Dr Paul L Chazot - Society of Chemical Industry
Transcript of Dr Paul L Chazot - Society of Chemical Industry
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Dr Paul L ChazotSchool of Biological & Biomedical Sciences/
h i h i l S i i / Durham Biophysical Sciences Institute/ NE Medicinal Plant Research Group (MPRG)
Medicinal plants: new uses & new mechanisms
Medicinal Plants: From crop to cureFrom crop to cure29th March 2011
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Recent positive projects in MPRG with various plant extracts
DementiasDementias• GINKGO (Ginko biloba) • SAGE (Salvia officinalis) • SAGE (Salvia officinalis) • LEMON BALM (Melissa officinalis) • GREEN TEA (Camellia sinensis)• GREEN TEA (Camellia sinensis)
DepressionDepression• ST JOHNS WORT (Hypericum perforatum)
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Alzheimers DiseaseAlzheimers Disease• Approx. 5% of people over 65 and 20% over 80 have
dementia (700 000 UK)dementia (700,000 UK)• Many severely demented patients develop symptoms of
agitation (severe restlessness, irritability and aggression) • Antipsychotic drugs have been widely used to treat this
e.g. risperidone, donezepil, major tranquilizersSid ff tSide-effects:
Over-sedation Social withdrawal/Reduced wellbeingSocial withdrawal/Reduced wellbeingFallsFurther decrement in cognitionPossibly risk of stroke
URGENT NEED FOR ALTERNATIVEApart from cogniti e therap increasing clinical e idence for Apart from cognitive therapy, increasing clinical evidence for
efficacy and safety of plant essential oils for symptoms such as agitation
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AROMATIC ESSENTIAL OIL THERAPYCONTROLLED CLINICAL TRIALS
IN PEOPLE WITH DEMENTIA –up to 4 weeks
Mitchell et al 1993 - 12 people, Melissa and lavender v placebo.
Holmes et al 2001 - 21 people with agitation, lavender, cross-over
Smallwood et al 2001 – 21 people, lavender, randomised Smallwood et al 2001 21 people, lavender, randomised comparison massage
Ballard et al 2002 72 people with agitation Melissa double blind Ballard et al 2002- 72 people with agitation, Melissa, double blind placebo controlled
L t l 2005 L d j j b Lee et al 2005 - Lavender versus jojoba massage
Lai et al 2007 - 70 people Lavender versus sunflower oil inhalation
SIGNIFICANT REDUCTION IN AGITATION/ AGGRESSION
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PHARMACOLOGICAL DISSECTION ESSENTIAL OILS
• Essential oils Melissa officinalis (lemon balm) and Lavendula angustifolia (lavender) have calming and Lavendula angustifolia (lavender) have calming and sedative properties.
• Both essential oils (EO) may help in agitationBoth essential oils (EO) may help in agitation• Melissa may also increase attention (pro-cognitive)• 3 trials: BMJ Editorial suggests improvement in 3 trials: BMJ Editorial suggests improvement in
behaviour, quality of life, as well as social and constructive activities
Burns et al (2002) BMJ;325(7376):1312-3
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Ion Channels Investigated to probe mechanism
• GABAA receptor major inhibitory receptor in the brainmajor inhibitory receptor in the brain
V lt G t d S di Ch l• Voltage-Gated Sodium Channelfundamental for neurotransmission
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Radioligand binding pharmacological screens
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Both oils contain a GABA-A receptor blocker
80
100
ndin
g
IC50 is 0.071mg/ml. Data are mean ± s.e.m.
60
80
BPS
bi
Predicts GABAA channel blockingactivity
40
60
[35 S]
T %
activity
20
0
peci
fic
0 001 0 01 0 1 10
Sp
0.001 0.01 0.1 1Melissa oil conc in mg/ml
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Melissa oil Batch supplier comparison
100
120MG (George Baldwins)MB ( B ld i )di
ng %
80
100 MB ( Baldwins) MP (Pranarom)MQ ( Quintessence)B
PS b
ind
40
60 MF( Fytosan)
[35S]
TB
MG MB MP MQ MF0
20
Spec
ific
MG MB MP MQ MFMelissa oil concs
0.01 mg/ml, 0.1 mg/ml, 1mg/ml
S
0.001,0.01, 0.1mg/ml
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Melissa & Lavender EO Combination effectMelissa & Lavender EO Combination effect
[3H] flunitrazepam binding to adult rat[ H] flunitrazepam binding to adult ratforebrain
120
140 MelissaLavenderM+LH
]bi
ndin
g
80
100 M+L
ecifi
c [3
zepa
m b
40
60
Spe
luni
traz
0.001 0.01 0.10
20fl
oil concs in mg/ml
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Electrophysiology
Recording electrodeRecording electrode
Primary rat cortical culturesDIV 21 28DIV 21-28
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Both oils completely blocked evoked GABA inhibitory currents
Effect of MO on GABA-mediated currents in primary cortical neurons.
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Both EOs contain a sodium channel modulator
550600650
400450500550
H] B
TX(%
)
250300350400
fic
[3 Hnd
ing
(
100150200250
Spec
i b
i n
0.001 0.01 0.1 0.5 10
50
Conc.mg/mL
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Both oils are profoundly depressant despite di i hibi idisinhibition
0.1mgml Melissa EO
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Compounds detected in essential oils from b th L v d l tif li d M li both Lavandula angustifolia and Melissa,
obtained from GC-MS
CompoundM.offinalis
OilComposition
(%)a
M.offinalisOil
Composition(%)b
L.angustifoliaOil
Composition(%)a
L.angustifoliaOil
Composition(%)b
Oct-1-en-3-ol 0.6 0.6 0.2 0.2
3-Octanone tr tr 0.3 tr
Myrcene 0.1 tr 0.5 0.4y
p-Cymene tr tr 0.2 0.3
Limonene tr tr 0.3 0.2
(E)-β-Ocimene 0 5 0 6 1 3 1 6(E) β Ocimene 0.5 0.6 1.3 1.6
Linalool 0.8 0.8 30.8 31.1
α-Terpineol 0.1 tr* 1.3 1.7*
N l 0 9 1 1 0 2 0 3Nerol 0.9 1.1 0.2 0.3
Geranyl acetate 3.3 3.6 1.0 1.1
(E)-Caryophyllene 12.3 12.9 3.6 3.6
Caryophyllene oxide 3.9 3.7 0.6 0.7
aChromatography performed on a DB-5MS phase ;bchromatography performed on a ZB-WAX phase ;tr: < 0.1 % ;* co-elution of α-terpineol and germacrene D.
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Identification of new GABAA receptor antagonistMelissa Oil
Oct-1-en-3-ol
100
120 3-Octanone
Myrcene
S
40
60
80 P-Cymene
Limonene
LinaloolSpec
ific[
35S]
TBPS
bind
ing(
%)
0
20
40 Linalool
Nerol
Geranyl acetate
S
-3.5 -3.0 -2.5 -2.0 -1.5 -1.0 -0.5Caryophyllene
Caryophyllene Oxide
Log.Conc (mg/mL)
Ocimene
Alpha-Terpineol
The effects of Melissa officinalis essential oil constituents on TBPS binding to well-washed rat forebrain membranes. All data are expressed as the mean ±SEM from at least three separate experiments.
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Effects of Melissa oil constituents on the channel binding site of the GABAAR
35labelled by [35S] TBPS Chemical name Molecular formula IC50(mg/ml
)IC50(µM)
)
Melissa Oil - 0.019 -
Geranyl acetate C12H2OO2 17.11 87166.9
(E)-Caryophyllene C H 0 028 137 7(E)-Caryophyllene C15H24 0.028 137.7
Caryophyllene oxide C15H24O 0.584 2650.3
Limonene C10H16 0.256 1878.3
Myrcene C10H16 6.840 50205.5
Ocimene* C H 0 006 40 5Ocimene C10H16 0.006 40.5p-Cymene CH3C6H4CH(CH3)2 0.035 261.6
3-Octanone C H O 0 038 298 63-Octanone C8H16O 0.038 298.6
Linalool C10H18O 0.900 5836.6
Nerol C10H18O 0.079 513.3
Oct-1-en-3-ol C8H16O 1.140 8890.9
α-Terpineol C10H18O 0.211 1369.2
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Effects of Melissa oil constituents on the sodiumh l bi di l b ll d b [3 ] T
Chemical name Molecular formula EC50(mg/mL) EC50(µM)
channel binding labelled by [3H] BTX-B
Geranyl acetate C12H2OO2 >1 -(E)-Caryophyllene C15H24 >1 -Caryophyllene oxide C15H24O >1 -y p y 15 24
Limonene C10H16 >1 -Myrcene C10H16 >1 -Ocimene C10H16 >0.5 -
C CH C H CH(CH ) 1p-Cymene CH3C6H4CH(CH3)2 >1 -3-Octanone C8H16O >1 -Linalool C10H18O >1 -Nerol C10H18O >1 -10 18
Oct-1-en-3-ol C8H16O >1 -Α-Terpineol C10H18O >1 -
•No potent effects (< 0.5 mg/ml) of major components upon [3H] BTX binding
•The sodium channel modulator is likely to be a minor componentThe sodium channel modulator is likely to be a minor component
•Fractionation and more sensitive detection methods required
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Biochemical pharmacology screenp gy
Sunflower oil (control) v Melissa essential oilSunflower oil (control) v Melissa essential oil
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Melissa oil is mildly neurotoxic at high concentrationsconcentrations
0 70.8
0.50.60.7
05
0.20.30.4
Abs 4
ol O l) l) l) l)
0.00.10.2
Control
SO
g/ml)
μ
LB (0.1
g/ml)
μ
LB (0.01
g/ml)
μ
LB (0.00
1
g/ml)
μ
B(0.
0001
L LB
LB
LB (
Treatment
Mild toxicity at high concentrations consistent with low affinity GABAAR inhibitory properties/modest protection at 0.001mg/ml
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Dominating neuronal depressant activityDominating neuronal depressant activity
Effect of MO (24h exposure) on cfos expression
C SO MO
Exposure to MO (0 01 mg/ml)Exposure to MO (0.01 mg/ml)reduces cfos expression (marker of neuronal activation
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Approach to separate the various pharmacological components:pharmacological components:
Solid Phase Fractionation of Essential oils
Whole Fraction 1 Fraction 2 Fraction 3
Hydrocarbons Carbonyls Primary alcoholsy(incl. ocimene
yEthersesters
Diols Acids
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N P t ti l f M li i il d New Potential of Melissa in epilepsy and hyperalgesia
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Reversible depressant effects of MO on evoked bursts of action potentials &
Concentration-dependent suppression of the secondary spikes in the burst profile
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MO can reversibly reverse ictal-like epileptiform activity in a 4 AP induced brain epileptiform activity in a 4-AP-induced brain
slice (visual cortex) model for epilepsy
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The modulated site for anticonvulsants on the VGSC complexon the VGSC complex
?
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Melissa and Lavender EO Ph l gi l di tiPharmacological dissection
• The volatile oils contain a novel GABAA receptor The volatile oils contain a novel GABAA receptor antagonist lead compound, namely ocimene
• The dominant net effect of the essential oils (lavender & Melissa) on neurons is depressant
• The mechanism for depressant defined as inhibition of membrane excitability/electrogenesis via a VGSC
• Fractionation protocols & computer modelling strategies under development for identifying VGSC g p y gmodulators
W h id tifi d l th ti f f • We have identified novel therapeutic arenas for use of Melissa and Lavender essential oils, namely epilepsy & hyperalgesia
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Acknowledgements
Durham University: Dr Paul L Chazot, Rushdie AbuhamdahD S Ab h d h ( U i i f J d A )Dr Sawsan Abuhamdah (now University of Jordan, Amman)
Formally Otago University Medical School, NZ: y g y ,Prof George Lees, Ihaia Hoskin, Dr Liping Huang
Sunderland Pharmacy School: Sunderland Pharmacy School: Dr Abdel Ennaceur, Mwajuma Mahita
N l U i iNewcastle University:Prof Elaine Perry, Dr Mike Carroll
Kew Gardens: Dr Melanie-Jayne Howes
FINANCIAL SUPPORTFINANCIAL SUPPORTALZHEIMERS SOCIETY UK, Durham Wolfson Institute, Islamic Development Bank