Karin Woelkart1

Wei Xu2
Ying Pei2
Alexandros Makriyannis2
Robert P. Picone2
The Endocannabinoid System as a Target for Alkamides
Rudolf Bauer1 from Echinacea angustifolia Roots

Rapid Communication
Abstract FAAH: Fatty acid amidohydrolase

Downloaded by: Collections and Technical Services Department. Copyrighted material.

PMSF: phenylmethanesulfonylfluoride
Alkamides are the major lipophilic constituents of Echinacea AEA: arachidonylethanolamide, anandamide
angustifolia roots. Due to their structural similarity with ananda- AA: arachidonic acid
mide, we have evaluated their ability to bind to rodent cannabi- IL: interleukin
noid receptors CB1 and CB2 by a standard receptor binding assay MCP: monocyte chemoattractant protein
using [3H]CP-55,940 as a radioligand. The alkamides exhibited BSA: bovine serum albumin
selective affinity especially to CB2 receptors and can therefore TNF-a: tumor necrosis factor-alpha
be considered as CB ligands. Most of the alkamides showed MAPK: mitogen-activated protein kinase
good metabolic stability as indicated by the similarity between JNK: jun N-terminal kinase
affinity to CB1 determined in the presence/absence of the pro- cAMP: cyclic adenosine monophosphate
tease inhibitor PMSF. It is suggested that CB2 interactions may CREB-1: responsive element binding protein-1
be the molecular mode of action of Echinacea alkamides as im- ATF-2: activating transcription factor-2
munomodulators. NF-kB: nuclear factor kappa B
Tris: Tris-(hydroxymethyl)-amino-methane
Abbreviations CP55940: (±)-cis-3-[2-hydroxy-4(1,1-dimethyl-heptyl)phenyl]- 701
CB1: cannabinoid receptor 1 trans-4-(3-hydroxypropyl)cyclohexanol
CB2: cannabinoid receptor 2

Echinacea species are used as herbal immunomodulators world-
wide [1]. Despite the popularity of the herb and the many phar-
macological and clinical studies, the molecular mechanism of ac-
tion of Echinacea remains poorly understood [2]. So far, com-
pounds from the classes of caffeic acid derivatives, flavonoids,
polyacetylenes, alkamides, pyrrolizidine alkaloids, polysacchar-
ides and glycoproteins have been isolated from Echinacea spe-
cies. Alkamides, the major lipophilic constituents; can be found
in high concentrations in the aerial parts of E. purpurea, E.
pallida and E. angustifolia, and in the roots of E. purpurea and E.
angustifolia [1], [3]. Recently Gertsch et al. [4] have shown a
modulation of TNF-a gene expression and multiple signal trans-
duction pathways for Echinacea alkamides and postulated a
mechanism related to cannabinoid receptors. In parallel, due to
the structural similarity of those alkamides with anandamide
(Fig. 1), we have investigated the receptor binding and now re-

Affiliation
1
Institute of Pharmaceutical Sciences, Department of Pharmacognosy, Karl-Franzens-University Graz, Graz,
Austria
2
Center for Drug Discovery, University of Connecticut, CT, USA

Correspondence
Rudolf Bauer ´ Institute of Pharmaceutical Sciences ´ Department of Pharmacognosy ´
Karl-Franzens-University Graz ´ Universitätsplatz 4/I ´ 8010 Graz ´ Austria ´ Phone: +43-316-380-8700 ´
Fax: +43-316-380-9860 ´ E-mail: rudolf.bauer@uni-graz.at

Received March 29, 2005 ´ Accepted May 25, 2005

Bibliography
Planta Med 2005; 71: 701±705 ´  Georg Thieme Verlag KG Stuttgart ´ New York
DOI 10.1055/s-2005-871290 ´ Published online August 11, 2005
ISSN 0032-0943

Fig. 1 Structure of anandamide, the endogenous ligand of CB recep-
tors.
port the affinities of 12 alkamides isolated from E. angustifolia
roots to CB1 and CB2 receptors and metabolic stability to fatty
acid amidohydrolase (FAAH).
Twelve alkamides have been isolated from the roots of E.
angustifolia and were identified as tetradeca-2E-ene-10,12-diy-
noic acid isobutylamide (1), undeca-2E/Z,4Z/E-diene-8,10-diyno-
Rapid Communication
ic acid isobutylamides (2), undeca-2E/Z-ene-8,10-diynoic acid iso-
butylamides (3), dodeca-2E,4Z-diene-8,10-diynoic acid isobutyl-
amide (4), dodeca-2E-ene-8,10-diynoic acid isobutylamide (5), do-
deca-2E,4Z-diene-8,10-diynoic acid 2-methylbutylamide (6), do-
deca-2E,4Z,10Z-triene-8-ynoic acid isobutylamide (7), dodeca-
2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides (8), pentadeca-
2E,9Z-diene-12,14-diynoic acid isobutylamide (9), dodeca-2E-
ene-8,10-diynoic acid 2-methylbutylamide (10), dodeca-2E,4E,
8Z-trienoic acid isobutylamide (11), and dodeca-2E,4E-dienoic
acid isobutylamide (12) by their chromatographic behaviour and
MS data by comparison with the literature [3], [5] (Fig. 2). The al-
kamides were tested for their ability to bind to CB1 and CB2 can-
nabinoid receptors. A well-described filtration assay was em-
ployed in which specific binding of the tritiated ligand (±)-cis-3-
[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxy-
propyl)cyclohexanol (CP-55,940) to cannabinoid binding sites was
displaced by increasing concentrations of the ligand under inves-
tigation [6]. The results of the standard receptor binding assay
702 using [3H]CP-55,940 as the radioligand are summarized in Table 1.
Fig. 3 and 4 depict the affinities of the alkamides from E.
angustifolia for the two cannabinoid receptors. Concerning se-
lectivity, pentadeca-2E,9Z-diene-12,14-diynoic acid isobutyl-
amide (9) showed the highest affinity for CB1 with a Ki of
2.0 mM, followed by 10 with a Ki of 4.1 mM, while compound 1
with a Ki of 1.9 mM was the most selective and most affine li-
gand for CB2.
Table 1 Affinities of Echinacea alkamides to CB1 and CB2 receptors, reported as Ki values (95 % confidence intervals), obtained by a standard
receptor binding assay using a radioligand
Compound CB1 rat Ki (mM) CB 1 rat + PMSF Ki (mM)
1 24.644 (18.9 ± 32.1) not tested
2 42.107 (33.0 ± 53.6) 17.596 (6.3 ± 49.2)
3 79.372 (53.1 ± 118.6) 48.775 (24.1 ± 98.8)
4 16.7 (12.47 ± 22.28) 13.688 (8.9 ± 21.1)
5 61.109 (41.2 ± 90.7) 27.064 (10.2 ± 71.9)
6 25.172 (18.1 ± 34.9) 15.818 (5.9 ± 42.7)
7 41.697 (31.8 ± 54.7) 24.76 (11.4 ± 53.6)
8 10.475 (8.4 ± 13.1) 11.271 (6.5 ± 19.6)
9 not tested 2.035 (1.6 ± 2.7)
10 6.731 (4.7 ± 9.6) 4.116 (2.8 ± 6.1)
11 10.994 (6.9 ± 17.3) 8.379 (5.5 ± 12.7)
12 7.554 (4.2 ± 13.5) 6.147 (4.8 ± 7.8)
Woelkart K et al. The Endocannabinoid System ¼ Planta Med 2005; 71: 701 ± 705
As a second series of testing, membranes were treated with
10 mM phenylmethanesulfonyl fluoride (PMSF), a common pro-
tease inhibitor. By measuring the affinities of the ligands for the
receptor in the presence and absence of PMSF, we were able to
obtain a relative measure of their metabolic stability. Ananda-
mide possesses a short metabolic half-life, which has been
shown to be improved in the presence of PMSF [9]. The metabolic
stability of Echinacea alkamides are shown in Table 1 and Fig. 3.
In most cases, affinity to CB1 receptors was higher in the pres-
ence of PMSF indicating that alkamides are susceptible to hydro-
lysis via serine-dependent proteases. Furthermore we tested the
ability of Echinacea alkamides to inhibit fatty acid amidohydro-
lase (FAAH) in a previously described enzymatic assay. FAAH is
another endogenous enzyme responsible for the hydrolysis and
therefore inactivation of anandamide. Thus, inhibition of FAAH
would prolong the activity of CB1 and CB2 ligands. At the con-
centration of 25 nanomoles modest inhibition of FAAH was de-
tected with compound 1 at a level of nearly 23 % and only weak
Downloaded by: Collections and Technical Services Department. Copyrighted material.
inhibition was noted with 11 and 12 at a range of 14.4 % and
11.7 %, respectively.
Interestingly enough, compound 1 displayed the best affinity for
CB2, had the highest degree of CB2 selectivity and the best inhi-
bition of FAAH. Collectively, this is intriguing in light of the re-
ported colocalization of FAAH and CB2 on human dendritic cells
which are known to be involved in eliciting potent immune re-
sponses [10].
The CB1 receptor is ubiquitous but is very densely distributed
throughout various regions of the brain, where it is responsible
for the characteristic effects of cannabis, including catalepsy, de-
pression of motor activity, analgesia and feelings of relaxation
and well being. In the periphery, the CB2 receptor is known to oc-
cur mostly in immune cells, including T cells, B cells, natural kill-
er cells, macrophages, neutrophils, and mast cells [11], [12]. The
four cannabinergic membrane-bound proteins, including the
CB1 and CB2 receptors, fatty acid amidohydrolase (FAAH) and
the anandamide transporter, are current targets for the develop-
ment of novel medications for pain, immunosuppression, per-
ipheral vascular disease, appetite enhancement or suppression,
and mental illness [13]. Most of the Echinacea alkamides showed
CB 2 mouse Ki (mM)
1.867 (1.0 ± 3.4)
16.680 (11.7 ± 23.8)
66.395 (36.6 ± 120.5)
20.027 (12.5 ± 32.1)
47.224 (31.7 ± 70.5)
12.743 (9 ± 17)
18.191 (12.3 ± 26.9)
5.499 (3.7 ± 8.2)
18.96 (12.9 ± 27.9)
7.204 (4.6 ± 11.4)
4.569 (3.3 ± 6.3)
9.694 (6.8 ± 13.8)

Fig. 2 Structures of alkamides isolated from the roots of Echinacea an-
gustifolia.
affinities to CB2 receptors with Ki values lower than 20 mM, some
only five times less active than the endogenous ligand ananda-
Woelkart K et al. The Endocannabinoid System ¼ Planta Med 2005; 71: 701 ± 705
mide (Ki = 0.371 mM). Similar activities have been found for syn-
thetic alkamides [14].
Gertsch et al. [4] recently described a significant upregulation of
constitutive cAMP levels and a moderate inhibition of forskolin-
stimulated cAMP measured with dodeca-2E,4E,8Z,10E/Z-tetra-
enoic acid isobutylamide (8) at a concentration of 1 mM which
approximates our experimentally determined Ki value (5 mM)
for this alkamide. Echinacea alkamides also upregulate tumor
necrosis factor-a mRNA via activation of CB2 receptors without
expression of the protein in the early phase demonstrated on ki-
netic experiments and increased p38/mitogen-activated protein
kinase (MAPK) and jun N-terminal kinase (JNK) signaling, as well
as NF-kappa B and activating transcription factor 2/responsive
Rapid Communication
element binding protein-1 (ATF-2/CREB-1) activation. Combined
with the results presented here we can conclude that alkamides
can interact functionally as agonists with CB2 receptors. As de-
picted by Jbilo et al. [15] stimulation of CB2 receptors with the
Downloaded by: Collections and Technical Services Department. Copyrighted material.
cannabinoid ligand CP-55 940 leads to an increased expression
of two proinflammatory chemokine genes: monocyte chemoat-
tractant protein (MCP-1) and IL-8 in human promyelocytic leu-
703
Fig. 3 Selectivity of alkamides from Echinacea angustifolia for CB 1 re-
ceptor from rat membranes with and without PMSF, obtained by a
standard receptor binding assay using a [3H]CP-55,940 as the radioli-
gand and reported as mean Ki values [mM] with corresponding 95 %
confidence intervals determined from at least three independent ex-
periments.
Fig. 4 Selectivity of alkamides from Echinacea angustifolia for CB 2 re-
ceptor from mouse membranes, obtained by a standard receptor bind-
ing assay using a [3H]CP-55,940 as the radioligand and reported as
mean Ki values [mM] with corresponding 95 % confidence intervals de-
termined from at least three independent experiments.
 kaemia HL-60 cells. Sugiura [16] and Kishimoto [17] also pub-
lished an accelerated production of IL-8 and MCP-1 after induc-
tion with 2-arachidonoylglycerol (2-AG), whereas anandamide
failed to augment the production of chemokines. The observa-
tion that CB2 receptors modulate the expression of these two
major inflammatory chemokines and the fact that the alkamides
were found not only to be CB2 ligands but that they were able to
inhibit FAAH and thereby able to enhance endogenous cannabi-
nergic signalling by preventing metabolism of anandamide
might provide new insights towards a possible mode of action
of Echinacea preparations. As for other types of cytokines, several
investigators reported that 2-AG induces the inhibition of the
production of IL-6 in J774 macrophage-like cells [18] and the in-
hibition of TNF-a production in lipopolysaccharide (LPS)-stimu-
Rapid Communication
lated mouse macrophages [19]. It was also observed that endo-
genous cannabinoids like 2-AG and anandamide, as well as the
synthetic cannabinoids (+)WIN 55,212 ± 2, CP-55,940, and HU210,
inhibited the LPS-induced TNF-a release from rat microglial cells
in a concentration-dependent manner [20]. Furthermore by
stimulation of human monocytic leukaemia cells (THP-1 cells)
cannabinoids inhibited secretion of Il-1b and TNF-a via CB2 re-
ceptors. Specific CB2 receptor ligands therefore could be useful
anti-inflammatory agents, avoiding the neurotoxic and psychoac-
tive effects of CB1 receptor ligands [21]. Recently endocannabi-
noids have already been considered as immunomodulatory com-
pounds by Parolaro et al. [22]. Studies in human volunteers have
established an association between IL-8 and common cold symp-
toms. In rhinovirus infections there is a direct correlation between
the severity of common cold symptoms and the concentration of
IL-8 in the nasal secretion [23]. Therefore Echinacea alkamides
might mimic a rhinovirus infection and modulate the immune
system. There is also evidence that alkamide containing Echina-
704 cea preparations trigger effects on the pro-inflammatory cyto-
kine TNF-a and chemokine IL-8 in an ex vivo study and therefore
not only bind but also activate these CB2 receptors (data not
shown). Since bioavailability and fast absorption of alkamides
from Echinacea preparations after oral application have recently
been demonstrated [24], the suggested mechanism is reasonable
also for the in vivo situation.
Material and Methods
Roots from two-year-old plants of Echinacea angustifolia DC., As-
teraceae, were obtained from Heilpflanzen Sandfort GmbH & Co
KG, Olfen, Germany. The roots were identified by their morpholo-
gical and chemical markers. A voucher specimen (No. 2 030 817) is
deposited at the Institute of Pharmaceutical Sciences, Department
of Pharmacognosy, University of Graz. The roots have been extract-
ed with supercritical CO2 by Finzelberg, Andernach, Germany
(drug:extract ratio 77: 1, yield 1.30 %).
The alkamides were isolated from this CO2 extract by semipre-
parative HPLC using a Merck Hitachi L-6200A Intelligent Pump,
Merck Hitachi L-4500 Diode Array Detector with UV detection
at 254 nm and fitted with a LiChroCART 250 ± 10, 10 mm RP-18
LiChrospher 100 column. The compounds were eluted with a
gradient from 60/40 to 90/10 acetonitrile/water in 40 min (2.0
mL/min). Yields obtained were 0.7 mg (1), 1.2 mg (2), 7.2 mg (3),
11.04 mg (4), 15.76 mg (5), 0.9 mg (6), 0.8 mg (7), 23.1 mg (8),
Woelkart K et al. The Endocannabinoid System ¼ Planta Med 2005; 71: 701 ± 705
3.1 mg (9), 0.7 mg (10), 1.7 mg (11), 27.1 mg (12). Identification
was performed by LC-MS (LCQ Deca XP Plus) according to the lit-
erature data [3], [5].
For the binding assays rat brain and mouse spleen membranes
containing either CB1 or CB2, respectively, were prepared as de-
scribed previously by Lan et al. [6]. Novel probes were tested for
their ability to bind to CB1 or CB2 receptors via competition-
equilibrium binding using [3H]-CP55940 (specific activity 100
Ci/mmol; NIDA). Where included, pretreatment with PMSF re-
quired addition of thawed membrane suspensions to 5 volumes
of 25 mM Tris-HCl, pH 7.4, 5 mM MgCl2 with 1 mM EDTA (TME)
containing 10 mM PMSF followed by 30 min incubation. The
treated suspension was sedimented and washed three times in
TME to remove unreacted PMSF. Alkamides were dissolved in
DMSO to a final concentration of 10 mM and stored at ±20 8C. Li-
gands were diluted into TME containing 0.1 % BSA for assay pur-
poses. Non-specific binding was assessed in the presence of 100
Downloaded by: Collections and Technical Services Department. Copyrighted material.
nM CP55940. Diluted compounds were transferred to 96 well
plates containing a final concentration of [3H]-CP55940 equiva-
lent to 0.8 nM. Binding was initiated with the addition of mem-
brane suspension (~50 mg membrane protein). Plates were incu-
bated at 30 8C in a shaking water bath for 60 minutes and binding
terminated by rapid filtration of the membrane suspension over
Unifilter GF/B-96 Well Filter Plates (Packard Instruments) using
a Packard Filtermate-196 Cell Harvester. The filter plates were
washed four times with ice-cold wash buffer (50 mM Tris-base,
5 mM MgCl2 with 0.5 % BSA) and bound radioactivity was deter-
mined by scintillation counting. Data were fitted using a four-
parameter logistic equation to determine the actual IC50 of the li-
gand using GraphPad Prism. A Ki value was calculated from the
IC50 [7]. All data were collected as points in duplicate with IC50
and Ki values determined from at least three independent ex-
periments.
The ability of novel alkamides to inhibit FAAH activity was assess-
ed as described previously [8]. In brief, 25 nanomoles of alka-
mide were incubated with 25 nanomoles of arachidonylethanol-
amide (anandamide, AEA) and 75 mg membrane suspension in
TME buffer with 0.1 % BSA (25 mM Tris base, 5 mM MgCl2, 1 mM
EDTA, pH 7.4; final volume of 250 mL) for 15 min at 37 8C. Samples
(100 mL) were removed at the start of the assay and after 15 min,
diluted 1 : 5 with acetonitrile and centrifuged to precipitate pro-
teins. The resulting supernatant was injected onto the HPLC.
Chromatographic separation was achieved using a Beckman Ul-
trasphere ODS Pre-column (4.6 ” 45 mm). The mobile phase con-
sisted of 8.5 % orthophosphoric acid:acetonitrile (3 : 7), run iso-
cratically at a rate of 1 mL/min and detection at 204 nm with a
Waters Millennium HPLC system using a 20 mL injection loop.
The total run time was 8 min with AEA eluting at 2.2 minutes
and arachidonic acid (AA) at 6.0 minutes. Percent inhibition of
FAAH was calculated as the peak area ratios for arachidonic acid
formed by the hydrolysis of AEA in the presence and absence of
novel alkamides.
Acknowledgements
The study (KW) was supported by NIH Grant Prime Award No. 5
R01 AT001146 ± 03 from the National Center for Complementary
and Alternative Medicine as well as (AM) DA09158, DA7215,
DA3801 and DA7312 from the National Institutes on Drug Abuse.
References
1
Bauer R, Wagner H. Echinacea species as potential immunostimula-
tory drugs. In: Wagner H, Farnsworth NR, editors. Economic and Med-
icinal Plant Research London: Academic Press, 1991: pp 253 ± 321
2
Bauer R. Chemistry, analysis and immunological investigations of
Echinacea phytopharmaceuticals. In: Wagner H, editor. Immunomo-
dulatory Agents from Plants, Basel: Birkhäuser Verlag, 1999: pp 41 ±
88
3
Bauer R, Remiger P, Wagner H. Alkamides from the roots of Echinacea
angustifolia. Phytochemistry 1989; 28: 505 ± 8
4
Gertsch J, Schoop R, Kuenzle U, Suter A. Echinacea alkylamides modu-
late TNF-a gene expression via cannabinoid receptor CB2 and multiple
signal transduction pathways. FEBS Lett 2004; 577: 563 ± 9
5
Remiger P. Zur Chemie und Immunologie neuer Alkylamide und an-
derer Inhaltsstoffe aus Echinacea purpurea, Echinacea angustifolia
und Echinacea pallida. PhD thesis, University of Munich: 1988
6
Lan R, Liu Q, Fan P, Lin S, Fernando SR, McCallion D et al. Structure-ac-
tivity relationships of pyrazole derivatives as cannabinoid receptor
antagonists. J Med Chem 1999; 42: 769 ± 76
7
Cheng YC, Prusoff WH. Relationship between the inhibition constant
(Ki) and the concentration of inhibitor which causes 50 per cent inhi-
biton (IC50) of an enzymatic reaction. Biochem Pharmacol 1973; 22:
3099 ± 108
8
Lang W, Qin C, Hill WA, Lin S, Khanolkar AD, Makriyannis A. High-per-
formance liquid chromatographic determination of anandamide ami-
dase activity in rat brain microsomes. Anal Biochem 1996; 238: 40 ± 5
9
Pertwee RG, Fernando SR, Griffin G, Abadji V, Makriyannis A. Effect of
phenylmethylsulphonyl fluoride on the potency of anandamide as an
inhibitor of electrically evoked contractions in two isolated tissue pre-
parations. Eur J Pharmacol 1995; 272: 73 ± 8
10
Matias I, Pochard P, Orlando P, Salzet M, Pestel J, Di Marzo V. Presence
and regulation of the endocannabinoid system in human dendritic
cells. Eur J Biochem 2002; 269: 3771 ± 8
11
Pertwee RG. Pharmacology of cannabinoid CB1 and CB2 receptors.
Pharmacol Ther 1997; 74: 129 ± 80
Woelkart K et al. The Endocannabinoid System ¼ Planta Med 2005; 71: 701 ± 705
12
Klein TW, Newton C, Friedman H. Cannabinoid receptors and immu-
nity. Immunol Today 1998; 19: 373 ± 81
13
Goutopoulos A, Makriyannis A. From cannabis to cannabinergics new
therapeutic opportunities. Pharmacol Ther 2002; 95: 103 ± 17
14
Lin S, Khanolkar AD, Fan P, Goutopoulos A, Qin C, Paphadjis D et al. No-
vel analogues of arachidonylethanolamide (anandamide): Affinities
for the CB1 and CB2 cannabinoid receptors and metabolic stability. J
Med Chem 1998; 41: 5353 ± 61
15
Jbilo O, Derocq JM, Segui M, Fur GL, Casellas P. Stimulation of periph-
eral cannabinoid receptor CB2 induces MCP-1 and IL-8 gene expres-
sion in human promyelocytic cell line HL60. FEBS Lett 1999; 448:
273 ± 7
16
Sugiura T, Oka S, Gokoh M, Kishimoto S, Waku K. New perspectives in
the studies on endocannabinoid and cannabis: 2-arachidonoylglycer-
ol as a possible novel mediator of inflammation. J Pharmacol Sci 2004;
96: 367 ± 75
17
Kishimoto S, Kobayashi Y, Oka S, Gokoh M, Waku K, Sugiura T. 2-Ara-
Rapid Communication
chidonoylglycerol, an endogenous cannabinoid receptor ligand, indu-
ces accelerated production of chemokines in HL-60 cells. J Biochem
2004; 135: 4517 ± 24
18
Chang YH, Lee ST, Lin WW. Effects of cannabinoids on LPS-stimulated
inflammatory mediator release from macrophages: involvement of ei-
cosanoids. J Cell Biochem 2001; 81: 715 ± 23
Downloaded by: Collections and Technical Services Department. Copyrighted material.
19
Gallily R, Breuer A, Mechoulam R. 2-Arachidonylglycerol, an endogen-
ous cannabinoid, inhibits tumor necrosis factor-a production in mur-
ine macrophages, and in mice. Eur J Pharmacol 2000; 406: R5R7
20
Facchinetti F, Del Giudice E, Furegato S, Passerotto M, Leon A. Cannabi-
noids ablate release of TNFa in rat microglial cells stimulated with li-
popolysaccharide. Glia 2003; 41: 161 ± 8
21
Klegeris A, Bissonnette CJ, McGeer PL. Reduction of human monocytic
cell neurotoxicity and cytokine secretion by ligands of the cannabi-
noid-type CB2 receptor. Br J Pharmacol 2003; 139: 775 ± 86
22
Parolaro D, Massi P, Rubino T, Monti E. Endocannabinoids in the im-
mune system and cancer. Prostaglandins Leukot Essent Fatty Acids
2002; 66: 319 ± 32
23
Turner RB. The treatment of rhinovirus infections: progress and po-
tential. Antiviral Res 2001; 49: 1 ± 14
24
Woelkart K, Koidl C, Grisold A, Gangemi JD, Turner RB, Marth E, Bauer
R. Bioavailability and pharmacokinetics of alkamides from the roots of
Echinacea angustifolia in humans. J Clin Pharmacol, 2005; 45: 683 ± 9
705