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
1930s - 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 vectorborne 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
Autonomic Sympathetic
ENS

Somatic Para-Sympathetic

Anatomic Classification
CNS PNS
Skull and Spinal cord 12 pairs of cranial nerves 31 pairs of spinal nerves

afferent

CNS
efferent

PNS

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

Nerves

Afferent Efferent Neurons Mixed

Spinal nerves are mixed for major length

then divide

Afferent dorsal root

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 Sympathetic - adrenergic Ach Adr/NA Ach

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

O CH3C-O-CH2-CH2-N+(CH3)3
Acetylcholine

Choline Acetylase

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

a d g

b
a

Ion Channel

Na+
a

d
a

Na+

Muscarinic receptor
Agonist
Outside

Inside G protein

Muscarinic receptor
G- proteins Excitatory action Phosholipase C

Inhibitory action
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 Nerve Terminal

2 DA, Nepi

Tyrosine

3
mitochondria contains MAO, oxidizes amines

1
transport

exocytosis

stored with ATP

in granules

5
re-uptake

a,b Receptors

Adr (NA) synthesis

Adrenergic Receptors
Alpha a a1 most effector cells a2 presynaptic, lipocytes, platelets, some smooth muscle Beta b b1 effector cells (*heart), brain, lipocytes, presynaptic b2 smooth muscle and myocardium b3 lipocytes

Adrenergic receptors: a2 and b

Adenylyl cyclase

b receptor

a2 receptor

Adrenergic receptors:

a1

Phospholipase C

a1 receptor

Pesticides
Organochlorines ChE inhibitors Organophosphates Carbamates Phenoxyherbicides Pyrethroids Bromine-based Phenol- derivatives Dipyridyl derivatives

Organochlorine insecticides

Organochlorine insecticides
DDT
first commercially produced insecticide (1940s) banned in the US in the 1970s 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 Bioaccumulates Persistent Lipophilic

Non-selective Endocrine disrupter Reproductive toxins 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 R2

O P
X
Parathion Malathion Soman Ecothiophate

R1-2 = aliphatic

X = e- withdrawing

Organophosphates
R1 R2

O P X

Strong Covalent Bond, Inactivates Enzyme (stable >100h) 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)

a-naphthyl thiourea (ANTU)

must be metabolically activated --> resistance

Anticoagulants (coumadin, warfarin) - antagonist of vit. K in

synthesis of clotting factors; requires multiple doses; resistance