Life Sciences, Vol. 29, pp.

861-865 Pergamon Press

Printed in the U.S.A.

SEROTONIN RECEPTOR INTERACTIONS OF HARMALINE

AND SEVERAL RELATED 8-CARBOLINES

Richard A. Glennon

Department of Pharmaceutical Chemistry

Medical College of Virginia
Virginia Commonwealth University
Richmond, Virginia 23298

(Received in final form June 22, 1981)

Summary

The isolated rat fundus preparation was used to determine the ser-
otonin (5-HT) receptor affinities (pA 2 values) of harmaline, hat-
mine and several other related B-carbolines. Replacement of a 7-
methoxy group by a hydroxy group decreases affinity; the 6-methoxy
B-carbolines possess two to three times the affinity of their
7-methoxy isomers. A new model is proposed for the interaction of
these B-carbolines with 5-HT receptors.

Harmala alkaloids derivatives of 8-carboline, are psychoactive constitu-

ents of a variety of plants and vines growing throughout various regions of
the world (1-3). Perhaps the best known genus is Banisteriopsi s which is indi-
genous to tropical and subtropical climates of, for example, the Americas.
South American natives prepare a hallucinogenic brew from species of Banister-
iopsis, or from closely related plants, which is variously called ayahuasca,
yage and caapi (i-3). A number of alkaloidal substances have been isolated
from these plants, and many of these have now been prepared by total synthesis.

Although a number of these compounds are inhibitors of monoamine oxidase,

for example, see Buckholtz and Boggan (4), human studies reveal that several
derivatives are also hallucinogenic (5). Such compounds have been shown to
have an effect on serotonergic systems (6-10) and McIsaac (ii) has hypothe-
sized that certain of them may be formed in vivo from the metabolism of endo-
genous indolealkylamines. Though there is now some evidence that this is pos-
sible (e.g. see references 12-16) the significance of these findings remains
unclear.

We have recently reported on the structure activity relationships for

the interaction of indolealkylamine derivatives with the serotonin (5-HT) re-
ceptors of the isolated rat fundus preparation (17). Those indolealkylamines
considered most hallucinogenic had the highest 5-HT receptor affinities (17,
18). Because the B-carbolines are composed of an indolealkylamine backbone,
and because several derivatives are moderately hallucinogenic, it was of in-
terest to examine their 5-HT receptor interactions using the isolated rat fun-
dus preparation.

0024-3205/81/080861-05502.00/0
Copyright (c) 1981 Pergamon Press Ltd.
862 B-Carbolines: 5-HT Receptor Interactions Vol. 29, No. 8, 1981

Methods

Using the rat stomach fundus preparation of Vane (19), dose response
curves were obtained for serotonin both in the absence and in the presence of
increasing concentrations of test compound, as we have previously reported in
greater detail (17). Affinities were calculated as pA 2 values by the method
of Arunlakshana and Schild (20). Four or five dose response curves were used
for each p ~ determination. The carboline derivatives were obtained from
Sigma Chemical Co. and were used as hydrochloride salts, or, were dissolved in
an equivalent amount of hydrochloric acid prior to final dilution.

Results and Discussion

All carbolines examined appear to interact with the 5-HT receptors in a

competitive manner (Schild plot slopes range from -0.98 to -1.14), and, affin-
ities fall within a narrow range (PA2 values = 5.28 to 6.15). Gryglewski et
al (i0) previously examined a small series of tetrahydro-~-carbolines, using
the rat fundus preparation, and also report a competitive interaction. Reduc-
tion of one or both of the pyrido ring double-bonds reduces affinity approxi-
mately 5-fold. This effect is not unlike the effects observed in various
animal assays by Gunn (21). Comparing the affinity of harmaline (or harmine)
with that of 6-methoxyharmalan (or 6-methoxyharman), i.e. replacing the car-
boline 7-methoxy group with a 6-methoxy group, results in a 2-fold (3.8-fold)
increase in affinity. When the corresponding indolealkylamines are examined,
5-methoxy-N,N-dimethyltryptamine (5-OMe DMT) is found to possess twenty times
the affinity of 6-OMe DMT (17). The affinity of the hydroxylated derivatives
harmol and harmalol, is two to three times less than that of harmine or harma-
line.

Several of the compounds in Table I are hallucinogenic (5), although

none is significantly more potent than DMT (pA 2 = 6.00) (17). Very few human
studies have been conducted with the carbolines; nevertheless, the scant re-
sults which are available are quite interesting when compared with the affini-
ty data. Naranjo (5) has found harmaline to be approximately twice as active,
as a hallucinogenic agent, as harmine but only half as active as 6-methoxyhar-
malan. With respect to affinity, the affinity of harmaline is approximately
twice that of harmine and one half that of 6-methoxyharmalan. Whereas reduc-
tion of the double bond of the latter compound, to give 6-methoxytetrahydro-
harman, decreases hallucinogenic potency by a factor of three, affinity is
decreased by five-fold. Thus, parallelisms appear to exist between activity
and affinity. It should be noted, however, that the differences in affinity,
as well as in activity, cover a very narrow range. The observed parallelism
between activity and affinity may be fortuitous; further human and/or behav-
ioral studies on additional analogues are necessary. Furthermore, although
5-HT has been previously implicated as playing a role in the mechanism of ac-
tion of the hallucinogenic carbolines, other neurotransmitter systems may also
be involved (22).

How might carbolines interact with 5-HT receptors? Although the 5-HT
receptors may possess more than one "amine site", tryptamines probably inter-
act with 5-HT receptors in such a manner that the side-chain is fully extend-
ed (e.g. see reference 23). If carbolines interact with 5-HT receptors in a
manner similar to that of the tryptamines (i.e. if the indole rings are super-
imposed), the pyrido nitrogen atom probably does not interact with the same
amine site at which the fully-extended tryptamine side-chain amine does. A
second amine site may be involved; alternatively, the interaction of the car-
bolines and tryptamlnes may be dissimilar (i.e. non-superimposed indole rings).
Vol° 29, No. 8, 1981 B-Carbolines: 5HT Receptor Interactions 863

TABLE I

Serotonin Receptor Affinities

CH 3
pA~ Slope~ Nc
Compound R
5.27 (~0.34) io00 (_+0.20) 3d
Harmol 7-OH

Harmine 7-OCH 3 5.47 (+0.i0) 1.02 (_+0.21) 4

6-Methoxyharman 6-OCH 3 6.05 (_+O,18) 0.98 (_+0.08) 6

CH 3

Harmalol 7-OH 5.28 (_+0.02) 1.14 (_+0.04) 2

Harmaline 7-OCH 3 5.82 (_+O.16) 1.01 (_+0°06) 4

6-Methoxyharmalan 6-OCH 3 6o15 (_+0.25) 1.02 (_+0.03) 6

H CH 3

6-Methoxytetra- 6-OCH 3 5.45 (_+0°06) 0.98 (_+0.15) 4

hydroharman

a b
-- The pA 2 value is followed by standard deviation° - Negative slope of
Schild plot followed by standard deviation. ! Number of pA 2 determina-
tions. ~ In two additional determinations, non-competitive slopes (i.eo
-0°69 and -0o71) were obtained°
864 B-Carbolines: 5HT Receptor Interactions Vol. 29, No. 8, 198

FIGURE i: Model depicting a

7-methoxytryptamine molecule
superimposed over the struc-
ture of harmalineo (Several
of the hydrogen atoms have
been supressed)o

MeO

As mentioned earlier, the affinity of 5-OMe DMT is 20-fold greater than

that of 6-OMe DMT; further repositioning of the methoxy group from the 5- to
4-position, to give 4-OMe DMT, results in an 8-fold decrease in affinity
(17,18). Interestingly, 6-OMe DMT possesses twice the affinity of 7-OMe DMT;
this is similar to the difference in affinity comparing 6-methoxyharmalan and
harmaline. With respect to a 5-HT receptor interaction, it might be that the
7-position of the tryptamines (i.e. indolealkylamines) is congruous with the
7-position of the carbolines (see Figure i). With this type of interaction,
the indole nucleus of the tryptamines would not interact in a manner similar
to the indole portion of the carbolines; however, it has already been demon-
strated that compounds (e.g. phenalkylamines) devoid of the pyrrole portion
of the indole ring possess affinity for 5-HT receptors (24). If such is the
case, the CI-N 2 region of the carbolines would correspond to the terminal
amine of the tryptamines. Finally, whereas 4-OH DMT and 5-OH DMT possess a
greater affinity than their corresponding methoxy derivatives, the affinity
of 7-OH DMT is 2.8-times less than that of 7-OMe DMT (17). The affinities of
harmol and harmalol are 2- to 3-times less than that of harmine and harmaline.
This alternative mode of binding for the carbolines offers a novel perspec-
tive from which to view these compounds, and, furthermore, suggests that the
5-methoxy derivatives of the carbolines (which would now correspond to the
5-methoxy derivatives of the tryptamines) should have a greater 5~HT receptor
affinity than either their 6- or 7-methoxy isomers.

Acknowledgments

This work was supported, in part, by U. S. Public Health Service Grant

DA-01642. The author also wishes to thank D. Leming Doot for her able assis-
tance.

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