Pesticides

Principles of Toxicology
 Pesticides

• EPA definition: “substances or mixtures

of substances intended for preventing,
destroying, repelling or mitigating any
pest…”
 A bit of history…
•Sulfur
– Chinese - 1000BC
– Europe - 1800s
– CA - today !

•Arsenic-containing
•Strychnine
•Nicotine (tobacco leaves extracts - 1690)
•Pyrethrum (chrysanthemum extract)
•Bordeaux mix: copper, lime (Ca(OH)2), water
 Later…
• 1930’s - modern era chemistry
– Alkylthiocyanate
– Dithiocarbamate
– Bromide compounds
• WWII -
– DDT
– Dinitrocresol
– 2,4 D
• Since then, synthesis with goal improved specificity,
reduced toxicity…

• No such a thing as “safe pesticide”

Integral part of crop and health
protection
Poisonings are anticipated…

3mil acute cases annually (ww)

220,000 deaths

CA - 25,000 pesticide related

illnesses, annually

USA - 80,000
Efficacy of crop protection
 Pesticide
poisonings by
occupational
activity
 Medical successes
• DDT
– Typhus in Naples, Italy
– River blindness, West Africa
– Malaria - Africa, Asia, Middle East

There are still many parasitic and vector-

borne diseases…
 Regulations…
• 1906 - First Federal Food and Drugs Act
• 1938 - Federal Food, Drugs and Cosmetics Act
• 1958 amend. - Delaney clause: “no additive shall be
deemed safe if …found to induce cancer…”
• 1947 - FIFRA: Federal Insecticide, Fungicide and
Rodenticide Act: all pest control products under one law -
by USDA
• 1972 - FIFRA reorganized and passed to EPA
• FIFRA Amendments - 1975, ‘78, ‘80, 84
• 1996 - Food Quality protection Act (children)
• Developing countries adapt or lack regulations
 Nervous System

Central Peripheral

Afferent Efferent

Somatic Autonomic

Para-Sympathetic Sympathetic

ENS
 Anatomic Classification

CNS  Skull and Spinal cord

PNS  12 pairs of cranial nerves

31 pairs of spinal nerves

afferent
CNS PNS
efferent
Sympathetic ANS

Fight or flight

Parasympathetic ANS

Rest and digest

Parasympathetic: Cholinergic
cranio-sacral

Sympathetic: Adrenergic
thoraco-lumbar
Parasympathetic nerves
4 Cranial
III oculomotor
VII facial
IX glossopharyngial
X vagus

Sacral
S2
S3
S4
Sympathetic nerves
T1
.
.
.
.
T12
L1
L2
L3
 Afferent
Nerves Efferent Neurons
Mixed

Spinal nerves are mixed for major length

Afferent  dorsal root
– then divide
Efferent  ventral root
Structure of a typical neuron
Pre-ganglionic neuron
Post-ganglionic neuron
organ
Ganglion
Multi-neuron synapse

Neuroeffector junction
 Somatic

Ach
Parasympathetic -cholinergic

Ach Ach
Sympathetic - adrenergic

Ach Adr/NA
Feature Sympathetic Parasympathetic

Start point thoraco-lumbar cranio-sacral

Ganglion near spine on organ (terminal)
Ganglion synapse Ach Ach
Pregangl. neuron short long
Postgangl. neuron long short
Effector synapse Adr Ach
Effector organs throughout body limited
Typical Synapse
 Cholinergic
Transmission
 Acetylcholine Synthesis
O
CH3C-O- + HO-CH2-CH2-N+(CH3)3
Acetate Choline

Coenzyme A Choline
Acetylase
O
CH3C-O-CH2-CH2-N+(CH3)3
Acetylcholine
Acetylcholine Catabolism
O
CH3C-O-CH2-CH2-N+(CH3)3
Acetylcholine

Cholinesterase

O
CH3C-O- + HO-CH2-CH2-N+(CH3)3
Acetate Choline
 Cholinergic Receptors
Muscarinic:
M1 CNS, sympathetic (exceptions), presynaptic
M2 Smooth muscle, heart, presynaptic
M3 Exocrine glands, blood vessels

Nicotinic:
NM skeletal muscle
NN ganglia (post-), presynaptic
Acetylcholine Nicotinic Receptor
Nicotinic Receptor structure





Ion Channel
 Na+


 


Na+
Muscarinic receptor

Agonist

Outside

Inside
G
protein
 Muscarinic receptor

G- proteins

Excitatory action Inhibitory action

Phosholipase C Adenylic cyclase

K+ channels
Muscarinic receptor stimulation

Gi
-
-
Signal transduction cascade
involving Adenylic cyclase -
Protein Kinase A
-
Muscarinic receptor stimulation

Gq

Signal transduction cascade

involving Phospholipase C
Cholinergic agonists
 Adrenergic
Transmission
Sympathetic Tyrosine 3
1 mitochondria contains
Nerve
transport
MAO, oxidizes amines
Terminal

4 exocytosis
2

DA, Nepi
stored with ATP 5
in granules 
re-uptake
Receptors
Adr (NA)
synthesis
 Adrenergic Receptors
Alpha 
most effector cells
presynaptic, lipocytes, platelets, some smooth muscle
Beta
effector cells (*heart), brain, lipocytes, presynaptic
smooth muscle and myocardium
lipocytes
Adrenergic receptors: 2 and 
Adenylyl
cyclase

+ -
 receptor  receptor
Adrenergic receptors: 1
Phospholipase C

+
 receptor
 Pesticides
•Organochlorines
•ChE inhibitors
•Organophosphates
•Carbamates
•Phenoxyherbicides
•Pyrethroids
•Bromine-based
•Phenol- derivatives
•Dipyridyl derivatives
Organochlorine insecticides
 Organochlorine insecticides
• DDT
– first commercially produced insecticide (1940’s)
– banned in the US in the 1970’s but is still manufactured and exported (1 ton/day)
• Cyclodienes
– Most toxic (CNS) and persistent pesticides known
• HCH and Cl-benzene
– Mixtures of isomers
– Medicinal use (lice shampoo) (lindane)

•t1/2 = 7-30y •Non-selective

•Bioaccumulates •Endocrine disrupter
•Persistent •Reproductive toxins
•Lipophilic •Neurotoxins (Lindane)
 Observed effects
• DDT
– Enzyme induction
– Competes with estradiol for receptor
• Cyclodienes
– Reproductive toxicity (reduced fertility, loss of pups,
teratogenic)
– CNS toxicity
• HCH and Cl-benzene
– CNS toxicity
– Increased hepatocellular tumors (mice)
 Mechanisms of action
• DDT
– Peripheral sensory neurons
– prolonged negative afterpotential in neurons
– K+ transport, inactivate Na+ channel closure, inhibit Na+ /K+
and Ca2+ /Mg2+ ATPases, inhibit calmodulin-transport of Ca2+
(fig. 22-4)
• Cyclodienes
– CNS localized
– GABAA receptor/channel antagonists, inhibit Cl--uptake and
Na+ /K+ and Ca2+ /Mg2+ ATPases
• HCH and Cl-benzene
– Suggested similar to cyclodienes but unknown
 Cholinesterase inhibitors
• Organophosphates (OP) and Carbamates
– Strong Acute neurotoxicity - AChE inhibition
(cholinergic effects)

– Nervous system toxins - nerve gas (sarin)

 1st OP: TEPP
(tetraethylpyrophosphate),
followed by parathion
1st carbamic: 1930

WWII chemical warfare

1988 Iraq- against Kurds
1994 Japan
1995 Tokyo subway
Cholinesterase
O
+
CH3 C - O - CH2 - CH2 - N(CH3)3

Esteratic Anionic

O
+
CH3C-O- + HO-CH2-CH2-N(CH3)3
 Organophosphates

R1 O
P Parathion
R2
X
Malathion
Soman
Ecothiophate

R1-2 = aliphatic X = e- withdrawing

Organophosphates
Strong Covalent Bond,
R1 O Inactivates Enzyme
(stable >100h)
R2 P X Aging of complex

Esteratic Anionic
Organophosphates are
slower to release from
AchE - “also aging effect”

Carbamates are faster:

reversible

Phase I metabolic
activation
Multiple metabolic
reactions
Neurobehavioral,
muscular and cognitive
effects
Delayed Neuropathy
(OPIDN) - ginger jake

WHO has a battery of

neuropsychological tests
OPs causing OPIDN
Phase I and II biotransformation
Antidote pralidoxime, removing OP from enzyme site
 Pyrethroids
Newer (1980) but were 30% of all use by 1982

Extensive agricultural use

Indoor use
Pet flee control
Household plants

Modify Na+ channel kinetics

Abnormal repetitive discharges
Type A shorter action than type B
Avermectins
New generation pesticides
 Herbicides
• 1.9% increase/year between 1980-1985 (x2 of
insecticides) due to:
– Monoculture
– Mechanization of agricultural processes

• Categories by application:
– Pre-planting
– Pre-emergent
– Post-emergent
• Low mammal toxicity
• Suspected mutagens, carcinogens, teratogens
• Skin irritants
 Phenoxyherbicides
• Introduced in 1946
• 2,4Dichloro- and
2,4,5Trichloro phenoxy acetic
acids
• Defoliants (Vietnam war) -
Forestry
• Nerve toxicity, peripheral
neuropathy
• Controversy about NHL and
HL
• Contaminants may be
responsible for toxicity
 Dipyridyl derivatives
“startling human toxicity”
Banned in many countries but still in use in 130 others
Lung is the most susceptible target organ
Highly polar- poor GI absorption (5-10%)

LD50=22-262mg/kg

LD50=100-400mg/kg
 Chloroacetanilides
Only slight acute toxicity but
Carcinogens of category 2B
Metabolic activation to
mutagenic metabolite (DEBQ1)
1985 Canada incident (well
water contamination)
 Phosphomonomethyl
aminoacids
•Non-selective systemic
herbicides
•Free acids or salts -
ocular and mucus
membrane irritants
•Class E carcinogens
(EPA)
•Solvent may be the
toxic compound (POEA)
 Fungicides
• Lipophilic, accumulate
• 90% are carcinogenic in animals --
> 75 mil pounds produced
annually
• 10% acreage but 60% of total
dietary carcinogenic risk
• Contaminants are dioxins and
furans

• Hexachlorobenzene (banned)
• Pentachlorophenol (banned)
• Phthalimides
• Dithiocarbamates
 Fungicides
• Dithiocarbamates
– Ferbam, ziram, maneb, zineb, nabam (metal-based names)
– Some reported as teratogenic
– Degradation to ethylene thiourea (ETU): a known mutagen,
carcinogen, teratogen and antithyroid compound.
– Some neurotoxicity at high doses
– May cross into CNS if bound to divalent metals
 Fumigants
• Very volatile - inhalation exposure
• Non-selective, highly reactive and cytotoxic
– acrylonitrile
– carbon disulfide
– carbon tetrachloride
– ethylene dibromide (gastric carcinomas, sterility)
– ethylene oxide (carcinogen, developmental tox.)
– phosphine (PH3) released from aluminum phosphide (AlP) in
moist conditions (grain storage)
 Rodenticides
• Rodents: vectors of disease
– Zinc phosphide - PH3 (cell toxicity,
necrosis, GI, liver, kidneys)
– Fluoroacetic acid and derivatives
(Fluoroacetyl-CoA --> fluorocitrate:
Krebs cycle collapse)
 -naphthyl thiourea (ANTU)
must be metabolically activated -->
resistance
– Anticoagulants (coumadin,
warfarin) - antagonist of vit. K in
synthesis of clotting factors; requires
multiple doses; resistance