Acetylcholine. Neuroscience with Pharmacology 2. Neuromuscular Junction 2: Pharmacology. Quaternary nitrogen. Neuromuscular Junction - Pharmacology
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1 Neuroscience with Pharmacology 2 Acetylcholine Neuromuscular Junction 2: Pharmacology + Sir Henry Dale Quaternary nitrogen Neuromuscular Junction - Pharmacology 1. Principles and methods for studying pharmacology at the NMJ 2. Targets for drug action at the NMJ 3. Drugs affecting Receptors (activation) 1. Principles and methods 4. Drugs affecting Esterase (inactivation) 5. Drugs affecting release 6. Drugs affecting synthesis and storage. When doing drugs a good pharmacologist needs to know: Drugs acting at the NMJ are useful as: Eperimental tools Clinical treatments Target specificity/ligand specificity Agonist/Antagonist Dose/Response Efficacy (EC50/IC50)/Affinity (K D ) Competitive/Non-competitive Clinical/Non-clinical uses Side-effects 1
2 Drug assays at the NMJ are based on: Measuring muscle contractions in response to release of neurotransmitter by nerve stimulation. Repetitive stimulus : Tetanus Biological response (->efficacy, EC50) Ligand binding (->affinity, K D ) Single nerve stimulus : Twitch 1 sec Specificity and dose-response of antagonists Atropine d-tubocurarine EC 50 Ligand binding visualised But normally measured chemically (eg radioactively-labelled ligand) Control α-bungarotoin 10µm α-btx 2
3 Ch.2 2 mv ms s Testing drugs on receptors Measuring EPP s. Iontophoresis I 10 µm X Action potential add µ-conotoin add d-tubocurarine V 2 mv 60 s Bath application Patch clamp Targets in neuromuscular pharmacology : Synthesis 2. Targets for drug action at the NMJ Storage Release Action Inactivation Targets of drug action at the neuromuscular junction vesamicol 3. Drugs affecting Receptors (activation) hemicholinium botulinium (A-D) α-bungarotoin d-tubocurarine atracurium suamethonium edrophonium neostigmine sarin 3
![[R] = ~ 80 µm [-R] Stereospecifity of skew ( gauche ) configuration of /receptor binding.](/docs-images/83/88891389/images/4-1.jpg "(View as magic eye stereo pair.")
4 General scheme for Agonist-Receptor Interaction (illustrated by binding to its Receptors) The nicotinic Receptor at NMJ 8 nm + R -R -R* bound activated -R 0 desensitized /ε K D = [].[R] = ~ 80 µm [-R] Stereospecifity of skew ( gauche ) configuration of /receptor binding. (View as magic eye stereo pair.) Anionic site Esteric site ε α Anionic site ε α Esteric site R/ E n Receptor Pharmacology - Neuromuscular Junction Agonist nicotine acetylcholine carbachol decamethonium suamethonium (I) Antagonist/blocker d-tubocurarine (curare) α-bungarotoin atracurium pancuronium suamethonium(ii) 4
5 Reversible competitive antagonism of the receptor. 1 Na + dtc Decamethonium Pancuronium K + Reversible competitive antagonism of the receptor. 2 Drugs that block Receptors (antagonists) dtc Eample: d-tubocurarine dtc dtc Eample: α-bungarotoin Irreversible competitive antagonism of Receptor Suamethonium Acetylcholine 2 αbtx 2 αbtx Not broken down by E (nmj) Broken down by BuChE (blood plasma) Short lasting Not counteracted by cholinesterase inhibitor Used clinically for brief muscle relaation Depolarising blocker 5
6 Dual effect of suamethonium (succinyl choline) dtc dtc neo su su su neo direct 1. Depolarisation Na + Na + Su + + K + Dual effect of suamethonium (succinyl choline) 2a. Sodium inactivation (depolarising block) Na + Na + Dual effect of suamethonium (succinyl choline) 2b. Desensitization of R (depolarising block) Su Su + K + Non-competitive antagonists of the nicotinic receptor 4. Drugs affecting Esterase (inactivation) Phencyclidine (PCP, angel dust ) Methyl violet 10B (heamethyl violet; crystal violet) 6
7 AC mv s Drugs that block esterase 5. Drugs affecting release Antidote: K channel blockers and some toins enhance transmitter release Drugs that inhibit transmitter release P/Q Ca 2+ channel blockers Botulinum toins = SNARE blockers ω-agatoin/ ω-conotoin α-latrotoin Ca 2+ BEFORE During AFTER Counteracting the depression of release by low Ca 2+ or high Mg 2+ using 4-AP 0 Ca Direct +Ca +4AP +Mg +4AP DirectTTX 6. Drugs affecting synthesis and storage 7
8 Drugs that inhibit transmitter synthesis/storage Choline acetyltransferase Inhibitor Choline uptake X Vesicle filling Summary 1. Every step in the cycle - synthesis, storage release, activation and inactivation - is a potential target for drug action at the NMJ. 2. Clinical uses of drugs acting at the NMJ: - muscle relaants as adjunct to anaesthesia in surgery - treatment of neuromuscular disease 3. Drugs affecting Receptors: - carbachol, decamethonium, suamethonium (agonists) - tubocurarine, α-bungarotoin, atracurium (antagonists) 4. Drugs that block Esterase: - edrophonium, prostigmine, neostigmine, sarin 5. Drugs affecting release: - TEA, 4-AP, α-latrotoin (agonists) - botulinum toin, ω-agatoin (antagonists) 6. Drugs that block uptake, storage and synthesis : - hemicholinium-3, vesamicol, AF64A 8
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