NEUSCI 404 Notes

Lecture 1 (3.29)
1.3) Pharmacology Review: Different levels of investigation
 Pharmacokinetics (PK): how molecule travels within the body
o Absorption, distribution, metabolism, excretion
 Pharmacodynamics (PD): how molecule interacts with other molecules
o Receptor binding, signaling mechanism, agonist& antagonist, dose-response
curve, physiological effect
1.4) Bioactivity
 Psychoactive drugs: molecule that modifies mood, thought process, behavior
 Ligands: endogenous, synthetic, plant-derived
 Receptors: post/pre, glial/immune cells
 Additional targets: hydrolyzing enzymes (ex. AchH), transporter

1.5) Drugs and receptors: diverse size & types

 Range from small to large
o Majority of drugs: MW btwn 100-1000
 Too small-> INSUFFICIENT SELECTIVITY FOR TARGET SITES
 TOO LARGE -> POOR ABOSRPTION AND DISTRIBUTION IN ORGNAISM
 Proteins: most common
o Bind hormones or NT
o Ion channels and transport proteins
o Enzymes
 Nucleic acids: DNA (target for many anticancer ), RNA
 Membrane lipids: cholesterol, phosphatidylserine
 “nonreceptor” drugs
o Antacids: chemically neutralize stomach acid
o Class of diuretics: directly increase osmolarity of nephron w(draws water from
blood to the lumen)
1.6) Properties of Drug-receptor Interactions: Types
 Covalent: not very common
 Reversible noncovalent the most common
o Multiple sites, short range interactions
o Specific interactions btwn chemical groups on drug and R
o Only R with selective drug-binding properties ahv clinical value
o Bindings of drug initiates a chain of events (signaling) leading to observed effect
o cause small conformational change in the protein
1.7) Effectors: main recept-orsignaling systems

1.8) Timescale of receptor-Effector Singaling Pathways

1.9) G Proteins: Beta-Gamma Subunit Fxn

 alpha and Beta/gamma subunit of G Proteins activate distinct effector proteins
1.10) cAMP singaling pathway: PKA
Ligand-> GCPR-> conformation change -> activation of cyclase activate PKA -> catalytic effect

1.11) Rapid Synthesis & Degradation of cAMP: overview

ATP —(Adenylyl cyclase)—> cAMP —(phosphodiesterase)—> 5’-AMP

1.12)Phospholipase C dependent signaling: Overview: (also alternate pathways of G protein)

1.13: singal transduction: Amplification

B. If ligand is tightly and bound for long, you can change the phenotype of the cell (more
fundamental effect)

1.14) Nitric Oxide Signaling:

 volatile gas, freely diffuse across cell membranes
 very-short half life in circulation
 major signaling molecule in neurons, immune system, and vasculature
 1.15) V-0Gates ion channel superfamily: See slides
1.16) Ion channel R: transmembrane proteins that regulate the flow of ions across
membranes
 A. Ligand0gated channels
o Opens closed channel in response to binding of extracellular NT
o Drug mimic (agonist) or block (antagonist) the action of endogenous ligands
 B. V-gated Channels:
o Transient opening in response to changes in mp
o Time elapsed between channel opening and cellular response often is
millisec
1.17) Nuclear Receptors:
 Intracellular proteins
 Bound by lipid-soluble agents estrogen thyroid hormone that can cross the cell
membrane
 Bound R enters the nuc and alters gene expression
 Slow onset but response is longer in duration

1.18) R Tyrosine Kinase: EGFR & Cytokine Rs (see sldies)

EGFR
 Extracellular ligand binding domain & intracellular kinase domain
 EGF binding: inactive monomers -> active dimer
 Cytoplasmic kinase domains are activated and become phosphorylated
 Activated kinase phosphorylates other downstream proteins
Cytokine Rs
 Tyrosine kinase activity (JAK) not intrinsic to R
 Cytokine binding promotes receptor dimerization and TYP-P
 Activation of associated JAK Kinase
 P of effect and activation of transcription (STAT)
 STATS dimerize, go to the nuc and regulate gene expression

1.19) EGF R Signaling pathways: parallel signaling (see slides)

1.20) Signal Transduction: Parallel pathways/cross-talk

Lecture 2 (3/31)
2.1) Membrane receptors: Drugs interacting with Receptors
 Aim: find a compound that fits tightly to the binding pocket of a receptor target (key-
lock notion)

2.2) Membrane R: The selectivity of a drug

 The drug is selective when
o Binds selectively only to one target (1000x less potent at other targets)
o Target is expressed by select tissue or cell type
  Ex. GABAergic neurons not glutamergic-> modulates inhibitory neuronal
networks
 Diseases tissue, not healthy tissue -> anti-tumor agent that will nto kill
neurons

2.3) Membrane R: The nature of the drug

 Covalent bonds are rarely formed; rather ionic interactions (e.g. hydrogen and
hydrophobic interactions that mkae it reversible) -> equilibrium
 Enantiomer (chiral) & stereoisomer; ex. S(+) can be 100 times more potent than R(-)
 Appropriate size and shape to fit binding site
 Concept: one protein can bind a set of structurally diverse molecules with similar
affinities, whereas a number of closely-related analogues may display different affinities
for that target

2.4) Pharmacodynamics: Dose-response relationship

 Higher potency drug act at lower dose
 Maximal effect: all receptors saturated and no more receptors to affect

2.5) Membrane R: Law of mass action

 To produce an effect, a drug (e.g. agonist) has to physically interact with a target (e.g.
receptor) and favor a specific conformational state (e.g. active) and modulate signal
transduction (e.g. increase)
 The bigger the number of target are occupied -> the bigger the biological response is

2.6) Pharmacodynamics: Drug-R Theory: Model of drug-r action

 Study the action of drugs on biological function: bioactivity
 Agonists: full, partial, inverse
 Law of mass action
 Occupancy model: Kd = EC50
 Different receptor conformations
o Ri = inactive form that produces no effect when bound by agonist
o Ra* = active form that produces a small effect

2.7) MembraneR: Agonists: Potency and partial efficacy

 Inverse agonist: ligand binds and receptor stabilizes the inactive form

2.8-9) Membrane R: Antagonist: competitive and non-competitive

 Competitive inhibitor interferes with the activity site of enzyme so substrate cannot
bind
o Maximal response does not change
o Just requires more agonist
 Noncompetitive inhibitor changes shape of enzyme so it cannot bind to substrate
o Maximal effect changes
 o Binds the receptor and does not allow to reach the fully active conformation

2.10) Membrane R: Antagonists & Modulators : see slides

2.11) Modulation of receptor/enzyme activity: orthosteric & allosteric

 Classes of allosteric modulators independent of MOA and dependent on response
o Positive allosteric modulators (PAM)
o Negative allosteric modulators (NAM)

2.12) target signal transduction pathways

 Target select molecular step?
 Antagonists (receptors) vs inhibitors (enzymes/transporters)
 Genetic controls: KO and overexpression

2.13-14) membrane transporters: competition for substrate

2.15) Bioassays: advantages and limitations

 Postulate: develop and validate biological assays that ebst recapitulate biological
functions and disease states
o Biochemical assays (e.g. radioligand binding and changes in cAMP)
 +: high throughput
 -: non cellular
o Cells in culture (e.g. mouse neurons in culture expressing receptor or transfected
gene to express receptor)
 +: high throughput
 -: reductionist model
o Rodent models (e.g. genetic mouse model with KO or OE receptor)
 +: better recapitulate neuronal function and brain disease
 -: low throughput and significant specifics differences

Lecture 3 (4/2)
3.1) Pharmacokinetics: Overview
3.2) Different routes of administration
 PK: Time course of onset and duration of the drug’s action both of which depend on
bioavailability
 Concept: total amount of drug injected in body will determine the intensity and duration
of both the therapeutic effect and side effects -> therapeutic index
3.3)
 Flux through membranes
o Hydrophilic molecule : channels and transporters
o Hydrophobic drugs: cross cell membrane depends on gradient
3.4) Liver
  First organ to encounter when taken orally; when injected, can bypass lvier and go
directly to the brain
 Hepatic biodegradation: Enzyme metabolize (oxidize) molecules (cytochrome P450 is
the major family)
 Decrease compound’s pharmacological activity by altering the chemical structure of the
drug (e/g/ add chemical moiety) -> doesn’t bind to receptor anymore
 Transporters: filtering metabolites by transport: drug carried by these mechanisms
 P450: oxidizing enzyme to process drugs
3.5) BBB
 Astrocyte & pericyte & neurons control the BBB
 Special organ and expresses special type transporters (ex. GLUT1);
3.6) Drug excretion: half-life
 Elimination of the drugs through kidneys (major route of excretion of drug metabolites),
lungs (only volatile) and skin
 Drug half-life: time required to reach 50%
drug free: subject has eliminated 98% of the drug (6 half lives)
 Concept: the time required to be drug free depends on drugs
3.7) Regiment treatment
 Treat before half-life of drug to reach steady state levels
 Concept: avoid overshooting and inducing side effects
 Clamp at therapeutic dose
3.8) therapeutic index: the art of heling without harming
 Wide vs narrow therapeutic index
o Wide ex. Ibuprofen; narrow ex. Chemotherapy
3.9) why does drug response vary?
 CYP polymorphism in p450 enzyme
o PM, IM, EM, UM
3.10) Pharmacogenetics
 Goal: find targets/enzymes that have amino acid sequence differences that change drug
activity and adapt treatment regimen
 Polymorphism: genetic differences between individuals. True for receptors and enzyme
that metabolize drugs
 P450: a change in one critical amino acid at active site can affect enzymatic activity and
metabolism. Although very small amounts of genetic differences occur, thy are
important when considering large populations
 Oncogenes: not exactly the same AA seq between WT and oncogene -> use drug that
targets mutant pop
3.11) Novel medical approaches
 Personalized medicine: a medical model that proposes the customization of
healthcare
 Pharmacoeconomics: evaluates whether the incremental cost of a drug is justified by
the incremental gains in quality of life and life expectancy, compared to the current
standard of care
Quiz 1:
#1)
Full agonist: active site; Vmax does not change
Competitive neutral antagonist: active site; Vmax does not change and has no biological effect 
inverse agonist: active site (stabilizes the inactive form); Vmax decreases below "0"  
non-competitive antagonist: other sites other than active site (changes the conformation so it
decreases the chance of a substrate binding to the active site ); Vmax decreases

Full agonist, competitive neutral antagonist, inverse agonist = same orthosteric

binding site
Non-competitive = different binding site
Full agonist = 100% Vmax, Competitive neutral antagonist has 0 Vmax
Inverse agonist has – VmaxNon-competitive (allosteric) can reduces Vmax and Kd
of agonist

#2)
P450s are a family of oxidizing proteins that are mainly involved in drug metabolism in liver. The
P450 impairment will cause greater side effects because the patient will not be able to degrade
the drug properly. In order to address this and avoid overshooting, I would deliver the drug in
lower doses but for a longer period.

Enzyme that metabolizes drugs/toxins in the liver. Low P450 activity will reduce
metabolism and increase drug bioavailability which can lead to greater side effects
since the drug is circulating in the body for longer. Reduce dosage, deliver orally.

TOPIC II: Mind Altering Drugs

Lecture 4 (4/5)
1.1) Difference btwn THC and CBD: accepted by scientific community
 THC and CBD diff by closure of Aromatic ring but v similar overall
 Difference: THC >0.3%, THC <0.3%
 THC:
o psychoactive-enhances sensory awareness
o medical: analgesic/apetite
o side effect: impact brain development, high not always wanted
 CBD:
o No high tames THC properties
o Medical: anti-epileptic, anti inflammatory
o Side effects: different than CBD?
1.2) Cannabis Plant and Cannabinoids: Poly-pharmacology
1.3) Biological effects: Molecular to Human Physiology
  THC : CB1/CB2; CBD: GPR55
 Long term THC take reduces immune response
 Endogenous cannabinoids = same fxn?
o Can study endogenous pathway using exogenous drugs
1.4) Behavioral effects: Responses to mailed questionaries
 Poly model responses
 Most common reported effects:
o Sensory effects: visual, auditory and touch
o Perception: space and time
o Interpersonal relationship: shy/talkative/paranoia
o Thought process: attention, focus & short-term memory
o Upper/downer/sleepy
 Positive self: enjoyment, altered perception, experimentation and sleep; other:
celebration
 Negative self: coping, boredom, social anxiety, sleep; other: conformity
1.5) Biological effects: Acute effects
 Classified as mind altering drug bc it doesn’t fir in classic simulant/depressant category
o Increases sensitivity to external stimuli, Anxiety, Time distortion, Attention
deficit/paranoia
 Concept: could cannabinoid agonist/antagonist be used to treat anxiety, attention
deficit, paranoia?
1.6) Biological effects: Long-term use (7 “hits” a day)
 Addiction
o Human: significant; more of a psychological addiction (thinking about the drug);
cannabis use disorder
o Rats/monkey: self administer CP55940, rarely with THC
o Withdrawal: anxiety & insomnia
o Tolerance: humans/animals more sensitive during initial use
o Cross-interactions: EtOH
o Major long term conseq.:
 Young adults: psychosis, can exacerbate/precipitate the psychosis
 Short term memory impairment
1.7) ID of cannabinoids binding site: overview
 Major psychoactive ingredient = ∆9THC = hydrophobic
 Controversy: are these biological/behavioral effects produced through membrane
(lipophilic) or protein/receptor
o Synthesized a new selective and high affinity cannabinoid ligand: compound
Pfizer
1.8) Cannabinoid compounds: chemical structure
1.9) Binding theory: Overview
 Law of mass action: D + R <- - > DR* <- - > biological effect
1. Need a selective and high affinity radioactive ligand
2. Need high affinity because number of R in prep is low
 3. 3H radiolabeled: best bc no change biological characteristic (no I bc bulky)
4. Drug produces behavioral effects -> thus incubation with brain homogenate
5. This is membrane bound receptors -> centrifuge to recuperate P2 fraction = membrane
with receptors
 Concept: increase specific binding by eliminating non-specific binding
1.10) -11 Binding theory: experiment
 Add radioactive ligand -> add radioactive +non-radioactive ligand -> check how much
radioactive ligand goes
1.12) Binding theory:
 Filter: eliminate the unbound radioligand and keeps radioligand bound to R and
membranes
 Total binding = specific + non-specific
 Add excess cold ligna (non radioactive): non-specific only 1 chance per 10.000 to bind R
 Total minus nonspecific = specific
 None specific= lignad binds to the intert bidnign sites ex. plasma membranes
(accumulation of hydrophobic molecule) or ionic attractant (glass/plastic)
 Increasing cold ligand does NOT displace the non-specific binding of radioactive ligand ->
infinte amounts of binding sites
1.13) Binding theory: results
1.15) Cannabinoid binding site: ;landmark paper!
 Saturation of specific binding = 1st criteria
 Adding increasing amount of radioactive ligand reaches a max binding site = Bmax
 Finite number of R and all are bound; if increase amount of homogenate -> increase the
number of R -> increase the Bmax
 Slope of curve = affinity = rate at which saturation is reached
1.16) Cannabinoid binding site: time course of association
 Incubate enough time for the ligand to bind receptors (~30-40 min)
 Equil. Reached when: rate of receptor-ligand binding = rate of receptor-ligand
dissociation
 Filter at increase time
o total and non-specific -> calculate specific immediate equilibrium (equal weak
on/off rate)
o predominant on rate progressively saturates
o time dependent equilibrium -> shake to increase probability of interacting with
the receptor
1.17) Cannabinoid binding site: dissociation/displacement
 dissociation/displacement = 2nd criteria
 as the radioligand dissociates, the large excess of cold ligand is more likely to bind to the
R
 compare rate of association and dissociation -> gives an indication of affinity
o ex. Rapid binding with slow dissociation = high affinity
 saturate with radioactive ligand (90min), add excess cold, filter at increase time; total
and non-specific -> calculate specific
1.18) cannabinoid binding site: competition
 competition experiments: 3rd criteria
 shows competition of bound radioligands with analogues; measure ki= concentration
that leads to 50% of competition of radioligand
 asymmetric carbon (enantiomer): (-) isomer is prevalent configuration for naturally
occurring agonists. (-) and (+) bind differently to R. ionic attractant similar absorb/bind
(membrane area)
o – usually binds with higher affinity
 Concept: structure activity relationship studies demonstrate a correlation between
receptor binding and in vitro activities

Lecture 5 (4/7)
2.1) Cannabinoid binding site: autoradiobiography
2.2) Cannabinoid receptor: cloning
2.3) Cannbainoid binding site: competition

2.5)
 Symmetric structure -> GABAergic
2.7) Crystallography and protein structures: CB1R
 Easier to stabilize in inactive form
2.8) Cannabinoid receptors: presynaptic channel regulation
 Neuronal cell express Cb1R endogenously
2.9) Cannabinoid receptors: signal transduction
-> CBD prevents the inhibition of cAMP accumulation by binding to Cb1 receptor
2.10) Receptor biased signaling: Molecular mechanism
2.11) Receptor signaling: biased signaling dose responses

Lecture 6 (4/9)
3.1) Endocannabinoids: Identification
 AEA is in brain -> bind to receptor -> activate or inhibits signal coupled to receptor ->
produced in an activity-dependent manner by neurons -> inactivated THEN physiological
role
3.2) Endocannabinoid: Quantification by GC/MS and LC/MS
 Lipid extraction from brain (homogenize, add methanol & chloroform)

3.3) Technic: chromatography

 Hydrophobic/philic Beads; hydrophobic beads will yield hydrophilic molecules first
 Size

3.4) Endocannabinoids: Functional activation of cannabinoid receptors

 CHO cells don’t express CB1 but transfected to express CB1; heterologous expression
(compare to endogenous expression); CHO only would be a genetic control
 AEA inhibits cAMP expression
 PEA (enantiomer of AEA?)
 Positive control = HU210 (THC analogue)

3.5) Endocannabinoid: Retrograde Messengers

 (activate and then have negative feedback) As activate post synaptic calcium increase
and the life pahse swills tart producing endogenous cannabinoid will regulate CB1R
  Presynaptic terminal active postsynaptic, calcium influx, active lipase, produce
endocannabinoid, regulated CB1 and 2 and do presynaptic inhibition
 Hydrolyzing enzymes will degrade endocannabinoid

3.6) Inhibition eCB inactivation: Avoid tolerance

 Acute inactivation: agonist – CB -> response
 Desensitization/tolerance: agonist – CB -> response reduced; to avoid, increase agonist
but this results in side effects
 to desensitize, you increase the number of endogenous ligand by inhibiting hydrolysis
of them

3.7) inhibition eCB inactivation: regulate synaptic plasticity

 MGL = hydrolyzing enzyme

3.8) Cb1R heterosynaptic regualtion: both inhibitory and excitatory neurons express CB1R
 Additional difference between Cb1R on Glut and GABA: coupling

QUIZ 2:
#1)
Ligand & tissues for specific binding: For the ligand, I would exogenous ligands such as THC or
CP that binds to the CB1 receptor. For tissues, I would need brain tissue (specifically the parts of
high glial cellular area and expression GABAergic neurons).
3 tested criteria: association time course (time to reach equilibrium), dissociation/displacement
time, and competition with analogues.
Importance of homogenizing tissue and using centrifuge: it’s important to homogenize the
tissue and use centrifuge the samples in order to first separate away the organelle pellet from
the cytosol and membranes components and to further separate away the cytosol supernatant
and keep the membrane components (main area of interest for receptors and membrane).
Affinity and Bmax values change after partial KO of CB1-R: after the partial KO of CB1-R, the the
affinity would remain relativelt the same while the Vmax would decrease as there’s less
receptors available.
Time course and association change in colder temp: time course and association change would
both be prolonged in colder temperature as molecules move have lower kinetic energy in lower
temp.

Radioactive cool, non-radioactive cool, a known CB1R ligand (e.g., CP or THC), and
brain tissue.Saturation, Dissociation, Competition.Centrifuge to concentrate
receptors, eliminate non-specific binding.No change in affinity, lower Bmax. Slower
association at colder temp.

#2)
CB1 & CB2 diff: CB1 and CB2 are pretty homologous, but CB1 has longer AA seq and are
expressed more widely through the brain and the body (CB2 is more selectively expressed in
the immune system). THC can bind to both CB1 and CB2 while CBN can only bind to CB2.
Experiment design: use antobody probes that bind to CB1-R and validate the results using
techniques such as immunohistology, electron microscopy, and/or high-resolution microscopy.
Then, I would check to see that these CB1R expressed in presynaptic terminal by using voltage
clamps and exogenous ligand and measure how the presence of ligand affects the post synaptic
neuron.

Similar structure, small diff in NT and CT, and in 3rd intracellular loop.CB1 neuronal,
CB2 immune, CB1 - THC, CB2 - THC, CBN
Perform immunohistology (antibody staining for CB1R, fluo microscopy for brain and
EM or high-resolution microscopy for presyn). Not autoradiography!CB 1R-KO brain
tissue

Lecture 7 (4/12)

4.1) Model system: from mice to men

 Biological effects produced cannabinoids in humans?
 Biological effects measured in mice?
 Basic vs complex physiological and behavioral effects

4.2) Cannabimimetic effects: tetrad in mice

 Locomotion
 Hypothermia
 Analgesia
 Catalepsy: cognitive dissociation

4.3) Cannabimimetic effects: drug discrimination

 Mice continuously trained to discriminate unknown drugs from THC

4.4) Receptor antagonist: identification of the first CB1R antagonist

 Positive control: CP,, WIN, THC

4.5) Cannabimimetic effects: genetic evidence for involvement of CB1R

 With KO, the effects of hypothermia, catalepsy, are gone
 Analgesic effect not so much affected

4.6) Cannabimimetic effects in Cb1RKO: evidence for novel receptors

 Potency of WIN very high
o Win inhibiting IPSCs
  WIN receptor responds to high concentration of WIN and is antagonized by high
concentration of SR1
 -> there’s a WIN receptor on Glut terminal; CB1R on GABA terminals

5.1) Pharmacokinetics: Multiple parameters control PK profile

 IV highest, orally much slower

5.2) PK in human: Blood levels and bioactivity

 Most common route: smoke > oral
 Smoking results loss of bioactive ingredient -> fraction of THC in smoke ends up in
mainstream (lost in side-stream smoke and pyrolytic degradation)
 Smoking not efficient delivery system
 Takes time bc THC need to travel, pass the bbb, activate the receptors and start signal
transduction mechanism, and change neuronal activity

5.3) Different routes of administration: PK in rats

 Injection
 Smoking
 Oral
 Injection & smoking very similar PK profile
 Error bars larger for oral; most likely bc liver p450 difference and digestive system
difference

5.4) Different routes of administration: specifics

 Smoking (lungs) = same PK as IV
o Binds to carrier proteins (albumin proteins)
o First effects occur within 10-30 minutes
o Half life 2-4 hours
 Oral
o Degradation by acid present in stomach
o THC = hydrophobic -> difficult to reach the blood bc trapped in food
o Degradation by first hepatic pass -> produces initial metabolites
o First effect within 4-12 hours
o Half life 20-30 hours

5.5) THC metabolism: portion of drugs metabolized by P450

 CYP3A4/5
 THC mainly metabolized by CYP2C/3A

5.6) excretion: THC metabolites in human urine

 THC: Eliminated through the kidney (after smoking)
 11OH THC: processed by liver
 THCCOOH: eliminated by urine (more hydrophilic)
5.7) bioavailability and excretion: specifics
 First THC enter, binds to albumin in blood then…
1. Smoking: Accumulation in adipose tissue
2. Orally: Eliminated by feces 2/3 and 1/3 by urine
3. Find over 20 metabolites, some of which psychoactive
4. Metabolites accumulate in hair
5. Not fully cleared from the body after 5 days; depends on
a. P450 expression
b. Analytical technique sensitivity

Lecture 8 (4/14)
5.8) Regimented administration: PK of smoking
5.9) Regimented administration: Delivery and select responses
 Burn plant, vape oil, dab resin; capsules, eatables, drinks
 Concerns: Higher potency products, quicker onset devices, cannabis use disorder,
vulnerable populations, toxicity of add-ons

5.10) Route and form of administration

 Subcutaneous, intramuscular, intraperitoneal; oral, rectal, inhalation; IV; intrathecal
 Change the route or form to have acute/long-term effects

5.11) Administration: Different devises

 Liquid, tablet, or capsule (slow release), and patch
5.12) Molecular imaging: PK/PD
 Reduction TI -> solution = PK/PD
 How long it takes to reach the target AND how much it is actually bound to the receptor

5.13) Molecular imaging: different imaging modalities

 PWT, SPECT, OPTICAL, MRI
 Ligand-linker
 By conducting an outcompeting experiment, you can check how much is reaching to the
targeted areas in the body

Lecture 9 (4/16)

Primary fxn of opioid = pain control

 Endogenous: Beta endorphin, enkephalin, dynorphin

Opioid affecs mood as well

 Mu & kappa receptors
o MU: euphoria
 Disinhibition of dopamine release
 o Kappa: dysphoria
 Inhibition of dopamine release

 Brain/spinal cord areas that underlie pain differ from those that underlie
reward/aversion
 Challenge of opioids is disconnecting the pain relieving properties from the effects on
mood

Signaling diversity at the kappa receptor

 Dynorphon binds, GDP & GTP exchange occurs
 Gai lowers adenynyl cyclase activity
 Bb/y opens K+ channels (go out) and closes Ca++channels -> hyperpol.
1. G protein effects: inhibition of excitability -> analgesia
2. Arrestin internalization: termination of G protein signaling
3. Arrestin signaling: p38AK -> dysphoria

Biased Signaling
 G protein and arrestin response -> ligand can prefer one over the other
 Ex. Nalfurafine: kappa receptor
o G-> analgesia; no arrestin so no dysphoria

Biological sex can “bias” signaling

 Estrogen can inhibit g protein signaling for kappa receptor

 Response of GPCR affected by many factors:

o Kind of agonist and how it changes receptor conformation
o Amounts of interacting partners within the cell
o Active regulation of those partners by other proteins

Overdose
 Characteristics of opiates and the MOR
o Euphoria: PK – route of administration
o Tolerance: PK – withdrawal
o Side effects: respiratory depression

Euphoria: PK
 Higher drug conc = more GABA inhibition = increased dopamine release = greater high
 What goes up must come down
o Upregulation of cAMP

Mu receptor and PK & PD

 Kinetics closely linked with mood when a drug alters mood
 Dynamics determine the continued ability of drug/receptor system to signal
 Therapeutic index (LD/ED) determines the safety profile of a drug
 MOR is a dramatic example of system that changes mood but also undergoes tolerance,
with low safety profile
 Biggest challenge is disconnecting pain relief from mood changes
 KOR (decouple two)
 MOR more complicated, and generating analgesia without either elevated mood or
breathing suppression difficult