Beruflich Dokumente
Kultur Dokumente
HALOGENATED AMPHETAMINES
Ray W. Fuller
The Lilly Research Laboratories
Eli Lilly and Company
Indianapolis, Indiana 46206
MULTIPLE
ACTIONS
OF p-CA ON SEROTONIN
NEURONS
occurs in vivo, though such inhibition would not result in depletion of serotonin
levels.
The actions of p-CA on serotonin neurons are summarized in TABLE 1. The
initial effects probably include inhibition of tryptophan hydroxylation, release
of serotonin from intraneuronal storage granules, inhibition of serotonin reup-
take by the neuronal membrane pump, and inhibition of serotonin oxidation by
monoamine oxidase. Lowering of serotonin by p-CA may occur largely through
inhibition of tryptophan hydroxylation, but these actions seem to vary in
prominence among various p-CA analogs, as will be discussed. Lowering of
5-HIAA may occur via inhibition of tryptophan hydroxylation, inhibition of
monoamine oxidase, and inhibition of serotonin reuptake.
Not only does p-CA produce rapid depletion of brain serotonin, but the
effects are also of extremely long duration. Sanders-Bush er d . 1 5 first recog-
TABLE
1
ACTIONSOF p-CA THATMAYINFLUENCE 5-HYDROWINDOLE CONCENTRATION
IN BRAINAND THATARE POTENTIALLY DISSOCIABLE
Influence on Concentra.tionof
Action Serotonin 5-HIAA
Inhibition of tryptophan hydroxylation decrease decrease
Release of serotonin decrease increase
Inhibition of serotonin reuptake no change decrease
Inhibition of monoamine oxidase increase decrease
Irreversible damage to
serotonin neuronal function * decrease decrease
nized that serotonin levels were still depressed for as long as 4 months after
a single injection of p-CA in rats. All parameters of serotonin neural function
(tryptophan hydroxylase,l5 serotonin concentration 15, l6 and turnover,I6 5-
H1AA,l5# and serotonin uptake l7) are depressed at these long time intervals,
which suggests that some serotonin neurons have been destroyed, and Harvey
et al. have published histologic evidence for selective neurotoxic effects of
p-CA.l8,l8 The initial effects of p-CA are not permanent, however, and can
be readily reversed for a few hours after p-CA administration *OJQ (TABLE 2).
Thus, the rapid initial effects and the long-lasting destructive effects of p-CA
on serotonin neurons can be readily distinguished.
By studying structural analogs of p-CA, we have been able to dissociate
these initial and long-term effects and have further been able to distinguish the
four multiple initial actions listed above. The structures of several of these
analogs are shown in FIGURE 1.
Fuller: Structure-Activity Relationships 149
FIGURE 1. Structures of p-CA and some of its analogs. All of the compounds used
were racemic mixtures of optical isomers.
INFLUENCE
OF CHLORINE
POSITION
ON THE RING
TABLE 2
DURATION
OF REVERSIBILITY
OF SEROTONIN DEPLETION
BY p-CA AND
p-BROMOAMPHETAMINE (p-BA)
Reversal ( % )
Hours After Injection ~~
*Uptake inhibitor (fluoxetine hydrochloride) was injected at this time, and sero-
tonin concentration was measured 24 hr later. p-CA data were calculated from
Reference 21, and p-BA data were from similar unpublished studies.=
150 Annals New York Academy of Sciences
serotonin in rats, particularly at longer times, could be attributed to their failure
to remain in the brain at effective concentrations. The metabolic differences
between these chlorinated amphetamines in rats can be eliminated hy pretreat-
ment with desmethylimipramine z 2 or iprindole 2 4 to block para-hydroxylation.
When that is done, m-chloroamphetamine lowers brain serotonin in exactly the
same manner as does p-CA.22s2 4 And, in the guinea pig, a species in which
para-hydroxylation of amphetamines does not occur, m-chloroamphetamine acts
like p-CA in lowering brain serotonin.z5 o-Chloroamphetamine, on the other
hand, does not lower but, instead, increases serotonin concentration in des-
methylimipramine-treated rats 2 2 and guinea pigs.2e Furthermore, the addition
of an o-chloro substituent to p-CA markedly diminishes its ability to lower brain
serotonin, although the reduction of 5-HIAA was perhaps even greater with
the dichloro compound.2e Since the dichloro compound is a more potent
rnonoamine oxidase inhibitor,ze this finding supports the idea that inhibition of
monoamine oxidase does contribute to the decrease of 5-HIAA caused by
these compounds.
OTHERTHAN
SUBSTITUENTS CHLORINEON THE RING
OF SUBSTITUENTS
INFLUENCE ON THE NITROGEN
all act via conversion to p-CA. W e have been unable to block this metabolic
conversion by any drug to determine whether the N-alkyl compounds are
capable of depleting serotonin o n their own.
N-Hydroxyl-p-CA was an analog of interest because hydroxylamines are
unstable and chemically reactive. We thought that N-hydroxyl-p-CA might be
a (the) neurotoxic metabolite that has been considered as a possible mediator
of the long-term effects of p-CA. However, we found that N-hydroxy-p-CA
was, in fact, rapidly and quantitatively converted to p-CA, so that its effects on
brain serotonin were indistinguishable from those of P-CA.~'
N-Cyclopropylarylalkylaminesare irreversible inhibitors of monoamine oxi-
~ ?general, and N-cyclopropyl-pCA is therefore a unique analog of
d a ~ e , ~ in
p-CA in that the monoamine oxidase-inhibiting component of its activity plays
TABLE 4
SEROTONIN
DEPLETIONAND p-CA CONCENTRATIONIN RAT BRAINAFTER
INJECTIONOF p-CA OR p-CMA *
* Drugs were injected intraperitoneally at 0.1 mmol/kg. Means and standard errors
for five rats per group are shown.
t p < 0.01,different from control group.
152 Annals New York Academy of Sciences
INFLUENCE
OF SIDE CHAIN
LENGTH
When the @-carbon of p-CA contains two fluorine atoms in place of hy-
drogen atoms, the basicity of the amine group is greatly reduced due to the
strong electron-withdrawing property of This unique p-CA analog
has a pK, of 6.8 and so exists at physiologic pH mainly in the neutral form
instead of being virtually completely ionized, as is p C A itself (pK, 9.3). Pre-
Fuller: Structure-Activity Relationships 153
WITH RIGID
p-CA ANALOGS CONFORMATION
OF LONG-TERM
POSSIBLEMECHANISMS EFFECTSOF p-CA
SUMMARY
ACKNOWLEDGMENTS
I am grateful to Drs. Jack Mills and Bryan B. Molloy and their associates
for the synthesis of the p-CA analogs; to Drs. David T. Wong, C. John Parli,
and E. Martin G i l for collaborative assistance in various phases of our studies;
and to Harold D. Snoddy, Kenneth W. Perry, John C. Baker, and Susan
Hemrick for their experimental work.
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H. BRUDERER
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156 Annals New York Academy of Sciences
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Fuller: Structure-Activity Relationships 157
DISCUSSION
DR. A. S. HORN: Have you studied the various isomers of these com-
pounds? If so, do they have differential actions?
DR. FULLER: All of the compounds that we have studied were administered
as mixtures of the D- and the L-isomers. It would be interesting to know if they
have differential effects; however, we havent given this a high priority, because
several years ago we found that the two isomers of p-chloroamphetamine
appeared to be identical with respect to their effects on brain serotonergic
neurons.
DR. E. SANDERS-BUSH: Have you looked at other possible antagonist effects
of fluoxetine on p-chloroamphetamine or its derivatives? I ask this question
because there is some debate about the mechanisms underlying the initial de-
crease in tryptophan hydroxylase activity seen after p-chloroamphetamine injec-
tions. Can this effect be reversed if animals are pretreated with fluoxetine?
DR. FULLER: In fact, all parameters that are depressed after p-chloro-
amphetamine treatment can be reversed by the administration of fluoxetine.
For example, brain serotonin levels and tryptophan hydroxylase activity are
normal within 4 hr after fluoxetine treatment. If the reversal in tryptophan
hydroxylase activity is dependent on the synthesis of new enzyme protein, that
implies that there is a more rapid rate of synthesis than one would anticipate
on the basis of values for tryptophan hydroxylase enzyme turnover described
previously in the literature. However, there is also no evidence that the reversal
of p-chloroamphetamine-inducedinhibition of tryptophan hydroxylase is related
to differential inhibition by p-chloroamphetamine or its metabolites. In short,
then, I really dont know what mechanisms might account for the ability of
fluoxetine to rapidly reverse the effects of p-chloroamphetamine.
Some indirect evidence indicates to us that p-chloroamphetamine might act
directly on serotonin-containing neurons, rather than via a neurotoxic metabolite
of the compound. For example, in one study, we reasoned that if the metabolite
was formed via a change in the ring structure of p-chloroamphetamine, a com-
pound like p-bromoamphetamine might be converted to a neurotoxic metabolite
at a rate different from that of p-chloroamphetamine. Since the rate of develop-
ment of irreversibility of p-chloroamphetamine and p-bromoamphetamine were
identical, we now think that p-chloroamphetamine exerts its effects directly on
the neuron.
DR. E. M. GAL: We have some evidence that pchloroamphetamine pro-
duces a strong inhibition of tryptophan hydroxylase activity either in vivo or
in v i m , if you use the natural cofactor for the enzyme in the assay. Have YOU
or Dr. Sanders-Bush tried this cofactor?
DR. FULLER:The fact that p-chloroamphetamine-induced inhibition of
tryptophan hydroxylase activity cannot be reversed by dialyzing tryptophan
hydroxylase obtained from brain homogenates would argue against the possi-
bility that p-chloroamphetamines effects only occur in the presence of a natural
cofactor but not an unnatural one. Hence, it seems unlikely that there is any
direct inhibition of tryptophan hydroxylase by either p-chloroamphetamine or
a metabolite that would be present in our preparation.
DR.SANDERS-BUSH:We have also found that p-chloroamphetamine de-
creases tryptophan hydroxylase activity in vivo whether we use Dr. Fullers
assay or the procedure developed by Dr. GBI.
Fuller: Structure-Activity Relationships 159
DR. GAL: There is some evidence that the more active form of the trypto-
phan hydroxylase enzyme molecule must be phosphorylated. Is there any possi-
bility that this process might be interfered with shortly after p-chloroamphet-
amine administration?
DR.FULLER: I think that something like your hypothesis might be possible.
DR. J. H. GROWDON: Immediately after injection of p-chloroamphetamine
in normal animals, one sees a rather complex set of changes in motor behavior
that can be reversed up to 30-45 min later by injecting these animals with
fluoxetine.