SYNTHESIS AND CHEMICAL PROPERTIES OF SOME

INDOLE DERIVATIVES

H. G. Schlossberger
Department of Organic Chemistry and Spectroscopy
Max Planck Institute for Biochemistry
8033 Martinsried, Munich, Federal Republic of Germany

Ortho-dihydroxytryptamines (0-DHT) have been claimed to occur in bio-

logic tissues (e.g., in the crustacean heart l ) and hypothesized to represent
“active” metabolites of serotonin.‘ These assumptions prompted us to synthesize
5,6-DHT R. and 4.5-DHT.5 More recent work by Baumgarten et al.Gp renders
these earlier assumptions unlikely, since both drugs damage central serotonin
(5-HT) neurons when introduced into the cerebrospinal fluid. Both 5,6-DHT
and 4,5-DHT have limited applicability as tools for induction of serotonin fiber
degeneration because of unspecific cytotoxicity. These findings stimulated a
search for neurotoxic indole derivatives with lesser side effects. Isomeric dihy-
droxytryptamines, in addition to Nu-monomethylated, Nu-dimethylated, a-C-
alkylated, and 0-methylated derivatives, and otherwise substituted dihydroxy-
tryptamines appeared to be of special interest in this

SYNTHESES

The synthesis of tryptamines modified with two substituents on the benzene

nucleus of the indole ring is generally performed according to methods for
synthesis of simple or nonsubstituted tryptamines; that is, in general, the side
chain is attached to an indole derivative.” For the synthesis of dihydroxy- and
hydroxymethoxytryptamines, the indoles required were the corresponding di-
benzyloxy- resp. benzyloxymethoxyindoles. The free hydroxyl groups in the
tryptamines to be synthesized must be protected in this stage of synthesis. We
obtained these indoles by two well-established methods: reductive cyclization
of o,w-dinitrostyrenes l:’ or according to the method of Reissert,” namely, con-
densation of an o-nitrotoluene with diethyl oxalate to the corresponding ethyl
o-nitrophenylpyruvate and, after cyclization and saponification, decarboxylation
of the indole-2-carboxylic acid.
The accessibility of the starting material, either o,o-dinitrostyrene or o-
nitrotoluene, is of importance for the choice of the method of synthesis of the
indole derivative. The last step in the synthesis, according to Reissert, decar-
boxylation, is the critical step; depending on the substitution, the yield of indole
varies greatly.15--“
FIGURE 1 shows the different methods we used to synthesize 4,5-dibenzyl-
oxyindole, the starting indole for the preparation of 4,5-DHT. This synthesis
has not been published before but will be outlined below.
6-Nitrovanillin (11) l R obtained from o-vanillin (I) could not be demethyl-
ated to 6-nitro-2,3-dihydroxybenzaldehyde (111) , so that 6-nitro-2,3-dibenzyl-
oxybenzaldehyde (XV) was not accessible by this method. Reduction of I
yields 2-hydroxy-3-methoxytoluene(VII) , the 6-nitro derivative of which, VIII,
25
0077-8923/78/0305-0025 $01.7510 @ 1978, The New York Academy of Sciences
26 Annals New York Academy of Sciences

FIGURE 1. Synthesis of 4,5-dibenzyloxyindole (XII), and 4-benzyloxy-5-methoxy-

indole (VI).

gives 6-nitro-2,3-dihydroxytoluene(M) during demethylation. From 6-nitro-

2,3-dibenzyloxytoluene (X), ethyl 6-nitro-2,3-dibenzyloxyphenylpyruvateis ob-
tained, and by its cyclization, ethyl 4,5-~dibenzyloxyindole-2-carboxylateis
produced. The yield of 4,5-dibenzyloxyinJole (XII) by decarboxylation of
4,5-dibenzyloxyindole-2-carboxylicacid (XI)lo is so poor that for the prepara-
tion of XII in larger quantities, another method had to be chosen. Compound
I was demethylated to 2,3-dihydroxybenzaldehyde (XIII) ,?O the dibenzyl
derivative (XW)of which could be nitrated to 6-nitro-2,3-dibenzyloxybemal-
dehyde (XV) in a yield of 40%. Condensation with nitromethane and sub-
sequent reductive cyclization give XII in an overall yield of 10%. Compound
II reacts with benzylchloride to 6-nitro-2-benzyloxy-3-methoxybenzaldehyde
(IV); via the stage of the o,odinitrostyreIie V, 4-benzyloxy-5-methoxyindole
(M)l8 is obtained, the indole derivative for the synthesis of 4-hydroxy-5-
methoxytryptamine.
 Schlossberger: Some Indole Derivatives 27
Among the different methods that exist for attaching side chains to an
indole, we chose the oxalyl chloride method of Speeter and Anthony Z1 for the
preparation of Nw-monomethylated and Nw-dimethylated tryptamine deriva-
tives and also for tryptamines with a free amino group. For a-alkylated deriva-
tives, the synthesis must proceed via the indole-3-carboxaldehyde and the
corresponding nitrovinylindole,?? but a-unsubstituted tryptamines are also avail-
able by this method.
FIGURE 2 shows all of the reaction steps that proceed via the gramine
(XVI)L‘3 and that we used successfully for preparation of numerous dihydroxy-
and hydroxymethoxyindole derivatives. The corresponding tryptophan ( X M )
is obtained according to Snyder and Smith.2.* Via the stages of the nitrile
(XVII),the tryptamines 2 5 , ?6 (XVIII),indole acetic acids 2 7 (XX),and tryp-
tophols ?i (XXI)are accessible. By means of catalytic debenzylation, the end
products are obtained. The stage of indoleacetaldehyde (XXII)would be the
intermediate for dihydroxy- and hydroxymethoxyindoleacetaldehydes.

2. Syntheses of indole derivatives via the stage of gramine.

FIGURE
28 Annals New York Academy of Sciences
In FIGURE 3, the synthesis of 4,5-DHr is shown. From 4,5-dibenzyloxy-
gramine (XXIII), 4,5-dibenzyloxyindoleace:tonitrile(XXIV) is obtained. HOW-
ever, attempts to reduce this compound to 4,5-dibenzyloxytryptamine (XXV)
with lithium aluminum hydride were unsuccessful at first; because large quan-
tities of dark-colored products were always obtained; it was not possible to
isolate XXV from the reaction mixture. Therefore, we condensed XXIII with
ethyl nitroacetate to ethyl a-nitro-&4,5-dibenzyloxyindole-3-propionate(XXVI)
according to Lyttle and Weisblat; 2 8 after saponification, the nitropropionic
acid derivative is decarboxylated to give 1-nitro-2- (4,5-dibenzyloxyindole-3)-
ethane (XXVII). After many attempts to reduce XXVII to XXV with lithium
aluminum hydride, we found it necessary to keep the reaction temperature at

P 3

-
OBZ CHz-N, OBz Cb-CN

H
XI1

Bz CH CH-COOR OBZ CH2-$H-COOR OBZ CHz-CH2-NH2

Bzo&-' N%- Bzo@.$\ NO2 8 \2 0 ~
\

xxx I Bzoe / ,,
- C H ? - N + BZOs ,
- C H 2 - N H HCI
2

HOOC-COOH \ \
H

xxxl I XXVll XXVlIl

I"
FIGURE3. Synthesis of 4,SDH'T hydrochloride (XXIX).

0" C to produce a reasonable yield of XXV; under these conditions, XXIV can
also be reduced with lithium aluminum hydride to XXV. For the catalytic
debenzylation, XXV must be transformed into a salt, for example, the hydro-
chloride (XXVIII). 4,5-DHT hydrochloride (XXIX) is then obtained as a
colorless substance, which, however, in the presence of oxygen becomes blue
within a few hours. Analogously, 4-hydroxy-5-methoxytryptamine 28 hydro- 5s

chloride is obtained from 4-benzyloxy-5-n1ethoxyindole(VI)via the stages of

the gramine, nitrile, and the 4-benzyloxy-5-methoxytryptamine.This compound
is somewhat more stable than X X M ;however, with time, it also becomes blue
when exposed to air. The ultraviolet absorption spectrum of 4-hydroxy-5-
 Schlossberger : Some Indole Derivatives 29

methoxytryptamine is seen in FIGURE 4; the spectra of XXV and 4-benzyloxy-5-

methoxytryptamine are shown in FIGURE 5 . Compound XXVI is reduced, ac-
cording to Lyttle and Weisblat,Zx to ethyl-~,~-4,5-dibenzyloxytryptophan, the
hydrogen oxalate, of which XXXI is debenzylated to ethyl-~,~-4,5-dihydroxy-
tryptophan hydrogen oxalate (XXXII).
The rapid autoxidation of both 4,5-DHT and 4-hydroxy-5-methoxytrypt-
amine in a buffer at physiologic pH prevents the application of these two trypt-
amines as selective neurotoxic pharmacologic agents.
The most potent and useful indole neurotoxins are 5,6-DHT and 5,7-
DHT.". :I1 5,7-DHT, first synthesized by Lee et a/.,'" exhibits effects on sero-
tonergic neurons that are qualitatively similar to those produced by 5,6-DHT.
For autoradiographic studies and for in vivo and in virro metabolism, both
5,6-DHT and 5,7-DHT were synthesized as l.lC-labeled compounds. The reac-
tion of 5,6- and 5,7-dibenzyloxygramine methiodide with [14C]NaCN gives the
two 'IC-labeled dibenzyloxyindoleacetonitriles. This reaction, which is normally
performed with a great excess of NaCN,25 is achieved with stoichiometric
cyanide quantities in a yield of approximately 60%. [d4C]-5,6-DHT and
[o-14C]-5,7-DHT and [curboxy-W]-5,6- and [c~rboxy-~~C]-5,7-dihydroxyindole-
acetic acid are obtained from the nitriles. The specific activities of the 14C-
labeled DHTs amounted to up to 25 mCi/mmol.

CHEMICAL PROPERTIES OF 5,6- AND 5,7-DIHYDROXYTRYPTAMINE

Both 5,6-DHT and 5,7-DHT are capable of damaging serotonergic neurons,

but the chemical properties of the two compounds differ markedly in some
respects.
One hour after intraventricular administration of [14C]-5,6-DHT, twice as
much radioactivity is found in the insoluble residue of perchloric acid-homoge-
nized brain tissue than in the case of animals treated with [14C]-5,7-DHT.32
In in vitro studies, Rotman et aL3"found even a higher proportion of 5,6-DHT-
derived radioactivity irreversibly bound to bovine serum albumin.
Only after intraventricular injection of 5,6-DHT are melanin-like pigments
deposited in the brain;34 this finding can be interpreted as an indication for
the in vivo formation of quinoid intermediates in the case of 5,6-DHT metabo-
lism. However, the possibility also exists that 5,7-DHT is oxidized to quinoid
33

In addition, spectroscopic investigations point to another form of reactivity

in the case of 5,7-DHT. Unlike 5,6-DHT, 5,7-DHT shows in buffer at pH 7.0
a relatively long-wave ultraviolet absorption maximum at 350 nm (FIGURE 4);
this absorption does not occur at low pH and if the two hydroxyl groups are
etherified (FIGURE 5 ) . The intensity of the maximum at 350 nm increases with
pH (FIGURE6 ) . The existence of isosbestic points can be explained by a
pH-dependent equilibrium between different keto-enol tautomeric forms of
5,7-DHT (FIGURE 7 ) . The mesomeric fi-diketo anion, which is formed from
the tautomeric species b and d through proton removal, can be expected to
undergo nucleophilic reactions. 5,6-DHT does not exhibit such spectroscopic
behavior, because it has no resorcinol-like structure. The concept of keto-enol
tautomerism in 5,7-DHT is supported by nuclear magnetic resonance spectro-
scopic data. In deuterium oxide, the two aromatic protons in positions 4 and
6 of 5,7-DHT creatinine sulfate are immediately replaced by deuterium; only
30 Annals New York Academy of Sciences

FIGURE4. Ultraviolet spectra of 5,6-DHT creatinine sulfate.2H20 (I), 4-hydroxy-

5-methoxytryptamine hydrochloride (11), and 5,7-DHT creatinine sulfate (III) , in
buffer at pH 7.0.
 Schlossberger: Some Indole Derivatives 31

3
w
m
-
0

k
2 Oh

1
220 240 260 280 300 320 340 mp
FIGURE 5 . Ultraviolet spectra of 5,6-dibenzyloxytryptaminehydrogen oxalate (I),
4-benzyloxy-5-methoxytryptamineformiate (II), 4,5-dibenzyloxytryptamine hydrogen
oxalate (In),and 5,7-dibenzyloxytryptaminehydrogen oxalate (IV) in ethanol.
32 Annals New York Academy of Sciences

I
I 1 I 1
400 350 300 210
nm
FIGURE 6. Changes in ultraviolet absorption of 5,7-DHT creatinine sulfate with pH:
2,1;pH 4, II;pH 5, In; pH 6, IV; pH 7, V; pH 8, VI.

H:Qq -
C%CH?NH2 :)wcH2-cH2-NH2

H H
0 0
C d
7. Keto-enol tautomeric forms of 5,7-DHT.
FIGURE
 Schlossberger : Some Indole Derivatives 33

the proton in position 2 is detectable. Neither with 5,6-DHT nor with 5-HT
creatinine sulfate was any exchange of aromatic protons detected in deuterium
oxide, even after 24 hr.
Another difference in the behavior of the two dihydroxytryptamines is
observed when the two compounds are dissolved in buffer solutions at physio-
logic pH at 37" C. While a dark precipitate becomes visible in the 5,6-DHT-
containing solution, the 5,7-DHT-containing solution slowly attains a blue-violet
color. The same result is observed when oxygen is passed through such solu-
tions in buffer at p H 7.4. When the ultraviolet absorption of these solutions

FJGURE 8. Reaction of 5.7-DHT creatinine sulfate in buffer at pH 7.4 with oxygen:

ultraviolet absorption spectra after 0 rnin (I), 40 rnin (II), 80 rnin (III), 160 rnin
(IV), and 10 hr (V)reaction times.

is studied over a longer period of time, initially a decrease of the extinction

of the absorption maximum at 299 nm is seen in case of 5,6-DHT, followed
by a broadening and leveling off of the absorption curve. After a few hours,
only the spectrum of creatinine is visible. In the case of 5,7-DHT, the absorp-
tion maximum at 350 nm disappears with time, whereas the intensity of the
maximum at 300 nm increases (FIGURE 8). and a low and flat maximum is
formed in the visible range of the spectrum at 520 nm. No precipitate can be
seen in such solutions. The ultraviolet spectra of 5,7-DHT measured at different
times show isosbestic points. We are now testing to determine whether this
reaction leads to defined products.
34 Annals New York Academy of Sciences

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