IRREVERSIBLE ENZYME INHIBITORS Sazan J. Ali Pharmacutical chemistry department 1
Jul 15, 2015
IRREVERSIBLE ENZYME
INHIBITORS
Sazan J. Ali
Pharmacutical chemistry
department1
Irreversible enzyme inhibitors : Are
binding covalently or non covalently to the
enzyme and permanently inhibit it. there is
no reversal of inhibition on decreasing the
inhibitor concentration.
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Types of irreversible inhibition
Group-specific covalent modifying
agents
Affinity labels
Suicide inhibitors
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1.Group specific covalent modifying agents: react with
specific type of enzyme functional group (e.g., Ser-OH, or
Cys-SH, or His imidazole) on any enzyme/protein
React with amino acid side chains.
Lead to inhibition by interfering with the catalysis (e.g. by reacting with side-chains important for the catalysis)
E.g. diisopropylfluorophosphate(DFP),Nerve gas.
Inhibits acetylcholine esterase (and many other proteases with Ser at the active site).
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2.Affinity labeling agents(reactive
based analog)
Active-site directed irreversible inhibitors.
Recognized by the enzyme (reversible,
specific binding) followed by covalent
bond formation.
structurally similar to substrate, transition
state or product allowing for specific
interaction between the compound and
target enzyme.
E+I E I E-IKD Kinactivation
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Affinity labels(reactive based
analog)
Structural similarity to substrate
guides reagent to the active site of
enzyme.
It is a common and powerful tool to
isolate and characterize the enzymes
More specific than group specific
reagents.
Tosyl phenylalanin chloromethylketon(TPCK)
irreversibly in activate chymotrypsin
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3.Mechanism-based enzyme inactivators(Suicide Inhibitors)
• Active-site directed reagent (unreactive) binds to the enzyme active site transformed to a reactive form. Once activated, a covalent bond between the inhibitor and the enzyme forms
• This approach minimizes side reactions (non specific covalent bond formation) which may occur with an affinity reagent
E+I E..I E..I E-I
E+P
• 1) inhibitor binds to active site 2) converted to reactive compound via enzyme's catalytic capabilities 3) covalently reacts with the enzyme
• Inactivation (covalent bond formation, k4) must occur prior to dissociation (k3) otherwise the now reactive inhibitor is released into the environment
KD K2K4
K
3
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Bind like normal substrate
Begine, but unable to complete the
reaction
Get stuck in active site
TSInactive
enzyme
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• Allopurinol - the anti-gout drug - is a suicidal irreversible
mechanism-based inhibitor of the enzyme xanthine oxidase that
works as oxidase or dehydrogenase. The enzyme commits suicide
by initial activating allopurinol into a transition state analog -
oxypurinol - that bind very tightly to molybdenum-sulfide (Mo-S)
complex at the active site . The molybdenum-sulfide (Mo-S)
complex binds the substrates
and transfers the electrons required for the oxidation reactions.
(Mo-S)
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Suicide inhibitors - penicillin
Interfere with the synthesis of the
bacterial cell wall.
Makes bacteria much less resistant to
stress.
Penicillin blocks the formation of the
link between the tetra peptide and the
pentaGly bridge.
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13Inactive trans peptidase enzyme
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