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Cannabidivarin is anticonvulsant in mouse and rat

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BACKGROUND AND PURPOSE: Phytocannabinoids in Cannabis sativa have diverse pharmacological targets extending beyond cannabinoid receptors and several exert notable anticonvulsant effects. For the first time, we investigated the anticonvulsant profile of the phytocannabinoid cannabidivarin (CBDV) in vitro and in in vivo seizure models. EXPERIMENTAL APPROACH: The effect of CBDV (1-100μM) on epileptiform local field potentials (LFPs) induced in rat hippocampal brain slices by 4-AP application or Mg(2+) -free conditions was assessed by in vitro multi-electrode array recordings. Additionally, the anticonvulsant profile of CBDV (50-200 mg kg(-1) ) in vivo was investigated in four rodent seizure models: maximal electroshock (mES) and audiogenic seizures in mice, and pentylenetetrazole (PTZ) and pilocarpine-induced seizures in rat. CBDV effects in combination with commonly-used antiepileptic drugs were investigated in rat seizures. Finally, the motor side effect profile of CBDV was investigated using static beam and grip-strength assays. KEY RESULTS: CDBV significantly attenuated status epilepticus-like epileptiform LFPs induced by 4-AP and Mg(2+) -free conditions. CDBV had significant anticonvulsant effects in mES (≥100 mg kg(-1) ), audiogenic (≥50 mg kg(-1) ) and PTZ-induced seizures (≥100 mg kg(-1) ). CDBV alone had no effect against pilocarpine-induced seizures, but significantly attenuated these seizures when administered with valproate or phenobarbital at 200 mg kg(-1) CDBV. CDBV had no effect on motor function. CONCLUSIONS AND IMPLICATIONS: These results indicate that CDBV is an effective anticonvulsant across a broad range of seizure models, does not significantly affect normal motor function and therefore merits further investigation in chronic epilepsy models to justify human trials.
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Themed Section: Cannabinoids 2012
RESEARCH PAPER
Cannabidivarin is
anticonvulsant in mouse
and rat
AJ Hill1,2, MS Mercier1*, TDM Hill1, SE Glyn1, NA Jones1,2,
Y Yamasaki1,2,3, T Futamura3, M Duncan4, CG Stott4, GJ Stephens1,
CM Williams2and BJ Whalley1
1Reading School of Pharmacy,University of Reading,Whiteknights, Reading, UK, 2School of
Psychology and Clinical Language Sciences,University of Reading,Reading, UK, 3Otsuka
Pharmaceutical, Co. Ltd,Tokushima, Japan, and 4GW Pharmaceuticals plc,Porton Down
Science Park,Salisbury, Wiltshire, UK
Correspondence
Andrew Hill, Reading School
of Pharmacy and School of
Psychology and Clinical
Language Sciences, University
of Reading, Whiteknights,
Reading, RG6 6AJ, UK. E-mail:
a.j.hill@reading.ac.uk
----------------------------------------------------------------
*Present address: MRC Centre
for Synaptic Plasticity, University
of Bristol, Medical Sciences
Building, University Walk,
Bristol, BS8 1TD, UK.
----------------------------------------------------------------
Keywords
epilepsy; cannabinoid;
cannabidivarin; seizure; side
effect; hippocampus
----------------------------------------------------------------
Received
23 April 2012
Revised
17 August 2012
Accepted
28 August 2012
BACKGROUND AND PURPOSE
Phytocannabinoids in Cannabis sativa have diverse pharmacological targets extending beyond cannabinoid receptors and
several exert notable anticonvulsant effects. For the first time, we investigated the anticonvulsant profile of the
phytocannabinoid cannabidivarin (CBDV) in vitro and in in vivo seizure models.
EXPERIMENTAL APPROACH
The effect of CBDV (1–100 mM) on epileptiform local field potentials (LFPs) induced in rat hippocampal brain slices by
4-aminopyridine (4-AP) application or Mg2+-free conditions was assessed by in vitro multi-electrode array recordings.
Additionally, the anticonvulsant profile of CBDV (50–200 mg·kg-1)in vivo was investigated in four rodent seizure models:
maximal electroshock (mES) and audiogenic seizures in mice, and pentylenetetrazole (PTZ) and pilocarpine-induced seizures
in rats. The effects of CBDV in combination with commonly used antiepileptic drugs on rat seizures were investigated. Finally,
the motor side effect profile of CBDV was investigated using static beam and grip strength assays.
KEY RESULTS
CBDV significantly attenuated status epilepticus-like epileptiform LFPs induced by 4-AP and Mg2+-free conditions. CBDV
had significant anticonvulsant effects on the mES (100 mg·kg-1), audiogenic (50 mg·kg-1) and PTZ-induced seizures
(100 mg·kg-1). CBDV (200 mg·kg-1) alone had no effect against pilocarpine-induced seizures, but significantly attenuated
these seizures when administered with valproate or phenobarbital at this dose. CBDV had no effect on motor function.
CONCLUSIONS AND IMPLICATIONS
These results indicate that CBDV is an effective anticonvulsant in a broad range of seizure models. Also it did not significantly
affect normal motor function and, therefore, merits further investigation as a novel anti-epileptic in chronic epilepsy models.
LINKED ARTICLES
This article is part of a themed section on Cannabinoids. To view the other articles in this section visit
http://dx.doi.org/10.1111/bph.2012.167.issue-8
Abbreviations
AED, antiepileptic drugs; 4-AP, 4-aminopyridine; CBD, cannabidiol; CBDV, cannabidivarin; DG, dentate gyrus;
ESM, ethosuximide; LFP, local field potential; MEA, multi-electrode array; mES, maximal electroshock; PTZ,
pentylenetetrazole; D9-THC, D9-tetrahydrocannabinol; TRP, transient receptor potential; VPA, valproate
BJP British Journal of
Pharmacology
DOI:10.1111/j.1476-5381.2012.02207.x
www.brjpharmacol.org
British Journal of Pharmacology (2012) 167 1629–1642 1629
© 2012 The Authors
British Journal of Pharmacology © 2012 The British Pharmacological Society
Introduction
Epilepsy is a CNS disorder affecting ~1% of the global popula-
tion, and is symptomatically characterized by chronic, recur-
rent seizures. A range of treatments are available, although
there is still a need for more effective and better-tolerated
antiepileptic drugs (AEDs) as illustrated by the pharmacologi-
cal intractability of ~30% of cases and the poor side effect
profile of currently available AEDs (Kwan and Brodie, 2007).
Cannabis sativa has a long history of use for the control of hu-
man seizures (O’Shaughnessy, 1843; Mechoulam, 1986), and is
legally used for this in some countries (Sirven and Berg, 2004).
There are >100 phytocannabinoids present in C. sativa,of
which D9-tetrahydrocannabinol (D9-THC) is the most abun-
dant (Elsohly and Slade, 2005; Mehmedic et al., 2010) and,
via partial agonism of the CB1cannabinoid receptor, is
responsible for the classical psychoactive effects of cannabis
(Pertwee, 2008). Although CB1cannabinoid receptor agonism
can exert anticonvulsant effects in in vitro and in vivo models
(Chesher and Jackson, 1974; Wallace et al., 2001; 2003; Desh-
pande et al., 2007), the most promising non-psychoactive
anticonvulsant phytocannabinoid investigated to date is can-
nabidiol (CBD), which exerts anticonvulsant actions via an,
as yet unknown, non-CB1cannabinoid receptor mecha-
nism(s) in animal models in vitro,in vivo and in humans
(Cunha et al., 1980; Consroe et al., 1982; Wallace et al., 2001;
Jones et al., 2010); CBD’s notable anticonvulsant properties
led us to investigate the anticonvulsant potential of its propyl
analogue, cannabidivarin (CBDV).
CBDV was first isolated in 1969 (Vollner et al., 1969). At
present, little is known about the pharmacological properties
of CBDV (Izzo et al., 2009), although Scutt and Williamson
reported that CBDV acts via CB2cannabinoid receptor-
dependent mechanisms (Scutt and Williamson, 2007). More
recently, De Petrocellis and co-workers reported differential
CBDV effects at transient receptor potential (TRP) channels in
vitro, where it acted as a human TRPA1, TRPV1 and TRPV2
agonist (EC50 values: 0.42, 3.6 and 7.3 mM, respectively) and
a TRPM8 antagonist (IC50: 0.90 mM) (De Petrocellis et al.,
2011a,b). Additionally, CBDV has been shown to inhibit
the primary synthetic enzyme of the endocannabinoid,
2-arachidonoylglycerol (Bisogno et al., 2003), diacylglycerol
lipase a(IC50 16.6 mM) in vitro (De Petrocellis et al., 2011a).
While the pharmacological relevance of these effects has not
been confirmed in vivo, they further illustrate the diversity of
non-D9-THC phytocannabinoid pharmacology and support
the emergent role of multiple non-CB receptor targets
(Pertwee, 2010; Hill et al., 2012).
Here, we identified anticonvulsant effects of CBDV for the
first time; CBDV suppressed in vitro epileptiform activity in
brain slices and acted as an anticonvulsant in vivo. However,
normal motor function was not significantly affected by
CBDV, therefore, further investigations into the clinical
development of CBDV as a novel AED are warranted.
Methods
In vitro electrophysiology
Tissue preparation. All studies involving animals are reported
in accordance with the ARRIVE guidelines for reporting
experiments involving animals (Kilkenny et al., 2010;
McGrath et al., 2010) and all experiments were carried out in
accordance with Home Office regulations [Animals (Scientific
Procedures) Act, 1986]. Transverse hippocampal slices
(~450-mm thick) for multi-electrode array (MEA) recordings
were prepared from female and male adult Wistar Kyoto rats
(P>21; Harlan, Bicester, UK) using a Vibroslice 725 M
(Campden Instruments Ltd., Loughborough, UK) as previ-
ously described (Jones et al., 2010).
MEA recordings. MEA recordings and analyses were con-
ducted as described in Hill et al. (2010). Once established [by
addition of either 100 mM 4-aminopyridine (4-AP) or omission
of MgSO4.7H2O without substitution], epileptiform activity
was permitted to continue for 30 min (control bursting) before
sequential addition of 1, 10 and 100 mM CBDV (30 min each).
Epileptiform activity was characterized by spontaneous local
field potentials (LFPs) recorded simultaneously from 59 elec-
trodes covering the majority of the hippocampal slice prepa-
ration. The amplitude and duration of epileptiform LFPs were
analysed for each electrode. Data from individual electrodes,
based on their position in each hippocampal subregion, were
pooled to provide mean results for each subregion across n
5 slices from n5 animals per model. Matlab 6.5 and 7.0.4
(Mathworks, Natick, MA, USA), Microsoft Excel (Microsoft,
Redmond, WA, USA), MC_DataTool and MC_Rack (Multi
Channel Systems GmbH, Reutlingen, Germany) were used to
process and present data as described in Hill et al. (2010).
Inherent changes in LFP amplitude and frequency were cor-
rected for, as described previously (Hill et al., 2010). For refer-
ence, the extent of amplitude rundown correction applied is
illustrated in Figure 1C and D. LFP frequency was calculated
per slice (n5 for each model) and represents the number of
LFP bursts per unit time. Examples of single bursts from each
model can be seen in Figure 1A and B. Drug-induced changes
in burst duration, amplitude and frequency are expressed as
normalized proportions of control values SEM, corrected
where necessary, and were analysed by Wilcoxon’s paired test
with Holm’s sequential Bonferroni correction.
In vivo seizure models
Animals. In all cases before seizure induction, animals were
maintained on a 12 h light/dark cycle with free access to food
and water (with the exception of rats that received oral CBDV,
see later). Audiogenic seizure experiments with dilute, brown,
non-Agouti (DBA/2) mice (3–4 weeks old; Elevage Janvier, Le
Genest-Saint-Isle, France) were performed at Porsolt Research
Laboratory (Le Genest-Saint-Isle, France) in accordance with
French legislation and under licence from the French Minis-
try for Agriculture and Fisheries. mES experiments with
ICR (CD-1) mice (5 weeks old; SLC Japan Inc., Shizuoka,
Japan) were performed at Otsuka Pharmaceuticals Co, Ltd.
(Tokushima, Japan) in accordance with the guidelines of the
Physiological Society of Japan. In total, 80 mice were used.
Seizure studies in male Wistar Kyoto rats (Harlan, 3–4 weeks
old; in total, 640 rats were used) were performed at the Uni-
versity of Reading, UK; all experiments were carried out in
accordance with UK Home Office regulations [Animals (Sci-
entific Procedures) Act 1986].
CBDV administration. CBDV (50, 100 or 200 mg·kg-1;GW
Pharmaceuticals Ltd., Salisbury, UK) in an ethanol : Cremo-
BJP AJ Hill et al.
1630 British Journal of Pharmacology (2012) 167 1629–1642
phor: saline (0.9% w v-1NaCl vehicle; 2:1:17; all Sigma, Poole,
UK) was administered by an i.p. injection 1 h before seizure
induction in all the models, with the exception of mES where
it was administered 30 min before seizure induction. All
experiments included a control group, which received
volume-matched vehicle, against which other groups were
assessed. In mice experiments, n=10 per group and in rat,
n=15 per group. In experiments where CBDV was adminis-
tered p.o. (gavage), 400 mg·kg-1CBDV or volume-matched
vehicle [20% solutol (Sigma) in 0.9% w v-1NaCl] was admin-
istered after the animals had been deprived of food for 13.5 h
and 3.5 h before i.p. administration of pentylenetetrazole
(PTZ), n=15 for both groups (see Supporting Information
Appendix S1 for details on oral dose levels).
Seizure induction. mES seizures were induced in mice by a
stimulator (Ugo Basile ECT, Comerio, Italy) via earlap
clamps at a current of 30 mA delivered at 100 Hz for
200 ms. DBA/2 mice were placed in a Plexiglas jar 1 h after
CBDV/vehicle administration. A mounted bell (110–
120 dB) was activated until occurrence of a tonic audiogenic
seizure or for a maximum of 60 s. To induce generalized
seizures in rats, 85 mg·kg-1PTZ was injected i.p. Status epi-
lepticus with a temporal lobe focus was induced in rats by
injecting pilocarpine hydrochloride (Sigma; in 0.9% w v-1
NaCl) 380 mg·kg-1i.p., 45 min after pretreating the rats
with methylscopolamine (Sigma; in 0.9% w v-1NaCl)
1 mg·kg-1i.p., which blocks the peripheral effects of
pilocarpine.
Seizure analysis. In mES experiments, mice were observed for
10 s during electroshock, tonic hindlimb extension occur-
rence was noted and expressed as a percentage of the total
number of animals for each group. Audiogenic seizure behav-
iour was observed visually, while rat seizures were video
recorded (Farrimond et al., 2009). For audiogenic seizures, the
incidence (as a percentage) of the most severe (tonic–clonic)
seizures, mortality and seizure-free animals were calculated
for each group. These parameters, as well as seizure duration
and severity, were also determined for rat seizures. Rat behav-
iour was coded blind offline using The Observer Pro software
(Noldus, Wageningen, The Netherlands) and seizure severity
scales appropriate to each seizure type (Table 1). Values are
expressed as mean SEM throughout.
Co-administration experiments. The effect of co-
administration of clinically-used AEDs with 200 mg·kg-1
CBDV on PTZ- and pilocarpine-induced seizures was investi-
gated. For details, see Supporting Information Appendix S1.
Briefly, in each experiment, an AED was administered i.p., at
either ~20, ~40 or ~70% maximal effective dose, in the
absence or presence of 200 mg·kg-1CBDV (n=15 per group,
120 per experiment); the convulsant (PTZ or pilocarpine) was
administered 1 h after CBDV or its vehicle. The experimental
design is illustrated and summarized in Table 2. In the PTZ
model, CBDV was co-administered with valproate (VPA) or
ethosuximide (ESM) before PTZ, and with VPA or phenobar-
bital (PB) before pilocarpine. These AEDs were chosen based
on their clinical profile and their reported efficacy in the
models used here, with VPA suppressing both seizure types
and ESM and phenobarbital suppressing PTZ and pilocarpine
respectively (Loscher et al., 1991; Sofia et al., 1993; Shantilal
et al., 1999; Lindekens et al., 2000; Loscher, 2011). In co-
administration experiments, seven (2.9%) rats exhibited a
fatal reaction to CBDV administration. Behaviourally, this
manifested as rapid development (within 300 s) of lethargic
convulsive movements followed by death. Overall, across all
PTZ and pilocarpine experiments, this effect was seen in 2.6%
of all rats that received 200 mg·kg-1CBDV, but not at all in
side effect tests. No adverse effects of other CBDV doses were
observed in rats, and none at any dose in mice. The animals
that died were omitted from all analyses.
Statistics. In experiments where i.p. CBDV alone was admin-
istered, the effects of CBDV on seizure severity, onset latency
and seizure duration were assessed by one-way ANOVA with
post hoc Tukey’s tests as appropriate. Chi-squared tests fol-
lowed by post hoc Fisher’s exact tests were used where appro-
Table 1
Seizure behaviour scoring scales for PTZ and pilocarpine-induced seizures
Score PTZ-induced seizures Pilocarpine-induced seizures
0 Normal behaviour Normal behaviour
1 Isolated myoclonic jerks Mouth clonus
2 Atypical clonic seizure Unilateral forelimb clonus
3 Fully developed bilateral forelimb clonus Bilateral forelimb clonus
3.5 Forelimb clonus with tonic component and body twist NA
4 Tonic–clonic seizure with suppressed tonic phase* Bilateral forelimb clonus with rearing and falling
4.5 NA Tonic–clonic seizure with postural control retained
5 Fully developed tonic–clonic seizure* Tonic–clonic seizure*
Seizure severity scoring scales are shown for each model, although no equivalency of severity should be assumed between scales for different
models.
*Indicates a loss of righting reflex.
NA =not applicable.
BJP
Cannabidivarin as an anticonvulsant
British Journal of Pharmacology (2012) 167 1629–1642 1631
priate to assess differences in incidence parameters. Where
CBDV was co-administered with an AED, two-way ANOVA or
log-linear modelling was used to analyse the effects of CBDV
and AEDs. Log-linear modelling was used to model the inter-
actions between drug co-administration and incidence
parameters (e.g. mortality, % seizure-free). If the model indi-
cated a significant effect of drug treatment, further analysis to
determine the contribution of CBDV, the relevant AED and
any drug ¥drug interaction was performed; these analyses are
given in the text and in Supporting Information Tables S1
and S2.
Motor function assays
The effects of CBDV (50, 100 and 200 mg·kg-1) and VPA (125,
250 and 350 mg·kg-1) on normal rat motor function were
assessed on a 1 m raised static beam and by a grip strength
test (see Supporting Information Appendix S1 for details).
All receptor and ion channel nomenclature conforms to
BJP’s Guide to Receptors and Channels (Alexander et al., 2011).
Results
Effects of pure CBDV in the Mg2+-free and
4-AP in vitro models of epileptiform activity
The effects of CBDV (1–100 mM) on epileptiform activity,
induced by Mg2+-free aCSF (Figure 1A) or 100 mM 4-AP
(Figure 1B), in rat acute hippocampal slices were examined.
CBDV significantly decreased the amplitude and duration of
epileptiform LFPs induced by Mg2+-free aCSF (Figure 1C and
D; P0.05); significant effects were seen at 10 mM, and the
CA3 region was more resistant to the effects of CBDV than
the dentate gyrus (DG) or CA1 (Figure 1C and D). Conversely,
CBDV significantly increased Mg2+-free-induced LFP fre-
quency (10 mM; Figure 1E; P0.05).
An anti-epileptiform effect of 100 mM CBDV on the
amplitude of 4-AP-induced epileptiform LFPs was observed in
the CA1 region alone (Figure 1F; P0.05), whereas LFP
duration was significantly lowered in all hippocampal regions
by 10 mM CBDV (Figure 1G) and, by contrast to the Mg2+-
free model, 4-AP-induced LFP frequency was significantly
decreased by all CBDV concentrations tested (Figure 1H;
P0.05). Thus, CBDV attenuated the duration of amplitude
of LFPs in both models, and had differential effects on
frequency.
Effects of CBDV on maximal electroshock
(mES) and audiogenic seizures in mice
The effects of CBDV (50–200 mg·kg-1) on mES convulsions
and audiogenic seizures in mice were investigated. CBDV had
a significant anticonvulsant effect on animals displaying
tonic hindlimb extension after mES [c2(3) =15.000; P
0.001; Figure 2A]; significantly fewer animals that received
100 or 200 mg·kg-1CBDV exhibited hindlimb extension
(both groups 30%) than those that received vehicle (90%,
Figure 2A; P0.001 vs. vehicle-treated group). Audiogenic
seizures were also significantly attenuated by CBDV
(Figure 2B–D). The incidence of tonic convulsions was signifi-
cantly lower after CBDV administration [c2(3) =19.436, P
0.001; Figure 2B]; 80% of vehicle-treated animals developed
tonic convulsions compared with only 20% (50 mg·kg-1
CBDV), 10% (100 mg·kg-1CBDV) and 0% (200 mg·kg-1
CBDV) after drug treatment (each P0.001 vs vehicle). The
percentage of animals that remained seizure-free was signifi-
cantly higher after administration of 200 mg·kg-1CBDV
(90%) than vehicle [0%; c2(3) =27.461, P0.001; Figure 2C].
Finally, a statistical trend was observed for the mortality rate
[c2(3) =6.667, P0.1], with lower mortality after 100 and
200 mg·kg-1CBDV treatment than vehicle (0% vs 30%,
respectively; Figure 2D). Thus, CBDV exhibits strong and sig-
nificant anticonvulsant effects in two broad-screen mouse
Table 2
Experimental design and time course of co-administration experiments
60 min
CBDV/vehicle treatment (i.p.)
Time
A
(min) AED treatment (i.p.)
Time
B
(min)
Seizure
induction
and recording
PTZ
experiments
200 mg·kg-1CBDV (n=60) 30 VPA vehicle, 50, 100, 250 mg·kg-1VPA (n=15 each) 30 85 mg·kg-1PTZ
30-min
recording
CBDV vehicle (n=60) VPA vehicle, 50, 100, 250 mg·kg-1VPA (n=15 each)
200 mg·kg-1CBDV (n=60) 30 ESM vehicle, 60, 120, 175 mg·kg-1ESM (n=15 each) 30
CBDV vehicle (n=60) ESM vehicle, 60, 120, 175 mg·kg-1ESM (n=15 each)
Pilocarpine
experiments
200 mg·kg-1CBDV (n=60) 15 VPA vehicle, 62.5, 125, 250 mg·kg-1VPA (n=15 each) 45 380 mg·kg-1
pilocarpine
60-min
recording
CBDV vehicle (n=60) VPA vehicle, 62.5, 125, 250 mg·kg-1VPA (n=15 each)
200 mg·kg-1CBDV (n=60) 15 PB vehicle, 10, 20, 40 mg·kg-1PB (n=15 each) 45
CBDV vehicle (n=60) PB vehicle, 10, 20, 40 mg·kg-1PB (n=15 each)
‘Time A’ column: time between CBDV/CBDV vehicle and AED administration. ‘Time B’ column: time between AED/vehicle and convulsant.
The duration of the seizure recording is indicated in the final column. PB, phenobarbital, VPA, valproate, ESM, ethosuximide.
BJP AJ Hill et al.
1632 British Journal of Pharmacology (2012) 167 1629–1642
seizure models. Next, we investigated the anticonvulsant
potential of CBDV in two further models of seizure in rat that
emulate more specific seizure types.
Effects of CBDV on PTZ- and
pilocarpine-induced seizures in rats
CBDV significantly decreased PTZ seizure severity (F3,58 =
4.423, P0.05; Figure 3A); the median seizure severity after
vehicle administration was tonic–clonic convulsion score 5,
but after 200 mg·kg-1CBDV administration seizure severity
was significantly lowered to a median severity of bilateral
clonic convulsion score 3 (P0.05). CBDV also significantly
reduced mortality (c2(3) =10.356, P0.05; Figure 3B) at 100
and 200 mg·kg-1CBDV (P0.01). The percentage of animals
that remained seizure-free was significantly increased by
CBDV administration [c2(3) =7.809, P0.05; Figure 3C];
33.3% of animals that received 200 mg·kg-1CBDV exhibited
no signs of seizure compared with only 6.7% of animals that
received vehicle (P0.01). Furthermore, seizure onset was
significantly delayed by CBDV treatment (F3,50 =2.971, P
0.05; Figure 3D); mean onset latency was significantly longer
after administration of 200 mg·kg-1CBDV than vehicle (65
11 s and 40 4 s, respectively; P0.05). Thus, CBDV,
administered alone, exhibited strong and significant anticon-
vulsant effects on PTZ seizures at 200 mg·kg-1(Figure 3A–D)
with more limited, but significant, effects at 100 mg·kg-1
(Figure 3B).
Figure 1
Effects of CBDV on hippocampal epileptiform activity. (A and B) Example traces showing effects of 100 mM CBDV on epileptiform LFPs induced
by Mg2+-free conditions (A) or 100 mM 4-AP (B) in the CA1 region. The black bar represents amplitude as corrected for inherent rundown (see
Methods); the dotted line below represents control burst duration. Scale in (A): 100 mV/200 ms; (B): 150 mV/200 ms. (C–H) Effects of CBDV on
amplitude (C and F), duration (D and G) and frequency (E and H) of epileptiform LFPs induced by Mg2+-free conditions (C–E) or 100 mM 4-AP
(F–H). Data are presented as mean SEM normalized to control (pre-drug) conditions and corrected for background changes where appropriate
(see Methods). LFP amplitude and duration values are expressed for each hippocampal region as in the key. n=9–12. *P0.05, **P0.01 and
***P0.001.
BJP
Cannabidivarin as an anticonvulsant
British Journal of Pharmacology (2012) 167 1629–1642 1633
We extended our studies to investigate the effects of
CBDV (50–200 mg·kg-1) on the convulsions associated with
pilocarpine-induced status epilepticus (380 mg·kg-1). CBDV
(50–200 mg·kg-1) had no significant effect on the severity
(F3,59 =0.049, P>0.1; Figure 3E) or resultant mortality of
pilocarpine convulsions [c2(3) =1.779, P>0.1; Figure 3F].
Similarly, CBDV did not significantly affect the percentage of
animals that remained seizure-free [c2(3) =0.110, P>0.1;
Figure 3G] or the latency to the onset of convulsions (F3,53 =
0.404, P>0.1; Figure 3H).
Effect of co-administration of CBDV and
AEDs on PTZ- and pilocarpine-induced
seizures in rats
We investigated the effects of CBDV when co-administered
with AEDs before PTZ or pilocarpine treatment. The effects of
combined drug treatment (CBDV +AED) on seizure param-
eters are illustrated in Figures 4 and 5, as is the contribution
of CBDV to these effects. The contribution of AEDs is illus-
trated in Figures 4 and 5 while statistical analyses of AED
effects and any interaction between CBDV and AEDs are
shown in Supporting Information Tables S1 and S2.
CBDV 200 mg·kg-1was co-administered with VPA (50–
250 mg·kg-1) or ESM (60–175 mg·kg-1). In the CBDV +VPA
experiments, drug co-administration had significant anti-
convulsant effects on all seizure parameters except the per-
centage of animals remaining seizure-free. CBDV and VPA
co-administration significantly decreased seizure severity
(F7,112 =10.449, P0.001; Figure 4A). When modelled
by log-linear analyses, our data indicated that drug
co-administration decreased mortality (Figure 4B) and the
incidence of the most severe (tonic–clonic) seizures
(Figure 4C). Seizure onset was significantly delayed by drug
co-administration (F7,109 =13.285, P0.001; Figure 4D) and
the mean duration of seizures was increased (F7,103 =5.250,
P0.001). VPA contributed significantly to all these effects
(Figure 4A–D, Supporting Information Table S1). CBDV sig-
nificantly contributed to the overall decrease in severity
(F1,112 =5.748, P0.05; Figure 4A) and mortality [c2(1) =
6.639, P0.01; Figure 4B] and the increase in onset latency
(F1,109 =7.393, P0.01; Figure 4C). CBDV did not signifi-
cantly affect tonic–clonic seizure incidence (Figure 4D) or
seizure duration (P>0.1). No effect of drug treatment on the
number of seizure-free animals was observed [X2(14) =8.930,
P>0.1] and no significant positive or negative interactions
between the effects of 200 mg·kg-1CBDV and VPA were
observed (Supporting Information Tables S1, P>0.1).
Co-administration of 200 mg·kg-1CBDV and ESM (60–
175 mg·kg-1) had significant anticonvulsant effects on all
parameters of PTZ-induced seizures: CBDV and ESM
co-administration significantly decreased seizure severity
(F7,110 =12.556, P0.001; Figure 4E), when modelled with
log-linear analysis, our data indicated that co-administration
also decreased mortality (Figure 4F) and the incidence of the
most severe seizures (Figure 4G). Seizure onset latency was
significantly increased (F7,76 =7.885, P0.001; Figure 4H), as
was the percentage of animals that remained seizure-free (log-
linear model; Figure 4I); seizure duration was also signifi-
cantly decreased (F7,102 =6.934, P0.001). ESM significantly
Figure 2
Effects of CBDV on mES and audiogenic seizures in mice. (A) The effect of CBDV on the percentage of animals that exhibited tonic hindlimb
extension in response to mES. (B–D) The effect of CBDV (50–200 mg·kg-1) on the percentage of animals that displayed tonic convulsions (B),
remained seizure-free (C) or suffered mortality (D) as a result of audiogenic seizure induction. n=10 in all cases, ***P0.001.
BJP AJ Hill et al.
1634 British Journal of Pharmacology (2012) 167 1629–1642
contributed to all anticonvulsant effects (Figure 4E-I; Sup-
porting Information Table S1). CBDV contributed signifi-
cantly to the overall decreases in seizure severity (F1,112 =
7.474, P0.01; Figure 4E) and mortality [c2(1) =5.174, P
0.05; Figure 4F]; the contribution of CBDV to the increase in
onset latency showed a statistical trend (F1,76 =2.791, P0.1;
Figure 4H). CBDV did not significantly contribute to the
effects on seizure duration, the proportion of animals that
remained seizure-free (both P>0.1) or the incidence of the
most severe seizures (P>0.1; Figure 4G). No significant
positive or negative interactions between the effects of
200 mg·kg-1CBDV and ESM were observed (Supporting Infor-
mation Tables S1, P>0.1).
We next investigated whether 200 mg·kg-1CBDV affected
the anticonvulsant actions of VPA or phenobarbital
on pilocarpine-induced convulsions. Interestingly, these
co-administration experiments highlighted significant anti-
convulsant effects of 200 mg·kg-1CBDV not previously
observed when CBDV was administered alone. Co-
administration of VPA (50–250 mg·kg-1) with 200 mg·kg-1
CBDV had significant anticonvulsant effects on all the
parameters except the percentage of animals that remained
convulsion-free: CBDV and VPA co-administration signifi-
cantly decreased severity (F7,100 =16.477, P0.001;
Figure 5A); when modelled by log-linear analysis, our data
indicated that mortality (Figure 5B) and the incidence of the
most severe (tonic–clonic) convulsions (Figure 5C) were also
decreased by drug co-administration; onset latency was
significantly increased (F7,105 =8.649, P0.001; Figure 5D).
VPA contributed significantly to all anticonvulsant effects
(Figure 5A-D, Supporting Information Table S2) with the
interesting exception of mortality. Mortality was higher (but
not significantly so) when 62.5 and 125 mg·kg-1VPA were
co-administered with vehicle (Figure 5B); however, CBDV
had an anticonvulsant effect, significantly decreasing mortal-
ity compared with administration of its vehicle [c2(1) =4.010,
P0.05; Figure 5D]. CBDV also significantly contributed to
the overall anticonvulsant effects of treatment on severity
(F1,110 =22.711, P0.001; Figure 5A) and the incidence of
tonic–clonic convulsions [c2(1) =4.010, P0.01; Figure 5C],
although it had no significant effect on onset latency
(P>0.1; Figure 5D). The percentage of animals that remained
convulsion-free [c2(6) =1.564, P>0.1] was unaffected by
treatment. No significant interactions between CBDV and
VPA effects were observed (Supporting Information Tables S2,
P>0.1).
Co-administration of 200 mg·kg-1CBDV and phenobarbi-
tal (10–40 mg·kg-1) had significant anticonvulsant effects on
the severity of pilocarpine-induced convulsions (F7,108 =
19.352, P0.001; Figure 5E). When modelled with log-linear
analysis, our data indicated that there was no effect of treat-
ment on mortality (Figure 5F), whereas the percentage of
animals that developed tonic–clonic convulsions was signifi-
cantly decreased (Figure 5G). No effect of drug treatment
Figure 3
Effects of CBDV on PTZ- and pilocarpine-induced seizures in rats. (A–D) The effect of CBDV on PTZ-induced seizures: seizure severity (A), mortality
(B), the proportion of animals remaining seizure-free (C) and the onset latency (D). (E–H) The effect of CBDV on pilocarpine-induced convulsions:
severity (E), mortality (F), the percentage of animals remaining seizure-free (G) and the onset latency (H). In (D and H), onset latency is presented
as mean SEM In (A and E), median severity is represented by a thick horizontal line, the 25th and the 75th percentiles by the box and maxima
and minima are represented by ‘whiskers’. n=15 in all cases. *P0.05, **P0.01 and ***P0.001.
BJP
Cannabidivarin as an anticonvulsant
British Journal of Pharmacology (2012) 167 1629–1642 1635
was observed on seizure onset latency (P>0.1; Figure 5H);
however, when modelled with log-linear analysis, our data
indicated that the percentage of animals that remained
convulsion-free was significantly increased (Figure 5I). Phe-
nobarbital significantly contributed to all anticonvulsant
effects (Figure 5E–I; Supporting Information Table S2). CBDV
significantly contributed to the overall decrease seen in sever-
ity (F1,108 =4.480, P0.05), and the effects of CBDV and
phenobarbital interacted significantly due to a convergence
of the severity observed in the absence and presence of CBDV
(Figure 5F, Supporting Information Table S2; F3,108 =3.105,
P0.05), no further significant interactions between the
effects of CBDV and phenobarbital were observed (P>0.1;
Supporting Information Table S2).
Data from the co-administration experiments demon-
strate that the AEDs strongly suppress PTZ-induced seizures
and pilocarpine-induced convulsions in a dose-dependent
manner (Figures 4 and 5). From several, but not all, of the
parameters examined, 200 mg·kg-1CBDV significantly con-
tributed to the anticonvulsant effects observed in these
experiments. To more precisely assess the effect of CBDV on
AED actions in these studies, we performed pairwise compari-
sons at each dose of AED between groups that received CBDV
vehicle and groups that received 200 mg·kg-1CBDV; these
analyses were only performed if two-way ANOVA or log-
linear analysis results indicated an overall effect of CBDV
upon a given parameter. Based on these analyses and
Figure 5F–I, the effect of CBDV on the actions of phenobar-
bital in the pilocarpine model appears limited and is not
significant. Similarly, the effect of CBDV on the actions of
VPA in the PTZ model was limited (Figure 4A–D); the primary
effect of CBDV is on delaying seizure onset, as 200 mg·kg-1
CBDV significantly improved the effect of 50 mg·kg-1VPA
(P0.05; Figure 4D) and showed a statistical trend towards
the same effect with 100 mg·kg-1VPA (P<0.1). More notably,
CBDV significantly improved the effect of 60 mg·kg-1ESM on
PTZ-induced seizure severity and onset latency (P0.05;
Figure 4E and H) and also showed a statistical trend to
improvement of the 120 mg·kg-1ESM effect for both these
measures (P<0.1). Furthermore, when 200 mg·kg-1CBDV
was administered together with VPA before pilocarpine
administration, it significantly improved the effects of VPA
on severity (62.5 and 250 mg·kg-1;P0.05), mortality (62.5
and 125 mg·kg-1;P0.05) and the percentage of animals
that experienced the most severe seizures (all doses, P0.01;
Figure 5A–C).
Thus, CBDV is well-tolerated when co-administered with
AEDs and does not interact antagonistically with any of the
AEDs studied in either seizure model. Furthermore, CBDV has
significant anticonvulsant effects when co-administered with
ESM in the PTZ model and even greater effects when
co-administered with VPA in the pilocarpine model, where
beneficial effects were generally observed at low and medium
AED doses. CBDV did not affect the effects of phenobarbital
Figure 4
Effects of co-administration of CBDV and AEDs on PTZ-induced seizures in rats. The effects of CBDV co-administration with VPA (A–D) or ESM (E–I)
on PTZ-induced seizures: severity (A and E), mortality (B and F), the incidence of tonic–clonic seizures (C and G), onset latency (D and H) and (for
CBDV +ESM only) the percentage of animals that remained seizure-free. In (D and H), onset latency is presented as mean SEM. In (A and E),
median severity is represented by a thick horizontal line, the 25th and 75th percentiles by the box and maxima and minima are represented by
‘whiskers’. Significance of CBDV treatment is given in text. n=15 in all cases. *P0.05, **P0.01 and ***P0.001 for AED effects.
BJP AJ Hill et al.
1636 British Journal of Pharmacology (2012) 167 1629–1642
in the pilocarpine model and had only very limited effects on
the onset of seizures when co-administered with VPA before
PTZ treatment.
CBDV motor side effect profile and
anticonvulsant efficacy when
administered p.o.
To further determine the suitability of CBDV as a clinical
candidate, we assessed both its motor side effect profile and
whether it could suppress seizures when administered p.o.
before PTZ treatment. Many currently used AEDs have sig-
nificant side effects at clinically effective doses, particularly
on motor function (Schachter, 2007). Additionally, a prereq-
uisite for human epilepsy treatment is that a drug is effective
after oral administration.
We used two motor tasks to investigate the side effect
profile of CBDV (50–200 mg·kg-1): a static beam test to assess
motor coordination (Stanley et al., 2005; Roberts et al., 2006)
and a grip strength test to assess drug-induced muscle relaxa-
tion and functional neurotoxicity (Nevins et al., 1993;
Crofton et al., 1996). CBDV had no significant effects on
motor performance at any dose compared with vehicle treat-
ment (Figure 6A–D). In the static beam assay, the pass rate
[c2(3) =4.053; P>0.1; Figure 6A] and mean distance travelled
(F3,79 =1.335; P>0.1; data not shown) were both unaffected
by CBDV. CBDV had no significant overall effect on the mean
number of foot slips (F3,79 =0.858; P>0.1; Figure 6B),
although we did note a non-significant increase in foot slips
in animals treated with 200 mg·kg-1CBDV (0.70 0.25 slips,
compared with 0.30 0.11 slips after vehicle treatment).
CBDV had no effect on grip strength (F3,79 =0.465; P>0.1,
Figure 6C). To validate the tests’ ability to detect AED-
induced motor deficits, a second group of animals received
VPA (125–350 mg·kg-1) or saline vehicle. VPA significantly
affected the percentage of animals that successfully com-
pleted the static beam test [c2(3) =35.084; P0.001;
Figure 6A], with doses 250 mg·kg-1significantly decreasing
the pass rate (P0.01). Similarly, both the number of foot
slips (F3,78 =9.140; P0.001; Figure 6B) and the mean dis-
tance travelled (F3,78 =15.561; P0.001; data not shown)
were significantly, negatively and dose-dependently affected
by treatment with 250 mg·kg-1VPA (P0.01). VPA also
significantly affected the grip strength of animals (F3,79 =
3.175; P0.05; Figure 6C), with a small, but significant
decrease in mean strength induced by 350 mg·kg-1VPA
(P0.05).
Finally, we investigated the ability of 400 mg·kg-1CBDV
administered p.o. (see Supporting Information Appendix S1
for dose details) to suppress PTZ seizures (90 mg·kg-1);
Figure 5
Effects of co-administration of CBDV and AEDs on pilocarpine-induced convulsions in rats. The effects of CBDV co-administration with VPA (A–D)
or phenobarbital (E–I) on pilocarpine-induced convulsions: severity (A and E), mortality (B and F), the incidence of tonic–clonic convulsions (C and
G), onset latency (D and H) and (for CBDV +phenobarbital only) the percentage of animals that remained seizure-free. In (D and H), onset latency
is presented as mean SEM In (A and E), median severity is represented by a thick horizontal line, the 25th and 75th percentiles by the box and
maxima and minima are represented by ‘whiskers’. Significance of CBDV treatment is given in text. n=15 in all cases. *P0.05, **P0.01 and
***P0.001 for AED effects.
BJP
Cannabidivarin as an anticonvulsant
British Journal of Pharmacology (2012) 167 1629–1642 1637
400 mg·kg-1CBDV significantly lowered the severity of PTZ-
induced seizures (Figure 6D, P0.05) from 5 to 3.5. There
were no significant effects of CBDV on seizure onset latency
(vehicle 58.6 3.7 s; CBDV 61.8 5.2 s; P>0.1), percentage
mortality (vehicle 26.7%; CBDV 20%; P>0.1) or develop-
ment of tonic–clonic seizures (vehicle 53.3; CBDV 33.3; P>
0.1). Overall, we demonstrated that the anticonvulsant
effects of CBDV in rat are due to genuine anticonvulsant
properties and not motor suppression, and that CBDV is
anticonvulsant when administered p.o. as well as i.p. in the
PTZ model.
Discussion
This study demonstrates, for the first time, that CBDV has
anticonvulsant properties, and, to date, is the only study that
has investigated the effects of CBDV in whole animals. Our
main finding is that CBDV suppresses seizures in four in vivo
seizure models at doses 50 mg·kg-1. CBDV also did not affect
normal motor function and was well-tolerated when
co-administered with AEDs. Moreover, CBDV suppressed epi-
leptiform activity in vitro.
In vitro effects of CBDV
In both in vitro models of epileptiform activity, LFP duration
and amplitude were significantly decreased by CBDV, with
efficacy varying between hippocampal subregions and
models. The CA3 region was most resistant to CBDV effects,
potentially due to its role as the epileptiform focus (Perreault
and Avoli, 1992; Hill et al., 2010). It has also been reported
that a smaller proportion of neurons in the CA1 contribute to
burst activity than in the CA3 (Perreault and Avoli, 1992),
potentially rendering the CA1 region more sensitive to the
effects of anti-epileptiform drugs. CBDV effects on LFP fre-
quency in the two models were opposite; CBDV increased
Mg2+-free-induced LFP frequency, but decreased 4-AP-induced
LFP frequency. This may be due to a genuine, model-
dependent CBDV effect on LFP frequency; however, the
response of frequency in the Mg2+-free model is in direct
contrast to all other findings across both models, where
varying degrees of anti-epileptiform effects were observed. In
addition, we have observed that LFPs in the Mg2+-free model
exhibit greater variation in frequency than the 4-AP model;
sporadic bursts of LFPs occur with periods of relative quies-
cence between them (see Hill et al., 2010). Thus, while the
frequency of LFPs in this Mg2+-free model was corrected to
allow for inherent increases, it may be that the unpredictabil-
ity of LFP incidence limits the accuracy of this process.
Overall, the magnitude of the effects of CBDV on LFP ampli-
tude and duration are comparable with those observed with
both CBD and clinically used AEDs (Sagratella, 1998; Hill
et al., 2010; Jones et al., 2010).
In vivo effects of CBDV and
clinical implications
We demonstrated that CBDV has significant anticonvulsant
effects in four seizure models with different bases across two
species. CBDV was effective in three models of generalized
seizure – mES and audiogenic in mice and PTZ in rats. In
particular, CBDV (200 mg·kg-1) completely prevented tonic–
clonic convulsions in the audiogenic seizure model and had
robust effects in the mES model, in line with the reported
Figure 6
Effects of CBDV on performance in the static beam and forelimb grip strength assays in rat and as an orally administered anticonvulsant. (A and
B): static beam performance; including the pass rate (A) and foot slips (B). (C) Performance in the grip strength assay. (A) Pass rate is represented
as percentage; (B and C), represented as mean SEM n=20 for static beam data, 10 for grip strength. (D) Effect of orally administered
400 mg·kg-1CBDV on the severity of PTZ-induced seizures. (A–C): n=20, (D): n=15. *P0.05, **P0.01 and ***P0.001 respectively. (A–C):
V=CBDV vehicle; S =VPA vehicle (saline).
BJP AJ Hill et al.
1638 British Journal of Pharmacology (2012) 167 1629–1642
efficacy of VPA and other AEDs in these models (Gareri et al.,
2004; Luszczki et al., 2011; 2012). Moreover, positive findings
in the mES model – a primary screen for putative anticonvul-
sants (Loscher, 2011) – are predictive of clinical efficacy
against generalized human seizures (Loscher, 2011). Audio-
genic seizures, although providing limited predictive differ-
entiation of future efficacy against human seizure types
(Loscher, 2011), are also a useful model of generalized seizure
(Pitkanen et al., 2006). Attenuation of PTZ-induced seizures
can be predictive of efficacy against absence seizures, as well
as predicting effective suppression of generalized seizures in
humans (Veliskova, 2006). Hence, CBDV should also be
investigated in non-convulsive seizure models (e.g. WAG/Rjj
rats; Coenen and Van Luijtelaar, 2003). Importantly, p.o.
CBDV (400 mg·kg-1) also suppressed PTZ-induced seizures,
showing that CBDV can exert anticonvulsant effects when
administered orally.
Systemic administration of pilocarpine induces status epi-
lepticus with a temporal lobe focus that subsequently gener-
alizes and is associated with motor convulsions (Curia et al.,
2008). Interestingly, the anticonvulsant effects of CBDV only
became apparent when 200 mg·kg-1CBDV and AEDs were
co-administered. Thus, effects were observed only in higher-
power experiments in which 60, as opposed to 15, animals
received 200 mg·kg-1CBDV. These effects were limited (see
later), suggesting that CBDV is less effective in this model
than in the others studied here. However, our statistical
analyses revealed that the effects of CBDV in these experi-
ments were independent of, and separate from, the actions of
AED. Hence, it would be of interest to characterize the effects
of CBDV on pilocarpine-induced status epilepticus using direct
recordings of brain activity, for example via electroencepha-
lographic or electrocorticographic recordings in this model as
status epilepticus activity can persist in the absence of motor
activity.
Many AEDs exert significant motor side effects (Schachter,
2007), which can limit patient quality of life. To address this
and confirm that CBDV’s anticonvulsant actions were due to
direct actions on seizures and not motor suppression, we
investigated the effects of CBDV on the performance of rats in
the static beam and grip strength tasks. These tests assess
balance, coordination, muscle relaxation and drug-induced
functional neurotoxicity (Nevins et al., 1993; Crofton et al.,
1996; Stanley et al., 2005; Muller et al., 2008). CBDV did not
affect grip strength, and although the number of foot slips
did increase after 200 mg·kg-1CBDV treatment, this effect
was not significant. Our tests were validated by the finding
that, consistent with previous studies, VPA negatively
affected all motor parameters (Roks et al., 1999).
Our in vivo results showing that CBDV has comparatively
strong anticonvulsant effects in a range of seizure models,
indicate that CBDV has significant potential for the treat-
ment of generalized, human seizures and should be further
investigated against temporal lobe seizures. Furthermore,
data from the motor function assays indicate that CBDV does
not have significant adverse motor effects at anticonvulsant
doses. In the future, it will be of great interest to investigate
CBDV’s properties in models of chronic epilepsy and hyper-
excitability. The effect of chronic CBDV treatment on behav-
iour in healthy and epileptic animals is also worthy of
investigation.
Co-administration studies
Clinical investigation of new anticonvulsants is typically per-
formed using the candidate AED as an adjunctive treatment
to the patient’s current treatment regimen (French et al.,
2001). Therefore, we investigated the effects of CBDV
(200 mg·kg-1) when co-administered with clinically used anti-
convulsants. The three anticonvulsants used were chosen
based on their use as prescribed AEDs and, more pragmati-
cally, reported efficacy in the seizure models used (Loscher
et al., 1991; Sofia et al., 1993; Shantilal et al., 1999; Lindekens
et al., 2000; Loscher, 2011). No negative interactions between
CBDV and the AEDs were observed, indicating that CBDV is
well-tolerated when co-administered with the three clinically
used AEDs employed in these studies. The anticonvulsant
effect of CBDV beyond that of these AEDs was variable, in our
study. When administered with ESM before PTZ or VPA
before pilocarpine, CBDV contributed significantly to the
effects seen on severity (both cases), mortality (VPA in pilo-
carpine only), latency (ESM only) and the incidence of tonic–
clonic convulsions (VPA in pilocarpine only). The majority of
the significant facilitatory effects of CBDV were seen at the
lower two doses; this could be due to the greater potential for
anticonvulsant actions when the AED is not producing a
maximal effect itself. However, 200 mg·kg-1CBDV appeared
to have little effect on pilocarpine-induced convulsions when
administered with phenobarbital at any dose, although it
should be noted that all doses of phenobarbital strongly
suppressed seizure activity, probably limiting CBDV’s effect.
CBDV had limited effects on PTZ-induced seizures when
co-administered with VPA. Thus, CBDV had AED-dependent
effects in these experiments, producing notable improve-
ments over AED treatment alone in two of four experiments.
Based on these data, we postulate that CBDV is well-tolerated
when co-administered with three AEDs used in the clinic for
a variety of epileptic syndromes, but that further investiga-
tion of its anticonvulsant properties in combination with
other drugs is required, for example, using isobolographic
experimental design and analysis (e.g. Luszczki et al., 2010).
Anticonvulsant mechanisms of CBDV
This is the first investigation of CBDV effects in any in vivo
model or system; in vitro information on CBDV pharmaco-
logical properties, while growing, is limited (Scutt and Wil-
liamson, 2007; De Petrocellis et al., 2011a,b) and remains of
unknown in vivo or clinical relevance. For example, reported
effects of CBDV at recombinant TRP channels are as yet
unconfirmed in native tissue and it is unknown how such
TRP-based mechanisms of action could affect excitability in
epileptogenic areas. While TRPV1 expression in brain areas
including the hippocampus remain controversial (Mezey
et al., 2000; Cavanaugh et al., 2011), the functional expres-
sion of other TRP subtypes in relevant parts of the brain has
yet to be confirmed (Crawford et al., 2009; Hirata and Oku,
2010). CBDV has also been reported to inhibit diacylglycerol
lipase (DAGL) a(De Petrocellis et al., 2011a), the enzyme
responsible for the production of the endocannabinoid
2-arachidonoylglycerol (2-AG; Stella et al., 1997). The effect
of inhibiting 2-AG production is likely to be complex. The
initial effect would be to decrease 2-AG levels and subsequent
activation of CB1cannabinoid receptors. However, the overall
effect of this on seizure activity would depend on propor-
BJP
Cannabidivarin as an anticonvulsant
British Journal of Pharmacology (2012) 167 1629–1642 1639
tional CB1cannabinoid receptor expression and localization
on different presynapses (i.e. excitatory or inhibitory), and
the contribution of inhibitory GABAergic circuits in brain
areas crucial to epileptogenesis, as a decrease in 2-AG would
result in less suppression of both excitatory and inhibitory
synapses. Furthermore, over longer time courses, it has been
reported that CB1cannabinoid receptor levels can be affected
by changes in agonist levels, that is higher levels of CB1
cannabinoid receptor agonists can increase internalization of
the receptor (Coutts et al., 2001). Thus, reduced 2-AG levels
could cause increased the number of CB1cannabinoid recep-
tors at the membrane. In addition, in this study the effects of
CBDV were only investigated on acute seizures and CB1can-
nabinoid receptor expression changes during both animal
models (e.g. pilocarpine-induced spontaneous recurrent sei-
zures as a model of temporal lobe epilepsy) of chronic epi-
lepsy and in human epilepsy (Magloczky et al., 2010; Karlocai
et al., 2011), which could affect the consequences of changes
in endocannabinoid levels upon seizure activity. D9-THC has
been reported to have a direct anticonvulsant action via
CB1cannabinoid receptor agonism (Wallace et al., 2001).
However, the effects of CBDV on CB1cannabinoid receptors
have not been characterized. Furthermore, 200 mg·kg-1
CBDV had no significant effects in the motor function assays
used here, whereas CB1cannabinoid receptor agonists
produce significant motor deficits (Carlini et al., 1974), which
suggests that CBDV does not act via CB1cannabinoid recep-
tor agonism.
CBDV is the propyl analogue of CBD and a qualitative
comparison of the effects of CBD and CBDV on PTZ-induced
seizures showed that both compounds improve mortality and
severity. However, CBD produced these effects at 100 mg·kg-1,
a dose at which CBDV did not affect severity. CBD did not
appear to affect onset latency (100 mg·kg-1), whereas CBDV
delayed seizure onset in a dose-dependent manner that
reached significance at 200 mg·kg-1. The comparison between
CBD and CBDV in the pilocarpine model is less simple as
CBDV at 200 mg·kg-1had wider-ranging anticonvulsant
effects in our co-administration experiments (on severity,
mortality and latency as well as the proportion of animals
that developed tonic–clonic convulsions), but was not effec-
tive in initial experiments at any dose, whereas low-dose CBD
affected tonic–clonic convulsions, but no other measures.
Hence, it would be of interest to perform a direct experimen-
tal comparison both of efficacy and how similarly CBD and
CBDV affect seizures. Although assumptions of pharmaco-
logical similarity between plant cannabinoids on the basis of
structural homology should be made with caution (e.g. the
opposing effects of D9-THC and D9-THCV on CB1cannabinoid
receptors), CBD is anticonvulsant in animals and humans,
and more is known about CBD’s pharmacological properties,
if not its specific anticonvulsant mechanism(s) of action.
CBD has a wide range of known pharmacological targets,
which are unlikely to include CB1cannabinoid receptors, that
could underlie its anticonvulsant effects (Hill et al., 2012).
These include inhibition of T-type Ca2+channels (Ross et al.,
2008), inhibition of GPR55 in some tissues/preparations
(Ryberg et al., 2007), modulation of mitochondrial calcium
handling in neurons (Ryan et al., 2009) and increased activity
of inhibitory non-cannabinoid GPCRs including 5-HT1A
(direct agonism; Russo et al., 2005) and adenosine A1(via
effects on adenosine uptake; Carrier et al., 2006). Thus, if
CBDV shares some or all of CBD’s pharmacological targets, it
is possible that CBDV also acts via multiple mechanisms to
produce its overall anticonvulsant effect, as opposed to exert-
ing a high-efficacy action at a single target. However, there is
no a priori reason to assume a common target and there is
clearly some divergence between the properties of CBD and
CBDV, for example CBD, but not CBDV, inhibits FAAH (De
Petrocellis et al., 2011a).
In conclusion, our most important finding is that CBDV
possesses strong anticonvulsant properties in a range of in
vivo seizure models that parallel a variety of human seizure
types and pathologies; anticonvulsant effects were also seen
after oral, as well as i.p., administration. As with many clini-
cally used AEDs, further work is required to determine the
anticonvulsant mechanism of CBDV, but the significant anti-
convulsant effects and favourable motor side effect profile
demonstrated in this study identify CBDV as a potential
standalone AED or as a clinically useful adjunctive treatment
alongside other AEDs.
Acknowledgements
UoR authors thank GW Pharmaceuticals and Otsuka Pharma-
ceuticals for research sponsorship and the provision of CBDV
and thank Simon Marshall for technical assistance.
Conflict of interest
The work reported was funded by grants to BJW, CMW & GJS
from GW Pharmaceuticals and Otsuka Pharmaceuticals. BJW,
AJH, NAJ, CMW & GJS were responsible for experimental
design. YY and TF are employees of Otsuka Pharmaceuticals
and hold stocks in this company. MD and CGS are GW
Pharmaceuticals employees, and CGS is a stockholder.
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Supporting information
Additional Supporting Information may be found in the
online version of this article:
Appendix S1 Methods.
Tables S1 and S2 For each seizure parameter that was
affected by CBDV +AED treatment, the analysis of the indi-
vidual AED effect is given (either as ANOVA or Chi-squared).
The directions of significant effects are also given by an
upward or downward arrow (irrespective of the parameter, all
significant AED effects described are anticonvulsant). Addi-
tionally, the doses at which AEDs were significantly anticon-
vulsant are indicated with post hoc p values given after.
Finally, analyses of interactions between CBDV and AED
effects are given.
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1642 British Journal of Pharmacology (2012) 167 1629–1642
... CBDV, a structural homolog of CBD, possesses antiepileptic properties when tested in animals and humans. When examined in vitro in rat brain slices, CBDV attenuates epileptiform local field potentials induced by 4-amino pyridine (Hill, et al., 2012). In vivo, CBDV (200 mg/kg per day) significantly reduces PTZ-induced seizure activity in the rats (Hill, et al., 2012). ...
... When examined in vitro in rat brain slices, CBDV attenuates epileptiform local field potentials induced by 4-amino pyridine (Hill, et al., 2012). In vivo, CBDV (200 mg/kg per day) significantly reduces PTZ-induced seizure activity in the rats (Hill, et al., 2012). However, when used alone, CBDV has no effect on pilocarpine-induced seizures, but requires the coadministration of valproate or phenobarbital to be effective. ...
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The medicinal use of Cannabis sativa L. can be traced back thousands of years to ancient China and Egypt. While marijuana has recently shown promise in managing chronic pain and nausea, scientific investigation of cannabis has been restricted due its classification as a schedule 1 controlled substance. A major breakthrough in understanding the pharmacology of cannabis came with the isolation and characterization of the phytocannabinoids trans -Δ ⁹ -tetrahydrocannabinol (Δ ⁹ -THC) and cannabidiol (CBD). This was followed by the cloning of the cannabinoid CB1 and CB2 receptors in the 1990s and the subsequent discovery of the endocannabinoid system. In addition to the major phytocannabinoids, Δ ⁹ -THC and CBD, cannabis produces over 120 other cannabinoids that are referred to as minor and/or rare cannabinoids. These cannabinoids are produced in smaller amounts in the plant and are derived along with Δ ⁹ -THC and CBD from the parent cannabinoid cannabigerolic acid (CBGA). While our current knowledge of minor cannabinoid pharmacology is incomplete, studies demonstrate that they act as agonists and antagonists at multiple targets including CB1 and CB2 receptors, transient receptor potential (TRP) channels, peroxisome proliferator-activated receptors (PPARs), serotonin 5-HT 1a receptors and others. The resulting activation of multiple cell signaling pathways, combined with their putative synergistic activity, provides a mechanistic basis for their therapeutic actions. Initial clinical reports suggest that these cannabinoids may have potential benefits in the treatment of neuropathic pain, neurodegenerative diseases, epilepsy, cancer and skin disorders. This review focuses on the molecular pharmacology of the minor cannabinoids and highlights some important therapeutic uses of the compounds.
... It is a non-psychoactive homologue of cannabidiol (CBD,8), with a propyl side chain. It is biosynthetically derived from cannabigerovaric acid, and is usually present in small amounts, but more is found in plants growing mainly in northeastern India, Nepal, and Afghanistan (WLD type), that naturally produce high levels of CBD (8) (Hill et al. 2012). ...
... To date, however, so far only a small number of studies has been performed with 17, and its mechanism of action has not been satisfactorily elucidated. Like CBD (8), CBDV (17) has a physiologically insignificant affinity for CB1 and CB2 receptors (Hill et al. 2012) and is an antagonist on the GPR55 receptor (Iannotti et al. 2014). 17 activates and rapidly desensitizes TRPV1, TRPV2, and TRPA1 receptors (Iannotti et al. 2014). ...
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Cannabis spp. are some of the most controversial medicinal plants in the world. They contain great amounts of biologically active secondary metabolites, including the typical phenolic compounds called cannabinoids. Because of their low toxicity and complex biological activities, cannabinoids can be useful in the therapy of various diseases, but adverse psychological effects (of Δ9-THC in particular) raise concerns. This review summarizes the current knowledge of selected active C. indica compounds and their therapeutic potential. We summarize the main compounds contained in cannabis, the mechanisms of their effects, and their potential therapeutic applications. Further, we mention some of the clinical tests used to evaluate the efficacy of cannabinoids in therapy.
... Interestingly, the non-psychotropic phytocannabinoid cannabidiol (CBD) has demonstrated anticonvulsant and antiseizure effects in humans (Devinsky et al. 2018;Laux et al. 2019;Koo et al. 2020) and in animal models (Jones et al. 2010;Vilela et al. 2017;Kaplan et al. 2017;Gu et al. 2019) and has been approved for treatment of certain pediatric epilepsies by the US Food and Drug Administration. In addition, CBD has also been shown to attenuate cocaine-induced seizures in rodents (Gobira et al. 2015), while the structurally similar phytocannabinoid cannabidivarin (CBDV) also has anticonvulsant effects in rodents (Hill et al. 2012(Hill et al. , 2013Huizenga et al. 2019). It may be the case that these and other phytocannabinoids might be useful in the mitigation of convulsant effects of SCRAs, but no studies in this regard have yet been performed. ...
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Rationale Synthetic cannabinoid receptor agonists (SCRAs) are found in illicit smoking products, such as “K2” or “Spice.” Convulsions are commonly reported adverse effects of SCRAs but are poorly understood. Objectives We determined convulsant effects of SCRAs AB-PINACA, and 5F-ADB-PINACA in adult male NIH Swiss mice, and then determined if convulsant effects of AB-PINACA, 5F-AB-PINACA, 5F-ADB-PINACA, and JWH-018 elicited seizure-like effects using EEG. Methods Mice were administered SCRAs or pentylenetetrazole (PTZ) and placed in observation chambers where convulsant effects were scored. The capacity of the CB1R antagonist rimonabant, the benzodiazepine diazepam, or the non-specific CYP450 inhibitor 1-aminobenzotriazole (1-ABT) to attenuate convulsant effects was determined. Other mice were prepared with EEG headmounts to ascertain whether observed convulsions occurred concurrently with seizure-like effects by assessing root-mean-square (RMS) power, high amplitude EEG spike analysis, and videography. Results Mice receiving AB-PINACA or 5F-ADB-PINACA exhibited dose-dependent convulsant effects that were blocked by 10 mg/kg rimonabant pretreatment but not by pretreatment with 10 mg/kg diazepam; these convulsant effects were not altered in the presence of 100 mg/kg 1-ABT. Repeated administration of 10 mg/kg AB-PINACA and 3 mg/kg 5F-ADB-PINACA produced partial tolerance to convulsant effects but did not lead to cross-tolerance to PTZ-induced convulsions. In EEG studies, convulsant doses of AB-PINACA, 5F-AB-PINACA, 5F-ADB-PINACA, and JWH-018 did not produce seizures concomitantly with convulsions. Conclusions These data extend previous findings of convulsant effects of SCRAs and suggest that convulsant effects of AB-PINACA, 5F-AB-PINACA, 5F-ADB-PINACA, and JWH-018 are CB1R-mediated but are not associated with electroencephalographic seizures. These results further suggest that benzodiazepines may not effectively treat convulsions elicited by SCRA use in humans.
... This is not unexpected since CBDV has been detected by HPLC-UV in different hemp oils [107]. CBDV induces anticonvulsant effects without impairing motor function at 50-200 mg/kg in experimental models of seizures [108]. Studies support that oral administration of CBDV at 400 mg/kg in rats induces anticonvulsant effects in pentylenetetrazol-induced seizures [109]. ...
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This study aimed to determine if orally administered cannabidiol (CBD) lessens the cortical over-release of glutamate induced by a severe traumatic brain injury (TBI) and facilitates functional recovery. The short-term experiment focused on identifying the optimal oral pretreatment of CBD. Male Wistar rats were pretreated with oral administration of CBD (50, 100, or 200 mg/kg) daily for 7 days. Then, extracellular glutamate concentration was estimated by cortical microdialysis before and immediately after a severe TBI. The long-term experiment focused on evaluating the effect of the optimal treatment of CBD (pre- vs. pre- and post-TBI) 30 days after trauma. Sensorimotor function, body weight, and mortality rate were evaluated. In the short term, TBI induced a high release of glutamate (738% ± 173%; p < 0.001 vs. basal). Oral pretreatment with CBD at all doses tested reduced glutamate concentration but with higher potency at when animals received 100 mg/kg (222 ± 33%, p < 0.01 vs. TBI), an effect associated with a lower mortality rate (22%, p < 0.001 vs. TBI). In the long-term experiment, the TBI group showed a high glutamate concentration (149% p < 0.01 vs. SHAM). In contrast, animals receiving the optimal treatment of CBD (pre- and pre/post-TBI) showed glutamate concentrations like the SHAM group (p > 0.05). This effect was associated with high sensorimotor function improvement. CBD pretreatment, but not pre-/post-treatment, induced a higher body weight gain (39% ± 2.7%, p < 0.01 vs. TBI) and lower mortality rate (22%, p < 0.01 vs. TBI). These results support that orally administered CBD reduces short- and long-term TBI-induced excitotoxicity and facilitated functional recovery. Indeed, pretreatment with CBD was sufficient to lessen the adverse sequelae of TBI.
... Animal models have demonstrated an antiseizure effect and a favorable safety profile. 19,27 MECP2-308 mouse models treated with CBDV demonstrated improved sociability, brain weight, and overall health and partial improvement in motor function. 25,28 In an adult focal epilepsy cohort, Brodie et al. 29 demonstrated safety in humans and observed a 41% reduction in seizure frequency with CBDV. ...
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Objective: Rett Syndrome (RTT), commonly caused by methyl-CpG-binding protein 2 (MECP2) pathogenic variants, has many co-morbidities. 50-90% of children with RTT have epilepsy which is often drug resistant. Cannabidivarin (CBDV), a non-hallucinogenic phytocannabinoid has shown benefit in MECP2 animal models. This Phase I trial assessed the safety and tolerability of CBDV in female children with RTT and drug resistant epilepsy, as well as the effect on mean monthly seizure frequency (MMSF), the electroencephalogram (EEG), and non-epilepsy co-morbid symptoms. Methods: Five female children with drug resistant epilepsy and a pathogenic MECP2 variant were enrolled. Baseline clinical and laboratory assessments, including monthly seizure frequency, were recorded. CBDV oral solution (50mg/mL) was prescribed and titrated to 10mg/kg/day. Data collected over 15 months included pharmacokinetics, seizure type and frequency, adverse events, EEG, and responses to Rett syndrome behaviour questionnaire (RSBQ) and Rett syndrome symptom severity index, and was compared to baseline data. Results: All five children reached the maximum CBDV dose of 10mg/kg/day and had a reduction in MMSF (median 79% reduction). Three children had MMSF reduction >75%. This corresponded to an overall reduction in seizure frequency from 32 to 7.2 seizures per month. 91% of adverse events were mild or moderate and none required drug withdrawal. 62% were judged unrelated to CBDV. 31% of adverse events were identified as possibly related, of which nearly all were mild, and the remainder were later assessed as RTT symptoms. Hypersomnolence and drooling were identified as related to CBDV. No serious adverse events reported were related to CBDV. No significant change was noted in EEG or non-epilepsy related symptoms of RTT. Significance: 10mg/kg/day of CBDV is safe and well tolerated in a paediatric Rett syndrome cohort and suggests improved seizure control in children with MECP2-related Rett syndrome.
... More than 140 phytocannabinoids have been identified in Cannabis sativa and recent preclinical research shows a growing number with anticonvulsant properties across a variety of animal models of epilepsy (Anderson et al., 2019b(Anderson et al., , 2021aBenson et al. 2020;Chiu et al. 1979;Davis and Hatoum 1983;Hill et al. 2010Hill et al. , 2012Kaplan et al. 2017;Karler and Turkanis 1979). This includes phytocannabinoid acids that are synthesized enzymatically in the cannabis plant, as well as neutral phytocannabinoids that are formed from the non-enzymatic decarboxylation of phytocannabinoid acids via thermal degradation. ...
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Objective: Cannabigerolic acid (CBGA), a precursor cannabinoid in Cannabis sativa, has recently been found to have anticonvulsant properties in the Scn1a+/- mouse model of Dravet syndrome. Poor brain penetration and chemical instability of CBGA limits its potential as an anticonvulsant therapy. Here, we examined whether CBGA methyl ester, a more stable analogue of CBGA, might have superior pharmacokinetic and anticonvulsant properties. In addition, we examined whether olivetolic acid, the biosynthetic precursor to CBGA with a truncated (des-geranyl) form, might possess minimum structural requirements for anticonvulsant activity. We also examined whether olivetolic acid and CBGA methyl ester retain activity at the epilepsy-relevant drug targets of CBGA: G-protein-coupled receptor 55 (GPR55) and T-type calcium channels. Methods: The brain and plasma pharmacokinetic profiles of CBGA methyl ester and olivetolic acid were examined following 10 mg/kg intraperitoneal (i.p.) administration in mice (n = 4). The anticonvulsant potential of each was examined in male and female Scn1a+/- mice (n = 17-19) against hyperthermia-induced seizures (10-100 mg/kg, i.p.). CBGA methyl ester and olivetolic acid were also screened in vitro against T-type calcium channels and GPR55 using intracellular calcium and ERK phosphorylation assays, respectively. Results: CBGA methyl ester exhibited relatively limited brain penetration (13%), although somewhat superior to that of 2% for CBGA. No anticonvulsant effects were observed against thermally induced seizures in Scn1a+/- mice. Olivetolic acid also showed poor brain penetration (1%) but had a modest anticonvulsant effect in Scn1a+/- mice increasing the thermally induced seizure temperature threshold by approximately 0.4°C at a dose of 100 mg/kg. Neither CBGA methyl ester nor olivetolic acid displayed pharmacological activity at GPR55 or T-type calcium channels. Conclusions: Olivetolic acid displayed modest anticonvulsant activity against hyperthermia-induced seizures in the Scn1a+/- mouse model of Dravet syndrome despite poor brain penetration. The effect was, however, comparable to the known anticonvulsant cannabinoid cannabidiol in this model. Future studies could explore the anticonvulsant mechanism(s) of action of olivetolic acid and examine whether its anticonvulsant effect extends to other seizure types.
... Cannabidivarin (CBDV) was isolated in 1969 and is a propyl analog of CBD [78]. Little is still known about the mechanism of action of this phytocannabinoid, but the lack of psychotropic activity and its anticonvulsive properties have already been demonstrated, making CBDV a promising therapeutic agent [79]. Similar to CBD, CBDV presents very low affinity for the CBs [80,81]. ...
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Full-text available
Although cannabinoids have been used for centuries for diverse pathological conditions, recently, their clinical interest and application have emerged due to their diverse pharmacological properties. Indeed, it is well established that cannabinoids exert important actions on multiple sclerosis, epilepsy and pain relief. Regarding cancer, cannabinoids were first introduced to manage chemotherapy-related side effects, though several studies demonstrated that they could modulate the proliferation and death of different cancer cells, as well as angiogenesis, making them attractive agents for cancer treatment. In relation to breast cancer, it has been suggested that estrogen receptor-negative (ER−) cells are more sensitive to cannabinoids than estrogen receptor-positive (ER+) cells. In fact, most of the studies regarding their effects on breast tumors have been conducted on triple-negative breast cancer (TNBC). Nonetheless, the number of studies on human epidermal growth factor receptor 2-positive (HER2+) and ER+ breast tumors has been rising in recent years. However, besides the optimistic results obtained thus far, there is still a long way to go to fully understand the role of these molecules. This review intends to help clarify the clinical potential of cannabinoids for each breast cancer subtype.
... It is mostly found in Cannabis indica, a species predominantly found in Asian and African countries (Iannotti et al., 2014). In the context of the medicinal uses of CBDV, the anti-convulsant activity can make it an effective drug for the treatment of seizures (Hill et al., 2012). Besides the single occurring seizures, this phytocannabinoid is effective for the treatment of recurrent seizures like epilepsy and other neuro disorders (Whalley et al., 2015). ...
Article
Cannabis, a genus of perennial indigenous plants is well known for its recreational and medicinal activities. Cannabis and its derivatives have potential therapeutic activities to treat epilepsy, anxiety, depression, tumors, cancer, Alzheimer's disease, Parkinson's disease, to name a few. This article reviews some recent literature on the bioactive constituents of Cannabis, commonly known as phytocannabinoids, their interactions with the different cannabinoids and non-cannabinoid receptors as well as the significances of these interactions in treating various diseases and syndromes. The biochemistry of some notable cannabinoids such as tetrahydrocannabinol, cannabidiol, cannabinol, cannabigerol, cannabichromene and their carboxylic acid derivatives is explained in the context of therapeutic activities. The medicinal features of Cannabis-derived terpenes are elucidated for treating several neuro and non-neuro disorders. Different extraction techniques to recover cannabinoids are systematically discussed. Besides the medicinal activities, the traditional and recreational utilities of Cannabis and its derivatives are presented. A brief note on the legalization of Cannabis-derived products is provided. This review provides comprehensive knowledge about the medicinal properties, recreational usage, extraction techniques, legalization and some prospects of cannabinoids and terpenes extracted from Cannabis.
Chapter
Here in part two, a brief explanation in essential oil/ terpene administration as well as cover the medicinal effects of terpenes focusing on biphasic pharmacokinetics and possible paradoxical reactions and molecular sites of interest, including the medicinal properties of a specific flavonoid; an explanation into the paradoxical entourage and identifying common misconceptions from cannabis use and education; we finalize our look into the paradoxical location learning biphasic and paradoxical reactions from cannabis with an in-depth look into the cause of ASR/ATD following with a fundamental explanation how stress with the wrong medication can instigate the situation. The Multi Cultivar Entourage Effect Chart (MCEEC) directed goal was to unravel multiple cultivars bioavailability to then combine and create a more robust and stronger entourage being pulled from multiple cultivars with specific bioavailability of cannabinoids, terpenoids, and flavonoids necessary to treat any specific indication. Indirectly the chart also identified inter-entourages, more importantly, “antagonistic” inter-entourages. By helping a patient describe their reactions, understand, identify and track terpenes and cannabinoids that cause specific reactions, the patient will be able to identify a profile that works for them, which gives an explanation and solution to identifying how to manage cannabis medication for the patient along with conclusion and thoughts.
Chapter
This two-part section helps the reader to achieve a better understanding of how cannabis works as a viable medication for the endocannabinoid system (ECS) and central nervous system (CNS) in humans by identifying individual synergies between cannabinoids, or cannabinoids and terpenes in their journey through the ECS and CNS in various mammalian patient indicators to unmask this paradoxical effect. The specific biphasic/paradoxical manner in question was researched and inevitably identifies cannabis use that manipulates tryptophan uptake, serotonin release, and dopamine actuation. Therefore, a patient’s diet may demand a higher tryptophan and dopa-L supplementation to avoid a paradoxical agitation on the receptor level. This chapter explains the pathology of how cannabis consistently reacts in the ECS for every individual, only separated by metabolism and disruption/trauma in the ECS and CNS, implying that there was no found paradoxical effect existing in cannabis, but in the patient, and thus is perceived the same in every individual, only mediated by metabolism, environment (surroundings), and the exception for individuals who process stimulants and tryptophan and/or serotonin in a disrupted manner causing a perceived paradoxical effect or the build-up to and/or what will be referred to as ASR/ATD. The cannabis industry, growers/breeders, interpeners/cannabis sommeliers/bud tenders, and dispensaries need to continue to constantly strive for more knowledge, just as the researchers and FDA need to continue their work to understand the benefits of cannabis, and most importantly, all must work together to remove cannabis from the Schedule I and Schedule 2 classification.
Book
An understanding of mechanisms underlying seizure disorders depends critically on the insights provided by model systems. In particular with the development of cellular, molecular, and genetic investigative tools, there has been an explosion of basic epilepsy research. Models of Seizures and Epilepsy brings together, for the first time in 30 years, an overview of the most widely-used models of seizures and epilepsy. Chapters cover a broad range of experimental approaches (from in vitro to whole animal preparations), a variety of epileptiform phenomenology (including burst discharges and seizures), and suggestions for model characterization and validation, such as electrographic, morphologic, pharmacologic, and behavioral features. Experts in the field provide not only technical reviews of these models but also conceptual critiques - commenting on the strengths and limitations of these models, their relationship to clinical phenomenology, and their value in developing a better understanding and treatments. Models of Seizures and Epilepsy is a valuable, practical reference for investigators who are searching for the most appropriate laboratory models for addressing key questions in the field. It also provides an important background for physicians, fellows, and students, offering insight into the potential for advances in epilepsy research.
Article
Delta(9)-tetrahydrocannabinol binds cannabinoid (CB(1) and CB(2)) receptors, which are activated by endogenous compounds (endocannabinoids) and are involved in a wide range of physiopathological processes (e.g. modulation of neurotransmitter release, regulation of pain perception, and of cardiovascular, gastrointestinal and liver functions). The well-known psychotropic effects of Delta(9)-tetra hydrocannabinol, which are mediated by activation of brain CB(1) receptors, have greatly limited its clinical use. However, the plant Cannabis contains many cannabinoids with weak or no psychoactivity that, therapeutically, might be more promising than Delta(9)-tetra hydrocannabinol. Here, we provide an overview of the recent pharmacological advances, novel mechanisms of action, and potential therapeutic applications of such non-psychotropic plant-derived cannabinoids. Special emphasis is given to cannabidiol, the possible applications of which have recently emerged in inflammation, diabetes, cancer, affective and neurodegenerative diseases, and to Delta(9)-tetrahydrocannabivarin, a novel CB(1) antagonist which exerts potentially useful actions in the treatment of epilepsy and obesity.