Brit. J. Pharmacol. (1960), 15, I II.

PHARMACOLOGICAL PROPERTIES OF THALIDOMIDE

(a-PHTHALIMIDO GLUTARIMIDE), A NEW SEDATIVE
HYPNOTIC DRUG
BY
G. F. SOMERS
From the Pharmacological Research Laboratories, The Distillers Company (Biochemicals) Ltd.,
Speke, Liverpool 24
(RECEIVED SEPTEMBER 21, 1959)

Thalidomide (a-phthalimidoglutarimide, " Distaval," " Contergan ") is a new sedative hypnotic
drug which produces no toxic effects when administered orally to animals in massive doses. This
lack of toxicity may be due to a limited absorption. The drug has a quietening effect on the central
nervous system, reducing the voluntary activity of laboratory animals and promoting sleep. Unlike
the barbiturate drugs it does not cause an initial excitation in mice, incoordination or narcosis.
It potentiates the actions of other central nervous system depressants, in particular the barbiturates.
Its sedative effects are counteracted by central nervous system stimulants. It has no deleterious side
effects and does not affect the heart, respiration or autonomic nervous system.
Thalidomide is a derivative of glutamic acid, its Acute Toxicity
chemical structure being shown in Fig. 1. The acute toxicity of thalidomide was determined
by the oral, subcutaneous and intraperitoneal routes
011 in male albino mice weighing between 19 and 21 g.
Comparisons were made with phenobarbitone sodium
A X zN H CH2 and glutethimide. Where possible the regression of
N-HG CH2 percentage mortality as probits on the logarithm of
the dose was found, and the LD50s and limits
o Q NNHo of error (P=0.95) were calculated by the method of
Litchfield and Wilcoxon (1949).
FIG. 1 Subacute Toxicity
The subacute toxicity of thalidomide was deter-
It was first synthesized by Kunz, Keller, and mined in young rats which were dosed orally each
Muckter (1956), who also described its main day over four weeks. Their body weights were
pharmacological properties. It occurs as a pure recorded twice weekly and at the end of the period
the rats were killed and a post-mortem carried out.
white, tasteless crystalline powder with a melting Microscopic preparations of the major organs were
point of 271. The substance is sparingly soluble also examined.
in water, methanol, ethanol, acetone, and glacial The pharmacological actions of thalidomide on the
acetic acid but readily soluble in dioxane, central and autonomic nervous systems and on the
dimethyl formamide, pyridine and chloroform. It cardiovascular system were studied in mice, rats and
is insoluble in ether and benzene. Chemically anaesthetized cats, according to the following methods.
it is related to bemegride (a-ethyl-a-methyl- Central Nervous System
glutarimide) and to glutethimide (8-ethyl-8- Motor Activity.-The sedative effects of thalidomide
phenyl-glutarimide), but its pharmacological were determined on the voluntary motor activity of
properties are different. mice by the photobeam method of Dews (1953). The
apparatus was a perspex tank measuring 15 in. x 9 in.
METHODS A beam of light shone across the short axis on to a
phototransistor (Mullard OCP71). Interruption of the
Except where specifically stated thalidomide was light beam activated a Thorp impulse counter (C. F.
used as a suspension in a 1 % solution of carboxy- Palmer) and a magnetic digital counter. Groups of
methylcellulose. ten mice were placed in the tank 30 min. after dosing
112 G. F. SOMERS
and the counts recorded at 15 min. intervals over
45 min. For the calculations the counts between
15 and 45 min. were used, since the initial count
was affected by the exploratory hyperactivity of the
mice. A control group was always run concurrently
with a treated group and the response of the treated
group calculated as a percentage of the control value.
Narcotic Activity.-This was tested in mice by
placing them on their backs and recording the number
which had lost their righting reflex.
Movement Co-ordination and Holding Reflex.-
The effects of thalidomide on motor co-ordination
were compared with phenobarbitone and glutethimide
by the rotating rod method of Gross, Tripod and
Meier (1955). Groups of ten mice were treated with
the respective drugs and placed at 30 min. intervals
on to a wooden rod rotating at 15 revs./min. The
times the individual mice were able to remain on the
rod were measured and the average determined for
each group. Mice remaining on the rod for more
than 2 min. were regarded as unaffected.
Depression of the holding reflex was assessed by
suspending mice by the front legs from a horizontal
wire and observing their ability to bring up their
back legs.
Anticonvulsanlt Activity.-This was determined by
measuring the ability of thalidomide to protect mice
against the convulsant actions of leptazol and
strychnine.
Analgesic Activity.-Thalidomide was tested for
analgesic activity in mice and rats by the hot plate
method of Woolfe and Macdonald (1944), the tail
pinching method as described by Bianchi and
Francheschini (1954) and electrical stimulation of the
feet (Dodds, Lawson, Simpson and Williams, 1945)
and tail (Kraushaar, 1953). It was also tested for
its ability to potentiate the analgesic actions of
morphine and pethidine.
Hypothermic and Antipyretic Activities. - These
were tested in normal rabbits and in rabbits made
febrile by an injection of a suspension of inactivated
Escherichia coli bacteria.
Autonomic Nervous System, Heart and Respiration
The pharmacological effects of thalidomide on the
autonomic nervous system were studied in cats
anaesthetized with chloralose. Blood pressure was
recorded from the left carotid artery and respiration
from a tracheal cannula by the method described
by Paton (1949). The right preganglionic cervical
sympathetic nerve was severed and the rostral stump
stimulated with rectangular pulses of approximately
8 V and of 2.5 msec. duration for 5 sec. An
electrocardiograph was taken from lead II using
subcutaneously implanted needle electrodes. The
record was made with a Cambridge direct-writing
cardiograph. Thalidomide, because of its low water
solubility, could not be given intravenously. It was,
therefore, injected into the peritoneal cavity or
directly into the duodenum.
Rabbit isolated hearts were perfused with Ringer-
Locke solution containing 10 mg./l. of thalidomide
and records were made of the heart beat and coronary
flow.
Gastro-intestinal Tract
The effects of thalidomide on intestinal muscle were
studied in vitro on the isolated duodenum of the
rabbit and the isolated ileum of the guinea-pig.
In vivo the charcoal meal test was used as described
by Bryant, Felton and Krantz (1957).
Urinary System
Kidney effects of thalidomide were studied in rats
hydrated with an oral dose of water (50 ml./kg.)
and placed into individual metabolism cages.
Interactions with Other Drugs
Barbiturates.-The duration of hexobarbitone
anaesthesia (125 mg. /kg., intraperitoneally) was
compared between groups of control and thalidomide
treated mice.
Alcohol.-Possible potentiation of the toxic effects
of alcohol was studied by determining the LDSOs of
alcohol in groups of mice given different doses of
thalidomide.
Reserpine and Chlorpromazine. - These were
studied in two ways. Firstly a comparison was made
of the duration of catatonia in groups of mice given
these drugs with and without thalidomide. Secondly
the effects of thalidomide on the potentiating action
of these drugs on hexobarbitone were determined as
described by Kopmann and Hughes (1958).
Methlainphetamine and Methylphenidate.-These
stimulant drugs were administered to mice previously
treated with thalidomide and the effects on motor
activity observed. The LD50s of these stimulant
drugs were also determined in mice treated with
thalidomide and compared with the values for normal
mice.
RESULTS
Toxicity
Acute Toxicity.-Thalidomide proved to be
virtually non-toxic. As reported by Kunz et al.
(1956), mice tolerated the maximum oral dose
(5 g./kg.) that could be administered, without ill
effects. They showed sedation without a
preliminary excitation. Motility was considerably
reduced, the mice bunching in a corner of the
cage and going to sleep. There was no inco-
ordination in their movements, and narcosis,
catalepsy, and convulsions were absent. Sleeping
mice could be easily aroused, and then they
resumed normal but slow movements and showed
little interest in their surroundings. In contrast
both phenobarbitone and glutethimide caused
initial excitation, marked ataxia, and incoordina-
tion of movement followed by narcosis and death.
The LD5Os of these two drugs are compared with
thalidomide in Table I.
 THALIDOMIDE 113

TABLE I There were no abnormal constituents in the

THE ACUTE ORAL TOXICITIES OF THALIDO-
MIDE, GLUTETHIMIDE AND PHENOBARBI-
TONE IN MICE
Confidence
LD50 Limits
Compound (g./kg.) (P=O095;
g./kg.)
Thalidomide .. .. >5
Glutethimide 0... .. 52 0 46-0)58
Phenobarbitone sodium .. 027 025-0 29
Mice survived intraperitoneal doses as high as
4 g./kg. Guinea-pigs given 650 mg./kg. orally
became quiet and sedated, while 400 mg./kg.
injected intramuscularly produced no effects at
all, insoluble material remaining unabsorbed at
the site of the injection for over two days.
The low toxicity of thalidomide has been
observed in man. Twenty cases of accidental or
intentional overdosage have been reported (de
Souza, 1959; Burley, personal communication),
and all recovered uneventfully. It may well be
that the absence of toxicity is due to a limited
absorption, for the compound has a low solubility
in body fluids, and when administered parenter-
ally remains at the site of the injection. In the
absence of a suitable assay method absorption
studies have not yet been made.
Subacute Toxicity.-Two groups of ten rats
given daily oral doses of 250 mg. and 1,000
mg./kg. respectively over a period of 21 days grew
normally compared with untreated controls
(Table II). Blood examinations showed no
abnormal changes in tjhe red and white cell counts.
TABLE II
THE EFFECT OF REPEATED ADMINISTRA-
TION OF THALIDOMIDE ON THE GROWTH
OF MALE RATS
Average Body Weights
urine. When the rats were killed no significant
pathological changes were seen in the major
organs or glands. Histological sections of the
thyroid glands showed only a slight reduction in
the colloid material, suggesting a slight depression
of secretory activity. Murdoch and Campbell
(1958) reported that thalidomide depressed the
uptake of radioactive iodine by the thyroid in
humans, but this has been shown to be common to
tranquillizing drugs (Friedell, 1958). Greene and
Farran (1958) stated that the effects on the human
thyroid were negligible. The thyroid weights of
our rats were normal, and histological changes
in the glandular epithelium which one normally
associates with an antithyroid drug were absent.
A slight depression of glandular activity can be
expected from a sedative drug due to a decreased
demand for the thyroid hormone following the
reduction in body activity.
Central Nervous System
Motor Activity.-Thalidomide caused a marked
reduction in motor activity. The results obtained,
using eight groups of mice at each dose level, are
shown in Fig. 2. It proved impossible to compare
the activity of thalidomide with phenobarbitone
and glutethimide by this method, as mice varied
considerably in their response to the latter drugs
between hypermotility due to excitation and lack
of movement due to narcosis. Kunz et al. (1956)
using activity cages found the oral doses inducing
sleep in mice to be 100 mg. for thalidomide, 40
mg. for phenobarbitone sodium, and 75 mg. for
glutethimide, all per kg. body weight.
100
80
Ma
0 60
v
I--O
U
40
._

._

(in g.) at Stated Times by

Daily Dose from Beginning of 20
Treatment (mg./kg.) Experiment
0 7 14 21 0
Days Days Days Days 50 100 200 300 400
Controls .. 0 135 177 207 228 Thalidomide mg./kg.
Thalidomide 250 142 179 214 226 FiG. 2.-The effect of thalidomide on voluntary motor
9,, 1,000 138 171 197 226 activity in mice. Each point is the mean of eight
groups.
114 G. F. SOMERS

Narcotic Activity.-Thalidomide was devoid of remained the same. The drug therefore had no
a narcotic action. Even in maximum doses significant effects on the heart, respiration or
(5 g./kg.) it did not affect the righting reflex in autonomic nervous system, nor did it show any
mice. It behaved quite differently in this respect antihistamine activity.
from glutethimide and phenobarbitone, which both The absence of cardiotoxicity was confirmed on
had a strong narcotic effect. the rabbit isolated heart where perfusion with
Ringer-Locke solution containing 10 mg./I. of
Movement Co-ordination and Holding Reflex. thalidomide produced no change in the amplitude
-Even in very large doses thalidomide showed no and rate of the contractions. The coronary flow
effect on motor co-ordination. Mice given oral was also unaffected.
doses of 5 g. /kg. were able to stay on the
rotating rod for more than 2 min. With pheno- Gastro-intestinal Tract
barbitone and glutethimide the mice were unable In vitro thalidomide had a mild and short-lived
to maintain their locomotion and balance and fell spasmolytic action, a dose of 0.5 mg. in a 15 ml.
off the rod. bath reducing the spasmodic contractions of the
guinea-pig isolated ileum to acetylcholine and
Anticonvulsant Activity.-Thalidomide did not histamine.
antagonize leptazol-induced convulsions in mice, In vivo an oral dose of 500 mg./kg. of
even in very large doses (1.6 g./kg.). Against thalidomide given 30 min. before a charcoal meal
spinal convulsions induced by strychnine there was did not significantly reduce the rate of transport
a slight protection, probably due to the sedative through the stomach and intestines. The rate of
action of thalidomide in reducing sensory stimuli. travel was only reduced by 15%.
Similar results were obtained by Fincato (1957). Urinary System
Kunz et al. (1956) found it ineffective against Thalidomide did not affect urinary output, the
electroshock seizures in the rat. rate of excretion in water-loaded rats being the
Analgesic Activity.-Thalidomide showed no same after thalidomide as in the controls.
analgesic activity in the tests described. In this Interaction of Thalidomide with Other Drugs
respect it behaved similarly to glutethimide and Central Nervous System Depressants.-( I)
to phenobarbitone, which do not reduce sensitivity Barbiturates. Thalidomide, in common with
to pain. It slightly potentiated
the analgesic activities of 10
morphine and pethidine in doses
of 400 mg./kg. in mice.
Hypothermic and Antipyretic
Activities.-Thalidomide did not 8
reduce the basal temperature of -------
lI

rabbits in oral doses of 200

mg./ kg. It showed a slight anti- v ~ ~ ~ ~ A (B)
pyretic activity, lowering and 6 _ I

shortening the pyretic response c 1 __

to inactivated E. coli bacteria. c
Cardiovascular, Respiratory and 3 ~~.
Autonomic Nervous System ' 4
In the anaesthetized cat *'(C o 60 90 12 15 80
administration of 125 mg./kg. E
of thalidomide intraperitoneally 6 Mm
or direct into the duodenum did 2
not affect the heart rate, blood
pressure or respiration. The
blood pressure responses to
acetylcholine, adrenaline and 0
histamine were unaltered, and 3
the responses of the nictitating
membrane to adrenaline and to
stimulation of the preganglionic FIG. 3.-The effect of thalidomide (- - - (A) 400 mg./kg.: - - - (B) 1,600
cervical sympathetic nerve mg./kg.) on hexo]barbitone sodium ( - (C) 125 mg./kg.) narcosis in mice.
 THALIDOMIDE 115

tranquillizing drugs, potentiated the narcotic
action of barbiturates. Fig. 3 illustrates the results
from a typical experiment with hexobarbitone
showing that the anaesthetic action was consider-
ably prolonged. The mean duration of narcosis
in groups of the mice injected with 125 mg./kg.
of hexobarbitone intraperitoneally 30 min. after
oral doses of 400 and 1,600 mg./kg. of thalidomide
were respectively 85 and 98 min. compared with
27 min. for the controls
12
10
-9
E No. Thalidomide
c 8
0
(2) Alcohol. Thalidomide increased the oral
toxicity of ethyl alcohol in mice. The increase in
toxicity was linearly related to the dose of
thalidomide (Fig. 4).
(3) Reserpine and chlorpromazine. A single
oral dose of thalidomide increased the duration of
catatonia produced in mice by chlorpromazine or
reserpine (Tables III and IV). Similarly thali-
domide increased the potentiating action of
chlorpromazine and reserpine on hexobarbitone
TABLE IV
THE EFFECT OF THALIDOMIDE ON THE
DURATION OF CATATONIA IN MICE
GIVEN RESERPINE
There were ten mice in each group.
Intraperitoneal Percentage of
Group Oral Dose of Dose of Mice Showing
Reserpine Catatonia after
(mg.fkg.) (mg./kg.) 24 hr.

m
I Controls 2 0
0
2 100 2 20
Lo 7 3 200 2 50
J 4 400 2 60
5 800 2 70
6

5 TABLE V
THE EFFECT OF THALIDOMIDE ON THE
POTENTIATING ACTION OF CHLORPROMA-
ZINE ON HEXOBARBITAL NARCOSIS
0 200 400 600 800 IN MICE
Thalidomide (mg.,/kg.) There were ten mice in each group. Thalidomide was
given 2 hr. before, and chlorpromazine 1 hr. before,
FIG. 4.-The effect of thalidomide on the acute toxicity hexobarbital. The-estimated standard error of a single
of ethanol in mice. mean duration of narcosis is 130/,,.
TABLE I Il
THE EFFECT OF THALIDOMIDE ON THE Intra- Intra- Geometric
iOral peritoneal peritoneal Mean
DURATION OF CATATONIA IN MICE Group Dose of Dose of Dose of Duration
GIVEN CHLORPROMAZINE No. Thalido- Chlor- Hexo- of
There were ten mice in each group. mide promazine barbital Narcosis
(mg. 'kg.) (mg./kg.) (mg./kg.) (min.)
Intraperitonel |Percentage of
Oral Dose of traperitoneal Mice Showing 1 0 0 125 21-0
Group Thalidomide Dose ofa!eCatatonia after: 2 0 1-5 125 37-9
No. (mg./kg.) Chlorpromazg.
(mg/kg.)
ne
24 hr. 48 hr.
3
4
100
200
1-5
15
125
125
36-7
52-6
5 400 1-5 125 49-4
I 0 40 50 0 6 800 1-5 125 70 8
2 100 40 40 0 7 100 0 125 30 0
3 200 40 60 20 8 200 0 125 30 5
4 400 40 90 30 9 400 0 125 33-0
5 800 40 100 90 10 800 0 125 44'7
116 G. F. SOMERS
TABLE V I DISCUSSION
THE EFFECT OF THALIDOMIDE ON THE Thalidomide has been shown experimentally to
POTENTIATING ACTION OF RESERPINE be a sedative hypnotic drug acting differently
ON HEXOBARBITAL NARCOSIS IN MICE from the barbiturates. It does not cause inco-
There were ten mice in each group. Reserpine and ordination, respiratory depression or narcosis, and
thalidomide were given 2 hr. before hexobarbital. The it is virtually non-toxic, possibly due to a limited
estimated standard error of a single mean duration of absorption. The clinical value of this drug has
narcosis is 15%. been reported by Jung (1956), Stark (1956) and by
Burley, Dennison, and Harrison (1959). Its safety
| Intra- Intra- Geometric has been confirmed in a number of cases of
Oral peritoneal peritoneal Mean
accidental and suicidal overdoses (de Souza,
Group Dose of Dose Dose of Duration 1959; Burley, personal communication).
No. Thalido- of Hexo- of
mide Reserpine barbital Narcosis
(g/)(nig./kg.) (mg./kg.) (min.) The author thanks Dr. L. Pirbos for histological
examinations and Mrs. S. M. Holland, Miss A.
I 0 0 125 194 Kennedy and Mr. B. Benson for valuable assistance.
2 0 04 125 311
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