Anesthetics and mechanisms of general anesthesia

Anna Kuznetsova

Volatile anesthetics Morton 1846

And Elemental noble gases: He, Ar Anesthetics are a mainstay of modern medicine, but their molecular mechanism of action is still not known Anesthetics are structurally very different and have weak affinity for their targets However, the mechanism will provide the key to making the perfect anesthetic!

Nerve impulse is the key to sensation

impulse

im pu

Synapsecontact between nerve cells

ls e

http://www.doctorspiller.com/local_anesthetics_4.htm

Depolarisation process for impulse generation and conductance

Resting Depolarization potential (impulse)

http://www.doctorspiller.com/local_anesthetics_4.htm

Current theory suggests anesthetics block impulse by:

1. Inhibiting the depolarization by reducing the influ of sodium ions into cytoplasm so that threshold necessary for sensation is not reached Threshold of sensitivity

2. decreasing resting potential (hyperpolarization) by increase K+ conductance of membrane or increasing Cl- conductance

To affect ion channels, the anesthetic molecules must actually enter membrane of the nerve. And herein lies the difference in the potency, time of onset and duration of the various anesthetics.

The Meyer-Overton rule correlation for anesthetics

The more the drug is soluble in olive oil the more potent as an anesthetic!!!

1901

This correlation is accurate for a broad range of general anesthetics:alkanols, volatile agents, and barbiturates. Also, volatile anesthetics are generally additive in their effects: a mixture of a half dose of two different volatile anesthetics was in fact equal to a full dose of either drug in isolation.

http://www.anes.upmc.edu/anesnews/volume/2003winter_spring/articles/focus.html

Theoretical interpretations of Meyer-Overton empiric rule

1954

1) First Nonspecific unitary theory of narcosis: distortion of the lipid bilayer is caused by accumulation of anesthetic molecules in the neuronal cell membrane providing anesthetic effect. Chemical structure of anesthetics per se doesnt matter!!

Expansion of the membrane concept: Meyer-Overton rule can be improved if molecular volumes that anesthetics occupy in the membrane are taken into consideration. The more space within membrane is occupied by anesthetic - the greater the anesthetic effect, because membrane thickening reversibly alters gating mechanisms of ion flux into the cell. The idea was supported by pressure reversal effect - Anesthetic effect is reversed by increases in atmospheric pressure.

1990s
2) Lipid Hypothesis: Inhibition of neuronal conduction by general anesthetics is due to changes in membrane density (critical volume hypothesis) and membrane fluidity (Suezaki, 1990)

Objections to Lipid Hypothesis:

Exception to Meyer-Overton rule: the anomalous lack of anesthetic potency of n-alkanols above a cutoff chain length

Cutoff
Anomaly can be explained by binding of anesthetics directly to protein hydrophobic pockets of well-defined volumes Thus the molecular volume of the alkanol at the cutoff length provides a measure of the volume of the putative binding site.

Changes in density and fluidity of membrane are so small that small increases in temperature can mimic these effects on membrane fluidity and density without causing anesthesia.

Theoretical interpretations of Meyer-Overton empiric rule:

3) Protein Hypothesis (specific interactions)

1998

GABA A receptors, glutamate receptors

When anesthetics bind to hydrophopic pockets on membrane proteins which undergo conformational changes and alter ion flux. So volatile anesthetics mediate their actions through direct interaction with membrane proteins rather than lipids (Franks and Lieb, 1998)
http://www.chemcases.com/alcohol/alc-13.htm

Currently no single protein or channel has emerged as the likely anesthetic target to the exclusion of all others. Receptors often exhibit differential modulation by the individual stereoisomers of optically active anesthetics
http://sophia.smith.edu/~ahall/research.html

4)Bilayer mediate interactions of anesthetic and channel (new)

Although the lipid and protein theories have not been proved or disproved, current evidence suggests that lipid solubility and/or protein perturbations are not the sole mechanisms involved

anesthetic Anesthetic is solubilized in the bilayer, causing redistribution of membrane lateral pressures, which shifts conformational equilibrium of integral proteins such as ligand-gated ion channels
It is largely the positional and orientational distribution within the bilayer of the segments and bonds of the anesthetic molecule that determines its potency

Anomalous lack of anesthetic potency of n-alkanols above a cutoff chain length (exception to Meyer-Overton rule):
In short alkanols, flexible hydrocarbon chain segments reside in

hydrocarbon region of bilayer but very close to hydroxyl group tethered to the aqueous interfacial region For alkanols with longer chain, the hydrocarbon tails penetrate deeper into the bilayer, reducing the magnitude of the effect until, for sufficiently long chains, no pressure redistribution is predicted and anesthetic potency is lost.

Short hydrocarbon chain redistributes lateral stresses from the bilayer interior toward the aqueous interface!!!

OH

OH

OH

OH

OH

OH

Polyhydric alkanols were suggested to have anesthetic effect similar to 1-alkanols. And indeed polyhydroxyalkanes 1,6,11,16hexadecanetetraol and 2,7,12,17-octadecanetetraol exhibited significant anesthetic potency.
J. Med. Chem., 2005, 48 (12), pp 41724176

Oleamide-induced closure of gap junction membrane channels

A mixture of phospholipids (green circles) and cholesterol (yellow squares) form a well-ordered lipid ring surrounding the gap junction connexon (blue).

On treatment with endogenous anesthetic oleamide (red triangles), this lipid ring is fluidized and becomes disordered, promoting a conformational change in the connexon oligomer and leads to closure.

Endogenous anesthetic oleamide (amide of oleic acid) potentiates sleep and lowers temperature through closing gap junction channels. Anesthetic Isoflurane was showed to cause flickering of the acetylcholine receptor
Richard A. Lerner PNAS Vol. 94, pp. 1337513377, 1997

Identification of genes of anesthetic targets in C. Elegance

The whole genome (~1000 genes) completely solved!!

http://www.nematodes.org/Caenorhabditis/C_elegans_genome/Celegans_acedb.shtml

Unraveling the mystery of general anesthesia mechanism Genes and gene sets that control the behavior of C. elegans in volatile anesthetics have been identified

Genetic data fail to identify a single protein or channel that is uniquely responsible for the effects of anesthetics!!
P.G. Morgan, E.-B. Kayser, M.M. Sedensky C. elegans and volatile anesthetics WormBook.org