Enhancement of Behavioral Effect and Acute Toxicity of Methamphetamine by Qu nine in Rats

Japan. J. Pharmacol. 35, 273-277

Enhancement

(1984)

273

of Behavioral

Effect

Methamphetamine

and

Acute

by Quinine

Toxicity

of

in Rats

Tsutomu SUZUKI, Horng-Jen FAN CHIANG, Saizo YANAURA, Takemi YOSHIDA* and Yukio KUROIWA* Departmentof Pharmacology,School of Pharmacy,HoshiUniversity, 2-4-41 Ebara,Shinagawa-ku,Tokyo142, Japan *Departmentof BiochemicalToxicology , School of PharmaceuticalSciences, Showa University,1-5-8 Hatanodai,Shinagawa-ku,Tokyo142, Japan AcceptedMarch 30,1984

Abstract•\The tested

behavioral

alone

increased

and

the

induced

in

effect

of

stereotyped

simultaneously

effect

of

enhancement be

due

of to

was

very

inhibition

the

of

the

when

behavior

effect

In our previous report (1), we demonstrated that quinine could markedly increase methamphetamine-induced anorexia in rats. This finding was based on the results that the inhibition of food consumption and the growth curve was greater in the rats given methamphetamine in combination with quinine than those given methamphetamine alone (1). These results suggest that quinine enhances effects of methamphetamine. Amphetamine-induced behavioral excitation in rats, consisting of increased exploratory behavior, grooming, forward locomotion or various stereotyped behaviors, had been described in detail by Randrup et al. (2). The interactions between amphetamine-like stimulants (amphetamine, methamphetamine, etc.) and various drugs have been reported by many authors in terms of the behavioral changes (3-6). Therefore, the behavioral study is thought to be useful for the investigation of combined effects of amphetaminelike stimulants and other drugs. The main purpose of this study was to investigate the interactions between methamphetamine and quinine in rats from the viewpoints of behavioral pharmacology. In

of

quinine the

was

not

of

the

was

addition, and

Animals:

(CA-1, available, the

ad

a

tested

and

was

Tokyo) fasting for

cellulose injection

was

1

in

light Food

was

the

fasting

Each

was Co.

animal

in (CMC)

volume and

quinine

quinine

Wako

Pure

Methamphetamine saline,

1%

and

carboxymethyl-

aqueous of

Meobtained

and

physiological

suspended

ml/kg

water

from Ltd.

a

always prior to

schedules:

Seiyaku

sodium

The

weight.

Tap

purchased

dissolved was

in

hr

was 18 hr

throughout

Industries,

quinine

12 08:30).

at

hydrochloride

Chemical

cages temperature

administration

was

were

only.

Dainippon

sulfate

wire

periods.

once

thamphetamine from

in

Tokyo)

constant

dosing.

post-administration

rats

Co.,

eight

libitum

was

methamphe-

examined.

Methods

at

Japan, for

and

Drugs

of also

of

experimental

supplied

The

quinine.

preprogrammed (light on

Clea except

The

Sprague-Dawley

groups

with cycles

or

me-

may

Animals

maintained

(22•}1•Ž) and dark

quinine.

and

Male

in

quinine.

were

Laboratory

housed

by

interactions

quinine Materials

(Tokyo

to

of

by

by

toxic

tamine

prior

onset

methamphetamine

methamphetamine

room

given for

affected

of

and

methamphetamine-

markedly

toxicity

were

prolonged

time

potentiated

and

metabolism

Quinine

enhancement

marked

also

methamphetamine

rats.

However,

was

behavioral

of

in

The

stereotyped methamphetamine

toxicity

quinine

methamphetamine.

thamphetamine-induced mortality

acute

with

methamphetamine. behavior

with

and

combination

solution.

methamphetamine was

5

ml/kg

body

274

T. Suzuki

Classification weighing

of

study

were steel

30

min

before to

In

this

study,

groups

the

that

quinine, 50 methamphetamine,

5

rats

s.c.,

and

(4)

after

methamphetamine,

for

8

hr

induced

by to

previously by

Table

and

The

individual

rats

in 10

a

30

latency

of

maximum

of

head

and

sitting

in

phase

(50 also

was

after

(7);

noted

for

8

mg/kg, studied

each

and

hr

Table1.

the

me-

parameters stereotyped

duration,

The

latency

was

continuous

follows: and of

asleep,

The

In

system

the

rat

evaluated or

by entering

addition,

administration or 1% CMC rats in each

of the

termination

resting,

hyperactivity.

the

repetitive

with

was

the

3)

by

sniffing,

posture.

1)

onset

determined

movements

Scoring

the

averaged

Three

of solution group).

used

statistical significance was determined using Student's t-test for the latency of onset and the duration of stereotyped behavior and Mann-Whitney's U-test for the maximum intensity of stereotyped behavior. Acute toxicity: Two hundred rats weighing 202.7±1.4 g were housed individually and divided into twenty groups with ten rats in each group. Ten doses (from 10.5 to 113.4 mg/kg, s.c.) of methamphetamine alone or in combination with a fixed dose (50 mg/kg, p.o.) of quinine were tested. All administrations were carried out between 1 3:00 and 14:00 to avoid diurnal variations in drug metabolism (8, 9). Deaths were recorded for three days after the drug administration. The lethal dose (LD50) and 95% confidence limits were determined by the LitchfieldWilcoxon procedure (10) in animals administered methamphetamine alone or in combination with quinine.

in

of

after

the

p.o.) (six

(6)

it is shown for

2)

single

of

Scheel-Kffiger

scores

behavior

of

behavior

was

behavioral

modification

as

falling

rat

The

by

a crouched

animal

After (given

slight

were

limb

min

s.c.

each

considered

stereotyped

p.o.; 60

methamphetamine

behavior

occurrence

a

mg/kg,

mg/kg,

of

a

intensity.

stereotyped

before simul-

p.o.,

longer.

min

onset,

(2)

min (3)

50

mg/kg,

injection.

were

(1)

alone;

methamphetamine-induced

behavior

four

methamphetamine,

Naylor

group

to

thamphetamine

of

60 s.c.;

used

Costall 1.

every

the

p.o., mg/kg,

or

according

that

into

methamphetamine

scored

of

5

behavior

change

ten

s.c.,

5

the

observed

to

follows:

mg/kg,

50

of

13:00),

and

as

quinine,

quinine,

administration at

divided

drugs

of

and

in 25•~15

situation.

5 mg/kg,

the

administration

were

administration

mg/kg,

(21 •~

experimental

received

rats of

placed

cages

drug the

Forty

beginning were

wire

methamphetamine,

taneous

at the

They

and

adaptation

allow

behavior:

g

used.

individual cm)

the

161.4•}1.2

et al.

the quinine was The

for estimation

Results Behavior: The time courses of behavioral change induced by methamphetamine alone or methamphetamine combined with quinine are shown in Fig. 1. The latency to onset, the duration and the maximum intensity of methamphetamine-induced stereotyped be havior are shown in Table 2. Methamphetamine (5 mg/kg, s.c.) caused a moderate behavioral excitation such as sniffing and head movement in all rats and continuous licking, gnawing, or biting in a few of them. The behavioral change lasted for 3-4 hr. The effect of methamphetamine was enhanced slightly when quinine (50 mg/kg, p.o.) was given 60 minafter the administration. Moreover, quinine, given 60 min prior to or simultaneously with methamphetof the

intensity

of behavioral

excitation

Fig.1. s.c.)

Time singly

● ― tration

course and

of

10

the

methamphetamine 1%

CMC

of Methamphetamine

behavioral

excitation

with of

●,administration

amine: •\•\, or

of

combined

Interaction

quinine

quinine

and

60

(50 min

after mg/kg, after

quinine. •¡•\•¡,

solution

only;

and Quinine

administration p.o.)

in

of

275

methamphetamine

rats. •›•\•›,

methamphetamine; • •\• , administration

, quinine

only.

(5

only;

simultaneous of

Each

mg/kg,

Methamphetamine

quinine symbol

60

min

adminis-

before

represents

methamphetthe

mean

of

6

rats.

Table2.

Effect

of quinine

on methamphetamine-induced

stereotyped

behavior

in rats

Methamphetamine (MA) was administered s.c. and quinine (0) was p.o.MA only (1); MA and Q administered simultaneously (2); 0, 60 min before MA (3); 0, 60 min after MA (4). Each value represents the mean±S.E. of 10 rats. aP<0.05, bP<0.01: Significant difference fromgroup 1. cP<0.01: Significant difference from group 2. dP <0.01 : Significant difference from group 3. Numerals following the drugs indicate their doses (mg/kg).

amine administration, potentiated very markedly the effect of methamphetamine both in degree and duration. However, the onset of methamphetamine-induced streotyped behavior was not altered by quinine. CMC solution (5 ml/kg, p.o.) or quinine (50 mg/kg, p.o.) alone did not produce any behavioral excitation during 6 hr of observation (Fig. 1). Acute toxicity: Deaths usually occured 3 hr after the injections of methamphetamine and were characterized by hyperpnea and convulsions. However, all data are presented as the number of deaths recorded 72 hr after the injection. Figure 2 shows the mortality

in rats after administration of methamphetamine singly and combined with a fixed dose (50 mg/kg, p.o.) of quinine. Methamphetamine killed the rats in a dose-dependent manner. The mortality of methamphetamine was markedly enhanced by quinine. The LD50 for methamphetamine alone was estimated to be 79.7 (61.8-120.3) mg/kg, but it was decreased to 21.3 (14.5-28.0) mg/kg by quinine. Discussion The present results indicated that quinine enhanced the behavioral excitation (such as increased locomotor activity or development

276

T. Suzuki

Fig. 2. Acute toxicity of methamphetamine (MA) given alone or in combination with a fixed dose (50 mg/kg) of quinine (Q). MA and Q were administered simultaneously. Each column represents the mortality of 10 rats during the 72 hr observation period.

of stereotyped behavior) induced by methamphetamine. It has been known that quinine at high doses can affect central nervous system functions (11). However, like the solvent (1% CMC solution), 50 mg/ kg, p.o. of quinine, tested in this study, it did not produce any change in the behavior. In addition, the onset time of methamphetamine-induced stereotyped behavior was not affected by quinine. These results suggest that the central effect of quinine is scarcely involved in the potentiation and prolongation of the behavior-stimulating effect of methamphetamine. Furthermore, it was shown that the duration and intensity of methamphetamineinduced behavioral excitation were more pronounced in the rats given the drug prior to or simultaneously with methamphetamine than those given the drug after methamphetamine. These findings indicate the possibility that the enhancing effect of quinine on methamphetamine-induced behavioral excitation is due to inhibition of methamphetamine metabolism. It has been reported that the excretion rate of methamphetamine in humans was dependent on urinary pH level (12, 13).

et al.

Methamphetamine is excreted in higher amounts at an acidic urinary pH than at an alkaline pH (12, 13). However, our recent study indicated that the urinary pH in rats was not changed by the combination of methamphetamine with quinine (1). These results suggest that the enhancing effect of quinine on methamphetamine-induced behavioral excitation is not due to a change in the urinary pH level. The present study also showed that mortality of methamphetamine was markedly increased by quinine at a dose of 50 mg/kg, p.o. However, it had been reported that 50 mg/kg, p.o., of quinine did not kill any rats even after a chronic administration (50 mg/ kg/day for 14 days) (1). Moreover, it had also been observed in mice that the mortality of methamphetamine was not affected by quinine (T. Suzuki et al., unpublished data). Such a species difference may be explained by the fact that aromatic hydroxylation of amphetamine at the para position is the major pathway of amphetamine 'metabolism in rats (14-16), but it is relatively minor in mice (16, 17). In addition, quinine itself is also hydroxylated by the liver microsomal enzymes (1820). Therefore, quinine may compete with the p-hydroxylation of methamphetamine and thereby delaysthe metabolism of methamphetamine. However, further studies are required to elucidate such a metabolic interaction between quinine and methamphetamine in relation to potentiation of the pharmacological or toxicological effects of the latter drug. References 1 Suzuki, T., Fan Chiang, H.-J., Abe, Y., Ohtani, K. and Yanaura, S.: The effect of quinine on methamphetamine-induced anorexia in rats. Folia Pharmacol. Japon. 82, 149-157 (1983) (Abs. in English) 2 Randrup, A., Munkvad, I. and Udsen, P.: Adrenergic mechanisms and amphetamine induced abnormal behaviour. Acta Pharmacol. Toxicol. 20, 145-157 (1963) 3 Quinton, R.M. and Halliwell, G.: Effects of amethyl-dopa and dopa on the amphetamine excitatory response in reserpinized rats. Nature 200, 178-179 (1963) 4 Munkvad, I. and Randrup, A.: The persistance of amphetamine stereotypies of rats in spite of strong sedation. Acta Psychiat. Scand. 42,

Interaction

of Methamphetamine

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