Insecticide Mode of Action

Training slide deck

IRAC MoA Workgroup

Version 1.0, April 2019

1 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 What is an Insecticide’s ‘Mode of
Action’?

The Mode of action defines the process

of how an insecticide works on an insect or mite at a
molecular level

Why is it good to know the Mode of Action of

an Insecticide?

Knowing the Mode of action of an

insecticide is key to managing resistance

The Insecticide Resistance Action Committee (IRAC) is a coordinated

industry response to resistance management

2 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 ADME is an important factor in an
insecticide’s bioavailability

n Absorption
q Through the cuticle
q Orally through consumption
q Inhaled through spiracles as vapor

n Distribution
Metabolic
Enzymes
q Through the body to target sites

n Metabolism (Break down)

q By insect defense mechanisms

n Excretion

3 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides act on key functional
proteins that regulate vital processes
Target protein
e.g. ion channel

binding
pocket
Small molecule
insecticide
Insecticides act on key
functional proteins that
regulate vital processes
Altered protein
function
1. A protein can have more than one binding
pocket for small molecules
Altered
2. Specific binding pockets can accommodate biological
chemicals with different structures response
4 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 The Pro-Insecticide concept

A Pro-Insecticide might have negligible activity

on the target protein due to protecting groups,
but can get metabolized in vivo to its active form.
This can be a tool to:
• Improve bioavailability
Pro-Insecticide Metabolic
• Enable selective toxicity
Enzyme(s)

Insecticide

Target
protein

Altered protein
function
5 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Major mechanisms of Insecticide
Resistance
Metabolic
Resistance Metabolic
Enzymes
and/or

Target
Insecticide protein

Target Site
Resistance
Insecticide

Genetic modifications of Altered

Target the target that interfere with target
protein binding of the Insecticide
protein

6 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Key functional proteins that are targets
for Insecticides

Ion channels Enzymes Receptors

essential for signal

biocatalysts
bioelectricity transducer

ligand

+
Na+ Ca2+ K+
or Cl-
Proteins accelerating
chemical reactions
Binding of a ligand
triggers a response

7 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticide Mode of Action
Major classes

Nerve & Muscle

Growth

Respiration

Midgut

Unknown or Non-Specific

8 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Ion channels are important targets of
neuromuscular disruptors

Na+ Ca2+ Cl-

Cells are surrounded by a cell membrane, which acts

- - - like an insulator separating two conducting media
K+
Large organic An asymmetric separation of charges across the cell
anions
membrane makes the inside negative compared to
the outside of the cell (membrane potential)

Ion channels are pore-forming membrane

Na+, Ca2+ proteins that transduce signals by controlling the flux
or Cl- of ions across the cell membrane. Their concentration
gradients together with the membrane potential
• Gating: Trigger for channel determine the direction of ion flux.
opening (voltage, ligand, etc.)
• Ion selectivity: Ion preference In electrically excitable cells (neurons, muscle cells)
of the channel ion channels play an important role in fast signal
transduction over long distances
9 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 The Insect Neuromuscular system:
Translating a stimulus into (muscle) action
Muscle

Stimulus

10 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Different types of neurons are involved
in signal transduction and fine tuning
Muscle

Stimulus CNS
modulatory Ganglion
neuron

The Central Nervous System

(CNS) is the insect’s control
center
sensory
neuron

interneuron

inhibitory
neuron

motoneuron

11 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Ion channels play diverse roles within
the neuromuscular system
Muscle

CNS
Ganglion

12 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

13 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 The TRPV channels exist in specialized
sensory neurons that detect stretch
Muscle

CNS
modulatory Ganglion
neuron

Na+, Ca2+

TRPV
sensory
neuron

interneuron

inhibitory
neuron

motoneuron

14 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 TRPV channels play an important role in
insect stretch receptor signaling
(Chordotonal Organ)

• Sight
Stimulus • Taste
• Smell
• Touch
• Hearing (antenna)
• Gravity (antenna)
Sensed by
• Proprioception* (joints) stretch receptors
• Others

sensory
neuron
*Proprioception is the detection of the relative
position and motion of body parts

When a doctor tests Insects also have stretch

reflexes with a rubber receptors that detect joint
hammer (knee jerk reflex) bending forces
local stretch receptors are
activated that signal to the
spinal cord and back to the
target muscle.
15 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on TRPV channels
interfere with stretch receptor signaling
n TRPV channel modulators affect Chordotonal organs (COs), stretch receptors in
the insect joints that sense relative position of body parts (proprioception)

Leg extension caused by a

TRPV modulator (can be
imagined as a ‘molecular’
rubber hammer)

n TRPV channels amplify the weak mechanosensory signal in chordotonal neurons

1. Signal
detection 2. Signal
amplification

Mechano-
sensory TRPV
channel

n Modulation of TRPV channels generates continuous chordotonal nerve signals

independent of joint movement. This leaves insects deaf and uncoordinated,
resulting in rapid feeding cessation, leading to starvation and ultimately death
16 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 TRPV channel and chordotonal organ
modulators

Group 9: Chordotonal Group 29:

organ TRPV channel modulators Chordotonal organ
modulators –
undefined target site

Pymetrozine

9B Pyridine Flonicamid
Pyrifluquinazon Afidopyropen
azomethine
derivatives 9D Pyropenes 29 Flonicamid

Flonicamid produces symptoms similar to TRPV channel modulators and like its
more active metabolite affects chordotonal organs. However, Flonicamid
appears not to act directly on TRPV channels, suggesting a different target site
in chordotonal organs.

17 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

18 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on Sodium channels
Muscle

CNS
modulatory Ganglion
neuron
Na+
Sodium
Channel

sensory
neuron

interneuron

inhibitory
neuron

motoneuron

19 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Sodium channels play a crucial role in
signal propagation in excitable cells

Axons are long cable-like parts of

the neuron that carry electrical
signals to a junction with another
Na+
cell Voltage-dependent sodium
Sodium channels contain a built-in
Channel voltage sensor which detects
local positive changes in
membrane potential

This triggers opening of the

channels and sodium entry

Entry of sodium results in a

more positive membrane
potential that in turn activates
adjacent sodium channels
thus propagating the signal
along the axon

20 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Sodium channel modulators & blockers

Sodium channel m
m odulators Sodium channel b lockers

Closed Open Open Closed

m b
m b

Na+ Na+
Na+

• prolonged opening of sodium channels • obstruction of the sodium channel

• prolonged action potentials pore
• restimulation of the nerve and repetitive firing • block of nerve action potentials
• hyperexcitation • paralysis

21 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on Sodium channels
Group 3: Sodium channel modulators

Esfenvalerate Permethrin
Bifenthrin
DDT

Deltamethrin lambda-
cyhalothrin
Methoxychlor
alpha-
Etofenprox
cypermethrin Tefluthrin
3B DDT, Methoxychlor
3A Pyrethroids Pyrethrins (Only major representatives of group 3A are shown)

Group 22: Voltage-dependent sodium channel blockers

Indoxacarb

Metaflumizone

22A Oxadiazines 22B Semicarbazones

22 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

23 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on nicotinic
Acetylcholine receptors (nAChR)
Muscle

External
stimulus CNS
modulatory Ganglion
neuron

sensory
neuron

Acetylcholine
interneuron

inhibitory
neuron

Na+, Ca2+
nAChR
motoneuron

24 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on nicotinic
Acetylcholine receptors (nAChR)

Neurotransmitters
such as Acetylcholine bridge the
signaling gap (synapse) between
excitable cells (neurons, muscle cells)
Acetylcholine-filled
vesicles

Ca 2+
Na +,
Na+, Ca2+
Na+, Ca2+
• Most fast excitatory synapses in the
insect CNS use Acetylcholine as the
neurotransmitter

• Synaptic vesicles store Acetylcholine

that can be released into the synapse in
response to nerve impulses and in turn
activate postsynaptic nAChRs
25 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on nAChR
Can cause
nAChR competitive modulators hyperexcitation
• Bind to the same site as acetylcholine
and/or
• desensitize nAChR
inhibitory paralysis

c c
nAChR allosteric nAChR channel
modulators m
b
blockers
• Bind to the acetylcholine- • Obstruct the pore and
opened channel form at a prevent ion flow
different (allosteric) site and • prevent acetylcholine
keep channels open signal transduction
Na+, Ca2+
Can cause hyperexcitation nAChR
and contractive paralysis Can cause
flaccid paralysis
26 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Summary of Insecticides acting on nicotinic
Acetylcholine receptors (nAChR)
Group 4: Nicotinic acetylcholine receptor (nAChR) competitive modulators

Dinotefuran

Nicotine 4B Sulfoxaflor 4C
Nicotine Sulfoximines
Acetamiprid Nitenpyram

Imidacloprid Thiamethoxam

Clothianidin
4A 4D 4E
Thiacloprid Flupyradifurone Triflumezopyrim
Neonicotinoids Butenolides Mesoionics

Group 5: Nicotinic Group 32: Nicotinic Group 14: Nicotinic acetylcholine receptor (nAChR) channel blockers
acetylcholine receptor acetylcholine receptor
Spinetoram (nAChR) allosteric (nAChR) allosteric
major component R = H 5,6 single
modulators - site I modulators - site II O S
minor component R = CH 5,6 double
3,
S
S
O
S S
N
Spinosad S O NH2 SO 3Na
N S
major component R = H O O S
minor component R = CH3 S
GS-omega/kappa Thiocyclam
Bensultap S
HXTX-Hv1a N
S NH 2
N SO3Na
peptide H Cl O

5 Spinosyns 14 Nereistoxin Cartap Thiosultap-

analogues hydrochloride sodium

27 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

28 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on GABA-gated
chloride channels (GABA-Cl)
Muscle

External
stimulus CNS
modulatory Ganglion
neuron

sensory
neuron

interneuron GABA

inhibitory
neuron

motoneuron Cl-

GABA-Cl

29 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on GABA-gated
chloride channels (GABA-Cl)
GABA

Cl -
Cl-

GABA-Cl inhibitory

Cl-
neuron

• GABA is a major inhibitory

Cl-
neurotransmitter in the CNS and
neuromuscular synapses

• Influx of the negatively charged

Cl- ions has an inhibitory effect,
counteracting excitatory signals

30 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on GABA-gated
chloride channel (GABA-Cl)

GABA-Cl antagonists GABA GABA GABA-Cl allosteric

modulators
• Block the pore and m
prevent chloride influx, b
• Binding of modulators
interfering with the negatively affects GABA-Cl,
channel’s inhibitory interfering with the channel’s
function Cl-
inhibitory function
GABA-Cl

Pore blockers and negative modulators both cause

convulsions and death
31 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Summary of Insecticides acting on
GABA-Cl
Group 2: GABA-gated chloride channel antagonists

Ethiprole Fipronil
Chlordane Endosulfan

2A Cyclodiene Organochlorines 2B Phenylpyrazoles (Fiproles)

Group 30: GABA-gated chloride channel allosteric modulators

Meta-diamides & Fluxametamide

Broflanilide
Isoxazolines

32 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

33 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on Glutamate-gated
chloride channels (GluCl)
Muscle

External
stimulus CNS
modulatory Ganglion
neuron

sensory
neuron

interneuron Glutamate

inhibitory
neuron

Cl-
motoneuron
GluCl

34 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Allosteric modulators of Glutamate-
gated chloride channels (GluCl)

Glutamate

Cl-
GluCl

• Inhibitory Glutamate-gated chloride channels (GluCl) are

widespread on insect nerve and muscle cells and likely
function in inhibitory neurotransmission
• GluCl modulators activate chloride influx, causing flaccid
paralysis

35 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Allosteric modulators of Glutamate-
gated chloride channels (GluCl)

Group 6: Glutamate-gated chloride channel (GluCl) allosteric modulators

major component R2 = Ethyl

Abamectin R1 = minor component R2 = Methyl
O

HO

O O

H
O O
H
O R
N Lepimectin H
Emamectin
O
benzoate R1 = N
O O
H
O
O O Milbemectin
O
OH H
O R
H
O

O O
O
OH H
H
OH

6 Avermectins & Milbemycins

O major component R = Ethyl
H
OH
minor component R = Methyl

36 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Overview of ion channels targeted by
neuromuscular disruptors
Muscle

CNS
Ganglion

37 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticides acting on the Ryanodine
Receptor (RyR)
Ca2+ Muscle

External
stimulus CNS
modulatory Ganglion
neuron

Ca2+
RyR

sensory
neuron

interneuron

inhibitory
neuron

motoneuron

38 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 The Ryanodine receptor (RyR) plays a
crucial role in insect muscle contraction
motoneuron • Ca2+ ions entering the muscle cell
terminal activate RyR located on the
muscle cell’s intracellular Ca2+
Glutamate-filled store to release Ca2+ into the
vesicle
cytoplasm
• The rise in Ca2+ activates more
RyRs, leading to massive Ca2+
Excitatory
glutamate release
receptors
• This in turn induces shortening of
Na+ Na +
Ca 2+ Voltage-gated
Ca 2+ calcium channels contractile filaments, leading to
Ca 2
Ca2+ Ca
2+ Ca2+
+
muscle cell contraction
Ca2+ Ca2+
Ca2+ • RyR modulators open the
Muscle filaments
2+
RyR ryanodine receptor independent
Ca 2+ Ca 2+
Ca 2+Ca Ca2+
Ca2+ Ca2+
of cytoplasmic calcium levels
Ca2+ Ca 2+

Ca2+
Ca2+
Ca2+ causing uncontrolled muscle
contraction, paralysis and death
muscle
cell
39 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 RyR modulators
Group 28: Ryanodine receptor modulators

Chlorantraniliprole R=Cl
Cyantraniliprole R=CN

Cyclaniliprole

Flubendiamide

28 Diamides Tetraniliprole

40 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Enzymes targeted by neuromuscular
disruptors
Muscle

CNS
modulatory Ganglion
neuron

Acetylcholine-
esterase
AChE

41 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Acetylcholinesterase (AChE) degrades
Acetylcholine in the synapse
ne
Acetylcholinesterases yl choli
Acet
degrade Acetylcholine in order to
terminate the signal after it has been AChE
sent across the synapse

ne
Acet
ate + choli

AChE
Acetylcholine-filled
vesicles
AChE inhibitors bind to the active site of the
Ca 2+

enzyme preventing Acetylcholine from

Na +,

binding
nAChR hol i n
e
Na+, Ca2+

yl c
Acet

-I
A C hE
Na+ , Ca2+

AChE
AChE

ne
Acet
ate + choli

42 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Acetylcholinesterase (AChE) inhibitors

Group 1: Acetylcholinesterase (AChE) inhibitors

Acephate Methamidophos
Aldicarb Oxamyl

Methiocarb Fenitrothion

Thiodicarb
Carbaryl
Chlorpyrifos Profenofos

Methomyl
Malathion

Carbofuran Pirimicarb (Only major representatives Dimethoate

of the groups are shown) Triazophos

1A Carbamates 1B Organophosphates

43 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Receptors targeted by neuromuscular
disruptors
Octopamine
receptor Muscle

CNS
modulatory Ganglion
neuron

44 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Octopamine receptor signalling
a-adrenergic-like
(OctaR)
• Octopamine is the major
modulatory neurotransmitter in
g PIP2
aq
b PLC DAG insects; it can increase the
general level of arousal like
IP3 Ca2+
adrenaline does in mammals
Ca2+
• Upon release, Octopamine
binds to G-protein-coupled
Ca2+ IP3R Receptors (GPCRs) on the
Octopamine
Ca2+
postsynaptic membrane, which
can be coupled via G-proteins
receptors
Ca2+
Ca2+ Ca2+
Ca2+

Ca2+
Ca2+
Ca2+
(abg) to Phospholipase C (PLC),
Ca2+
Ca2+
an enzyme catalyzing the
formation of IP3, which in turn
can activate intracellular Ca2+
ATP release channels. Other
cAMP
octopamine receptors are
b
as g AC coupled to adenylyl cyclase (AC)
to stimulate cAMP production
• Octopamine receptor agonists
b-adrenergic-like can mimic the action of this
(OctbR) neurotransmitter

45 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Octopamine receptor agonists

Group 19: Octopamine

receptor agonists

Amitraz

19 Amitraz

46 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Targets of Neuromuscular Disruptors &
Their key Physiological Roles

TRPV Sodium
nAChR GABA-Cl GluCl RyR
Channel Channel
Excitatory Inhibitory Inhibitory
Action potential Muscle
Proprioception neuro- neuro- neuro-
propagation contraction
transmission transmission transmission

Acetylcholin- Octopamine
esterase Receptor
Excitatory Excitatory
neurotransmission neurotransmission

47 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Insecticide Mode of Action
Major classes

Nerve & Muscle

Growth

Respiration

Midgut

Unknown or Non-Specific

48 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Growth & Development Disruptors
Background
The insect’s skeleton is external (exoskeleton) and contains chitin.
Since the exoskeleton cannot expand, it must be replaced with a larger one
by the process of molting as the insect grows, requiring chitin synthesis.
Molting is under strict hormonal control:

Ecdysone Juvenile
Hormone (JH)
Egg
Molt
Ecdysone releases

Ecdysone is released Molt

JH prevents molting to
by the prothoracic
a more mature stage

JH levels
Larval
gland in the thorax Molt Instars
JH production stops
Pulses of Ecdysone
during metamorphosis
induce molting Pupation
Pupa

Metamorphosis
Adult

49 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Targets of Growth & Development
Disruptors

Juvenile
Ecdysone
NHRs alter the expression of genes needed for Hormone
Receptor ecdysis (EcR) or to form adult structures (JHR) Receptor
Ecdysone JH

EcR JHR
mRNA
NHR
Pulses of Protein JH prevents
Ecdysone molting to a
induce more mature
molting stage

EcR agonists directly and persistently JH mimics activate the JH receptor and
activate Ecdysone receptors causing the strongly suppress the development of
insect to go into a precocious, incomplete adult characteristics keeping the insect in
molt, leading eventually to death an ‘immature’ state

50 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Juvenile hormone mimics
Group 7: Juvenile hormone mimics

Hydroprene R1 = ethyl, R2 = H
Methoprene R1 = isopropyl, R2 – OCH3

Fenoxycarb Pyriproxyfen
Kinoprene R1 = propargyl, R2 = H

7A Juvenile hormone analogues 7B Fenoxycarb 7C Pyriproxyfen

Ecdysone receptor agonists

Group 18: Ecdysone receptor agonists

Chromafenozide Methoxyfenozide

Halofenozide Tebufenozide
18 Diacylhydrazines

51 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Targets of Growth & Development
Disruptors

Chitin
synthase

• Chitin is a polymer of N-Acetyl glucosamine

(NAcGlc; )
ChS • Chitin synthase uses activated NAcGlc
(UDP-NAcGlc; UDP ) to extend the
growing chitin chain
UDP UDP • The nascent chain is released into the
extracellular space
• Interfering with chitin synthesis results in a
weak and soft exoskeleton as well as
deformed appendages and sexual organs

52 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Chitin synthesis inhibitors
Group 10: Mite growth inhibitors affecting CHS1

Diflovidazin

Etoxazole
Clofentezine Hexythiazox
10B Etoxazole
10A Clofentezine, Diflovidazin, Hexythiazox

Group 15: Inhibitors of chitin biosynthesis affecting CHS1

Group 16: Inhibitors of
chitin biosynthesis, type 1

Diflubenzuron Lufenuron Teflubenzuron

Buprofezin
Flufenoxuron Novaluron
16 Buprofezin
15 Benzoylureas (Only major representatives
of group are shown)

53 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Targets of Growth & Development
Disruptors

• ACCase (acetyl CoA

ACCase carboxylase) catalyzes the
first and rate-limiting step of
CO2
Acetyl Malonyl fatty acid biosynthesis
CoA CoA
CO2 Acetyl • ACCase inhibitors prevent
CoA biosynthesis of fats needed
COOH for growth and development
resulting in incomplete molts
and desiccation of the insect
Fatty acid

54 Details are accurate to the best of our knowledge but IRAC and its member companies cannot accept responsibility for how the information is used or interpreted. Protected by © Copyright
 Inhibitors of acetyl CoA carboxylase

Group 23: Inhibitors of acetyl CoA carboxylase

Spiromesifen

Spirodiclofen Spirotetramat

23 Tetronic & Tetramic acid derivatives

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 Overview Growth & Development
Targets
Juvenile
Ecdysone
Hormone
Receptor
Receptor
Ecdysone JH

EcR mRNA JHR

NHR
Protein
Pulses of JH prevents
Ecdysone molting to a
induce more mature
molting NHRs alter the expression of genes needed for stage
ecdysis (EcR) or to form adult structures (JHR)

Chitin Acetyl CoA

synthase ChS carboxylase
Chitin synthesis Fatty acid
synthesis

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 Insecticide Mode of Action
Major classes

Nerve & Muscle

Growth

Respiration

Midgut

Unknown or Non-Specific

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 Respiration and Energy Conservation
Background

Cells get the energy they need by "oxidizing" food such as

fat and sugars to produce carbon dioxide and water. In the
cell this process is conducted in many steps in which
oxygen is added to the fuel (from water) and hydrogen is
removed so that the fuel is converted to carbon dioxide.

The hydrogen is removed by adding it

to NAD (a form of vitamin B3) forming
NADH. The regeneration of NAD
requires oxygen, produces water and
releases a lot of energy.
Mitochondrion

Most of these steps are contained in

a special part of the cell called the
mitochondrion. The mitochondrion
has a closed membrane system that
allows the energy released to be
captured by generating ATP which
fuels many cellular processes.
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 A number of complex proteins are needed
to conserve the energy available
H+ H+ H+
H+
Outer mitochondrial membrane

H+ H+ H+ H+
H+ H+ H+

H+

e- e- e- e- e- e- e-
H+
Complex I Complex IV e -
Inner membrane

Complex III ATP

Complex II synthase (V)
H2O
NADH e- O2
NAD ADP
+Pi
H+
ATP

NADH oxidation at Complex I Electron flow e initiated by NADH-

The enzyme ATP
is dependent on an electron oxidation drives consumption of synthase (Complex V) is
transfer chain mostly oxygen at Complex IV and the driven by the proton
contained within Complex I, pumping of protons H across the +
gradient and catalyzes the
Complex III and Complex IV membrane formation of ATP

The enzyme succinate dehydrogenase (Complex II) feeds electrons into

the chain and is also required for the overall process of food oxidation.

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 How insecticides interfere with cellular
respiration:

H+ H+ H+
H +
Outer mitochondrial membrane

H+ H+ H+ H+
H+ H+ H+

H+

e- e- e- e- e- e- e-
H+
Complex I Complex IV e -
Inner membrane

Complex III ATP

Complex II synthase (V)
H2O
NADH e- O2
NAD ADP
+Pi
H+
ATP

Inhibitors of one of the electron transport complexes I-IV or the

mitochondrial ATP synthase (Complex V) starve cells of energy by
preventing ATP formation, resulting in paralysis

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 Inhibitors of Complexes I-V
Group 21: Mitochondrial complex I electron transport inhibitors Group 25: Mitochondrial complex II electron
transport inhibitors
CN N N

O O

Rotenone
Fenazaquin Pyrimidifen Cyenopyrafen 25A beta-
21B Rotenone F 3C
Ketonitrile
Pyridaben
O
derivatives
Tolfenpyrad NC
Pyflubumide
O

O
21A METI O
25B Carboxanilides
Fenpyroximate acaricides and Tebufenpyrad Cyflumetofen
insecticides

Group 20: Mitochondrial complex III electron transport inhibitors Group 24: Mitochondrial complex
AlP IV electron transport inhibitors
O Aluminium
Hydramethylnon O
CF3 O
O
phosphide Zn3P2
CN-
O HN NH
F 3C O
HN
N O O
N
N N
H
O N O
O
Zinc
Cyanide
O

phosphide
Acequinocyl Ca3P2 PH3 salts
CF 3
Fluacrypyrim Bifenazate
Calcium
phosphide 24A Phosphine 24B Cyanides
20A Hydramethylnon 20B Acequinocyl 20C Fluacrypyrim 20D Bifenazate Phosphides

Group 12: Inhibitors of mitochondrial ATP synthase

O
O O
S
O Sn O O O S Cl
S Sn Sn
O

N N N
H H N Cl Cl Cl
N

Azocyclotin Fenbutatin
Diafenthiuron Propargite Tetradifon
Sn
oxide
12A Diafenthiuron OH
12B Organotin 12C Propargite 12D Tetradifon
Cyhexatin miticides

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 How insecticides interfere with cellular
respiration:

H+
H +
Outer mitochondrial membrane

H+ H+
H+ H+ H+

e- e- e- e- e- e- e-
Complex I Complex IV e-
Inner membrane

Complex III H+
ATP
Complex II synthase (V)
H2O H+
NADH e- O2 H+
H+

NAD H+ ADP
+Pi ATP
H+

have the ability to carry protons across the inner mitochondrial

membrane, thus removing the proton gradient
è ATP synthase is no longer able to provide cellular ATP
è O2 consumption is accelerated

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 Uncouplers

Group 13: Uncouplers of oxidative phos-

phorylation via disruption of proton gradient

Br
CN

F3C
N

O Cl OH

O2N
Chlorfenapyr Sulfluramid

NO2

DNOC

13 Pyrroles, Dinitrophenols, Sulfluramid

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 Insecticide Mode of Action
Major classes

Nerve & Muscle

Growth

Respiration

Midgut

Unknown or Non-Specific

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 The insect midgut as a target for
insecticidal agents
• Released in the insect midgut upon
ingestion of ‘packaged forms’
Bacillus
• Cause lysis of midgut epithelial cells
thuringensis Baculoviruses
via different mechanisms
derived toxins
• Result in loss of midgut integrity and
ultimately death of the pest insect

Midgut
cells

Hindgut
Foregut

Midgut

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 Mode of Action of Bacillus thuringiensis
Cry Toxins on Midgut Epithelium
1. crystal solubilization
2. protoxin proteolytic activation
3. activated toxin monomer binding to cadherin & cleavage of helix α-1
4. pre-pore tetrameric structure formation
5. tetramer binding to aminopeptidase-N or alkaline phosphatase
6. pore formation.

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 Microbial disruptors of insect midgut

Group 11: Microbial disruptors of insect midgut membranes

Includes transgenic crops expressing Bacillus thuringiensis toxins (however, specific guidance
for resistance management of transgenic crops is not based on rotation of modes of action)

Bacillus thuringiensis
and the insecticidal proteins produced

B.t. israelensis, B.t. aitzawai, B.t. kurstaki, B.t. tenebrionis Bacillus sphaericus
Bt crop proteins*
Cry1Ab, Cry1Ac, Cry1Fa, Cry1A.105, Cry2Ab, Vip3A,
mCry3A, Cry3Ab, Cry 3Bb, Cry34Ab1/Cry35Ab1

11A Bacillus thuringiensis 11B Bacillus sphaericus

Different B.t. products that target different insect orders may be used
together without compromising their resistance management.

Rotation between certain specific B.t. microbial products may

provide resistance management benefits for some pests. Consult
product-specific recommendations.

* Where there are differences among the specific receptors within the
midguts of target insects, transgenic crops containing certain combinations
of these proteins provide resistance management benefits.

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 Mode of Action of Baculoviruses on
Midgut Epithelium
1. Viral occlusion body disintegrates in alkaline midgut
2. Released occlusion derived virus passes through the peritrophic membrane to
bind to species-specific receptors on the microvilli of midgut columnar
epithelial cells
3. Viral envelope fuses with cell membrane releasing nucleocapsids in the cells
4. Entry mediated by viral proteins called per os infection factors (PIF 0-8)

From Boogaard et al., 2018 Insects 9, 84

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 Baculoviruses acting on insect
midgut

31 Baculoviruses

Anticarsia
Cydia pomonella GV gemmatalis MNPV

Thaumatotibia Helicoverpa
leucotreta GV armigera NPV

Granuloviruses & Nucleopolyhedroviruses

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 Insecticide Mode of Action
Major classes

Nerve & Muscle

Growth

Respiration

Midgut

Unknown or Non-Specific

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 Non-specific (multi-site inhibitors)

Group 8: Miscellaneous non-specific (multi-site) inhibitors

Methyl bromide
Cryolite
Dazomet
Na2B4O7·10H2O
8A Alkyl halides
Borax Tartar emetic

8D Borates 8E Tartar emetic

Sulfuryl
fluoride Metam

8C Fluorides 8F Methyl isothiocyanate

Chloropicrin
generators
8B Chloropicrin

Unlike many other insecticides and miticides, non-specific or multi-site inhibitors do not act
on a distinct target site but likely disrupt a variety of important physiological functions.

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 Insecticidal agents of unknown MoA
Group UN: Compounds of unknown or uncertain MoA

O O
O

Dicofol
OH

Bromopropylate
O OH
O O Cl
O O
O O Cl
O
Cl Cl
H H N
Cl O Cl CF3
O OH OH
O
O
O

Azadirachtin CCl3
Pyridalyl

O N S
S
O N O

Cl
O CaSX
O
N S
(Lime sulfur)
O
Chinomethionat Sulfurs
Benzoximate Mancozeb

Group UNB: Group UNE: Group UNF: Group UNM:

Bacterial agents Botanical essence including Fungal agents of Non-specific
(non-Bt) of unknown or synthetic, extracts and unknown or uncertain mechanical disruptors
uncertain MoA unrefined oils with unknown MoA
or uncertain MoA

Chenopodium ambrosioides
Beauveria bassiana strains
near ambrosioides extract
Burkholderia spp, Metarhizium anisopliae strain F52
Fatty acid monoesters with Diatomaceous earth
Wolbachia pipientis (Zap) Paecilomyces fumosoroseus
glycerol or propanediol
Apopka strain 97
Neem oil

Compounds with the unknown designation may act on a distinct target site, but the mechanism of
action has not been conclusively determined. Because of their non-specific or unknown MoA, active
ingredients in IRAC MoA groups 8 (non-specific, multi-site inhibitors), 13 (uncouplers of oxidative
phosphorylation), and all UN groups (UN, UNB, UNE, UNF, UNM) are thought not to share a common
target site and therefore may be freely rotated with each other unless there is a reason to expect
cross-resistance.
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 Important links
https://www.irac-online.org/

Modes of
Action

Pests Crops Teams Test Methods News

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