Schedule Y for Toxicological Study

Presented By-Krushangi Shah Nirma University

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

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.

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.

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.

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

Schedule YREQUIREMENTS 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

Appendices in Schedule YI. Data required for import/manufacture/ conduct CT of new

drugsIA. Drugs approved in other country

II. Format for clinical study reports(ICH E6) III. Animal toxicology IV. Animal pharmacology V. Informed consentVI. FDC VII. Undertaking by the investigator VIII. Ethics committee IX. Stability testing X. Proposed protocol XI. SAE Reporting

Appendix IIIANIMAL 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)

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

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

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

parameters to be monitored in long-term

toxicity

Biochemical & microscopic observations of urine and blood

Reproductive Toxicity

• Male Fertility Study• Female Reproduction and Developmental

Toxicity Studies• Teratogenicity Study • Perinatal Study

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

Observations should be

carried out for

Post-partum health & gross

pathology of affected organ

Progress of gestation/ parturition

periods

Length of gestation

Parturition

Growth parameter

Survival

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.

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

Allergenicity/ Hypersensitivity

Local Lymph Node Assay

Guinea Pig Maximizati

on Test

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.

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.

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

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

Laboratory parameters to be included in toxicity studies.

• Hematological parameters • Urinalysis Parameters • Blood Biochemical Parameters • Gross and Microscopic Pathology

Hematological parametersHaemoglobin

General Blood

Picture

Coagulation Parameters

(Non-rodents only):

Coagulation Parameters: Bleeding Time, Coagulation Time, Prothrombin Time, Activated Partial Thromboplastin Time.

Urinalysis ParametersColour &

Appearance24-hour

urinary output

Specific Gravity

Sugar & Acetone

Albumin & Bile pigments

Reaction (pH)

Urobilinogen

Occult Blood

Microscopic examination of

urinary sediment.

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

Gross and Microscopic PathologyBrain*: Cerebrum,

cerebellum, Midbrain

Liver*

(Rectum)Urinary bladder

Uterus*

Epididymis

OvaryTestis*

Skin

Mammary gland

Mesenteric lymph node

Skeletal muscle

(Middle Ear)

Eye

(Spinal Cord)

(Parathyroid) Thyroid

Spleen*

(Trachea)

Lung*

Stomach

Oesophagus

AortaHeart*

(Pancreas)

Adrenal*

ThymusKidney*

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.

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

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

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)

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

Application Of Good Laboratory Practices (GLP)

• The animal studies be conducted in an accredited laboratory. Toxicology studies, also be conducted in an accredited laboratory.

Bibliography

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