Drugs and the Brain

Week 1, miniLecture 3
What is a drug?
Types of drug molecules

2014 California Institute of Technology

drug
noun

Origin: Middle English drogge

Date: 14th century

1a...
b : a substance used as a medication or in the preparation of medication
c : according to the Food, Drug, and Cosmetic Act
(1) : a substance recognized in an official pharmacopoeia or formulary
(2) : a substance intended for use in the diagnosis, cure, mitigation, treatment,
or prevention of disease
(3) : a substance other than food intended to affect the structure or function of
the body
2 ...
3 : something and often an illegal substance that causes addiction, habituation, or a
marked change in consciousness

http://www.merriam-webster.com/dictionary/drug

Merriam-Webster, Inc

Trivial names and Structural Formulas

nicotine
Pubchem 89594

lidocaine
Pubchem 3676

morphine
Pubchem 5288826

botulinum toxin
PDB 1S0G

Pubchem is an NIH database;

PDB (Protein Data Bank) files are curated by an international organization
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This sessions drugs exemplify the concepts in Drugs and the Brain

Drugs Activate (nicotine) and block (lidocaine) ion channels

Drugs act on G protein pathways (morphine)

Drugs activate genes (nicotine, morphine)

Protein drugs may become more useful for neuroscience diseases

(botulinum toxin)

Not treated in this session:

Drugs act on neurotransmitter transporters
4

Atomic-scale Structures
nicotine

lidocaine

morphine

botulinum toxin

http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=89594&loc=ec_rcs
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3676&loc=ec_rcs
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5288826&loc=ec_rcs
http://www.ncbi.nlm.nih.gov/Structure/mmdb/mmdbsrv.cgi?uid=26967

Each moiety in a drug molecule has importance.

Example: lidocaine
Alkyl substituents:
may adjust
charge density of amine.
Affects membrane
permeation.
Also affects binding to the
receptor.

Charged amine:
may bind to
charged groups or
electrons on the protein

Amide: hydrolyzed to
terminate drug action

Alky groups affect both

membrane permeation
and receptor binding

Aromatic: may bind to

nonpolar groups on
the receptor protein.
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Formulas and molecular weights (Da = g/mol ~ amu)

morphine

nicotine

lidocaine

C10H14N2

C14H22N2O

C17H19NO3

162.2

234.3

285.3

botulinum
toxin
~150,000

Ionization state and counterion

nicotine
pubchem nicotine

lidocaine-H+

usually uncharged

chloride

morphine-H+
sulfate,
chloride

botulinum
toxin
many counterions

We dont use trademarks; they vary by country, by preparation, and by salt

nicotine

lidocaine

morphine

Marlboro
Camel

Lignocaine
Xylocaine
Anestacon
Esracaine
Gravocain
Leostesin
Lidoderm
Maricaine
Cappicaine

Avinza
Kadian
Roxanol
MS Contin
Astramorph,
Kapanol,
Skenan
Duramorph
Oramorph

Blu eCig
Nicorette
Nicoderm

botulinum
toxin

Botox
Xeomin
Dysport
Neuronox
Myobloc

Type of Compound
morphine

nicotine
alkaloid

alkaloids are a group of nitrogen-containing drugs,

usually natural

Type of Compound
lidocaine
local anesthetic
(Synthetic organic compound)

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Type of Compound
botulinum
toxin
protein:
Chain of amino-acid residues joined by peptide bonds

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Drugs and the Brain

End of miniLecture 3, Week 1

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H+
RNH2

RNH3+
blood

H+
RNH2

RNH3+

Drugs and the Brain

Week 1, miniLecture 4
Drug entry into the nervous system

mouth,
stomach
or lungs

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Routes into the Body

nicotine

lidocaine

Smoked;
chewed;
skin patch

Injected

morphine

botulinum
toxin

Smoked;

Injected;

Injected;

eaten

suppository

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often the active

and predominant form

H+
RNH2

RNH3+

blood

Lipid barrier,
e. g. membrane(s)

mouth, stomach or lungs

H+
RNH2

RNH3+
higher pH

15

Nicotines path from the lungs to the blood and the brain
pKa = 8.0
H+
blood,
then brain
3 cells and 6 membranes
lungs
pKa = 8.0
H+
vaporized

Most cigarettes also contain ammonium hydroxide,

To maintain neutral pH
16

Blood nicotine concentrations during and after a cigarette

smoking

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Another example of neutral drug permeation.

In Parkinsons Disease: most neurons that make dopamine die
The challenge: replace the dopamine in the brain
catalytic protein
Greek, to leaven
enzyme:
decarboxylase

levodopa, L-dopa
zwitterionic
permeates into brain
via a transporter
(treated in a future miniLecture)

dopamine
does not enter brain

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Endothelial cells form the blood-brain barrier

Brain

Tight junction

Other organs

~ 10 m

Protein
Nonpolar molecule
Polar molecule (e. g., glucose)

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Endothelial cells form the blood-brain barrier

Blood vessel

Blood
Glial foot

Tight junction

Endothelial cells lining the capillary

Red blood cells

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The Structural Basis

of the
Blood-Brain Barrier
extracellular
space

Alberts et al., Essential Cell Biology, Garland Science

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Drugs in the Body and in Neurons

Acid-base equilibrium and permeability
Uptake from the stomach
Uptake from smoke
Prodrugs, storage, enzymatic breakdown, breakdown, and elimination
Pharmacokinetics
Blood-brain barrier:
molecular basis
an opportunity for drug specificity

More in later
sessions

a problem for drug delivery

Crossing the cell membrane
Neurotransmitter transport inhibitors
Drugs in organelles: subcellular pharmacokinetics
Short-circuiting synaptic vesicles
Pharmacological chaperoning and inside-out pharmacology
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H+
RNH2

RNH3+
blood

H+
RNH2

RNH3+

mouth,
stomach
or lungs
Drugs and the Brain
End of miniLecture 4, Week 1
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Drugs and the Brain

Week 1, miniLecture 5.

Introduction to Drug Receptors

Most Drug Receptors are Proteins: Parts of a Protein

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Receptor
a molecule on the cell surface or in the cell interior that has an affinity for a
specific molecule (the ligand or drug).
Latin,
to tie

Most drug receptors are proteins.

Greek, first

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shortest: 9
longest: 5500

peptide
or
amide bonds

20 types
side chains

link the
backbone
or
main chain
or
-carbons

Alberts et al., Essential Cell Biology, Garland Science

26

Proteins contain a few structural motifs:

helices

sheets

Hide side chains

Show H-bonds and distances
Show ribbons & arrows
Show side chains
Show Van der Waals radii
Show stereo view

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Drugs and the Brain

End of miniLecture 5, Week 1

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Drugs and the Brain

Week 1, miniLecture 6
Most Drug Receptors are Proteins: More about Receptor Proteins

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Nearly Complete Nicotinic Acetylcholine Receptor (PDB 2BG9)

~ 2200
amino acids
in 5 chains
(subunits),

Binding
region

MW
~ 2.5 x 106

Membrane
region

Colored by
secondary
structure

Colored by
subunit
(chain)

Cytosolic
region

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The AChBP interfacial aromatic box occupied by nicotine (PDB 1UW6)

Hydrogen bond

Y198
C2

Y93
A

W149
B

Cation- bond
Y190
C1

nonW55
D
(MuscleNicotinicnumbering)

31

Simple views of nicotinic acetylcholine receptor

5 subunits

each subunit has 4 -helices

in the membrane
(20 membrane helices total)

(extracellular)

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How does binding of a drug activate a receptor protein?

Well discuss this in a future session,
but here we note that most receptors are allosteric proteins
Ligand-binding
region

Greek, other + body

acetylcholine
or
nicotine

acetylcholine
or
nicotine

acetylcholine
or
nicotine

~ 100
(10 nm)

M1
M2
M3

M4

An allosteric protein binds a ligand at one

site, affecting the function of a different site
within the same protein.
33

Concepts associated with Allosteric Proteins

Conformational changes.

Several subunits.

In some allosteric proteins, all subunits undergo concerted transitions

between at least two states
(in a ligand-gated channel, open, closed, desensitized)
Some allosteric proteins undergo sequential transitions,
as though the ligand induces a fit of the protein.
The subunits may transition independently.
Shape-shifting protein is a more general term.
These concepts are being refined and complicated as we obtain
atomic-scale structural information about receptor proteins
in their various functional states.

34

We cannot yet predict the structure of a protein simply from knowing its
sequence.
This is especially true for membrane proteins like receptors.
But structure prediction techniques are improving.
Every two yr, theres a contest to predict the structure of an important protein:
http://predictioncenter.org/

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Drugs and the Brain

End of miniLecture 6, Week 1
Introduction to Drug Receptors:
More about Receptors as Proteins

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Drugs and the Brain

Week 1, miniLecture 7
Introduction to Mammalian Brains:
Neuronal circuits, neurons, and synapses

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The Central Nervous System

Front
rostral

Top dorsal

Back
caudal

Brain

Bottom ventral

Spinal cord

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Function
Functionisisoften
oftenlocalized
localizedtotospecific
specificbrain
brainregions
regions
movement

sensations

vision
rostral
Front

caudal
Back

judgement
reward
acetylcholine
memory
(nicotine)
and
dopamine

coordination

A typical pathway:
sensation of pain
and the reaction
to pain

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Spinal reflexes, such as the knee-jerk, involve just two neurons.

sensory neuron

motor neuron

the sensory neurons act like

strain gauges wrapped around
special muscle fibers.
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Parts of two neurons

Postsynaptic
neuron

Presynaptic
neuron
Excitatory Inhibitory
terminal terminal

Greek, axis

presynaptic
terminal

axon

dendrites
Greek, tree

cell
body
nucleus

direction of information flow

estler, Hyman, Malenka, Molecular Neuropharmacology,

McGraw-Hill Professional Publishing

presynaptic
terminal
synaptic
cleft

postsynaptic
dendrite

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The synapse is a point of information processing

Greek, connection, junction

presynaptic neuron

postsynaptic neuron

Nestler, Hyman, Malenka, Molecular Neuropharmacology,

McGraw-Hill Professional Publishing

Box 2-2 Figure A

An adult human brain contains ~ 1011 neurons,

and each of these might receive 103 synapses apiece,
for a total of 1014 synapses.
Most of these synapses form during the first 2 yr of life.
Thus 1014 synapses/108 s = 106 synapses/s form in a fetus and infant!
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Chemistry is a Language of the Brain, for Instance at Synapses

cytosol

synaptic cleft

cytosol

receptor
presynaptic
terminal

postsynaptic
dendrite
receptor

transmitter molecules
receptor

direction of information flow

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Drugs and the Brain

End of miniLecture 7, Week 1

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Drugs and the Brain

Week 1, miniLecture 8
Sample Recordings and Techniques for Studying the Brain:
The frequency of a neurons action potentials (nerve impulses) is the most
important parameter in brain signalling . . . and in drug effects.

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Electricity is also a Language of the Brain.

Intracellular recording with sharp glass electrodes.
1. A current applied by the experimenter increases firing rates

V, I

Prof. David McCormicks data

http://info.med.yale.edu/neurobio/mccormick/movies/rly_exp.avi

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Intracellular recording with sharp glass electrodes.

2. Artificially applied acetylcholine acts on muscarinic receptors to
change the membrane potential, increasing action potential
frequency.

Prof. David McCormicks data

http://info.med.yale.edu/neurobio/mccormick/movies/ach_fin.avi

(The spikes in these examples are about 100 mV in amplitude)

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Modern Neuroscience Techniques:

Time scales, Distance Scales, and Invasiveness
magnetoencephalography
+
event-related potentials

functional magnetic
resonance imaging

positron
emission tomography

Optical
Dyes
Silicon Array

Microlesions
2-deoxyglucose

Extracellular Single Unit or Tetrode

Intracellular
Patch/Sharp

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Drugs and the Brain

End of miniLecture 8, Week 1
Sample Recordings and Techniques for Studying the Brain

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Drugs and the Brain

Week 1, miniLecture 9 Botulinum Toxin, a Protein Drug
(drugs, proteins, drug entry, synapses, transmitter release)

nicotine
Pubchem 89594

lidocaine
Pubchem 3676

morphine
Pubchem 5288826

botulinum toxin
PDB 1S0G

botulinum toxin

2014 California Institute of Technology

51

Drugs and the Brain

Week 1, miniLecture 9 Botulinum Toxin, a Protein Drug
(drugs, proteins, drug entry, synapses, transmitter release)

nicotine
Pubchem 89594

lidocaine
Pubchem 3676

morphine
Pubchem 5288826

botulinum toxin
PDB 1S0G

botulinum toxin

2014 California Institute of Technology

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More about Botulinum Toxin, which has Become a Modern Protein Drug
Botulinum toxin is made by Clostridium botulinum, an anaerobic bacterium.
Botulism comes from a German physician who noticed cases of paralysis associated with
eating an uncooked smoked sausage in 1793. 13 people in Wildbad shared the sausage that
had been sitting for hours; all became ill and six died. (To describe their illness, the word
botulism was derived from the Latin botulus, for sausage.)
The conditions beneath the skin of the sausage had been anaerobic (i.e., there was very little
oxygen in the meat) and enough time had elapsed to allow the clostridial bacteria to multiply
and produce the toxin within the sausage.
Botulinum toxin is fatal in extremely low quantities (~ 1 molecule per synapse), because it
paralyzes muscles. 10-8 grams kills a mouse. The paralysis occurs because presynaptic
terminals cannot release transmitter.

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The bacterium synthesizes botulinum toxin as a single protein chain,

then cleaves the chain
S-S

S-S

S-S

S-S

The light chain enters cells,

then acts as an enzyme

54

Many diseases and discomforts are caused by excess muscle activity.

Botulinum toxin, injected in minute quantities,
blocks this excess activity and gives relief from squint and spasm.

Botulinum toxin also decreases frown lines (Botox).

55

What are some possible new therapeutic uses for botulinum toxin?

Lets examine the biomedical literature today:

http://www.ncbi.nlm.nih.gov/pubmed?term=botulinum%20toxin

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Drugs and the Brain

End of miniLecture 9, Week 1
botulinum toxin

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H2O

K+ ion

carbonyl

Drugs and the Brain

Week 1, miniLecture 10
Origin of the Resting Potential;
Electrical Aspects of Ion Channels
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Storing energy in a concentration gradient

without osmotic stress:
Simply reverse the ratio of Na+ and K+
Na

External
Monovalent cations:
High Na+
Low K+

K+

Na+

K+

Internal:
Low Na+
High K+

Na+

Na

Na+

Na+

K+
K+
K+

Na+

Na+

K+
59

Typical extracellular and cytosolic ion concentrations (mammalian cell)

Na+

major
monovalent K+
Ions
Cldivalent
cations

Other ions

Extracellular Intracellular
conc
(Cytosol)
145 mM
15 mM
4 mM

150 mM

110 mM

10 mM

Ca2+

2 mM

10-8 M

Mg2+

2 mM

0.5 mM

Pi-2

2 mM

40 mM

H+

10-7 M

10-7 M

Protein

0.2 mM

4 mM

60

Converting a concentration gradient

to an electrical potential:
Create permeability to one ionic species (K+)
Na

Na+

Na+

K+

Na+

Lost positive charge

leads to net negative
interior potential

K+

K+
K+

Na+

Na+

K+ channels

Na+

Na+
61

The Nernst potential:

the energy of discharging the concentration gradient for K+ ions
balances
the energy of moving the K+ ions through the potential difference

K+

K+

K+
K+

K+

62

Deriving the Nernst Potential (Chemistry Units)

63

Deriving the Nernst potential

(physics units)

64

Neurons are not Equilibrium Systems

Several types of channels of channels are open at once.

To analyze resting potentials under these circumstances,
well switch to electrical calculations.

65

Atomic-scale structure of (bacterial) Na+ channels

Views
from the
extracellular
solution
Payandeh et al,
Nature 2011;
Zhang et al,
Nature 2012;

electrically, open channel

conductor

Views
from the
membrane
plane

66

H2O

K+ ion

carbonyl

Drugs and the Brain

End of miniLecture 9, Week 1

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Na+

K+

Drugs and the Brain

Week 1, miniLecture 11
Electrical Aspects of Ion Channels
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The miniature single-channel conductors add in parallel

GNa = Na

GK = K

outside

GNa

GK
=

ENa

EK
(- 60 mV)

Na

(+60 mV)
cytosol = inside

Kirchhoffs Current Law

(Conservation of Charge)

Na
mostly Na+

mostly K+

69

The membrane potential at steady state

(not at equilibrium)

outside

Na+

K+

resting
potential:
K+ channels
open

Oversimplified view of
excitatory postsynaptic
responses
(see miniLecture 13):
Na+ channels open too

E K GK E NaGNa
V
GK GNa

cytosol = inside

70

Two Major Roles for Ion Channels in Drugs and the Brain

Drugs at synapses:

[neurotransmitter or agonist]
open

closed

Future lectures:
Drugs at axons and
cell bodies

electric field or drug

closed

open

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Na+

K+

Drugs and the Brain

End of miniLecture 10, Week 1

2014 California Institute of Technology

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