GABA

RECEPTOR

DR. NITIN SHINDE

FYR DEPARTMENT OF
PHARMACOLOGY
Lokmanya Tilak Municipal
Medical College & GH SION
MUMBAI-22
DATE:11-01-2014.

CONTENTS
Section-A :
1.
2.
3.
4.

(Basic Understanding Of GABA Receptors And Their Types)

Neurotransmitter :- GABA
GABA: SYNTHESIS, UPTAKE, AND METABOLISM
A,B,C , types of GABA receptors.
GABA : A receptor
a) GABA A- structure
b) GABA A- subunits
c) GABA A- MOA

5. Extra Synaptic GABA A receptors

6. GABA B receptors
a) Introduction
b) Molecular structure of GABA B receptor
c) M.O.A of GABA B receptor

Neurotransmitter :- GABA

4-aminobutanoic acid

Chief inhibitory neurotransmitter in mammalian central

nervous system.
Approximately 40 % neurons in mamalian CNS are purely
gabaminergic.
Synthesis of GABA

GABA: SYNTHESIS, UPTAKE,

AND
METABOLISM
Synthesized in brain GABA shunt

GABA: SYNTHESIS, UPTAKE,

AND
METABOLISM

A,B,CTypes of GABA
Receptor

Three main types: A, B, and C.

The GABAA receptor, is a ligand-gated Cl ion

channel,
an "ionotropic receptor.

The GABAB receptor is a GPCR.metabotropic.

The GABAC receptor is a transmitter-gated Cl
channel.Newly discovered.

Subunits at GABA A

SUBUNITS OF GABA

GABAA receptor structure

It is a multimeric transmembrane
receptor that consists of 5 subunits
arranged around a central Cl ion
pore.
Members of family of Cys-loop
ligand-gated ion channels (loop
formed by a disulfide bond betn 2
cysteine residues)
The receptor is usually located postsynaptically.
However, some isoforms may be found extrasynaptically

 Each subunit is composed of a polypeptide sequence of

approximately 450630 amino acids (4060 kDa) with large Nterminal and smaller C-terminal extracellular domains.

individual subunits contain four distinct transmembrane(TM)

domains, with the second transmembrane domain (TM2) lining the
channel lumen.

A large intracellular loop

connects the TM3 and TM4
regions providing sites for
phosphorylation
by a range of serine, threonine, aspargine & tyrosine
kinases

GABAA receptor MOA

Binding of GABA triggers opening of the Cl ion pore.
This drives the membrane potential towards the reversal
potential of the Cl ion which is about 65 mV in neurons,
inhibiting the firing of new action potentials.
This makes it more difficult for excitatory neurotransmitters to
depolarize the neuron.
The net effect is typically inhibitory, reducing the activity of
the neuron.

Video clip

Extrasynaptic GABAA Receptors

Synaptic GABAA receptors Underlie
phasic inhibition
By contrast, extra-synaptic receptors
are usually exposed to low but
persistent
GABA concentrations leading to
tonic inhibition.
Compared to their synaptic counterparts, these show
i. Increased sensitivity to GABA
ii. Reduced propensity to desensitize
iii. More rapid deactivation phase after removal of GABA

Extrasynaptic GABAA Receptors

The occurrence of tonic GABAA inhibition coincides with the
expression of relatively rare receptor subunits, articularly the
4, 6, and subunits, and as a general rule-of-thumb,
subunit-containing receptors are extrasynaptic.
The subunit can also govern receptor pharmacology:
extrasynaptic GABAARs are insensitive to benzodiazepine
agonists
but highly sensitive to the GABAAR super agonist
tetrahydroisoxazolopyridineol (THIP/gaboxadol)

Extrasynaptic GABAA Receptors

& drugs
Studies have begun to identify extrasynaptic GABAARs as
novel targets for a diverse array of endogenous and
clinically relevant agents, including :

Certain Neuroactive steroids

Amino acid Taurine
Ethanol
Several Anesthetic
Hypnotic agents
Analgesics
Some Anticonvulsant drugs.

SUBUNIT AND PHARMACOLOGICAL

ACTION

GABAB - discovery
Baclofen was first synthesized in 1962 by a
chemist Heinrich Keberle and was shown to exert
potent muscle-relaxant and analgesic properties
. A report from Bowery et al. holds an invaluable
pharmacological tool in elucidating the role of GABAB
receptors in several disorders including epilepsy, cognition
defect.
Bowery and Hudson described a bicuculline insensitive action
of GABA in beclofen which lead to the discovery of GABA B
later.

Molecular
Structure of
GABA
(B)Receptors

Heterodimer composed of two similar subunits each with a seven trans-membrane

-helix (7 TM) topology.
GABA (B) R1 and GABA (B) R2.
Each subunit comprises a large N terminal extracellular domain
followed by 7-transmembrane helicesand an intracellular C terminus.
GABA (B) R1 binds to ligand and initiates a
conformational change in the receptor complex,
GABA (B) R2 interacts with and transmits this
signal to the intracellular G-protein trimer.
Two GABA (B) R1 isoforms, GABA (B)R1a and GABA (B) R1b, are expressed in
the brain.
Two Sushi motifs are present GABA (B) R1a and absent in GABA (B) R1b.

GABA-B
RECEPTOR
MOA

GABA-B RECEPTOR MOA

GABA- subclass ( GABAC)

A subclass of ionotropic GABA receptors, insensitive to
typical allosteric modulators
GABA receptors are exclusively composed of (rho)
subunits that are related to GABAA receptor subunits
Designated as the subfamily of the GABAA receptors
(GABAA-).
These receptors are found in the retina, spinal cord, superior
colliculus, and pituitary.

 Three homologous
-subunits, 1 to 3,
have now been
identified.
There is only limited evidence that the -subunits coassemble with any of the other GABAA receptor subunits.
The genes encoding the 1- and 2-subunits are found on
chromosome 6 of man, and are thus distinct from the
clusters of receptor subunit genes which are found on
Chromosomes 4, 5, 15 and X with the exception of
, which is found on chromosome 1.

GABA C MOA
Its is ionotropic receptor with action
similar to GABA-A Receptor.
These receptors, are Cl- pores that are
insensitive to both bicuculline and
baclofen.
They are designated GABAC in
1984 but An IUPHAR
nomencleature the term GABAC
be avoided and classifies it as
bicuculline and baclofen-insensitive
GABA receptors as a minor group

PHARMACOLOGICAL
APPLICATION OF GABA-A

BZD RECEPTOR MODULATORS

ALCOHOL & GABA
ROLE IN ANTIEPILEPTICS.
Neuroactive steroid

BASICS...
Agonists: Bind to the main receptor site - also referred to as the
"active" or "orthosteric" site- and activate it
Antagonists: Bind to the main receptor site but do not activate it.
Positive Allosteric Modulators: Bind to allosteric sites on the
receptor complex and affect it in a positive manner, causing
increased efficiency of the main site and thus increase in Cl conductance.
Negative Allosteric Modulators: Bind to an allosteric site on the
receptor complex and affect it in a negative manner, causing
decreased efficiency of the main site.
Uncompetitive Channel Blockers: Bind to or near the central pore
of the receptor complex and directly block Cl- conductance.

GABA A Agonist

Ibotenic acid and Muscimol

Contained in Amanita muscaria/pantherina/gemmata
(hallucinogenic mushroom) with muscarine, muscazone.
GABAA agonist + potent partial GABAC agonist
Muscimol is as much as 10 times more potent
Effects are frequently compared to a lucid dream state.
Psychoactive dose of muscimol is around 1015 mg

GABOXADOL
Extrasynaptic GABAA agonist

Increases deep sleep (stage 4).

BZDs/Z drugs work on the 1 subtype of receptors for the

neurotransmitter GABA -- that's akin to an 'on/off switch' for
the central nervous system.

On the other hand, gaboxadol works on another subtype,

called 4 -- it's more of a 'dimmer switch' that might help
regulate sleep in a less disruptive way

GABAA Antagonist

Various GABAA antagonists

Drug

Detail

Bicuculline Competitive antagonist of GABAA receptors

Utilized in laboratories in the in vitro study of epilepsy
Routinely used to isolate glutamatergic (excitatory amino
acid) receptor function.
Gabazine

Antagonist at GABAA receptors

Used in scientific research and has no role in medicine
Phasic (synaptic) inhibition is gabazine- sensitive, tonic
(extrasynaptic) inhibition is relatively gabazine- insensitive

Cicutoxin & Found in various plants, most notably water hemlock (Cicuta
Oenantho- & Oenanthe species)
toxin
Potent noncompetitive GABA receptor antagonist
Nausea, emesis and abdominal pain within 60 mins of
ingestion. Can lead to tremors, seizures & death

Various GABAA antagonists

Drug

Detail

Thujone

Found in a number of plants, such as arborvitae

(Thuja)
Used in herbal medicine, mainly for their immunesystem stimulating effects
Reported to be toxic to both brain and liver cells
Side-effects include anxiety and sleeplessness,
seizures at high dose

Picrotoxin
(cocculin)

Poisonous crystalline plant compound

Found in the fruit (fishberry) of climbing
plant Anamirta cocculus.
Noncompetitive antagonist for GABAA receptor
- Channel blocker
Can be used to counter barbiturate poisoning

Positive allosteric
modulators

GABAA receptor allosteric MODULATORS

Benzodiazepines (BDZ)
Benzodiazepines act at GABAA
receptors by binding directly to a
specific site, distinct from that of
GABA.
They do not activate GABAA
receptors directly but rather require
GABA to express their effects; i.e.,
they only modulate the effects of
GABA.

Studies of cloned
GABAA receptors
have shown that the
coassembly of a
subunit with and
subunits confers
benzodiazepine
sensitivity to GABAA
receptors

BDZ BINDING SITE

Benzodiazepines bind across
the interface between the
and subunits but only to
receptors that contain 2 and
1, 2, 3 or 5
subunits.1

MOA.

Benzodiazepines (BDZs) bind to the gamma sub-unit of the

GABA-A receptor. Their binding causes an allosteric (structural)
modification of the receptor that results in an increase in GABA
A receptor activity. BDZs do not substitute for GABA, which
bind at the alpha sub-unit, but increase the frequency of channel
opening events which leads to an increase in chloride ion
conductance and inhibition of the action potential

Binding Affinity at various subunits

CLINICAL USES OF BDZ

MODULATORS

NON BENZODIAZEPINE GABA

MODULATORS
commonly referred to as "Z compounds".
They include: zolpidem (AMBIEN),
zaleplon(SONATA),
zopiclone (Not marketed in the U.S.), and
eszopiclone (LUNESTA), which is the S(+) enantiomer of
zopiclone

ZALEPLON
Pyrazolopyrimidine class of
compound.
Zaleplon preferentially binds to the
benzodiazepine-binding site on
GABAA receptors containing the
1receptor subunit.
Metabolised by aldehyde
P.K - Absorbed rapidly and reaches
oxidase.
peak plasma concentrations in ~1
Zaleplon (usually
hour. Its bioavailability is ~30%
administered in 5-, 10-, or
because of presystemic metabolism
20-mg doses) has been
volume of distribution of ~1.4 L/kg
studied in clinical trials of
and plasma-protein binding of
patients with chronic or
~60%.
transient insomnia.

ZOLPIDEM
Imidazopyridine
Although the actions of zolpidem are
due to agonist effects on GABAA
receptors and generally resemble those
of benzodiazepines, it produces weak zolpidem is approved
only for the short-term
anticonvulsant effects in experimental
treatment of insomnia.
animals.
During U.S. clinical trials, withdrawal Therapeutic doses (5 to
10 mg) zolpidem
effects within 48 hours of drug
infrequently produces
discontinuation occurred at an
residual daytime sedation
incidence of 1% or less. Postor amnesia adverse
marketing reports of abuse,
effects(e.g.GI complaints
dependence, and withdrawal have been
or dizziness) is also low.
recorded.

ESZOPICLONE
Active S(+) enantiomer of
zopiclone. Eszopiclone has no
structural similarity to
benzodiazepines, zolpidem, or
zaleplon.
Eszopiclone is used for the longterm treatment of insomnia and for
sleep maintenance. It is prescribed
to patients who have difficulty
falling asleep as well as those who
experience difficulty staying asleep,
and is available in (1-, 2-,or 3mg)tablets.

Eszopiclone received
FDA approval based on
six randomized placebocontrolled clinical trials
that showed it has
efficacy in treating
transient and chronic
insomnia

Inverse agonist at BZD Receptor

Inverse agonists at the BZD site act as negative allosteric

modulators of GABA-receptor function.
Their interaction with BZD sites on the GABAA receptor can
produce anxiety and seizures.

Negative allosteric modulators at

benzodiazepine
Sarmazenil

Partial inverse agonist at

the benzodiazepine site.
It is used in veterinary medicine to reverse
the effects of benzodiazepine sedative drugs
in order to rapidly re-awaken anaethetised
animals.

Carbolines

In addition to their direct actions, these

molecules can block the effects of
benzodiazepines.
Uses as convelsants.

BZD-site Antagonist

FLUMAZENIL: A BENZODIAZEPINE
RECEPTOR ANTAGONIST
Flumazenil (ROMAZICON, generic), the only member of this
class, is an imidazobenzodiazepine that behaves as a specific
benzodiazepine antagonist.
Flumazenil binds with high affinity to specific sites on the
GABA-A receptor, where it competitively antagonizes the
binding and allosteric effects of benzodiazepines and other
ligands.
Flumazenil antagonizes both the electrophysiological and
behavioral effects of agonist and inverse-agonist
benzodiazepines and B-carbolines.
Recommended dose 1mg IV . t1/2 of ~1 hour.dose repeated till
5 mg

FLUMAZENIL
Can be used to reverse the effect of benzodiazepine
overdosage , or to reverse the effect of benzodiazepines such
as midazolam used for minor surgical procedures.
It has been found to be effective in overdoses of nonbenzodiazepine sleep enhancers - zolpidem and zaleplon.
Use in hepatic encephalopathy & alcohol intoxication have
yielded mixed results.
Used as a PET radioligand labeled with carbon-11 to visualize
the distribution of GABAA receptors in brain.
Flumazenil is not effective in single dose overdoses with either
barbiturates or Tricyclic antidepressants rather it may cause
seizures in patients poisoned with TCAS.

Barbiturates:

Barbiturates also facilitate the actions of GABA -A at multiple sites in

the central nervous system, butin contrast to benzodiazepinesthey
appear to increase the duration of the GABA-gated chloride channel
openings. At Higher concentrations, barbiturates directly increase
chloride ion conductance.

Antiepileptic actions at
GABAA Receptors
Modulate GABAA receptor
activation
Phenobarbital
Clonazepam, Diazepam
Topiramate
Increase GABA biosynthesis
Valproate
Decrease GABA degradation
Tiagabine
Vigabatrin

GABAergic terminal

Inhibition of uptake increases GABA action

VALPROIC ACID
An analogue of valeric acid
Believed to affect the function of the neurotransmitter GABA in
the human brain by inhibition of GABA transaminase.
Other mechanisms of action
Blocks the voltage-gated Na+ channels
Blocks T-type Ca++ channels.
Inhibits the enzyme histone deacetylase 1

(anti convulsant
action)

VALPROIC ACID
Indications
Manic episodes associated with bipolar disorder.
As an anticonvulsant in
Absence seizures
Juvenile myoclonic epilepsy
Tonic -clonic seizures Posttraumatic epilepsy
Complex partial seizures Lennox-Gastaut syndrome
Valproic acid is cytotoxic to many different cancer types
through its action as a histone deacetylase inhibitor.
Being tried in multiple, myeloma, glioma, melanoma, breast
cancer, cervical cancer and ovarian cancer.

VIGABATRIN
VIGABATRIN is a suicide
inhibitor of GABA transaminase.
It blocks the conversion of GABA to
succinic semialdehyde,resulting in
high intracellular GABA
concentrations and increased
synaptic GABA release.
Primary indication for vigabatrin
treatment of simple and complex
partial seizures but it can also be
used for generalised seizures.

Other Uses Drug refractory

epilepsy and infantile spasm.
Dose:- 2 gm/ day .
Side Effects:-1.Behavioural
changes.
sedation ,Amnesia ,Weight gain
In 1% of cases visual field
defects due to peipheral retinal
atrophy.

TIAGABIN
Tiagabine (GABITRIL) is a
derivative of nipecotic acid
Approved by the FDA as
adjunct therapy for partial
seizures in adults.
Tiagabine inhibits the GABA
transporter, GAT-1, and thereby
reduces GABA uptake into
neurons and glia.
Paradoxically, tiagabine has
been associated with the
occurrence of seizures in
patients without epilepsy

Its contraindicated in
absence seizure.
Other side effects Sedation,
fatigue, tremors and confusion.
Dose:- 20-60 mg per day in 3
to 4 equally divided doses.

GABAPENTIN & PREGABALIN

Gabapentin (NEURONTIN, others) and
pregabalin (LYRICA) are anti-seizure drugs
that consist of a GABA molecule covalently
bound to a lipophilic cyclohexane ring or
isobutane respectively.
Pregabalin increases neuronal GABA levels
by producing a dose-dependent increase
in glutamic acid decarboxylase activity.
Gabapentin & pregabalin bind to the 2
(alpha2delta) subunit of the voltagedependent calcium channel in the central
nervous system and decrease the synaptic
release of glutamate..

Recent evidence
suggest that they
also function as
GABAB receptor
agonist.

 P.K - Gabapentin and

pregabalin are absorbed
after oral administration . It
is neither metabolised nor
bound to plasma proteins
and are excreted unchanged,
mainly in the urine.
Their half-lives,
approximate 6 hours.
These compounds have no
known interactions with
other anti-seizure drugs.

Therapeutic Uses: Drug resistant partial

seizures & G.T.C . also is being used for the
treatment of migraine, chronic pain, and
bipolar disorder. FDA Approves Lyrica
(Pregabalin) for Treatment of Neuropathic
Pain in 2005.
Side effects :- Somnolence, dizziness, ataxia,
and fatigue.
Dose 200-300 mg TDS.

Role OF GABA RECEPTORS IN

ANAESTHESIA

Anaesthetics primarily act by either enhancing inhibitory

signals or by blocking excitatory signals

GENERAL ANAESTHETICS

Currently, there are 5 major

inhalational and 5 intravenous
anesthetics used to induce or
maintain general anesthesia.
Inhalational:Nitrous Oxide,
Isoflurane, Sevoflurane, Desflurane
and Xenon.
Intravenous: Propofol, Etomidate,
Ketamine, Methohexital and
Thiopental.

Of these 10 general anesthetics, except

ketamine, nitrous oxide and xenon
other7 general anesthetics and
sedatives like midazolam, diazepam
and lorazepam share a common target
and mechanism of action,i.e. they all
enhance the function of GABA-A
receptors.

General anaesthetics
The extrasynapic 532 receptor in hippocampus is probably
assosciated with amnestic action of anaesthetic.
Those receptors having subunit s at ventrobasal thalamic
nucleus may be linked to intruding reversible loss of
consciousness during anaesthesia.
Since general anaesthetics are hydrophobic and need to access
the CNS they target hydrophobic pockets within the
transmembrane domains of the receptor.

Single amino acids determine i.v.

anaesthetic activity ?
If GABA-A receptors are important targets for general
anaesthetics,are all receptors equally sensitive to their effects?
or are certain subunit combinations more sensitive than
others?
Etomidate which demonstrates selective effects for receptors
containing certain subtypes of the subunit. For example,
receptors containing 2or 3, but not 1subunits are
particularly sensitive to the modulatory effects of this
anaesthetic.4 Studies using a molecular techniques
demonstrated that the selectivity of etomidate maps to a single
amino acid located within the second transmembrane domain
(TM2) of the b subunit (asparagine for 2 and 3, and serine
for 1 subunits). Exchanging the asparagine for serine at 2
increased the sensitivity of etomidate.

A binding site for volatile anaesthetics?

The effects of volatile agents on GABA-A receptor activity are also
influenced by a single amino acid. (serine) is located within the TM2
region in the subunit.
Subsequent studies demonstrated that the volume of this amino acid
correlated with the activity of volatile anaesthetics on GABA-A
receptors. Specifically, substitution with small volume amino acids
enhanced the potentiating activity of isoflurane,halothane, and
chloroform, whereas the presence of bulkier amino acids reduced
anaesthetic activity.
Use of volatile agents of differing size (isoflurane, 144 A ;
halothane, 110 A , and chloroform, 90 A ) led to the proposalthat
the four amino acid residues from the extracellular end of TM1, TM2,
TM3, and TM4 domains of the a subunit contribute towards an
anaesthetic binding pocket for volatile agents

Neuroactive steroids

Synthesized in the central and peripheral nervous system, especially

in myelinating glial cells, from cholesterol or steroidal precursors.
These include pregnenolone (PREG) and
dehydroepiandrosterone (DHEA), their sulfates, and reduced
metabolites.
Neurosteroids have a wide range of potential clinical applications
from sedation to treatment of epilepsy and traumatic brain injury.

NEUROACTIVE STEROIDS
Allopregnanolone and tetrahydrodeoxycorticosterone have
been surmised to enhance GABA-mediated Cl currents,
whereas pregnenolone sulfate & dehydroepiandrosterone
(DHEA) sulfate display functional antagonistic properties at
GABAA receptors.
Ganaxolone, an analog of the endogenous neurosteroid
allopregnanolone, is under investigation for the treatment of
epilepsy and was effective in the treatment of partial seizures
in adults and was tolerated.

This animation shows one model for how neurosteroids may increase
Cl flux through a GABA-A receptor. Binding of the neurosteroid
(ALLO) allows a protein kinase C (PKC) phosphorylation site to become
accessible. Phosphorylation of the channel increases flux through the
channel, allowing more Cl ions to flow through the channel in the
presence of GABA than when the channel is activated by GABA without
prior phosphorylation. The top of this animated gif shows how the
channel, the ligands, and G protein-activated PKC may be interacting at
the plasma membrane. The bottom of the animation shows how these
interactions affect the current trace recorded using electrophysiological
techniques. In the current trace, the size of the signal is smaller in the
absence of neurosteroid compared to the size of the signal after exposure
to neurosteroid and subsequent phosphorylation of the channel.

ALPHAXOLONE
Neurosteroid general anaesthetic.
A study 1987 found the primary
mechanism for the anaesthetic
action of alfaxalone to be
modulation of neuronal cell
membrane chloride ion transport,
induced by binding of alfaxalone
to GABAA cell surface receptors.
It is licensed for use in both dogs
and cats.
Unlike some of its predecessors
alfaxalone is not associated with
histamine release and anaphylaxis.

A 1994 study found that

alfaxalone binds to a
different region of this
receptor than the
benzodiazepines.
Alfaxalone is metabolised
rapidly in the liver.
Alfaxalone has a very short
plasma elimination half-life
in dogs and cats

OTHER NEUROACTIVE STEROIDS

Drug
Hydroxydione

Althesin
(alphaxolone +
alphadolone)
Minaxolone

Details
Proved to be a useful anaesthetic drug with a
good safety profile.
But was painful and irritating when injected
probably due to poor solubility
Withdrawn from human use due to rare but
serious toxic reactions, but is still used in
veterinary medicine
Around three times more potent than althesin
without the toxicity problems.
Withdrawn because animal studies suggested
potential carcinogenicity

ALCOHOL & GABA

RECPTOR

Alcohol
Ingestion of ethanol results in a
dose-dependent reduction of
central nervous system (CNS)
activity.
For several decades, it was
presumed that this CNS
depression was due to a major
effect of ethanol is through
Direct action of ethanol on
BZD receptors on GABA.
43 or 63 selectivity for
But, a lack of ethanols effect
separate GABA mimetic action
is proved today.
on GABA responsiveness from
isolated neurons with this
receptor subtype discontinued
this contention.

NEUROSTEROID INVOLVEMENT IN THE

GABAMIMETIC PROFILE OF ETHANOL

Following ethanol consumption there is activation of the hypothalamicpituitaryadrenal (HPA) axis (denoted by the darkened arrows), ethanol increases neurosteroid
precursors from the adrenal, which in turn results in
increased neurosteroids in brain . Since neuroactive steroids enhance GABA
responsiveness (Lambert et al, 2001, 2003; Paul and Purdy, 1992), it is proposed that
the ethanol-induced enhancement of neurosteroid presence in brain synergizes
theeffect of GABA released by ethanol (Criswell and Breese, 2005).

Alcohol

GLUTAMATE

GABA

Chronic Alcohol

GLUTAMATE

GABA

ROLE OF GABA IN ALCOHOL

WITHDRAWAL
Thaugh benzodiazepines have been the primary treatment
modality for alcohol withdrawal.
A growing number of preclinical and clinical studies suggest that
the anticonvulsants like gabapentin m
Two RCT First (n=60) and second (n=21) proved that
gabapentin was significantly more effective than placebo for
prevention of alcohol withdrawal symptoms
But except insomnia.
Other drugs which are currently investigated for alcohol
withdrawal effect are Baclofen and GHB.

GHB (Gamma hydroxy butyric acid)

Gamma-hydroxybutyric acid (GHB)
gammaaminobutyric
acid (GABA) that has been used in
research and clinical medicine for
many years.

is a naturally occurring analog of

GHB (Gamma hydroxy butyric acid)

GHB usually exists either as a free acid, or as a sodium salt. The sodium
salt is called Sodium Oxybate
and is soluble in water and methanol.
GHB as is used to treat cataplexy and excessive daytime sleepiness in
patients with narcolepsy.
MOA:
GHB has at least two distinct binding sites in the CNS.
Agonist at the newly-characterized GHB receptor, which is excitatory
Weak agonist at the GABAB receptor, which is inhibitory. Previously used
as an anesthetic in 1960s but
was withdrawn due to its side effects like seizures and
coma. Due to its euphoric effects, it is nowadays used as
POTENNTIAL ABUSIVE DRUG

EPIDEMIOLOGY OF GHB ABUSE

Effects

GHB Rage or reprive?

NEGATIVE POINTS POSITIVE POINTS
Rave drug
Can be used in narcolepsy
Date rape drug
Tried for alcohol withdrawal
Body builders
addiction.
Euphoria & crime

Excellent dilator of cervix

Can be used as an
aphrodesiac

GABAB Agonist

BACLOFEN
Derivative of GABA (parachlorophenyl GABA)
It is primarily used to
treat spasticity. In disorders like
multiple sclerosis,ALS,Spinal
injuries.
However its relatively
ineffective in stroke ,
parkinsonism
Cerebral palsy and rheumatic
muscle spasm.

Tolerance does not develop baclofen retains its therapeutic antispasmodic effects even after years
of use.

Baclofen
Primary site is possibly the spinal cord
where it depresses both polysynaptic
and monosynaptic reflexes
Also used in tardive dyskinesia and
alcohol withdrawal
ADR - Sedation (less than BZDs),
confusion, weakness.
Baclofen can be delivered directly into
the space around the spinal cord by use
of a surgically implanted pump and an
intrathecal catheter.

Excreted unchanged in
urine with t1/2- 3 to 4 hrs
Dose -10 to 25 mg
TDS.

Phenibut
-Phenyl--aminobutyric acid is a derivative of GABA.
Binds the GABAB metabotropic receptor.
The addition of a phenyl group in the position allows
phenibut to cross the blood brain barrier.
Only the R enantiomer is biologically active
Sold as a dietary supplement in the United States, while
in Russia it is sold as a neuropsychotropic drug.

Phenibut
Structurally similar to baclofen & phenylethylamine.
Pharmacological effects of phenibut are similar to baclofen,
but less potent.
Additionally, can function as a phenylethylamine receptor
antagonist.
Furthermore, phenibut has been shown to enhance levels of
dopamine.
Has anxiolytic effects in both animal models and in humans.

Phenibut
Used to treat a wide range of ailments including posttraumatic stress disorder, anxiety, and insomnia.
It has been reported by some to possess neurotropic actions for
its ability to improve neurological functions,
ADR - Sedation

GABAB Antagonist

Saclofen & Phaclofen

These are competitive antagonist at GABAB receptor.
This drug is an analogue of the GABAB agonist Baclofen.
The action of saclofen on the CNS is understandably modest,
because G-proteins rely on an enzyme cascade.
However, in animal experiments, saclofen is paradoxically
observed to have an antiepileptic effect.

GABA analogues

Progabide
Analog and prodrug of GABA used in the treatment of
epilepsy.
Agonist at both the GABAA and GABAB receptors.
Approved for either monotherapy or adjunctive use in the
treatment of epilepsyspecifically, generalized tonic-clonic,
myoclonic, partial, and Lennox-Gastaut syndrome seizures
in both children and adults

Progabide
Also being investigated for
Parkinson's disease
Schizophrenia
Depression
Anxiety disorder
Spasticity
with various levels of success.
Tolgabide - analogue of progabide and acts similarly to it as a
prodrug of GABA, and therefore as an indirect agonist of the
GABA receptors.

Picamilon
Dietary supplement formed by combining
niacin with GABA.It was developed in
the Soviet Union in 1969 by the All-Union
Vitamins Scientific Research Institute
Crosses BBB and is hydrolyzed into GABA
and niacin. The released GABA would
activate GABA receptors potentially
producing an anxiolytic response
The second released component, niacin acts
as a strong vasodilator, which might be useful
for the treatment of migraine headaches

Picamilon
In Russia, Picamilon is used for treatment of
these illness.
1.
2.
3.
4.
5.
6.
7.
8.
9.

Ischemic stroke
Asthenia
Depression
Senile psychosis
Alcohol intoxication
Migraine
Craniocerebral trauma
Neuroinfections
Primary open-angle glaucoma

GABA C AGONIST
CACA C Amino caproic acid . Weak agonist.
TACA Trans isomer of CACA.strongest agonist at GABA
C receptor.
R- CAMP:- Recemic mixture of C-AMP acts as agonist .

TPMPA, Selective GABA-C

Antagonist
Tetrahydropyridine ring of isoguvacine was
unfavourable for interactions with GABAB
receptors, while the
methylphosphinic moiety of 3-APMPA was
unfavourable for interactions with GABA-A
receptors led to the synthesisof TPMPA
which incorporates both the
tetrahydropyridine and methylphosphinic
acid moieties in the one compound.
TPMPA has been used to provide evidence
of functional GABAC receptors in the rat
superior colliculus

mediating
disinhibition , in
cells in the gut
involved in
neuroendocrine
secretion and in rat
spinal cord .
TPMPA has been
shown to enhance
memory in chicks .

GABA C
Recombinant receptor technology. The cloning of 1 and 2
cDNAs from a human retinal library enabled expression of
receptors in Xenopus oocytes that showed the characteristics
expected of GABAC receptors .
GABAC receptors are expected to mediate the lateral
inhibition of light responses and have been shown to inhibit
transmitter release at bipolar cell terminals.

CRITERIA

GABA-A

GABA-B

GABA-C

Distribution

Mammalian CNS

Cerebellum &
interpeduncular
nuclei

retina, spinal cord,

superior colliculus,
pituitary, and
gastrointestinal
tract.

CATEGORY

LGCC

GPCR

LGCC

AGONIST

GABA,
Muscimol,THIP

Baclofen
SKF 97541

CACA ,GABA
Isoguvacine(p)

MODULATOR

Benzodiazepines
Anaesthetics
Barbiturate
Alcohol
Neuroactive
steroids
.

ANTAGONIST

Bicucillin.
Picrotoxin
Gabazine

Zinc

Phaclofen
Saclofen

TPMPA
THIP
Picrotoxin
Low conc.Zinc ion

Section III Herbal Preperations and

GABA Receptors
Many herbal medicines are used to influence brain function in
order to treat anxiety, cognitive disorders, and insomnia
involving GABA receptor.
Usually Herbal products act as second order modulators
i.e.they enhance the effect of first order modulators.
GABA itself is an important plant constituent, and
thus it should not be surprising that plants contain a
range of other chemicals that can influence GABA
function

Bicuculline
First isolated from the plant Dicentra
cucullaria and subsequently from a
variety of Corydalis,Dicentra, and
Adlumia species.
Competitive antagonist of GABAA
receptor activation.
Utilized in laboratories in the in
vitro study of epilepsy
Routinely used to isolate glutamatergic
(excitatory amino acid) receptor
function.

Picrotoxin
Picrotoxin is an equimolar mixture of
picrotoxinin and picrotin isolated from
Anamirta cocculus and related poisonous
plants of the moonseed family.
Picrotoxinin is relatively nonspecific in that
it is a potent antagonist at GABA-A and
GABA-C, moderate at glycine, and weak at
5HT3 receptors.
Can be used to counter barbiturate poisoning
It is clear that bicuculline and picrotoxinin
act at different sites to antagonize GABA.

Muscimol
Muscimol is one of the most widely
used agonists in the investigation of
ionotropic GABA receptors.
It is a more potent agonist at GABAC receptors than at GABA-A
receptors.
The agonist action of muscimol at
GABA-C receptors is not blocked by
bicuculline but is sensitive to
picrotoxin
(Gaboxadol) is a conformationally
restricted analogue of muscimol with
analgesic and sleep-promoting
properties

It was investigated for the treatment

of insomnia, but was recently
withdrawn from phase III clinical
trials due to efficacy and side-effect
problems.

Apigenin GABA Receptor Modulator

Common flavonoid found in a range of

plants, including chamomile
tea(Matricariarecutita).
The chamomile tea used in treatment
for insomnia and anxiety led to
investigations of its active constituents,
including apigenin.Apigenin was found
to have anxiolytic properties,and it
competitively inhibited the binding of
flunitrazepam to brain membranes
Enhancement of the diazepam-induced
positive allosteric modulation of GABA
responses by lower concentrations of

apigenin , described as a
second-order modulation

Epigallocatechin gallate (EGCG)

Epigallocatechin gallate (EGCG) is the
major polyphenol in green tea (Camellia
sinensis).
Studies on 122GABA-A receptors
showed that EGCG at low
concentrations has a potent secondorder modulatory action on the firstorder modulation by diazepam ( more
than apegenin)
In well-controlled epidemiological
studies it alters brain-aging processes
and to serve as possible neuroprotective e.g. Parkinsons and
agents in progressive neurodegenerative Alzheimers diseases
disorders.

Hispidulin and Related Flavonoids

Hispidulin (the 6-methoxy derivative of
apigenin) was isolated together with apigenin from
Salvia officinalis (sage) recently using a benzodiazepine binding assayguided fractionation.
Hispidulin was some 30 times more potent than apigenin in displacing
flumazenil binding.
Used in herbal medicine to assist memory.
An extract of Salvia lavandulaefolia (Spanish sage) has
been shown to enhance memory in healthy young volunteers.

Section IV

CURRENT KNOWEDGE
SCENARIO & FUTURE
TRENDS

GAD 65 OR GAD 67 ?
Knockout of GAD65 has major impact on synaptic
GABA synthesized from astrocyte-derived
Glutamine.1
Two isoforms : GAD- 65 & GAD- 67
GAD65 is crucial for maintenance of biosynthesis of synaptic
GABA particularly by direct synthesis from astrocytic
glutamine via glutamate.
The GAD67 was found to be important for synthesis of
GABA from glutamine both via direct synthesis and via a
pathway involving mitochondrial metabolism.

GENE ORGANISATION OF
SUBUNITS
The majority of genes
coding for the
GABA-A receptor
subunits are organized
into four clusters on
chromosomes 4, 5, 15,
and X in the human
genome

GENE KNOCKOUT & KNOCK IN

TECHNIQUE
Pharmacological
analysis of GABA
receptor was simplified
after introduction of
animal models in which
particular GABA- A
receptor subunits are
either inactivated
(knock out) or
selectively point
mutated ( knock in).

KNOCK
OUT
SUBUNIT

RESPONSE

Motor impairing action of mice

on rota rod to daiazapine.

Die in neonatal period.

Epileptic seizure.

Sleep time was prolonged.

No action of
benzodiazapines.

Increased sleep time and

convulsions.No action of
alcohol.

GABA OLD RECEPTOR

NEW TARGETS

GABA AS IMMUNOMODULATOR
In lymphocytes,exposure to GABA reduced but did not
abolish the transient increase in the intracellular calcium
concentration that was associated with activation of the cells
(Alam et al.2006).
GABA activated GABA-A ionchannel currents in T cells and
macrophages (Bjurstom et al. 2008; Bhat et al. 2010;
GABA application resulted in decreased cytokine secretion
and T cells proliferation (Mendu et al. 2011)

GABA AS IMMUNOMODULATOR
GABA appears to have a role in autoimmune diseases
like MS, type 1 diabetes and rheumatoid arthritis and
maymodulate the immune response to infections
(Bhat et al. 2010; Mendu et al. 2011; Soltani et al. 2011;
Tian et al. 2011; Wheeler et al. 2011).
Has even a role in Alzheimer disease, stroke and
traumatic brain injury (Popovich and Longbrake 2008;
Schwartz and Shechter 2010)

Role of GABA IN TYPE- I DM

Currently a trial with hypothesis that GABA, a naturally
occurring substance, has the potential to reduce the
inflammation and protect the pancreatic beta cells from
autoimmune destruction.
GAD-alum may contribute to the preservation of residual
insulin secretion in patients with recent onset, Type I Diabetes.
condition :Type I Diabetes
Drug: Glutamic Acid Decarboxylase in alum formulation

Status :-Phase 1

GABA & SCHIZOPHRENIA

In subjects with schizophrenia, impairments in working
memory are associated with dysfunction of the dorsolateral
prefrontal cortex (DLPFC).
This dysfunction appears to be due, at least in part, to
abnormalities in Gamma-aminobutyric acid (GABA)-mediated
inhibitory circuitry.
Various experimental findings suggest that schizophrenia is
associated with alterations in inhibitory inputs from SST/NPYcontaining and CCK containing subpopulations of GABA
neurons and its signaling via certain GABA-A receptors that
mediate synaptic (phasic) or extrasynaptic (tonic) inhibition .

GABA-B Receptor Complex Target

for Tumor Therapy
A positive correlation between
GABA
B2 (minor b1)expression and that
of thyroid tumors is seen.
GABAB1 immunostaining of
human ductal breast cancer
tissues (from patents shows
Intensive b unit staining in the
cytosol and less frequently in the
nucli.

Trial scenario related to GABA

INTERVENTION CONDITION

STATUS

1.

GAD-Alum

Juvenile onset type-1

DM.

PHASE 1 A

2.

Merck L-830982
GABA-A Alpha2/3
Receptor Agonist

Schizophrenia

PHASE 2.

3.

Pregabalin

Pain after posteriar

spinal fusion.

PHASE - 4

4.

Vigabatrin

Coacaine abuse

PHASE -2 A.

5.

Gabapentin

Smoking

PHASE 1.

6.

Lomazenil

ALCOHOL ABUSE

PHASE -1.

Conclusion.
Despite the overwhelming representation of the
GPCRs in the human genome, it is the ionotropic
receptors which achieved most visibility till today.
The paucity of molecular heterogeneity exhibited by the
GABA-B receptors has proved problematic for specific drug
targeting.
Pharmacologists had to wait till gene knockout tecnique and
readymade animal models became available till early 2000.
The next decade will undoubtedly prove
Exciting with these recent genetic tools in hand...........

THANK YOU.