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EYPERFORIN AS A POSSIBLE ANTIDEPRESSANT COMPONENT OF

EIYPERICUM EXTRACTS

S.S. Chatterjee’, SK. Bhattacharya2, M. Wonnemann3, A. Singer3 and W.E. Mtiller3*

’ Pharmacology Department, Dr. Willmar Schwabe GmbH & Co., D-76227 Karlsruhe, Germany,
2 Pharmacology Department, Institute of Medical Sciences, Banaras Hindu University,
Varanasi, India, 3Department of Pharmacology, Biocenter University of Frankfurt,
D-60439 Frankfurt, Germany

(Rtccived in final form May 25,1!%8)

Summary

We demonstrate that the phloroglucinol derivative hyperforin is not only the major
lipophilic chemical constituent of the medicinal plant Hypericum perforatum (St.
John’s wort) but also a potent uptake inhibitor of serotonin (5-HT), dopamine
(DA), noradrenaline (NA), GABA and L-Glutamate with IC~Ovalues of about
0.05-0.10 &ml (5-HT, NA, DA, GABA) and about 0.5 @nl (L-glutamate) in
synaptosomal preparations. Furthermore, potencies of two different hypericum
extracts in two conventional pharmacological paradigms useful for the detection of
antidepressants (behavioral despair, learned helplessness), closely correlate with
their hyperforin contents. In addition, most till now known neuropharmacological
properties of the clinically used hypericum extracts can also be demonstrated with
pure hyperforin. It appears, therefore, that this non-nitrogenous constituent is a
possible major active principle responsible for the observed clinical efficacies of the
extract as an antidepressant and that it could also be a starting point for drug
discovery projects engaged in the search of psychoactive drugs with novel mode of
action.

Key Word: Hypericumextract,antidepressantactivity,hyperforin

Several herbal products widely used and recommended in Germany for treatment of nervous
disorders and sleeplessness contain hydroalcoholic extracts of the medicinal plant Hypericum
perforatum (St. John’s wort) (1). Such therapeutic uses of the extracts have long been known
and recent well-designed clinical trials have demonstrated their antidepressant activities (for
reviews see 2,3). A recent meta-analysis of the existing double-blind studies (3) demonstrates
that these extracts are more effective than placebo and similarly effective as some standard
antidepressant drugs like amitrityline, imipramine, and maprotiline for the treatment of mil’dto
moderately severe depressive disorders (2,3,4). Furthermore, it has also been shown that
treatments with such extracts seem to be devoid of major side effects typical for the tricyclic
antidepressants (TCA) or for the specific serotonin reuptake inhibitors (SSKI) and less
expensive (3,4,5,6,7). Although such reports clearly suggest that exploitation of this medicinal

l To whom correspondence should be addressed

500 Hyperforin as Antidepressant Agent Vol. 63, No. 6, 1998

plant could not only eventually lead to more rational uses of the hypericum extracts for the
treatment of depression, but also could be helpI% in the search of antidepressant molecules with
potentially novel mechanisms or mode of action Prerequisites for such goals are, however, the
knowledge of active antidepressant constituents of Hypericum perforatum and of the possible
mechanisms involved in their observed clinical effects.

Recent reports from our laboratories and elsewhere (89, IO) demonstrate that a
therapeutically used hypericum extract (Li 160) like conventional antidepressants, concentration
dependently inhibits uptake of serotonin (5HT), noradrenaline (NA), and dopamine (DA) in
synaptosomal preparations. The IGo values of the extract as inhibitor of monoamine uptake
determined in these studies were at least more than 100 times lower than those for the inhibition
of MAO-A or MAO-B (10). In addition, analogous to many other antidepressant drugs, chronic
oral treatment of rats with the extract also led to adaptive changes in B-adrenergic and 5-HT2-
receptor densities in their frontal cortex (10). Doses of the extract needed to observe such
effects were similar to those used by others for the demonstration of antidepressant like
activities of the same or other hydroalcoholic hypericum extracts in behavioral models
(11,12,13). Such observations not only suggested that the biochemical mechanisms of action of
hypericum extract might be similar to that of other antidepressant drugs like TCA’s or SSRI’s,
but also suggested that conventional in vitro and in vivo experimental models for
antidepressants could as well be uset?rl for proper characterization of the active constituents of
the extracts. Several attempts using activity guided fractionation procedures and a synaptosomal
uptake model were, however, unsuccessful. They demonstrated only that hypericines are not
synaptosomal uptake inhibitors and that after fractionations, the effects of the extracts
diminished and/or were almost evenly distributed in various fractions,

Such difficulties were, however, not encountered in another project dealing with the same
medicinal plant but searching for its active constituents responsible for other traditionally known
(14) and recommended (15) therapeutic uses, i.e., for the treatment of dyspepsia and as an
ointment for healing wounds and bacterial infections. Use of concoctions of the medicinal plant
in olive oil, commonly known as oleum hyperici or “Johanniskrautol” are recommended for such
purposes (15). For pharmacological studies, therefore, several types of lipophilic extracts were
prepared and analyzed for their known chemical constituents and possible biological activities.
Such efforts revealed that an oily extract obtainable by supercritical extraction with carbon
dioxide (CO& potently and concentration dependently inhibited contractions of isolated guinea
pig ileum preparations induced by 5-HT and several other spasmogens (16). Such extracts were
found to be devoid of hypericines and most other constituents of the hydroalcoholic extracts
and were concentrated in hyperforin (16) (for structure see fig. 1). Although antibacterial
properties of hyperforin have been known since long (for review see 17) no other biological
effects of this unique phloroglucinol derivative have been reported before. It was, therefore, of
interest to test whether this constituent of the hydroalcoholic extract was responsible for its
observed antidepressant like effects in experimental models. The observations reported here
demonstrate that such indeed is the case and suggest further that efforts should be made to
exploit the antidepressant potential of this naturally abundant compound.

Methods
Behavioral despair

The procedure described by Porsolt et al. (18) was followed with slight modifications. The
dimension of the rectangular polypropylene swim vessel was 45 x 40 x 30 cm, which was tilled
with tap water (23 % 2°C) up to a level of 20 cm. Rats were forced to swim individually for 10
Vol.63, No. 6, 1998 Hyperforinas AntidepressantAgent 501

min and thereafter the total period of complete cessation of swimming with the head floating
just above water level was determined during subsequent 5 min period. The data presented here
were generated with animals treated with the agents for three consecutive days whereupon the
last dose was given 1 h before the experiments. Such a dosing schedule was fixed after several
initial experiments in which the effects of a single and of repeated doses of the agent on body
temperature, muscle relaxation, locomotor behavior, apparent respiratory depression and swim
performance were studied. The maximal doses of the agents used in this study were well
tolerated i.e. no changes in these parameters, except the ones reported here, were observed. The
extracts and their vehicle (0.3 % carboxymethyl cellulose; 10 ml/kg) were administered orally
whereas imipramine - the reference drug - was applied intraperitoneaily. All data presented are
the mean values of four experiments conducted by the same observer in the same laboratory
within 6 weeks. Each experiment consisted of 3 animals in control group and 2 in each treated
group.

HYPERFORIN

Structure of the phloroglucinol derivative hyperforin identified as the main single chemical
constituent of Hypericum COz-extract.

Learned helplessness

Choice of experimental conditions and dosing schedule was based on the observations made
in the behavioral despair test and historical data on the behavior of the animals of our colony in
this test. Groups of rats were at first subjected to footshock (60 scrambled shocks; 0.8 mA, 15
set every min) in a two compartment jumping box (Techno, India) with the escape route to the
unelectrified safe chamber closed. The concurrently run control group, which in later days
received vehicle only, was kept similarly in the closed chamber but did not receive shocks. The
following days, i.e., on day 2,3 and 4 after start of the studies, the animals were treated with the
test agents or vehicle. On day 3, i.e., 48 h after the initial shock challenge and I h after dosing,
the animals were again subjected to the inescapable shock. Performance of the rats in the shock
avoidance paradigm was quantified on day 4 (1 h after last dosing) using the same chamber,
whereupon they were first allowed to acclimatize in the shock chamber for 5 min (without
shocks). In total 30 avoidance trials with inter-trialintervals of 30 set were used. During the first
502 Hyperforinas AntidepressantAgent Vol. 63, No. 6, 1998

3 set of a trial a conditioning buzzer stimulus (US) was presented which immediately followed a
0.8 mA electric footshock (US) for 3 sec. The avoidance response, characterized by an escape
to the safe chamber during CS was quantified and failure to exhibit escape response during US
was assessed as escape failure. In general, the guidelines described by Seligman and Beagley
(19,20) were followed and inhibition of the escape failure was used as the measure of
antidepressant activity. The study design was analogous to that described for behavioral despair
test, except that only two animals were included in the control group of each experiment. In
addition, one experiment with four control animals with shocks and 6 such without shocks was
conducted during the days between the first and last two experiments.

Synaptosomal uptake of neurotransmitters

Synaptosomal preparations from rat striatum (male Wistar rats, 180 g) were used for DA-
uptake studies. For other neurotransmitter uptake experiments similar preparations were
obtained from whole brain (for 5-HT and GABA), occipital cortex (for NA) or forebrain (for L-
glutamate) of mice (female NMRI, 18 g) (10). The tissues were homogenized in 15 ml (10 ml
for rat striatum) ice cold sucrose solution (0.32 M) and diluted with 10 ml (5 ml for rat
preparations) of the homogenizing medium. The nuclear fraction was eliminated by
centrifugation (10 min at 750 xg, 0-4°C) and the supematant was centrifuged (20 min at 17400
x g; 0-4’C) to obtain the crude synaptosomal pellets. It was then suspended in ice cold HEPES-
buffer (NaCI: 150; HEPES: 10; KCl: 6.2; Na&IPOd: 1.2; MgSO4: 1.2; glucose: 1OmM. pH 7.4
at 37°C) aliquoted in glass tubes and incubated at 37°C for 10 min and then cooled on ice. The
3H labeled tracers (S-HT: 2.94, NA: 3.86, DA: 2.0, GABA: 0.5, L-Glutamate: 1, final
concentrations in nM) were then added and the uptake started by incubation at 37°C (10 min for
NA and 4 min for all others). The probes were then cooled and immediately filtered and washed
twice with 3 ml (each time) cold buffer solution. The filters were then placed in plastic vials
containing 4 ml Quickszint 402 (Zinsser, Frankfurt, FRG) and radioactivity counted in a
Beckman liquid scintillation counter. Nonspecific uptake was estimated in parallel probes
containing unlabelled neurotransmitters (1 mM of 5-HT, DA, GABA or L-Glutamate) or 1 pM
desipramine (for NA). For 5-HT-, NA- and DA-uptake experiments 0.1 % ascorbic acid and 10
uM pargylin were added in the incubation medium.

AL4 O-assays

MAO-A and MAO-B activities were assayed in whole mouse brain homogenates (10) with
14C-5-HT (65 PM) and 14C-phenylethyl amine (10 PM) as substrates and according to the
methods described by Young et al. (21).

Materials and other details

Animals

For the behavioral studies conducted in the Pharmacology Department of Banaras Hindu
University, adult male rats (Charles Foster; body weight between 150-180 g) bred in the
institute animal house were used. The animals were maintained in groups of 5 per cage at 25 f
2°C and 45-55% relative humidity. They were brought to the laboratories at least one week
before the experiments and had always free access to commercial chow pellets and tap water,
which were removed only during experimental procedures. All in vivo studies were started at
least 3 h after the start of light period (in a 12 h dark / 12 h light cycle) and all experiments were
conducted between 10 h and 16 h. The animals used in biochemical studies conducted in
Germany, were purchased from Harlan-Winkelmann (Borchen, FRG).
Vol. 63, No. 6, l!XB Hyperforin as Antide~t Agent 503

All animals used in this study were maintained in our laboratories for at least one week in
groups of 5 per cage at 25 f 2°C and 45-55 % relative humidity. They had free access to
commercial chow pellets and tap water, which were removed only during experimental
procedures. Principles of laboratory animal care (NIH publication no. 85-23; revised 1985) were
always followed. All behavioral studies were started at least 3 h after the start of light period (in
a 12 h dark / 12 h light cycle) and all experiments were conducted between 10 h and 16 h.

Extracts and chemicals

The ethanolic and supercritical CO*-extracts of hypericum perforatum and pure hyperforin
were supplied by the Research Department of Wilmar Schwabe GmbH, Karlsruhe, FRG. The
methanolic extract used in some biochemical experiments was the same as described in our
earlier report (10). The hyperforin samples were stored in dark at 4°C in nitrogen atmosphere
whereas the ethanolic extract was stabilized by the addition of ascorbic acid (5 % by weight of
the extract). Stabilities of hyperforin and hypericines were checked by repetive quantitative
HPLC analysis of the samples during the course of the study. Hyperforin contents of the
methanolic, ethanolic and COz-extracts were 1.5; 4.5 and 38.8 % respectively. Hypericines were
not detectable in the CO*-extract and its contents in methanolic and ethanolic extracts were 0.3
and 0.23 % respectively.

The following radiochemicals were from NEN (Dreieich, PRG): [7,8-3H]Dopamine (specific
activity: 46 Unmol), [ 1,2-3H (N) ]5-Hydroxytryptamine creatine sulfate (30 Ci/mmol), Levo-
[7-3H (N)]norepinephrine (15 Ci/mmol), [2,3-3H (N)]y-Aminobutyricacid (90 Ci/mmol), L-
[2,3-3H (N)]-Glutamic acid (22 Ci/mmol), 5-Hydroxy[side-chain-2-“C]tryptamine creatine
sulfate (56 mCi/rnmol) and 13-[ethyl-l-‘4C]-phenylethy1amine HCl (42 mCi/mmol). All other
reagents and chemicals were of highest purity obtainable from Sigma (Munich, PRG).

Results

Behavioral despair

In initial experiments acute administration of even high doses of the ethanolic or of the COZ-
extract did not reveal any antidepressant like effects in this test. Repeated oral administration of
either of them for three consecutive days did, however, dose dependently reduce the immobility
time of rats. Such observed effects of the extracts are summarized in Table I. The dose response
relationships of the extracts were analysed using a closed testing procedure (22) with ANOVAs
at each step and a multiple error rate of a = 0.05 was controlled for every parameter.
Stastistical data in Table I refer to the results of the final step of the closed testing procedure
comparing the corresponding group with the vehicle treated group. Differences marked were
significant at the multiple error rate. Additionally, a non-parametric trend test (23) applied to
analyse the dose response relationship indicated p < 0.0001 for both extracts.

The dose-effect curves for the two extracts obtained from the data in Table I are shown in
fig. 2A. In this figure % inhibition of immobility was calculated using the data for the vehicle
treated control group as 100 %. Also included in the figure are the theoretical dose-effect
curves of hyperforin calculated on the concentration of hyperforin in the tested extracts and
assuming that it is their only active constituent. It can be seen that the curves of the two extracts
are almost parallel to each other and that the corresponding extrapolated ones for hyperforin are
almost superimposable.
 X escape failure % Inhibition( Immobility)
% 2) g 2 g 0 s !z $2 g s 8
L

m
Vol. 63, No. 6, 1998 Hyperforin as Antidepressaat Agent 505

TABLE I

Group (route of Dose

administration) (mgkglday)

Vehicle (p.0.) ---

Ethanolic
extract (p.0.) 133.7 f 1.91*** 15.0* 0.65*** 1 4.5 f 0.32***
115.4 *2.60*** 1 10.6* 0.68*** 1 5.2*0.24***

COz-extract I 5 153.9 f 2.43* 19.2* 0.75 1 3.5 f 0.32*** 1

(P.0.) 131.1 f 3.06*** 14.7* 0.75*** I 4.9*0.29***
119.43 f 3.01*** 10.9 f 0.69*** 1 5.2 f 0.24***

103.7 f 2.55*** I 9.9* 0.54*** 5.2*0.26***

Effects of ethanolic and COz-extracts of Hypericum perforatum and imipramine in the

behavioral despair test and the learned helplessness paradigm in rats. Vehicle treated
control groups consisted of 12 animals each whereas in all other groups 8 animals per
group and dose was used. The control group (not receiving inescapable shock) in the
learned helplessness model consisted of six animals. The mean (* SEM) number of escape
failures (EF) and avoidance responses (AR) in this group were 5.3 f 2 and 6.2 f 0,3
respectively. Statistically significant differences from the corresponding vehicle treated
groups are marked with *, ** or ***, representing p < 0.05; < 0.01 or < 0.001
respectively. All values given in the table are mean f SEM.

Learned helplessness

In the learned helplessness paradigm the animals are initially exposed to uncontrollable stress
by confining them in a shock chamber from which they cannot escape. When such animals are
later placed in a situation in which shock is avoidable via escape in a safe chamber, the animals
do not respond normally. It has been postulated that the escape failures in rats observed in the
escapable shock situation represent depressive behavior (19). Under the experimental conditions
used in this study control rats with prior experiences of inescapable shocks exhibited marked
increases in escape failures (72.5 %) as compared to those with no such prior experiences (22.4
%). As shown in Table I, the escape failures in the ethanolic extract treated groups significantly
and dose dependantly decreased after 150 and 300 mg/kg/day. Such effects of the CO*-extract
were also apparent after 15 or 30 mg/kg/day dose regimens. As in the behavioral despair test,
oral doses of 30 mg/kg/day of CO*-extract or 300 mg/kg/day of ethanolic extract was almost
equieffective to 10 mg/kg/day (ip.) dose of imipramine. For statistical and other analysis the
same procedures as described for behavioral despair was used. The dose-effect curves are
shown in Iig. 2B.

Synaptosomal uptake of neurotransmitters andMAO-inhibition

In earlier reports from our laboratories the in vitro ICSOvalues of a methanolic extract as
inhibitor of noradrenaline, serotonin or dopamine uptake in synaptosomal preparations were
506 Hyperforin as Antidepressant Agent Vol. 63, No. 6, 1998

shown to be between 0.85 to 4.47 ug/ml (810). The values for the COz-extract and hyperforin
determined under identical conditions are summarized in Table II. In addition, the in vitro
effects of the extracts and pure hyperforin on synaptosomal GABA- and L-Glutamate uptake
are also summarized in this table. It is apparent that hyperforin, like the two hypericum extracts,
is a potent uptake inhibitor of not only the conventional biogenic amines but also of amino acids
like GABA and L-Glutamate. Although the I& values of pure hyperforin and of both extracts
for L-Glutamate uptake were higher than the corresponding values for other neurotransmitters,
the rank orders of potencies of the extracts were in no way comparable to each other or to that
of the pure constituent. Such orders for hyperforin, COz-extract and methanolic extract were
NA 2 DA > GABA z 5HT >> Glut.; DA > GABA > NA = 5-HT >> Glut. respectively. As
shown in Table II, the efficacies of the extracts in none of the neurotransmitter uptake systems
were strictly correlated to their hyperforin contents and the ratios of the corresponding ICSO
values of the two extracts were never close to 25.9 i.e. to the ratio of hyperforin contents in
COz-extract and methanolic extract. Such observations demonstrate that hyperforin is a potent
inhibitor of neurotransmitter uptake present in the medicinal plant, but that it is not the only
active constituent, It is alternatively possible that other constituents present in the extracts
modulate the efficacy of hyperforin.

Neither hyperforin nor the C02-extract in concentrations up to 50 &ml inhibited MAO-A

or MAO-B activities (results not shown). In earlier studies (10) the 1% value of methanolic
extract for inhibiting these enzymes were much higher than 100 l&ml. Such results demonstrate
that although MAO-inhibitors are present in the medicinal plant, their inhibitory potencies or
concentrations are lower than those of the components responsible for the inhibition of neuro-
transmitter uptake in synaptosomes.

TABLE II

I KS0 @g/ml)

Uptake system Hyperforin I C02-extract Methanolic Extract

5-HT 0.11010.024 (205) 0.26 f 0.06 (188) 2.43 +0.40* (67)

NA 0.043 + 0.013 (80) 0.25 f 0.08 (181) 4.47+ 2.05* (123)
DA 0.055 k 0.010 (102) 0.056 i (41) 0.85 k 0.14* (24)
GABA 0.099* 0.022 (184) 0.12 f 0.04 (87) 1.11 + 0.056 (31)
L-Glutamate 0.445 + 0.369 (830) 2.83 f 1.8 (2045 21.25 + 10.47 (586)

Effects of hypericum extracts and hyperforin on synaptosomal uptake of various

neurotransmitters. The mean (* SD) half-maximal inhibitory concentrations (I&) in
&ml given in the table were obtained by log-probit analysis of inhibition curves obtained
with 6-8 different concentrations of the agents, The numbers in parentheses represent the
corresponding mean nM concentrations of hyperforin which for each extract were
calculated from its concentration in the test solution used and assuming that it is their only
active constituent. The COI-extract and the methanolic extract contained 38.8 and 1.5 %
hyperforin respectively. For comparison the values for the methanolic extract marked with
* are reproduced from our earlier report (10).
Vol.63, No. 6, 1998 Hyperforin as AntidepressantAgent 507

Discussion

Observations reported here demonstrate that hyperforin is a potent but unspecific inhibitor
of the neuronal uptake of several biogenic amines and amino acid neurotransmitters and that it
does not possess relevant MAO-inhibitory activities. In addition, we also demonstrate that
efficacies of two completely different types of hypericum extracts in two behavioral paradigms
commonly used for prediction of clinical efficacy of antidepressants correlate nicely with their
hyperforin contents and not with other known constituents of the medicinal plant.

Amongst a wide variety of proposed and critically assessed in vivo animal models of
depression (24,25) the two most commonly used paradigms are learned helplessness (19,20)
and behavioral despair forced swim tests (18). A few reports have also demonstrated the
efficacies of standardized hypericum extracts ins these two animal models (11,12,13,26).

Our present behavioral observations not only confirm that hypericum extracts possess
imipramine like antidepressant activities, but also indicate that hypericines and many other
constituents of the medicinal plant not present in the CO*-extract can not be their major active
constituents. Until now we have been able to quantify hyperforin only in the COz-extract and
most other constituents of this extract still remain undefined. Although the efficacy of the
extracts tested correlate well with their hyperforin contents, definitive statements concerning the
efficacy of this component in behavioral models for antidepressants can be made only when the
results of studies with the purified constituent are available. Unfortunately, due to lack of
sufficient quantities of the agent needed for such in vivo studies, these conclusive experiments
could not yet be conducted. It appears certain, however, that the presence of hypericine and
other constituents (not extractable from the medicinal plant by supercritical COZ extraction) are
not essential requirement for the antidepressant activity of the extracts,

Depressive disorders are now well recognized as major public health problems in primary
health care setting (27,28). Despite extensive efforts and considerable progress made during the
past five decades, successful treatment of clinical depression with currently available therapeutic
agents can be achieved only in 65 to 75 % of patients and only 40 to 50 % achieve complete
recovery (29,30). Such a situation necessitates the development of more effective
antidepressants (30,3 1). The first generation of antidepressants, the TCA, discovered only after
fortuitous clinical findings with imipramine (32), are still widely used because of their familiarity
and low cost. Although many newly developed second generation antidepressants, like the
SSRI’s, have reduced the risks of side effects of TCA’s, they have made little impact on
improving the effectiveness of treatment (30,3 1,33). Search for new molecules and targets for
antidepressant drug discovery projects remains, therefore, a continuing challenge for modem
psychiatry. In view of the situation, it has been pointed out that like in various other therapeutic
areas (34,35) investigation of purported therapeutic activity of traditional herbal products may
be a good chance for uncovering new mechanisms and novel treatments for affective and other
CNS disorders (36,37).

Extracts of Hypericum perforatum are the mostly prescribed herbal drugs for the treatment
of depressive disorders (1,2,3). Currently, most therapeutically used hypericum extracts are
standardized in their contents of hypericines, a group of characteristic components of the
medicinal plant. Such practices rely on recommendations of the German Health Authorities
dating from 1984 (15) and are based on reports suggesting monoamine oxidase (MAO)
inhibitory activities of such components (38,39). Since then, several laboratories could not
confirm such properties of hypericines and have demonstrated that moderate to mild MAO-
 Hyperforia as Antidepressant Agent Vol. 63, No. 6, 19%

inhibitory effects of the extracts observed in vitro are due to the presence of xanthiones or
flavonoids in the extracts (10,14,40,4 1,42,43). Such effects of the extracts or their components
have, however, never been demonstrated in experimental animals after oral dosing, and reports
of antidepressant like activities of these constituents in animal models are also not available.

A recent report demonstrates, though, that hypericines administered together with some yet
undefined flavonoid constituents of hypericum extracts reveal antidepressant like activities in
behavioral models (12). In view of the observations that the COz-extract (devoid of hypericines
and flavonoids) also possesses antidepressant activities, it can be speculated that several
constituents i.e. hyperforin, hypericines and some yet undefined flavonoids, could be involved in
the observed clinical efficacies of Hypericum extracts. The concentrations of hyperforin in
Hypericum perforatum is, however, well known to be much higher (> 2.5%) than those of
hypericines (< 0.4%) or other till now defined (and extractable) constituent of the herb (17). We
speculate, therefore, that this acylphloroglucinol derivative is the major active component of the
extracts. Further studies are, however, necessary to confirm this suggestion.

All pharmacological studies conducted with pure hyperforin have also been performed with
the different extracts described here. Except for MAO-inhibitory and GABA-A-receptor binding
activities (41,44) of the alcoholic extracts, no other in vitro effects of other constituents of
Hypericum perforatum were observed by such efforts. In most in vitro studies, however, the
efficacies of the extracts could never be accounted for only by their hyperforin contents. In view
of the situation we conclude that in addition to hyperforin, the medicinal plant contains some
not very lipophilic constituents with high affinity towards GABA-A receptors and weak MAO-
inhibitory properties. In addition, some lipophilic constituents structurally analogous to
hyperforin, either possess hyperforin like activities or modulate its pharmacological activities.

It has recently been pointed out (45) that although the majority of world’s health care
services uses herbal medicine, and the beneficial effects of not only hypericum extracts but also
of many other herbal drugs have been demonstrated in proper clinical trials, established medical
and research communities are skeptical about their therapeutic usefulness. To our judgement,
broader acceptance and proper or rational uses of such botanical medicines is possible only
when the active constituents and their modes of action are known. Our efforts to clarify the
situation with various medicinal plant extracts indicate, though, that almost in all cases several
components and mechanisms are involved in their therapeutic actions. The case of hypericum
extracts does not also seem to be much different. It must be pointed out though, that it was only
the effort made to clarify the mode(s) of action(s) of hypericum extracts that led us identify the
potential antidepressant activity of hyperforin. Our observations, thus, not only open up
possibilities for rationalizing therapeutic possibilities with hypericum extracts, but also can
facilitate the search of novel antidepressants with unique mechanisms of action.

Acknowledgements

We thank Dr. C. Erdelmeier and his colleagues for kindly supplying us the extracts, pure
hyperforin and for constantly helping us in solving the analytical and stability problems. Thanks
are also due to Prof. J. Holzl for supplying us the very first sample of hyperforin and to
Lichtwer Pharma (Berlin) for the methanolic extract. The expert secretarial assistance of E.
Nuding, the excellent technical assistance of C Schafer, and the support of the Fonds der
chemischen Industrie (Germany) are gratefully acknowledged.
Vol. 63, No. 6, 1598 Hyperforin
as Antidepressant
Agent 509

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