1. Schedule Y for Toxicological Study
Presented By-
Krushangi Shah
Nirma University

2. OVERVIEW OF PRESENTATION
• Indian Medical Organizations
• Drug Regulatory Laws
• The Drugs And Cosmetics Act, 1940
• Schedule Y –What it Covers And Associated
Rules
• Appendices Of Schedule Y
• Appendix III: Non-Clinical Animal Toxicity
Studies
• Bibliography

3. Indian Medical Organizations
To ensure uniform
quality of clinical
research by Good
Clinical Practices
throughout the country
and to generate data for
registration of new
drugs before use in the
Indian population.

4. CSIR
• Council of Scientific and Industrial Research
• is an autonomous body and India's largest Research and
Development (R&D) organization.
CDRI
• Central Drug Research Institute
• CDRI is the laboratory functioning under the aegis of the
council of scientific and Industrial Research of India.
SAFETY &
CLINICAL
DEVELOPMENT--
TOXICOLOGY
• Toxicology group is involved in profiling of candidate drugs
according to schedule Y guidelines. Systemic
toxicology, reproductive toxicology, genetic
toxicity, immunotoxicity, local toxicity and carcinogenicity
are being done for NCEs.

5. DRUG REGULATORY LAWS
• 1940 -Drugs and Cosmetic Act
• 1985 -Narcotic Drugs and Psychotropic
Substances Act
• 2000 -Ethical Guidelines for Biomedical Research
on Human Subjects, ICMR
• 2001 -Indian GCP Guidelines
• 2002 -Amendment to Drugs & Cosmetics Act
• 2005 -Revised Schedule Y
• Future :
– Guidelines for pre-clinical data for r-DNA
vaccines, diagnostics & biological.
– Draft Guidelines for Stem Cell Research/
Regulation, ICMR.

6. THE DRUGS AND COSMETICS
ACT, 1940
• An Act to regulate
import, manufacture, distribution and sale of
drugs and cosmetics.
• Passed by the Indian Parliament.
• It extends to the whole of India
• Both the Act and the Rules came into force
from April 1947
• Schedule(organized plan for matters to be
attended to) are from A to Y

7. Schedule Y
REQUIREMENTS AND GUIDELINES FOR
PERMISSION TO IMPORT AND / OR
MANUFACTURE OF NEW DRUGS FOR SALE OR
TO UNDERTAKE CLINICAL TRIALS
RIGHTS SAFETY WELL
BEING OF HUMAN
SUBJECTS
SCOPE
NEW DRUG
DEVELOPMENT
PROCESS IN INDIA
UTILITY

8. Appendices in Schedule Y
I. Data required for import/manufacture/ conduct CT of new
drugs
IA. Drugs approved in other country
II. Format for clinical study reports(ICH E6)
III. Animal toxicology
IV. Animal pharmacology
V. Informed consent
VI. FDC
VII. Undertaking by the investigator
VIII. Ethics committee
IX. Stability testing
X. Proposed protocol
XI. SAE Reporting

9. Appendix III
ANIMAL TOXICOLOGY
(NON-CLINICAL TOXICITY STUDIES)
General Principles:-
• Laboratory parameters to be included in
toxicity studies. GLP, Qualified staff.
• Calibrated standardized equipments
• SOPs
• Test substances and test systems-
characterized and standardized
• Documents - 5 years (Histo slides, reports)
• Toxicokinetic studies (– dose – tox)

10. Different Toxicity Studies mentioned
in Appendix-III
• Systemic Toxicity Studies
– Single-dose Toxicity Studies
– Repeated-dose Toxicity Studies
• Special Toxicity Studies
– Reproductive Toxicity
• Male Fertility Study
• Female Reproduction and Developmental Toxicity Studies
• Teratogenicity Study
• Perinatal Study
– Local toxicity
– Allergenicity/ Hypersensitivity
– Genotoxicity
– Carcinogenicity

12. Systemic Toxicity Studies
Single-dose Toxicity Studies
•2 rodent species.
•using the same route as
intended for humans.
•Target organ of toxicity.
•Mortality should be observed
for up to 7 days after parentral
administration and up to 14
days after oral administration.
•Symptoms, signs and mode of
death should be reported, with
appropriate macroscopic and
microscopic findings.
•LD 10 and LD 50 with 95% CI.
Repeated-dose Toxicity Studies
•2 mammalian species
(1 nonrodent)
•Duration of the study will depend
on the duration, therapeutic
indication and scale of the
proposed clinical trial.
•Drug should be administered 7
days a week by the route intended
for clinical use.
•Control group of animals given
the vehicle alone should be
included

13. Number of animals required for
repeated-dose toxicity studies
14-28 days 84-182 days
Group Rodent (Rat) Non-rodent
(Dog or
Monkey)
Rodent (Rat) Non-rodent (Dog
or Monkey)
M F M F M F M F
Control 6-10 6-10 2-3 2-3 15-30 15-30 4-6 4-6
Low dose 6-10 6-10 2-3 2-3 15-30 15-30 4-6 4-6
Intermediate
dose
6-10 6-10 2-3 2-3 15-30 15-30 4-6 4-6
High dose 6-10 6-10 2-3 2-3 15-30 15-30 4-6 4-6

14. parameters
to be
monitored in
long-term
toxicity
Physiological
Organ
Pathology:
Gross and
Microscopic
ECG and
funduscopy
(non-rodent)
Behavioral
Biochemical &
microscopic
observations of
urine and blood

15. Reproductive Toxicity
• Male Fertility Study
• Female Reproduction and Developmental
Toxicity Studies
• Teratogenicity Study
• Perinatal Study

16. Male Fertility Study
• Species: (One rodent)
• Dose selection: from the 14 or 28-day
toxicity study in rat.
• Groups: Three dose groups. 6 adult
male animals in each group.
• Dosing interval: Test substance by the
intended route of clinical use for min 28
days & max 70 days before they are
paired with female animals of proven
fertility. Drug treatment of the male
animals should continue during pairing.
• Parameters: (i) Females getting thus
pregnant should be examined for their
fertility index after day 13 of gestation.
(ii) Weights of each testis and
epididymis. (iii) Sperms from one
epididymis- their motility and
morphology.
Female Reproduction and
Developmental Toxicity Studies
• Carried out for all drugs to be used in
women of child bearing age.
• Species: (one rodent)
• Dose selection: administered to both
males and females, beginning a sufficient
number of days before mating
• Groups: 15 males and 15 females per
dose. Control and the treated groups
should be of similar size.
• Dosing interval: Three graded doses. The
route of administration should be the
same as intended for therapeutic use.
Drug treatment should continue during
mating and, subsequently, during the
gestation period.
• Dams should be allowed to litter and their
medication should be continued till the
weaning of pups.
Reproductive Toxicity

17. Observations
should be
carried out
for
Clinical signs
of
intoxication
Mating
behavior
Body
weight
Food
intake
Post-partum
health & gross
pathology of
affected organ
Progress of
gestation/
parturition
periods
Length of
gestation
Parturition
Growth
parameter
Survival

18. Teratogenicity Study
• Species: One rodent & a non-rodent (rabbit)
• Dose selection: Drug administered
throughout the period of organogenesis, using
three dose levels. The route of administration
should be the same as intended for human
therapeutic use.
• Groups: The control and the treated groups
should consist of at least 20 pregnant rats (or
mice) and 12 rabbits, on each dose level.
• All fetuses should to be subjected to gross
examination, Skeletal abnormalities and
visceral abnormalities. Observation
parameters should include:
• (Dams) signs of intoxication,
• effect on body weight,
• effect on food intake,
• examination of uterus, ovaries and uterine
contents,
• number of corpora lutea,
• implantation sites, resorptions (if any);
• the fetuses, the total number, gender, body
length, weight and gross/ visceral/ skeletal
abnormalities, if any.
Perinatal Study
• Carried out for the drugs to be given to
pregnant or nursing mothers for long periods
or if adverse effects on fetal development are
there.
• Species: One rodent species (preferably rat)
• Dose selection: should be administered
throughout the last trimester of pregnancy
and continued throughout lactation and
weaning.
• Groups: 4 groups, each having 15 dams.
• Animals should be sacrificed at the end of the
study and the observation parameters should
include
• (Dams) body weight,
• food intake,
• general signs of intoxication,
• progress of gestation/ parturition periods
and gross pathology (if any);
• pups, the clinical signs, sex-wise distribution
in dose groups, body weight, growth
parameters, gross examination, survival and
autopsy (if needed) and where
necessary, histopathology.

19. Local toxicity
• Required when route of
administration is some
special route (other than
oral) in humans.
• Applied to an appropriate
site (e.g., skin or vaginal
mucous membrane) to
determine local effects in a
suitable species.
• Typical study designs
includes three dose levels
and untreated and/ or
vehicle control, preferably
with use of 2 species.
•Dermal toxicity study
•Photo-allergy or dermal
photo-toxicity
•Vaginal Toxicity Test
•Rectal Tolerance Test
•Parentral Drugs
•Ocular toxicity studies
•Inhalation

20. Allergenicity/ Hypersensitivity
Local
Lymph
Node
Assay
Guinea Pig
Maximization
Test

21. Genotoxicity
• Genotoxic compounds, shall be presumed to be
trans-species carcinogens, implying a hazard to
humans.
• Such compounds need not be Subjected to long-
term carcinogenicity studies.
• However, if such a drug is intended to be
administered for chronic illnesses or otherwise
over a long period of time - a chronic toxicity
study (up to one year) may be necessary to
detect early tumorigenic effects.

22. Carcinogenicity
• More than 6 months
• Drugs used frequently in an intermittent
manner in the treatment of chronic or
recurrent conditions.
• Structure-activity relationship suggests
carcinogenic risk.

23. Route of
administration
Duration of proposed
human
administration
Human Phase(s) for
which study is
proposed to be
conducted
Long term toxicity
requirements
Systemic Toxicity Studies
Oral or Parentral or
Transdermal
Single dose or several
doses in one day,
Upto 1wk
I,II,III 2sp,2wk
> 1 wk but Upto 2wk I,II,III 2sp;4wk
> 2 wk but Upto 4wk I,II,III 2sp;12wk
Over 1mo I,II,III 2sp;24wk
Inhalation (general
anesthetics, aerosols)
Upto 2 wk I,II,III 2sp;1mo; (Exposure
time 3h/d, 5d/wk)
Upto 4wk I,II,III 2sp;12wk, (Exposure
time 6h/d, 5d/wk)
> 14wk I,II,III 2sp;24wk, (Exposure
time 6h/d, 5d/wk)
Animal Toxicity requirements for clinical
trials and marketing of a new drug

24. Local Toxicity Studies
Dermal Upto 2 wk I,II 1sp;4wk
III 2sp;4wk
> 2 wk I,II,III 2sp;12wk
Ocular or Otic or
Nasal
Upto 2 wk I,II 1sp;4wk
III 2sp;4wk
> 2 wk I,II,III 2sp;12wk
Vaginal or Rectal Upto 2 wk I,II 1sp;4wk
III 2sp;4wk
> 2 wk I,II,III 2sp;12wk

25. Laboratory parameters to be
included in toxicity studies.
• Hematological parameters
• Urinalysis Parameters
• Blood Biochemical Parameters
• Gross and Microscopic Pathology

26. Hematological parameters
Haemoglobin
Total
RBC
Count
Haematocrit
Differential
WBC Count
Total WBC
Count
Reticulocyte
Platelet
Count
Terminal
Bone
Marrow
Examination
ESR (Non-
rodents
only)
General
Blood
Picture
Coagulation
Parameters
(Non-
rodents
only):
Coagulation Parameters: Bleeding Time, Coagulation Time, Prothrombin Time,
Activated Partial Thromboplastin Time.

27. Urinalysis Parameters
Colour &
Appearance
24-hour urinary
output
Specific Gravity Sugar & Acetone
Albumin & Bile
pigments
Reaction (pH) Urobilinogen
Occult Blood
Microscopic
examination of
urinary sediment.

28. Blood Biochemical Parameters
Glucose
Cholesterol &
Triglycerides
HDL & LDL
(Non-rodents only)
Alkaline
Phosphatase (ALP) &
GGT (Non-rodents
only)
SGPT (ALT) & SGOT
(AST)
Bilirubin
Blood Urea Nitrogen
& Creatinine
Albumin & Globulin
Sodium, Potassium,
Phosphorus,
Calcium

29. Gross and Microscopic Pathology
Brain*: Cerebrum,
cerebellum, Midbrain
Liver*
(Rectum)
Urinary bladder
Uterus*
Epididymis
Ovary
Testis*
Skin
Mammary gland
Mesenteric
lymph node
Skeletal muscle
(Middle Ear)
Eye
(Spinal Cord)
(Parathyroid)
Thyroid
Spleen*
(Trachea)
Lung*
Stomach
Oesophagus
AortaHeart*
(Pancreas)
Adrenal*
Thymus
Kidney*
Colon
Terminal ileum
JejunumDuodenum
* Organs marked with an asterisk should be weighed.
() Organs listed in parenthesis should be examined if indicated by the nature of the
drug or observed effects.

30. For Phase I studies: -
• Systemic toxicity Studies
• Single dose toxicity studies
• Dose Ranging studies
• Repeated dose systemic studies of appropriate
duration to support the duration of proposed
human exposure.
– Male Fertility study
– In-vitro Genotoxicity tests
• Relevant local toxicity studies
• Allergenicity/hypersensitivity tests
• Photo-allergy or dermal photo-toxicity test

31. Phase II Clinical Trials
• non-clinical safety data (listed previously) already
submitted while obtaining the permissions for
phase I trial, with appropriate references.
• directly starting Phase II trial – complete details of
the non-clinical safety data needed for obtaining
permission for Phase-I trial
• Repeat dose systemic toxicity studies of
appropriate duration to support the duration of
proposed human exposure.
• In-vivo genotoxicity tests
- Segment II reproductive/developmental toxicity
study

32. Phase III Clinical Trials
• Summary of non-clinical safety and Phase I and Phase II
trials data already submitted while obtaining the
permissions
• Directly starting Phase III trial – complete details of the
non-clinical safety data needed for obtaining permission
for Phase-I trial and Phase II
• Repeat dose systemic toxicity studies of appropriate
duration to support the duration of proposed human
exposure
• Reproductive/developmental toxicity studies. (female
reproduction or teratogenic toxicity)
• Carcinogenicity studies ( when there is a cause for
concern or when the drug is to be used for more than 6
months)

33. Phase IV Clinical Trials
• Summary of all the non-clinical safety data
already submitted while obtaining the
permissions or Phase I and Phase II trials with
appropriate references

34. Application Of Good Laboratory
Practices (GLP)
• The animal studies be conducted in an
accredited laboratory. Toxicology studies, also
be conducted in an accredited laboratory.

35. Bibliography
• http://www.cdsco.nic.in/
• http://www.csir.res.in/
• http://www.cdriindia.org/