily obtained. Essentially, in our experience
the limitation of this vehicle system is the solubility of the
test compound in the 1:1 emulphor:ethanol and the
compound's potency.
Emulphor ( E L - 6 2 0 ) was the generous gift of the General
Analine and Film Corporation (GAF). ( + ) - a " ( 1 1 ) - T H C
and zx9 - T H C were obtained from Dr. R. E. Willette
(NIDA, Rockville, MD). ( + ) - P r o p o x y p h e n e HC1 and
(+)-propoxyphene napsylate were gifts of the Eli Lilly
Company (Indianapolis, Inc.). Cocaine HC1 was purchased
commercially.

G E N E R A L METHOD

EXPERIMENT 1: DEMONSTRATION OF THE

ANTINOCICEPTIVE ACTIVITY OF THE WATER-SOLU BLE
AND WATER--INSOLUBLE SALTS OF (+)--PROPOXYPHENE

Animals
The experimental animals were male albino mice (ICR,
2 0 - 3 0 g, Dublin Farms) and male rhesus monkeys (Macaca
mulatta). Monkeys were housed in individual primate cages
and had free access to water. Mice were housed 6 per cage
and had free access to food and water. All animals were
housed in temperature controlled rooms which had a 12 hr
light/dark cycle.

Synthesis
(3-HHC)

of

(+_)-9-nor-93-OH-HexahydrocannabinoI

ts-HHC was synthesized as described by Wilson and May

[ 33,34 ]. (+)-Ax 9 (11 )-Tetrahydrocannabinol was treated
with OsO 4 and NaI04 in T H F - H 20. The ketone formed in
this reaction was purified by silica gel column chromatography using ether:pet ether ( 1 : 2 0 - 3 : 1 ) . The ketone was
reduced with NaBH, and resulted in an essentially quantiative yield of the ( _ + ) - 9 - n o r - 9 - O H - H H C . The 9 3 - O H
and 9c,-OH isomers of the 9-nor-9-OH-HHC were separated by silica gel column chromatography with acetone:
pet ether (1 : 1) which resulted in a yield of the two isomers
in a ratio of about 12il (3:~). The purity of the ~ and ts
isomers was determined by TLC, GLC and IR analysis and
compared to the authentic samples obtained from Dr. E. L.
May (Department of Pharmacology, Medical College of
Virginia, Virginia Commonwealth University, Richmond,
VA 23298). Details of the synthesis, purification conditions, and spectral data for the HHC's are available upon
request.

Drugs
Water-soluble drugs were dissolved in 0.9% saline. Waterinsoluble drugs were first dissolved in emulphor:ethanol

Previous work has demonstrated that narcotic analgesics from a wide variety of different chemical classes have
antinociceptive effects when tested in rodents [12].
( + ) - P r o p o x y p h e n e HC1 has been reported to be about
10-times less potent than morphine in these antinociceptive
tests [12]. Because of its limited water solubility, the
napsylate salt of (+)-propoxyphene was selected as a
positive control to evaluate the applicability of the emulphor suspension system. This experiment represents the
first phase in the evaluation of the emulphor suspension
system for use in behavioral research. The bioavailability of
a water-insoluble narcotic was compared to its water-soluble
salt in the mouse tail-flick test for antinociceptive activity.

Method
The animals were male albino mice. They were housed six
per cage and kept in the central mouse facility until used.
All tail-flick tests were conducted on the same day between
10a.m. and 12 noon. Subcutaneous injections were administered 30 min before testing in a volume of 1.0 cc/kg.
The antinociceptive effects of the two propoxyphene salts
were assessed using a modification [11] of the tail-flick
method of D'Amour and Smith [9]. Three doses of each
salt as well as vehicle were tested. Antinociceptive potencies
were calculated as the percentage of the maximum possible
effect (% MPE) according to the method of Harris and
Pierson [20], with a 10 sec cut-off and a control latency of
2 - 4 sec. ED 50 values and confidence limits were determined by the method of Litchfield and Wilcoxon ['23].
( + ) - P r o p o x y p h e n e was delivered using the emulphor suspension system as described in the General Methods
section. A control group of mice was also tested with the

WATER-INSOLUBLE DRUG SELF-ADMINISTRATION IN MONKEYS

emulphor suspension system alone at the highest emulphor:ethanol concentration used.
R e s u l t s a n d Discussion

As shown in Table 1, both (+)-propoxyphene HC1 and

(+)-propoxyphene napsylate were active in the mouse
tail-flick test. ( + ) - P r o p o x y p h e n e base in the form of the
napsylate salt appeared to be slightly more potent than the
HC1 salt, however, this difference in potency was not
statistically significant. In addition to the similar potency
va]iues for the two salts, the slopes of the two dose-effect
curves were not significantly different.

TABLE 1
TAIL-FLICKACTIVITYOF (+)-PROPOXYPHENE SALTSIN MICE*
Drug
(+)-Propoxyphene HC1
(+)-Propoxyphene Napsylate

ED.~ot

Confidence
Limits

Slope (r)

7.63
4.92

4.5-12.8
3.26- 7.26

1.90 (0.99)
1.65 (0.99)

*Groups of 6 mice/dose with at least 8 doses tested for each salt

tmg/kg of the base
The comparable potency values in the mouse tail-flick
test for the two propoxyphene salts demonstrate that the
ernulphor suspension system is an effective vehicle for
testing a water-insoluble narcotic analgesic. The emulphor
suspension has been used in testing the behavioral and
biochemical effects of a number of water-insoluble cannabinoids [5,15]. Bloom et al. [5] reported that # - H H C was
equipotent to morphine in the mouse tail-flick test. Because
of its water-insolubility, ~ - H H C was tested using the same
emulphor suspension system as used in the present study.
In both the Bloom et al. study and the present study, the
emulphor vehicle was not active in the tail-flick test when
given alone.
EXPERIMENT 2: EFFECTS OF
(+)--PROPOXYPHENE SALTS AND/3--HHC ON
FOOD--REINFORCED RESPONDING IN MONKEYS
A wide variety of drugs affect schedule-controlled
re,;ponding in animals and humans when the drugs are given as
presession injections. Relatively low doses of narcotic
analgesics may increase the rate of responding and higher
doses produce decreases in responding under a number of
different schedules of reinforcement in laboratory animals
[25,32]. Dose-related increases and decreases in operant
re,;ponding also have been reported for cannabinoids
[ 15,24]. The present experiment was the second phase in
the evaluation of the usefulness of the emulphor suspension
system in behavioral research using rhesus monkeys. The
relative behavioral potencies of (+)-propoxyphene HC1,
(+)-propoxyphene napsylate, and of the 9t3-OH and 9 a OH isomers of HHC were determined using food-reinforced
responding under a chain fixed-interval fixed-ratio schedule
(chain F I - F R ) of food reinforcement.
Method

Four adult male rhesus monkeys which weighed between

359

5 and 8 kg were deprived to 80% of their ad lib feeding

weights and maintained at this weight by postsession
supplemental feeding in their home cages. The animals were
trained to respond under a chain FI 9 min, FR 10 schedule
of food reinforcement as previously described [ 1,2]. Under
this schedule the first lever-press response after 9 min (FI)
changed the stimulus light from orange to white and
advanced the schedule into the FR 10 component of the
chain. During the FR 10 component every tenth response
resulted in the delivery of a 1.0 g banana-flavored food
pellet (P. J. Noyes). After 1 min had elapsed in the FR
component, the white light was turned off, the orange light
was turned back on and the schedule was advanced into the
next chain F I - F R . The session ended after 8 chain F I FR's had occurred or after 2 hr had elapsed. IM injections
were given 5 min before the start of the session. Sessions
occurred daily and drug injections were given every fifth
day. Saline or emulphor vehicle injections were periodically
tested throughout the study. Baseline data was obtained
from the daily non-injection sessions which preceded
vehicle or drug injetions. Animals were tested once at each
dose in an unsystematic order of drug doses. The order of
drug testing in each monkey was # - H H C , ~ - H H C (+)-propoxyphene HC1 and (+)-propoxyphene napsylate. Data are
expressed as the percent of the non-injection baseline
response rate. Quarter-life values, a measure of the response
distributions within the FI, were calculated on the basis of
the average responding within successive 1 min segments of
the FI 9 min and obtained according to the method of
Gollub [18]. The quarter-life statistic represents the average amount of time, expressed as a proportion of the total
FI time, required for the emission of one-fourth of the total
FI responses.
R e s u l t s a n d Discussion

Responding under the chain F I F R schedule of food

reinforcement was relatively stable from day to day for
each animal. FI 9 min responding in the initial link of the
chain was characterized by a positively accelerated pattern
of responding (baseline quarter-life = 0.59 0.12 SD).
FR 10 responding in the terminal link of the chain occurred
at a relatively high and stable rate. The average rates of
responding in the FI and FR components were 0.57 ( 0.05
SD) and 1.92 ( 0.15 SD) responses per second, respectively.
The hydrochloride and napsylate salts of (+)-propoxyphene were equipotent in producing dose- related decreases
in responding (Fig. 1). Both salts of (+)-propoxyphene
appeared to have a similar rate of onset following IM
injection. Thus, it appears that the emulphor suspension of
(+)opropoxyphene napsylate had an absorption and distribution pattern to the CNS which was comparable to that of
the water-soluble hydrochloride salt.
13-HHC also produced decreases in FI and FR responding
at the higher test doses (Fig.2). One of the monkeys
showed substantial FI rate increases at the low O-HHC
doses. The other two monkeys showed only response rate
decreases across the o - H H C doses tested. At doses which
suppressed responding, ~-HHC had a prompt onset following injection. The 9 a - O H isomer of HHC was tested at
doses up to 10.0 mg/kg and had no behavioral effects.
Control injections of emulphor:ethanol:saline were not
different from saline and are presented together as the
saline vehicle control in Figs. 1 and 2.

360

CARNEY, UWAYDAH AND BALSTER

(+)-PROPOXYPHENE

,2~l"

9-NOR-9~-OH-HEXAHYDROCANNABINOL

FI

FR

FI
150

m 75

=_z
_J

LL
0
I--

125

~< qoo

z 5o

I-z

0
n~
UJ

z5

t~
to
[3_

25

5O
|

SALINE1.0

3.2

~1

I0.0 3;~.0

SALINE1.0

3.2

I0.0

32.0
25

mg/kg
FIG. 1. Effects of (+)-propoxyphene HCI (open symbols) and
(+)-propoxyphene napsylate (solid symbols) on food reinforced
responding under a chain FI FR schedule of reinforcement. The
three different symbols represent the data for three different
monkeys. Data points for the two propoxyphene salts represents a
single determination at each dose. Saline values are the mean (+_ SD)
of 8 observations in each monkey. Injections (IM) were given 5 min
before the start of the session.

C o m p a r e d to t h e effects of m o r p h i n e o n the c h a i n F I F R
r e s p o n d i n g (Balster a n d Harris, u n p u b l i s h e d d a t a ) ( + ) - p r o p o x y p h e n e is a b o u t 1/3 as p o t e n t in p r o d u c i n g d i s r u p t i o n
of s c h e d u l e - c o n t r o l l i n g r e s p o n d i n g . This relative p o t e n c y
value for ( + ) - p r o p o x y p h e n e is similar to t h a t observed for
c o d e i n e p r e t r e a t m e n t s , c o m p a r e d to m o r p h i n e , u n d e r a
multiple FR schedule of food and intravenous codeine
r e i n f o r c e m e n t in t h e rhesus m o n k e y [ 6 ] . T h e c o m p a r a b l e
b e h a v i o r a l p o t e n c i e s in t h e m o n k e y for ( + ) - p r o p o x y p h e n e
a n d c o d e i n e relative to m o r p h i n e agree well w i t h t h e
relative p o t e n c y e s t i m a t e s o b t a i n e d in r o d e n t a n t i n o c i c e p t i r e tests [ 1 2 ] .
C o m p a r e d to m o r p h i n e , t 3 - H H C was a b o u t 3 t i m e s m o r e
potent. The a-HHC
was inactive at doses up to
10.0 mg/kg. B l o o m e t a l . [5] a n d Wilson e t a l . [34] have
r e p o r t e d t h a t t h e ~-isomer of HHC is a p o t e n t a n t i n o c i c e p tive agent, while t h e a - i s o m e r is c o n s i d e r a b l y less p o t e n t or
inactive in t h e tail-flick t e s t for a n t i n o c i c e p t i v e activity.
z x 9 - T H C is also c o n s i d e r a b l y less active in this p r o c e d u r e

[51.
The data o b t a i n e d for t h e IM i n j e c t i o n o f t h e wateri n s o l u b l e c o m p o u n d s in m o n k e y s r e s p o n d i n g u n d e r a c h a i n
FI F R s c h e d u l e of f o o d r e i n f o r c e m e n t d e m o n s t r a t e t h a t t h e
e m u l p h o r s u s p e n s i o n s y s t e m was an effective vehicle for
evaluating t h e b e h a v i o r a l effects of w a t e r - i n s o l u b l e analgesics. ( + ) - P r o p o x y p h e n e n a p s y l a t e is p o o r l y w a t e r soluble,
less t h a n 1.5 m g / c c [ 1 9 ] , a n d t h e a- a n d #-isomers of HHC
are o f c o m p a r a b l e w a t e r i n s o l u b i l i t y to ~ 9 _ T H C ,
0.77 m g / l i t e r [ 1 6 ] . It w o u l d have b e e n impossible to test
these c o m p o u n d s w i t h o u t t h e use of a s u s p e n s i o n system.
In a d d i t i o n , u n d e r t h e s e test c o n d i t i o n s the e m u l p h o r
vehicle was w i t h o u t b e h a v i o r a l effects a n d did n o t a p p e a r
to affect the relative p o t e n c y of ( + ) - p r o p o x y p h e n e w h e n
t h e water-soluble a n d w a t e r - i n s o l u b l e salts were c o m p a r e d .
Thus, it appears t h a t t h e e m u l p h o r vehicle w o u l d be an

h
SALINE 0.03

0 1

0,32

1.0

..I__L
SALINE 0.03

0.1

0.32

I0

mg/kg

FIG. 2. Effects of (_+)-9-nor-9/3-OH-hexahydrocannabinol (t3-HHC)

on food-reinforced responding under a chain FI FR schedule of
reinforcement. Different symbols represent the data for three
different monkeys. The corresponding symbols for saline represent
the effects of saline plus the highest concentration of emulphor:
ethanol (1:1) used to suspend I3-HHC in the present study. Each
saline point is the mean (_+ SD) of 4 observations in each monkey.
Injections (IM) were given 5 min before the start of the session.

ideal vehicle for s t u d y i n g the s e l f - a d m i n i s t r a t i o n of these

a n d o t h e r water-insoluble c o m p o u n d s .
EXPERIMENT3:
WATER-SOLUBLE

SELF--ADMINISTRATION
AND

WATER--INSOLUBLE

RHESUS

OF
DRUGS

IN

MONKEYS

This e x p e r i m e n t was c o n d u c t e d to d e t e r m i n e t h e effectiveness o f the e m u l p h o r s u s p e n s i o n system in s t u d y i n g t h e

r e i n f o r c i n g p r o p e r t i e s of water-insoluble drugs. In o r d e r to
validate t h e use o f e m u l p h o r , the water-soluble a n d
water-insoluble salts of ( + ) - p r o p o x y p h e n e were used as
positive c o n t r o l s since the water-soluble h y d r o c h l o r i d e salt
has b e e n r e p o r t e d t o m a i n t a i n s e l f - a d m i n i s t r a t i o n b e h a v i o r
in the m o n k e y [ 2 2 , 3 1 ] . ~ 9 - T H C was i n c l u d e d to serve as
the negative c o n t r o l since p r e v i o u s studies have s h o w n t h a t
it does n o t f u n c t i o n as a r e i n f o r c e r of s e l f - a d m i n i s t r a t i o n
b e h a v i o r in t h e m o n k e y [ 2 1 ] . t 3 - H H C was also t e s t e d for
s e l f - a d m i n i s t r a t i o n because it is a p o t e n t a n t i n o c i c e p t i v e
a g e n t in t h e r o d e n t a n d has p o t e n t b e h a v i o r a l effects in t h e
monkey.
Method
T h e subjects were 5 a d u l t male rhesus m o n k e y s w h i c h
weighed b e t w e e n 5 and 8 kg. T h e animals h a d free access to
w a t e r a n d f o o d t h r o u g h o u t t h e study. T h e y were h o u s e d in
individual s e l f - a d m i n i s t r a t i o n cubicles w h i c h have b e e n
previously described [ 1 4 ] . U n d e r p h e n c y c l i d i n e - p e n t o b a r b i t a l anesthesia t h e a n i m a l s were surgically p r e p a r e d
with c h r o n i c indwelling IV c a t h e t e r s [ 1 0 ] . A f t e r surgery
t h e animals were r e s t r a i n e d b y a m e t a l h a r n e s s a t t a c h e d to
t h e rear wall of t h e cubicle b y a flexible m e t a l arm. T h e

W A T E R - I N S O L U B L E DRUG S E L F - A D M I N I S T R A T I O N IN MONKEYS
infusion catheter passed through the metal arm and was
connected to the implanted catheter at the back of the
monkey. IV drug injections (1.0 cc in 10 sec) were delivered by a peristaltic pump (Cole-Parmer). Solid-state
programming and control equipment, digital counters and
cumulative recorders (Ralph Gerbrand, Inc.) were located
in the adjoining room.
Daily self-administration studies were conducted in the
arfimals' home cages for 2 hr at the same time each day.
Monkeys were trained to respond under an FR 10 schedule
of IV cocaine reinforcement as previously described [3, 4,
14]. Under this schedule, every tenth response resulted in
the delivery of an intravenous injection of cocaine
(1 O0 ug/kg/inj). The availability of cocaine injections under
the FR 10 schedule was indicated by the illumination of
two orange lights immediately above the response lever.
Daring cocaine injections the orange lights were extinguished and a white light was illuminated over the lever.
At the end of the 2 hr session all of the lights over the lever
were extinguished. A substitution procedure [3, 4, 14] was
used to test for positive reinforcing properties of the two
salts of (+)-propoxyphene, ~ 9 - T H C and t3-HHC. Doses of
test compounds or vehicle were substituted for cocaine for
six consecutive daily sessions. Which of the 5 animals were
tested with each compound can be determined from the
subject identification numbers included in Figs. 3 - 5 . For
(+)-propoxyphene, equimolar doses of each salt were tested
in sequence before the next pair of doses. For the
cannabinoids, # - H H C was tested before zx~ - T H C . Blocks
of 6 cocaine self-administration sessions alternated with 6
sessions in which test compounds were substituted for
cocaine. The last 3 of the 6 days of drug self-administration
were compared to the last 3 of the 6 days of saline or
emulphor vehicle substitution. A drug was identified as a
positive reinforcer if response rates on the last 3 days of
drug self-administration were above and did not overlap
with rates on the last 3 days of vehicle self-administration.

Results and Discussion

Cocaine-reinforced responding under the FR 10 schedule
of reinforcement was fairly stable from day to day. The
range of cocaine injections per 2 hr session was between 25
and 55 for the group of 5 monkeys. Monkeys self-administered the greatest amount of cocaine in the first 30 rain
portion of the 2 hr sesssions and then progressively reduced
their intake of cocaine in the remaining 90 rain of the
session. This within-session pattern of responding was
similar to that previously reported for cocaine-reinforced
re.sponding at a unit dose of 100 #g/kg/inj [ 14].
Substitution of (+)-propoxyphene HC1 or (+)-propoxyphene napsylate for cocaine reliably maintained selfadministration responding in all three of the monkeys
tested (Fig. 3). Both the high and low doses of either
propoxyphene salt maintained responding above vehicle
control values. The low dose of both propoxyphene salts
resulted in substantially greater self-administration rates
then observed with cocaine at the 100 ~g/kg/inj dose. The
high dose of both propoxyphene salts also maintained
self-administration responding above saline rates and at
about the same level as the baseline cocaine self-administration rate. The two salts appeared to be equipotent as
reinforcers when the doses were calculated as gg/kg/inj of
base. At the high (+)-propoxyphene napsylate dose
(750 ug/kg/inj) the injection solution concentration ranged

361

(+)- PROPOXYPHENE
HYDROCHLORtDE
5 0 qg/kg/inj

~5o1

NAPSYLATE HYDROCHLORIDE NAPSYLATE

7 5 uq/kg/inj

(o.~6 mg/c)

500 qq/kg/inj

to.s~ mg,,~:)

7 5 0 u~/kg/inj

{~.1 ml,'e)

(S* mo,'~=)

14o1
130:
120

II0
I00
90
80
70
60
50
40
30

20
IO
B4107

120
(o~o ~g/~)

I10

(0.4W m,/cc)

(s.o ~Qtcc)

(4S mgtc)

I00

90
80
70
60
50
.

40
50
20
I0
~4116

IlO
(o.soz m~/1

I00

(3 ~em~/c)

(s,oz ~elc)

go
80
70
60
50
40
30

20
I0
I

45

~ 2

45

SUCCESSIVE

45

~ 2

345

SESSIONS

FIG. 3. Mean number of injections per session for 6 successive daily

sessions at two dose levels of (+)-propoxyphene HC1 and (+)-propoxyphene napsylate are represented by solid symbols. The horizontal dashed lines represent the range of baseline cocaine
(100 ~g/kg/inj) injection rates obtained from 30 cocaine injection
sessions for each monkey. Injection rates after substitution of the
injection vehicle alone are indicated by the open symbols. The
vehicle for (+)-propoxyphene HC1 was 0.9% saline and the vehicle
for (+)-propoxyphene napsylate was the emulphor suspension
system. The (+)-propoxyphene doses refer to the salt. The high and
low doses of (+)-propoxyphene represent equimolar doses of
(+)-propoxyphene base at the two dose levels. The concentrations of
the injected solutions are indicated in parenthesis for each monkey.

between 4.5 and 5.4 mg/cc. This exceeds the water-solubility of the compound (1.5mg/cc; ref. [19]). Thus,
without the use of the emulphor suspension system it
would not have been possible to test this high dose of
(+)-propoxyphene.
Neither ~ ~ - T H C nor ~ - H H C functioned as reinforcers
when substituted for cocaine (Figs. 4 and 5). In general, the
data for the two cannabinoids closely paralleled the vehicle
substitution (extinction) data across the 6 substitution
days. One exception was monkey B002 at the 3.0 and
30.0 ~g/kg/inj doses of t3-HHC. At the 3.0 ~g/kg/inj dose,
responding appeared to alternate across the days between
relatively high and relatively low self-administration rates
compared to vehicle rates, with a general trend downward

362

C A R N E Y , U W A Y D A H A N D BALSTElq
( - ) - A9 - TETRAMYDROCANNABINOL
3.0 qg/kg/inj

30 qg/kg/inj

9-NOR-9B-OH-HEXAHYDROCANNABINOL
300 qg/kg/inj
B002

(0.022 mg/cc)

(0.22 mg/c)

O. 3 0 q g / k g / i n j

3.0 qg/kg/inj

(0 o02m~llecJ

(0 022mglcc)

50 qg/kg/inj

60

(220 rag/co)

BOO2

50

(0 22mglc)

50
40
40
30

30
2O
20

,o
[0
t~

i
,

z
0
F-- 6 0
--

B4114
(o.o2o rag/co)

(o,2o m,;I/CC)

(2.oo rag/co)

6O

50

B4163

5o

40

(0 O02mglcc)

(0

020mglcc)

(0 O02mg/cc)

(o 020rag/co)

(0

20mg/cc)

4o
&

30

3o
o
I--

20

,Y, 2 o
z

I0

I0

4 5

SUCCESSIVE

3 4 5

SESSIONS

FIG. 4. Number of ~9-tetrahydrocannabinol (zX9 -THC) injections

(solid symbols) per session at three dose levels across the 6
successive days of substitution. Open symbols represent the number
of emulphor vehicle injections. Horizontal dashed lines represent the
control range of cocaine injections (100 ~g/kg/inj) for each monkey.
The concentration of Ag-THC at each of the dose levels are
indicated in parenthesis for each monkey.

B4H4
60

(0 20rag/co}

50

40

30

20

I0

t o t h e vehicle i n j e c t i o n s rates at the e n d of t h e 6 days. A t

t h e 30.0 ~g/kg/inj dose r e s p o n d i n g a p p e a r e d t o be m o r e
stable across days, w i t h t h e last 2 days slightly a b o v e
vehicle c o n t r o l values. However, even for m o n k e y B 0 0 2 ,
s e l f - a d m i n i s t r a t i o n b e h a v i o r did n o t satisfy t h e criterion o f
n o overlap of t h e drug i n j e c t i o n rate w i t h t h e vehicle
c o n t r o l values. B o t h of these c a n n a b i n o i d s are essentially
water-insoluble a n d c o u l d n o t have b e e n t e s t e d if a vehicle
was n o t e m p l o y e d . T h e e m u l p h o r s u s p e n s i o n vehicle did
n o t result in s e l f - a d m i n i s t r a t i o n rates t h a t were d i f f e r e n t
f r o m saline s u b s t i t u t i o n values. T h u s , u n d e r t h e p r e s e n t
e x p e r i m e n t a l c o n d i t i o n s n e i t h e r A g _ T H C n o r t3-HHC
f u n c t i o n e d as a positive r e i n f o r c e r of s e l f - a d m i n i s t r a t i o n
r e s p o n d i n g in m o n k e y s . In a d d i t i o n , t h e use of t h e
e m u l p h o r s u s p e n s i o n s y s t e m did n o t i n t e r f e r e w i t h t h e
d e t e r m i n a t i o n o f t h e r e i n f o r c i n g p r o p e r t i e s o f drugs w h i c h
f u n c t i o n as positive reinforcers.
DISCUSSION
T h e p r e s e n t s t u d y d e m o n s t r a t e s t h a t t h e e m u l p h o r susp e n s i o n s y s t e m is an effective vehicle for s t u d y i n g t h e
s e l f - a d m i n i s t r a t i o n a n d o t h e r b e h a v i o r a l effects o f waterinsoluble compounds. Under the substitution paradigm

SUCCESSIVE

SESSIONS

FIG. 5. Number of (+)-9-nor-9~-OH-hexahydrocannabinol (j3-HHC)

injections (solid symbols) per session at three dose levels across the
6 successive days of substitution. Open symbols represent the
number of emulphor vehicle injections. Horizontal dashed lines
represent the control range of cocaine injections (100 #g/kg/inj) for
each monkey. The concentrations of ~3-HHC at each of the dose
levels are indicated in parenthesis for each of the monkeys.
b o t h t h e HC1 a n d n a p s y l a t e salts of ( + ) - p r o p o x y p h e n e
m a i n t a i n e d s e l f - a d m i n i s t r a t i o n b e h a v i o r a b o v e vehicle control. Moreover, t h e t w o salts a p p e a r e d to be equi-active in
t e r m s of t h e a m o u n t of b e h a v i o r m a i n t a i n e d at the doses
tested. T h e data for ( + ) - p r o p o x y p h e n e HC1 are similar to
t h a t r e p o r t e d in a s t u d y of H o f f m e i s t e r a n d S c h l i c h t i n g
[2"2]. In t h e i r s t u d y t h e m a x i m u m n u m b e r of ( + ) - p r o p o x y p h e n e injections i n c u r r e d at 50ug/kg/inj; higher doses
resulted in c o r r e s p o n d i n g l y less s e l f - a d m i n i s t r a t i o n behavior. A similar r e l a t i o n b e t w e e n doses a n d r e s p o n s e rate

WATER-INSOLUBLE

DRUG SELF-ADMINISTRATION

was o b s e r v e d in t h e p r e s e n t s t u d y f o r b o t h p r o p o x y p h e n e
salts a n d has b e e n d e m o n s t r a t e d f o r a wide v a r i e t y o f
s e l f - a d m i n i s t e r e d drugs at doses a b o v e t h o s e w h i c h m a i n t a i n
m a x i m u m rates [3, 4, 7, 14, 17, 22, 3 5 ] . T h e decline in
r e s p o n d i n g o b s e r v e d at t h e h i g h p r o p o x y p h e n e dose m a y
have b e e n d u e t o t h e r e s p o n s e - r a t e decreasing effects of
( + ) - p r o p o x y p h e n e , since pre-session i n j e c t i o n s o f p r o p o x y p h e n e did p r o d u c e dose-related decreases in f o o d - r e i n f o r c e d
r e s p o n d i n g . As s h o w n in Fig. 1, decreases in food-reinf o r c e d r e s p o n d i n g o c c u r r e d at or a b o v e doses o f 1.0 m g / k g
(HC1) or 1.5 m g / k g ( n a p s y l a t e ) . S u c h a p r o p o x y p h e n e dose
w o u l d b e a c h i e v e d a f t e r t h e s e c o n d i n j e c t i o n at t h e h i g h
p r o p o x y p h e n e dose in t h e s e l f - a d m i n i s t r a t i o n e x p e r i m e n t .
W h e t h e r t h e decrease in t h e n u m b e r o f p r o p o x y p h e n e
i n j e c t i o n s / s e s s i o n at t h e h i g h dose was t h e result o f a
generalized d i s r u p t i o n o f o p e r a n t b e h a v i o r o r if it was t h e
result of a r e i n f o r c e r - s p e c i f i c satiation-like effect [6] o f
propoxyphene self-administration cannot be determined
f r o m t h e p r e s e n t l y available data.
N e i t h e r zx9 - T H C n o r ~ - H H C f u n c t i o n e d as r e i n f o r c e r s
w h e n s u b s t i t u t e d for c o c a i n e at t h e doses tested. Similar
negative data f o r zx9 - T H C s e l f - a d m i n i s t r a t i o n was r e p o r t e d
b y Harris etal. [ 2 1 ] . In t h e i r s t u d y , A 9 - T H C failed to
m a i n t a i n s e l f - a d m i n i s t r a t i o n r e s p o n d i n g w h e n it was substit u t e d for cocaine, w h e n naive m o n k e y s were allowed
12 h r access t o THC i n j e c t i o n s , or a f t e r m o n k e y s h a d b e e n
e x p o s e d for 3 0 days to p r o g r a m m e d THC infusions. Pickens
e t a l . [26] r e p o r t e d THC s e l f - a d m i n i s t r a t i o n in r h e s u s

IN M O N K E Y S

363

m o n k e y s t h a t h a d b e e n t r a i n e d to r e s p o n d u n d e r a
p h e n c y c l i d i n e s e l f - a d m i n i s t r a t i o n baseline. H o w e v e r , res p o n d i n g f o r THC was n o t c o n s i s t e n t l y m a i n t a i n e d across
sessions and, as suggested b y Harris etal. [ 2 1 ] , t h e
a p p a r e n t THC s e l f - a d m i n i s t r a t i o n m a y have b e e n due to
s o m e s t i m u l u s p r o p e r t y o f THC w h i c h was c o m m o n t o t h a t
of p h e n c y c l i d i n e a n d f u n c t i o n e d as a s e c o n d a r y r e i n f o r c e r ,
w h i c h w o u l d e x t i n g u i s h at a slower rate t h a n saline
s u b s t i t u t i o n . T h e failure of # - H H C to m a i n t a i n self-adminis t r a t i o n r e s p o n d i n g is o f i n t e r e s t since it is a p o t e n t
a n t i n o c i c e p t i v e c o m p o u n d in t h e r o d e n t [ 5 ] . # - H H C is
a b o u t 3 t i m e s m o r e p o t e n t t h a n m o r p h i n e in d i s r u p t i n g
o p e r a n t b e h a v i o r . T h u s , t h e failure o f #-HHC t o f u n c t i o n as
a r e i n f o r c e r was n o t due t o a relatively w e a k b e h a v i o r a l
p o t e n c y . It a p p e a r s t h a t c a n n a b i n o i d s like # - H H C m a y
r e p r e s e n t a class o f c e n t r a l l y active analgesics w h i c h have
p o t e n t a n t i n o c i c e p t i v e effects a n d p o t e n t effects in disrupting schedule-controlled responding, but which do not
f u n c t i o n as positive r e i n f o r c e r s in IV s e l f - a d m i n i s t r a t i o n
studies in m o n k e y s .

ACKNOWLEDGEMENTS
We would like to thank Dr. William L. Dewey for conducting the
antinociception studies. The technical assistance of George W.
King III, J. Marshall Newbern and Dr. Thomas G. Aigner in
conducting the behavioral studies is gratefully acknowledged.

REFERENCES
1. Balster, R. L. and L. D. Chair. The behavioral pharmacology of
phencyclidine. Clin. Toxic. 9: 5 1 3 - 5 2 8 , 1976.
2. Balster, R. L., L. D. Chair and G. W. King III. The effects of
acute and chronic phencyclidine on schedule-controlled behavior in the rhesus monkey. Fedn Proc. 3 5 : 5 6 4 (abstract),
1976.
3. Balster, R. L., C. E. Johanson, R. T. Harris and C. R. Schuster.
Phencyclidine self-administration in the rhesus monkey. Pharmac. Biochem. Behav. 1: 167-172, 1973.
4. Balster, R. L. and C. R. Schuster. A comparison of d-amphetamine, ~-amphetamine and methamphetamine self-administration in rhesus monkeys. Pharmac. Biochem. Behav. 1:
2 7 6 - 2 8 1 , 1973.
5. Bloom, A. S., W. L. Dewey, L. S. Harris and K. K. Brosius.
9-nor-9/3-Hydroxyhexahydrocannabinol, a cannabinoid with
potent and antinociceptive activity: Comparisons with morphine. J. Pharmac. exp. Ther. 200: 2 6 3 - 2 7 0 , 1977.
6. Carney, J. M. Selective modulation of codeine-reinforced
responding in rhesus monkeys. Doctoral Thesis submitted to
faculty of the University of Michigan, Ann Arbor, 1976.
7. Carney, J. M., M. E. Llewellyn and J. H. Woods. Variable
interval responding maintained by intravenous codeine and
ethanol injection in the rhesus monkey. Pharmac. Biochem.
Behav. 5: 5 7 7 - 5 8 2 , 1976.
8. Cradock, J. C., J. P. Davignon, C. L. Litterst and A. M.
Guarino. An intravenous formulation of A9-tetrahydrocan nabinol using a nonionic surfactant. J. Pharrn. Pharmac. 25:
345, 1973.
9. D'Amour, F. E. and D. L. Smith. A method for determining
loss of pain sensations. J. Pharrnac. exp. Ther. 72: 7 4 - 7 9 ,
1941.
10. Deneau, G., T. Yanagita and M. H. Seevers. Self-administration
of psychoactive substances by the rhesus monkey. A measure
of psychological dependence. Psychopharmacologia
16:
3 0 - 4 8 , 1969.

11. Dewey, W. L., L. S. Harris, J. F. Howes and J. A. Nuite. The

effects of various neurohumoral modulators on the activity of
morphine and the narcotic antagonists in the tail-flick and
phenylquinone tetst. J. Pharmac. exp. Ther. 175: 4 3 5 - 4 4 2 ,
1970.
12. Eddy, N. B., H. Friebel, K.-J. Hahn and H. Halbach. Codeine
and lts Alternatives for Pain and Cough Relief. WHO Publication, 1970.
13. Fenimore, D. C. and P. R. Lory. Injectible dispersion of
delta-9-tetrahydrocannabinol in saline using polyvinylpyrrolidone. J. Pharm. Pharmac. 23: 31 O, 1971.
14. Ford, R. D. and R. L. Balster. Reinforcing properties of
intravenous procaine in rhesus monkeys. Pharmac. Biochern.
Behav. 6: 2 8 9 - 2 9 6 , 1977.
15. Ford, R. D., R. L. Balster, W. L. Dewey and J. S. Beckner.
A9_THC and 11-OH - A g - T H C : Behavioral effects and relationship to plasma and brain levels. Life Sci. 20: 1993-2004,
1977.
16. Garret, E. R. and C. A. Hunt. Physiochemical properties,
solubility and protein binding of A 9-tetrahydrocannabinol. J.
Pharrnac. Sci. 63: 1056-1964, 1974.
17. Goldberg, S. R., Hoffmeister, U. U. Schlichting and W. Wuttke.
A comparison of pentobarbital and cocaine self-administration
in rhesus monkeys. J. Pharmac. exp. Ther. 197: 2 7 7 - 2 8 3 ,
1971.
18. Gollub, L. R. The relation among measures of performance on
fixed-interval schedules. J. exp. Anal. Behav. 7: 3 3 7 - 3 4 3 ,
1964.
19. Gruber, C. M., V. C. Stephens and P. M. Terrill. Propoxyphene
napsylate: Chemistry and experimental design. Toxic. appl.
Pharmac. 19: 4 2 3 - 4 2 6 , 1971.
20. Harris, L. S. and A. K. Pierson. Some narcotic antagonists in
the benzomorphan series..L Pharmac. exp. Ther. 143:
141-148, 1964.

364
21. Harris, R. T., W. Waters and D. McLendon. Evaluation of
reinforcing capability of delta-9-tetrahydrocannabinol in rhesus
monkeys. Psychopharmacologia 37: 23-29, 1974.
22. Hoffmeister, F. and U. U. Schlichting. Reinforcing properties
of some opiates and opioids in rhesus monkeys with histories
of cocaine and codeine self-administration. Psychopharmacologia 23: 5 5 - 7 4 , 1972.
23. Litchfield, J. T., Jr. and F. Wilcoxon. A simplified method of
evaluating dose effect experiments. J. Pharmac. exp. Ther. 96:
99-113, 1949.
24. McMillan, D. E. Behavioral pharmacology of the tetrahydrocannabinols. In: Advances in Behavioral Pharmacology, Vol. I,
edited by T. Thompson and C. B. Dews. New York: Academic
Press, 1977, pp. 1-34.
25. McMillan, D. E. and W. H. Morse. Some effects of morphine
and morphine antagonists on schedule-controlled behavior. J.
Pharmac. exp. Ther. 157: 175-184, 1957.
26. Pickens, R., T. Thompson and D. Muchow. Cannabis and
phencyclidine self-administration by animals. In: Bayer
Symposium IV: Psychic-Dependence, edited by L. Goldberg
and F. Hoffmeister. New York: Springer-Verlag, 1973, pp.
78-86.
27. Ravin, H. A., A. M. Seligman and J. Fine. Polyvinylpyrrolidone
as a plasma expander. Studies on its excretion distribution and
metabolism.New Engl. J. Med. 247: 921-929, 1952.
28. Schaeppi, U. and R. S. Phelan. Emulphor (EL-620) vehicle:
Blood pressure effects of IV infusion in dogs and rhesus
monkeys. U.S. Department of Commerce National Technical
Information Service, PB-220 052, 1972.

C A R N E Y , U W A Y D A H AND B A L S T E R
29. Schuster, C. R. and C. E. Johanson. The use of animal models
for the study of drug abuse. In: Research Advances in Alcohol
and Drug Problems, edited by R. S. Gibbons, Y. Israel, H.
Kalant, R. E. Popham, W. Schmidt and R. G. Smart. New
York: John Wiley and Sons, 1974, pp. 1-31.
30. Sofia, R. D., R. K. Kubena and H. Barry III. Comparison of
four vehicles for intraperitoneal administration of xl-tetrahydrocannabinol. J. Pharm. Pharmac. 2 3 : 8 8 9 - 8 9 1 , 1971.
31. Talley, W. H. and I. Rosenblum. Self-administration of
dextropropoxyphene by rhesus monkeys to the point of
toxicity. Psychopharmacologia 27: 179-182, 1972.
32. Thompson, T., J. Trombley, D. Luke and D. Lott. Effects of
morphine on behavior maintained by four simple food-reinforcement schedules. Psychopharmacologia 17: 182-192,
1970.
33. Wilson, R. S. and E. L. May. Analgesics properties of the
tetrahydrocannabinols, their metabolites and analogs. J. Med.
Chem. 18: 700-703, 1975.
34. Wilson, R. S., E. L. May, B. R. Martin and W. L. Dewey. The
9-nor-9-hydroxy-hexahydrocannabinols. Synthesis, some behavioral and analgesic properties and comparison with tetrahydrocannabinols. J. Med. Chem. 19:1165-1167, 1976.
35. Winger, G. D. and J. H. Woods. The reinforcing property of
ethanol in the rhesus monkey: I. Initiation, maintenance and
termination of intravenous ethanol-reinforced responding. Ann
N.Y. Acad. SoL U.S.A. 215: 162-175, 1973.