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Central Nervous System Drug Portfolio: Vigabatrin

Central Nervous System Drug Portfolio: Vigabatrin

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An essay for the 2012 Undergraduate Awards Competition by Anna-Mieke Bishop. Originally submitted for Undenominated Science, Pharmacology at National University of Ireland Galway, with lecturer Eilis Dowd in the category of Life Sciences
An essay for the 2012 Undergraduate Awards Competition by Anna-Mieke Bishop. Originally submitted for Undenominated Science, Pharmacology at National University of Ireland Galway, with lecturer Eilis Dowd in the category of Life Sciences

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Published by: Undergraduate Awards on Aug 31, 2012
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CNS Drug Portfolio:
Anna-Mieke Bishop
Vigabatrin is an irreversible gamma-amino-butyric acid transaminase inhibitor and acts to increase the amount of GABA in the brain. It is used in the treatment of epilepsy, a disorder characterised by abnormal electrical dischargesin the brain, resulting in seizures.
: GABA; gamma-amino butyrate. GABA-T; gamma-amino butyrate transferase. VGB; Vigabatrin. ECF;extracellular fluid. IC50; concentration at which 50% of target is inhibited. Ki; dissociation constant, measure of affinity.
Table 1: Drug Targeting
Cells/ Tissues examined Intervention Effect Ref:
- Rat brain-Pseudomonas fluorescensEnzyme purification from tissuesamples and incubation with 4-amino-hex-5-enoic-acid (VGB)(concentration range of 0.05
1mM).Complete irreversible inhibition of GABA-T at 0.1 mM VGB; T
of 6 minutesApparent selectivity
no inhibition of GABA-T from Pseudomonas fluorescens.Lippert
et al 
.,1977- Cultured neurons- Cultured astrocytesCells cultured in presence of VGB toobtain IC
valuesGABA-T inhibition: IC
; 24 uMGABA-T inhibition: IC
; 89 uMLarsson
et al 
.,1986- Cerebral cortex of newbornmice- Cultured neurons of embryonic mice embryos- Cultured astrocytes of newborn mice; cortexGABA-T activity assay: 14
-GABAused. VGB and its separate isomerswere tested at variousconcentrations to obtain IC
resultsfor inhibition.IC50 for GABA-T inhibition by:(S)-isomer: 51uM (R)-isomer: 5453uM(S)-isomer: 43uM (R)-isomer: 5162uM(S)-isomer): 91uM (R)-isomer: 4678GABA-T levels increased to control levelsfollowing VGB removal.Gram
et al 
.,1989- Human brain: Ocipital lobe 3-6 g/day VGB administered toepileptic patients. GABA levelsmeasured via
H nuclear magneticresonance spectroscopyGABA levels increased to 2.6umol/gfollowing VGB therapy, versus 0.9umol/g,following no treatment.Petroff 
et al 
.,1995- Rat hippocampal neurons GABA release measured using patch-clamp recordings followingadministration of 0.05
100uM VGBVGB enhances depolarisation-evokednon-vesicular GABA releaseWu
et al 
., 2001- Pig liver GABA-T The crystal structure of GABA-T wassolved in its native form and incomplex with vigabatrinFormation of a covalent ternary adductwith the active site Lys-329 and the
pyridoxal 5’
- phosphate cofactor.Storici
et a
l.,2003- Pseudomonas fluorescens In vitro inhibition of GABA-T with 51mM VGBGABA-T inhibition at 51 mM. Ki; 26 +- 3mMSulaiman
et al 
.,2003- Blood and microdialysatesfrom brain of Sprague-Dawleyrats500 & 1000 mg/kg VGB administeredIP. Jugular vein catheter (bloodsampling) and microdialysis probes infrontal cortex and hippocampus (ECFsampling). VGB & GABA measuredwith HPLC1000mg/kg: Accumulation of VGB infrontal cortex. Increase in ECF GABAconcentrations in the frontal cortex butnot hippocampus. These elevations didnot reflect the concentration of VGB,rather its distribution.Tong
et al 
., 2009
et al 
. (1977) showed that 0.1 mM VGB, when incubated with rat brain GABA-T, resulted in a rapid decrease inenzyme activity (T
of inhibition: 6 minutes). Selectivity for mammalian GABA-T was confirmed, as no inhibition of GABA-T from Pseudomonas fluorescens occurred, at the same concentration of VGB. The protocol and results wereclear and succinct. However, the exact source/location of the enzymes was not clear
(‘rat brain’).
Future studieswould need to be carried out on human tissue to confirm that VGB has the same effect here also; enzymaticdifferences could alter results. The year of publication indicates that little was known of VGB at this time and so thisstudy would have been one of the first of its kind. Larsson
et al 
. (1986) and Gram
et al 
. (1989) aimed to investigatethe differential effects of VGB on neuronal and glial cell GABA-T. Their findings imply a direct GABA-ergic mechanismof action of VGB, as can be seen by the inhibitory effect and IC
s of VGB (and, in the case of Gram et al., theseparate isomers), preferentially in neuronal cells. Both studies demonstrated, after stopping treatment, GABA-Tactivity returned to control levels in less than 6 days. Limitations of both studies were that assumptions were madeon the levels of VGB present in the brain as no studies at that time had examined this aspect. The researchersextrapolated from results on GVG blood levels obtained from patients and from brain levels in mice (Gram
.,1983; Seiler
et al 
., 1987). These indirect measurements may have been an under or over-estimation. The subsequentstudies are from a decade later, when much more information about vigabatrin was known. Wu et al. (2001)demonstrated that VGB caused GABA efflux in neighbouring neurons and subsequent GABA
receptor activation of the neuron being studied
the use of electrophysiological recordings to measure receptor activation as an assay of GABA release allowed for detection of tiny quantities of GABA in synapses, compared with assays using HPLC orradiolabled GABA, which are not as sensitive. Storici
et al 
., (2003) managed to determine the crystal structure of GABA-T, alone and in combination with vigabatrin, providing support for the specific inactivation mechanismsproposed. This method provided precise adduct structure information, and did not require processes like radioligandsynthesis and peptide mapping. Sulaiman
et al.
(2003) studied the kinetics of GABA-T inhibition in Pseudomonasfluorescens, with complete inhibition occurring at 51mM with a Ki of 26mM. They found that VGB had no effect on
-ketoglutarate, transamination of which results in glutamate, indicating that VGB did not just block transamination
in general 
. It could be argued that P.fluorescens is not a valid model for human GABA-T, particularly as previouspapers have indicated human GABA-T specificity. Petroff 
et al 
. (2005) took advantage of the recent developments in
H nuclear magnetic resonance imaging, and used it to demonstrate the VGB-induced rise in GABA levels comparedto controls, in epileptic patients. Drug targeting using human patients has a clear advantage due to the particular invivo environment
it allows for more accurate measurements of a drugs effects and kinetics. Tong
et al 
. (2008) usedrat blood and ECF samples to examine the relationship between vigabatrin in blood and the different brain areas andto examine the relationship between vigabatrin and GABA concentrations. Vigabatrin had linear and dose-dependent pharmacokinetics in the blood. Brain-region specificity was also noted. The use of a living rat model hassignificant advantages over cell studies but as always, the data must be extrapolated to humans in order to be of relevance.
Table 2: Animal Testing
Species Strain Model & Test Effect of VGB Reference
Non-humanprimateGuinea baboon;
‘Papio papio’
 Photosensitive baboon; Photically-inducedmyoclonusComplete protection againstgeneralised myoclonus induced byphotic stimulation.Meldrum &Horton, 1978Cat - Amygdala & hippocampal-kindled seizures;model of partial onset and secondary generalisedlimbic seizuresDecrease in seizure stage andprolongation of after-dischargeduration. 50-200 mg/kg, IPMorimoto
et al 
.,1993Rat Wistar Kainic acid; model for status epilepticus. Prevention of kainic acid-inducedneuronal damage; no effect onreduction of convulsionsHalonen
et al 
.,1995Rat Wistar Systemic administration of pilocarpine, for amodel of intractable epilepsy. Focally-evokedpilocarpine-induced seizures to model thegeneration of complex partial seizuresSeizures ompletely prevented byVGB administeredintraperitoneally. Only partialprevention/protection followingintrahippocampal and intranigraladministrationSmolders
et al 
.,1997Mouse Swiss Pentylenetetrazole (PTZ) -induced clonicconvulsionsInhibited clonic seizures; ED
of 879mg/kg and potentiation of anticonvulsant activity of ethosuxamideSwiader
et al 
.,2003Rat Sprague-Dawley Amygdala-stimulation model of human temporallobe epilepsyDecrease in mean seizurefrequency by 60 +- 15 %. No effecton seizure durationNissinen &Pitkänen, 2006Mouse Ts65Dn Down Syndrome model of infantile spasms 65% reduction in GABAB-receptorinduced acute epileptic extensorspasmsCortez
et al 
., 2009Rat Sprague-Dawley Cryptogenic infantile spasms; prenatal primingwith betamethasone and postnatal trigger withNMDADelayed latency to onset anddecreased frequency of spasms(250mg/kg)Chachua
et al 

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