STUDIES IN TOLERANCE, I-NICOTINE AND LOBELINE

B CHARLES WALLIS EDMUNDS. M.D.

(From the Pharmacological Laboratory of the University of Michigan)

(Received for publication March 21, 1909)

The question of the production of tolerance to certain chemical

substances when they are introduced into the body is one of consider-
able interest not only from the theoretical but also from the practical
standpoint. The manner by which the body gains such tolerance has
by no means been satisfactorily explained as yet and indeed the process
may not be the same for all drugs. Probably in the case of some we
have certain tissues in which the faculty for destroying or rendering
inert these poisons becomes developed through the constant presence
in the fluids of the body of small quantities of the drug. In the case
of other drugs it is probable that the tissues, being constantly bathed
in fluid containing the drug, become accustomed to its presence, it
becoming now practically a normal constituent of the body so that if
the drug should be withdrawn marked symptoms of poisoning may
appear.
A slightly different form of tolerance is that described by Morawitz
and Pratt,’ who, in their studies upon experimental anemia, found that
phenylhydrazin, after a few days’ administration, had not as marked
an effect upon the blood as at first and that this was due to some alter-
ation in the red blood cells themselves which had become resistant
not only to it but also to various other hemolytic agents. The case
of arsenic illustrates another form of this local tolerance. Cloetta2
found that in giving this drug to animals by the mouth for a long period
of time, less and less was excreted by the kidneys while increasing
amounts were found in the feces. This he explains is due to the fact

1 Morawitz u. Pratt: Munch. med. Wochen., lv, 1817, 1908.

2 Cloetta: Arch. f. exp. Path. u. Pharm.. liv, 196, 1906.
28 CHARLES WALLIS EDMUNDS

that the intestinal mucous membrane becomes less able to absorb the
drug and that the apparent tolerance is merely due to local changes
in the intestine by which less of the arsenic is taken up into the tissues.
The practical aspect of the question needs hardly to be mentioned
as the tolerance which is gained to such drugs as morphine, cocaine
and tobacco is a matter of every day experience.
Also of a good deal of interest is the so-called crossed tolerance in
which tolerance to one drug may be carried over so that the body is
insusceptible to ordinary doses of drugs which are closely related to
the first. The most familiar example is that shown by persons addicted
to the use of alcoholics who are notoriously insusceptible to the general
ana?sthetics.
Tolerance to tobacco is so easily obtained in man and of such com-
mon occurrence as scarcely to excite comment, much less scientific
inquiry, and there are therefore remarkably few references in literature
to work which has been carried out with this special object in view.
Within the past two or three years references to nicotine tolerance in
animals are more numerous, as it has been discovered that the injec-
tion of nicotine will, like adrenalin, produce sclerotic changes in the
blood vessels, and in the course of experiments upon this effect of
nicotine, the question of reaction to the drug when it is frequently
administered has naturally excited attention. The reports of such
experiments, while fairly uniform among themselves, are so utterly
opposed to the general view of the question and also to references in the
literature that some time ago I took up the question of the production
of tolerance to nicotine in animals
and the closely allied question as
to whether animals which have
been rendered tolerant to nicotine
would also exhibit any tolerance to the active principle of Indian
tobacco, lobeline, which as I showed some time ago possesses essen-
tially the same physiological action as nicotine. This is not the place
nor is it hardly necessary to go into the question as to whether nicotine
is the injurious constituent of tobacco, and as to whether tobacco
depends upon this substance alone for its effects as the recent works
of Ratner and of Lehmann, as well as of many others leave little room
for doubt.

Edmunds: Amer. Journ. of Physiol., xi, 79, 1904.

 NICOTINE AND LOBELINE 29

Among the writers upon the subject of tolerance to nicotine is

Kobert,4 who says that it is produced in animals by frequent adminis-
tration. Esser5 in his work upon the relation of the vagus to diseases
of the heart and lungs had occasion to observe the effects of nicotine
when it was given frequently to animals. He worked with dogs, to
which he gave nicotine tartrate subcutaneously, commencing with
daily doses of 10 mg. which at weekly intervals were increased 5 mg.
until a dose of 50 mg., per day was reached. At the time of the earlier
injections the animal (Experiment I)showed pulse and respiration irreg-
ularities, restlessness, vomiting and purgation. During the latter part
of the first week the vomiting and purgation were absent and no more
poisoning symptoms appeared when the dose was increased to 15 mg.
or 20 mg. The animal gained such a degree of tolerance that doses as
high as 50 mg. gave no symptoms except on the part of the pulse and
respiration, vomiting occurring rarely. The dose was then doubled
(100 mg.), but these amounts were followed by convulsions which
recurred daily after the injections so that the dose was reduced to
75 mg. Still later it was increased so that at times the animal received
as much as 200 mg., these larger doses being followed by marked acute
symptoms, especially at first, but on subsequent administrations the
animal showed some degree of tolerance. Esser treated four other
dogs and some rabbits in the same way with nicotine tartrate, giving
it to the rabbits by mouth. In all these animals he got practically
the same results as with the animal the protocol of which is abstracted
above. In each case as the dose was increased symptoms of poisoning
were obtained which on subsequent injections were not as severe. In
his conclusions upon this phase of the work he says that animals differ
in respect to the time at which tolerance appears, and, that in general,
young animals do not tolerate the drug as well as old.
In the following year Hatcher8 published his work upon nicotine
tolerance in rabbits. He found that a tolerance was gained, but only
with difficulty and to a limited degree. In one animal (D), which
reacted feebly to the nicotine sulphate given subcutaneous, a dose of
25 mg. per kg. of body weight nearly proved fatal on the 11th day, but

Kobert: Lehrbuch d Intoxicationen, 1064, 1906.

Esser: Arch. f. exp. Path. u. Pharm., xlix, 190, 1903.
6 Hatcher: Amer. Journ. of Physiol., xi, 17, 1904.
30 CHARLES WALLIS EDMUNDS

it finally survived 36.5 mg. on the 28th day and died from 40 mg. on
the 33d day. It seemed thus to have gained a slight degree of tolerance
from 14 injections. Some of the rabbits showed practically none
even when treated at short intervals for a month. Hatcher concludes
that tolerance is better established by large doses given at intervals
of three days than by frequently repeated moderate doses.
Gouget,7 in the cpurse of his studies on experimental tobacco poison-
ing, administered to rabbits an infusion of tobacco. When it was
given by the vein he found each injection was followed by convulsions
which were always about the same when equal doses were given. He
says he got no apparent tolerance in spite of the fact that nicotine is
considered one of the poisons to which tolerance is obtained most
easily.
Adler and Hensel,8 in their studies upon experimental arterio-
sclerosis in rabbits administered nicotine in 1 mg. doses, making the
injections into an ear vein. They could get no tolerance whatsoever
and could not increase the dose. Convulsions came on immediately
after the drug was given and they were always of the same intensity
whether 10 or 100 injections had been given.
Clark,9 writing upon the effects of tobacco upon the nervous system,
quotes Adler and Hensel’s work showing they could obtain no toler-
ance, and he then adds that in feeding animals with crude tobacco or
the pure nicotine mixed with food tolerance is easily and early estab-
lished, requiring hut two or three days. From his article it seems
plain that he carried out no experiments himself, but apparently
referred to the statements of Kobert cited above, as the remainder of
his article shows free quotations from the same source.
Several French writers have compared the effects upon animals of
the ordinary tobacco with the denicotiized variety when they are
administered for long periods. One of these workers, Lesieur,’#{176}says
each injection of tobacco was follosed by epileptiform convulsions
and transient paralysis. It appears from the statement as if no great
degree of tolerance was gained.

Gouget: Presse Medicale, xiv, 533, 1906.

8 Adler and Hensel: Journ. Med. Research, xv, 229, 1906.
Clark: Medical Record, lxxi, 1072, 1907.
10 Lesieur: Comp. rend. Soc. d. biol., lxii, 430, 1907.
 NICOTINE AND LOBELINE 31

Guillain and Gy11 evidently reach about the same conclusions as

Lesieur upon the question of tolerance, as they say that after intraven-
ous injections of infusions of denicotinized tobacco rabbits suffer from
dyspnoea and more or less severe convulsions followed by transient
paralysis.
Fleig and Vismeu compared the effects of the habitual use of tobacco
in both its clinical and experimental aspects. They report that they.
were able to accustom animals to the smoke by massive and repeated
inhalations, but that a graphic study of the cardiovascular phenomena
has shown them that in spite of this tolerance the same reactions are pro-
duced (qualitatively at least) at the inhalations of a first few puffs of
smoke. That “custom” implies not the necessity of absence of reac-
tion but simply the absence of intolerance; that to be “accustomed”
oannot signify anything else than to tolerate a reaction. Richon and
Perrin,’3 studying the effect of tobacco upon the development of
animals, employed an infusion which they injected into rabbits. They
say that the injections (especially at the beginning of the intoxication)
have been followed by spasmodic phenomena. This statement would
indicate that a certain degree of tolerance was obtained.
Finally Dixon and Malden14 say that in the Pharmacological Labora-
tory of Cambridge it has been shown that nicotine and morphine
tolerance is but the capacity of the tissues for the destruction of the
poison.
A study of these few references in the literature upon the subject
of nicotine tolerance in animals emphasizes the very peculiar fact of
the great difficulty with which they are rendered tolerant of the poison.
Esser has certainly been the most successful, giving massive and
frequently repeated doses of the drug regardless of the symptoms of
poisoning, being careful only that a lethal dose was not given. But
that this should be necessary is surprising when it is remembered with
what ease man develops a tolerance, as it is a matter of common
knowledge that after the first trial or two all the symptoms of acute
poisoning, especially the nausea and associated phenomena, usually

Guillain et Gy: Comp. rend. Soc. d. biol., lxiii, 684, 1907.

12 Fleig and Visme: Comp. rend. Soc. d. biol., lxiv, 114, 1908.
13 Richon and Perrin: Comp. rend. Soc. d. biol., lxiv, 563, 1908.
and Maiden: Amer. Journ. of Physiol., xxxvii, 75, 1908.
32 CHARLES WALLIS EDMTJNDS

disappear and the smoking becomes a pleasure. It is perfectly evident

that in such cases there is no such massive dosage as it has been found
necessary to administer to animals. I therefore decided to try
whether it was not possible to gain tolerance in animals by administer-
ing to them minimal doses, giving an amount slightly smaller than
had been found necessary to produce vomiting at the time of the
initial trials. I hoped in this way to approach as closely as possible
to the dosage which is taken by man when he develops his tolerance
by means of one or two trials.
My first attempts were made upon cats, giving nicotine chloride’5
to one animal and lobeline chloride to a second, in each case giving the
drug by the stomach to the fasting animal.

Experiment I. Cat. 6OO G. Nicotine chloride given by the stomach tube.

November 18 20 mg. Vomiting in 5 minutes.

19 a No vomiting.
20 15 Vomiting in hour. Wt. 2650 G.
21 12
23,24 12 No vomiting.
25, 27 12 a Vomiting.
28 10
December 2
1, 10 No vomiting.
3, 4, 5 10 Vomiting each day. Wt. 2350 G.
7 8
8 6 lo vomiting.
9 7a
10,11 7 Vomiting each day. Wt. 2310 G
12, 14, 15 No vomiting.
16 8 a
17 io a Vomited. Wt. 2360 G.

‘5The nicotine chloride was prepared by neutralizing pure nicotine with hydro-
chloric acid. Each cubic centimeter represented about 4 mg. of nicotine.
The lobeiine chloride was the same as I prepared for use in my earlier work.
(Edmunds: Amer. Journ. of Physiol., xi, 80, 1904.) It had been preserved since then
in a sealed bottle and was intensely active-0.5 mg. given subcutaneously to a
large cat causing marked vomiting and purgation.
 1

NICOTINE AND LOBELINE 33

Experiment II. Cal. 4O G. Lobeline chloride given by stomach tube.

November 21, 23 5 mg. Vomited each day.

24, 25, 27, 28 4 “ No vomiting. Wt. 2470 G.
December 1 a Vomited.
2 4
3 4 No vomiting.
4, 5 a Vomited. Wt.2550G.
7, 8 a No vomiting.

9, 10 a Vomited.
11 a No vomiting. Wt. 2540 G.
12, 14 a Vomited.
15 a No vomiting.’

16 a Vomited.
17 a a Wt.2550G.

The results of these experiments show that not only was no tolerance
gained to either drug, but that each animal became more susceptible,
reacting to smaller doses than at the beginning of the experiment.

Experiment JJJ17 Small dog. Nicotine chloride given subcutaneously.

March 24 12 mg. Vomited.

28 10 a a
29, 31 8 a No vomiting.

April 1 8 60 Vomited.
4 a a
5, 6, 7 6 “ No vomiting.
8 6 a Vomited.
11, 12, 13, 14 5 “ No vomiting.
25 5 “ Vomited.
April 26 to May 11 (12 injections
given) 5 “ No vomiting.
May 12 766 a a

13 8 a a
16, 17 9 a

18 10
19 iia

20 12 a Vomited.
23 12 “ No vomiting.
25 13 “

27 14
31 16 “ Vomited.
June 1 15 “ No vomiting.
3, 7,10
13 16f 16 Vomited.
20 16 No vomiting.

17 This animal was injected by Dr. Leroy W. Childs and Dr. Jas. A. Work, Jr.
34 CHARLES WALLIS EDMUNDS

During the first six or seven weeks the animal showed the same
increase in susceptibility which the two cats had shown, but at the end
of this time it began to gain a slight tolerance so that at the end of
another six weeks it was able not only to take once more the original
dose but practically double the original amount. This tolerance,
which is really slight, was only obtained after three month’s careful
administration of the drug.
As the results in Experiment III had shown that a certain degree
of tolerance to nicotine could be gained in dogs I decided to make
another attempt, giving to one dog nicotine and to a second lobeline,
obtaining the minimum vomiting dose for each drug with each animal
so that should tolerance be gained to either lobeline or nicotine it
would be easy to ascertain whether cross-tolerance was also gained.
In order to avoid any cumulative action of the drug in the body which
might possibly explain the increase in susceptibility which the previous
animals had shown, the injections were made only every second or
third day so as to give ample opportunities for the complete excretion
of the drugs. As a matter of fact this lengthened interval between
injections hardly seemed necessary, as the animals appeared to vomit
just as frequently after a three or four-day interval as after a rest of
only one or two days.
As it will hardly be necessary to give the complete protocols of these
experiments; only so much will be given as is required for clearness.
Experiment IV. Shepherd dog was given nicotine chloride for
three months, when it was taken ill and died of enteritis and broncho-
pneumonia.
At the beginning of the experiment it took 5 mg. without vomiting
while 6 mg. caused emesis. During the next 90 days it showed the
same increase in susceptibility as the previous animals. It soon began
to vomit after injections of 5 mg. and then 4 mg. and finally vomited
several times on doses of 3 mg. When this stage had been reached
the dog developed a severe enteritis from which it died in two days
as stated above.
 NICOTINE AND LOBELINE 35

Experiment V. Hound. Nicotine chloride given subcutaneously.

March 10 15 m g. nicotine Vomited.

a
11 12 a a
9, 13, 16 10 a a No vomiting.
18 a lobeline. “ a
20 2 a a Vomited.

March 23-April 13-dose gradually increased from 10mg. nicotine chloride to 20 mg.
without vomiting.

April 15 20 mg. nicotine Vomited.

a a
17,20 15 a
a a
15 No vomiting.
24, 27, 29 5 a Vomited each day.

The dose was then increased rapidly to see whether tolerance would
be gained more quickly by that method because in the six weeks from
March 10 to April 29 none seemed to have been gained.

May 4 20 mg. nicotine Vomiting.

5,6 25” “ a
.
8 25 a No vomiting.
9 30 “ “ Vomited.
11 20 “ “ No vomiting.
60
12 30 a a
13 40 “ “ Vomited.
14 50 “ “ No vomiting.
15 50 “ “ Vomited.
18 25 “ “ No vomiting.
19 2 “ iobeline. a
20 4 “ “ Vomited.

A very definite tolerance to nicotine had been gained and also some
degree of crossed tolerance, so the experiment was continued.

May 21 50 m g. nicotine. Vomited.

22 60 “ “ “

66
23 70
o
24 80 a a
25 100 “ “ Did not vomit; trembled.
26 50 “ “ Salivation but no vomiting.
27 4 “ lobeline. “ “ “

28 8 “ “ No vomiting.
29 16 “ “ Vomited.
36 CHARLES WALLIS EDMUNDS

Experiment VI. Black and white dog. Lobeline chloride injected subcutaneously.

December 14 1 mg. lobeline. Vomited.

16 .... 0.6 a a No vomiting.
18 6 “ nicotine. Vomited.
20 a a No vomiting.

December 24 to January 6, 0.8 mg. lobeline given three times weekly

did not cause vomiting.
January 8 to February 6. 1 mg. lobeline given in the same way
rarely caused vomiting.
February 8 to March 13. Dose was increased to 1.2 mg., which did
not cause vomiting at the first fewinjections, but finally caused vomit-
ing every time; so dose was reduced to 1 mg. and continued between
1 mg. and 1.2 mg. until April 15, when it was gradually increased until
by April 24 1.8 mg. lobeline caused no vomiting. April 27 7 mg.
nicotine caused vomiting, showing no crossed tolerance.
The dose was then rapidly increased as in Experiment V.

April 29 2 mg. lobeline. Vomited.

May 1, 4 2” a
a
5
a a
6
5” a
8
a
9
11 a
12 8” a
13 10 a a Very restless, dyspnaa.
14 10 a a
15 10 a a
a a oo a
18 2.5” a
19 1.5 a a No vomiting.
20 2” a
7 60 nicotine. Vomiting.
23

My results certainly show the extreme difficulty with which animals

gain tolerance to small doses of nicotine, as it is only after weeks of
careful administration that a limited tolerance is attained. On the
other hand with massive doses, following somewhat after Esser’s
method, a tolerance is apparently quite easily gained and this imparts
at the same time a tolerance to the lobeline. With the latter drug
 NICOTINE AND LOBELINE 37

only a very limited degree of tolerance could be obtained. Also with

this drug massive dosage produced no different results as it had with
nicotine, and in so far the results agree with those found in man, in
whom use of the Indian tobacco is always followed by nausea.
Some further points of interest are brought out by the research
especially as regards the broad question of the causation of tolerance.
It would seem proved conclusively that Dixon and Malden’s explana-
tion will not suffice. If tolerance is nothing but the ability of the
tissues to destroy the poison, why are they practically unable to destroy
the small doses ranging from 1 mg. in Adler and Hensel’s work to the
8 or 10 mg. doses in my experiments and yet they very promptly take
care of the drug when it is given in massive doses?
Exactly the same objection can be made to the explanation that
some change takes place in the tissues by which they adapt them-
selves to the foreign substance so that the new conditions become
normal. Indeed it may be stated at once that no satisfactory expla-
nation can now be offered for this peculiarity.
Another point which might be. considered is the great difference
in the development of tolerance in man and in the dog, though it
is possible that this difference is not as great as appears at first sight
and in this connection a few facts might be pointed out merely as
suggestive.
It is clear that a true tolerance to nicotine is gained by man and
that Fleig and Visme’s theory cannot be considered as a sufficient
explanation. Man certainly does not merely “tolerate the reaction”
of nausea and vomiting which would be necessary if their theory
was correct.
However, it may be pointed out that very many habitual smokers
really possess very little tolerance-” a stronger” cigar than usual
being frequently the cause of nausea and even of emesis. The same
is true of many if smoking is indulged in before breakfast or when
the individual is not feeling quite “up to the mark.” It is also true
that some men have never learned to smoke because they have been
unable to get over the feeling of nausea.
These and other facts which might be cited seem to point to the
fact that the tolerance to nicotine in man is in many cases not very
great, slight disturbing factors being sufficient to determine symp-
toms of acute poisoning.
38 CHARLES WALLIS EDMUNDS

It is possible that in man a partial explanation of his tolerance

may be due to local changes in the mucous membrane lining the
respiratory passages by which they become less able to absorb the
products contained in the smoke, a change comparable to that de-
scribed by Cloetta to occur in the production of arsenic tolerance.
This theory would receive weight from the fact that habitual smokers
very frequently vomit or are at least nauseated by even very small
amounts of tobacco when taken in any other way than by smoking.
This cannot account entirely for man’s tolerance, because it fails to
take into consideration his changing susceptibility which as pointed
out above is a factor of common knowledge.

CONCLUSIONS

(1) Tolerance to nicotine or tobacco can be obtained in animals

only with very great difficulty when the drug is administered in
small doses.
(2) To large toxic doses dogs develop a resistance quite quickly
and when rendered tolerant by such means they exhibit a crossed
tolerance to the related alkaloid of Indian tobacco, lobeline.
(3) Only a very limited tolerance could be gained in dogs to lobeline
itself either by the method using small doses or by massive injections.