Female Reproductive Toxicology Studies segment 1 & 3 Acc to schedule Y and ICH guidelines

1. Naveen K L
M Pharma (2ND Sem)
Dept. of Pharmacology
Srinivas College of Pharmacy
Valachil, Mangaluru
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2.  Reproductive toxicology refers to structural and functional alterations that
affects reproductive system in sexually matured male and females.
 Reproductive toxicology include effects on male fertility, female fertility and
lactation.
 The Globally Harmonized System of Classification and Labelling of
Chemicals (GHS) separates reproductive toxicity from germ cell
mutagenicity and carcinogenicity, even though both these hazards may also
affect fertility.
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3.  Reproductive toxicity:
Effects on sexual behaviour and fertility in males and non-pregnant
females.
 Developmental toxicity:
Abnormal structure or functional development following exposure of
pregnant or lactating females.
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4.  Reproductive toxins are hazards associated with some chemical
substances, which interfere with normal reproduction; such as substance
called reprotoxic. They may adversely affects sexual function and fertility
in adult males and females, as well as causing developmental toxicity in
the offspring.
Eg. Bisulfan, Chlorambucil, Cyclophosphamide, Nitrogen mustard.
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5.  Drugs administered to both males (28 days) and female (14 days) before
mating
•Fertility and general reproductive
performance studySegment I
•TeratogenicitySegment II
•Prenatal and post-natal study: fertility
and early embryonic developmentSegment III
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6.  The study should be done in one rodent species (rat preferred).
 The drug should be administered to both males and females, beginning a
sufficient number of days (28 days in males and 14 days in females) before
mating.
 Drug treatment should be continue during mating and subsequently during
the gestation period.
 Three graded dose should be used, the highest dose (usually MTD obtained
from previous systemic toxicity studies) shouldn’t affects general health of
the parent animals, at least 15 males and 15 females should be used per
dose group.
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7.  Control and the treated groups should be of similar size.
 The route of administration should be the same as intended for
therapeutic use.
 Dams should be allowed to litter and their medication should be
continued till the weaning of pups.
 Observation on following is done:
Body weight, food intake clinical symptoms of intoxication, mating
behaviour, progress of gestation, parturition, length of gestation, gross
pathology (and affected organ histopathology) of dams should be recorded.
 The pups from both treated and control groups should be observed for
general signs of intoxication, sex-wise distribution in different treatment
groups, BW, growth parameter, survival, gross examination and autopsy.
 Histopathology is also done.
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8. Study Design: Group Size = 22 males & 22 females
Day G0 Day G6 Day G13
dosing period
G = Day of gestation
14 days 14 days
Starting
dose
Mating
Female
last dose
Caesarean &
uterine
examination
Male
necropsy
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9.  This study is specially recommended if the drug is to be given to pregnant
or nursing mothers for longer periods or where there are indications of
possible adverse effects on foetal development.
 One rodent species is needed. Dosing at levels comparable to multiples of
human dose should be done by the intended clinical route.
 At least 4 groups (including control), each consisting of 15 dams should be
used.
 The drug should be administered throughout the last trimester of
pregnancy (from day 15 of gestation) and then dose that cause low foetal
loss should be continued throughout the lactation and weaning.
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10.  One male and female from each litter of F1 generation (total 15 males and
females in each group) should be selected at weaning and treated with vehicle
or test substance (dose level described above)throughout their periods of
growth to sexual maturity, pairing, gestation, parturition and lactation.
 mating performance and fertility of F1 generation should be evaluated to
obtain F2 generation whose growth parameter should be monitored till
weaning.
 At the end of the study the animal should be scarified and the criteria of
evaluation as below
DAMS = body weight, food intake, signs of intoxication, progress of gestation,
parturition period and gross pathology.
PUPS = sex wise distribution in dose group, body weight, growth parameter,
gross examination, survival and autopsy (if needed) and where
necessary histopathology.
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11. Study design:
G6 G22 Day 22 pp
Day 0 pp
F1 survival, growth and
F1 mate
behavioural tests
F1 generation
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12.  Estrus female rats at 10 to 11 weeks of age were cohabitated overnight
with a single dose.
 The next morning, females with sperm in their vaginal smears were
regarded as pregnant, and this day was designated as day 0 of gestation.
 Once insemination was confirmed, the females were weighed and
randomly allocated to six experimental group.
 The dams were allowed to deliver naturally and nurse their pups until
postnatal day (PND) 21.
 On PND 0 (the day of birth), all pups were weighed and their sex was
determined, and litters were culled randomly to eight (four pups/sex/litter
when possible).
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13.  In the present study, three to five males and three to five females per
litter were used.
 All neonates were given daily gavage administration of 12.5,25,50 or 100
mg/kg Genistein.
 The measurement of sexual organ weight and histopathological
observation of the reproductive tracts were performed.
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14.  Eaton, D. L., and Klaassen, C.D., (1996) Section on Developmental
and Reproductive Toxicity, p. 31 in Chapter 2, Principles of
Toxicology, in Cassarett and Doull’s Toxicology, Fifth Ed., McGraw
Hill
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