This slide is useful to the life science student. It include the detailed information of toxicology and guidelines for conducting toxicity study.

1. TOXICOLOGY & REGULATORY
GUIDELINES FOR CONDCTING
TOXICITY STUDY
Presented By:- Janhavi Yashwant Burade
M.Pharm 2nd sem
(Pharmacology)
Vidyabharti College of Pharmacy Amravati

2. Content
■ Introduction
■ Besic definition
■ Types of toxicology
1. General
2. Mechanistic
3. Regulatory
4. Descriptive
■ Regulatory guidelines for conducting toxicity studies
1. OECD
2. ICH
3. EPA
4. Schedule Y

3. Introduction
■ The study of how natural or man-made poisons cause adverse
effects in living organisms is called as toxicology
■ Phillip von Hohenheim (Paracelcius) is known as the“ father of
toxicology“
■ “All things are poison and nothing is without poison, only the dose
permits something not to be poisonous.”

4. Basic Definitions
■ Toxin
Toxic substances that are produced naturally (natureorigin)
■ Toxicants
Any chemical that can injure or kill humans, animals,or plants; (poison)
■ Toxicity
Describes the degree to which a substance is poisonous or can cause
injury.
■ Factors: dose, duration and route of exposure, shape and structure
of the chemical itself, and individual human factors.

5. Toxicology
■ The study of how natural or man-made poisons cause adverse effects in
living organisms.
■ It involves observing and reporting symptoms, mechanisms, detection and
treatments of toxic substances.
■ It includes environmental agents and chemical compounds, as well as
pharmaceutical compounds that are synthesized for medical use. These
substances may produce toxic effects leading to , discomfort, disease and
even death in living organisms.

6. Purpose of Toxicology
■ It provides protection to humans and environment from toxic effects
of toxicants.
■ This study will ultimately lead toward the development of newer,
innovative and more selective drug therapies to treat different
diseases such as cancer having reduced toxic potential to human
body.

7. Types of Toxicology
1. General Toxicology
2. Mechanistic Toxicology
3. Regulatory Toxicology
4. Descriptive Toxicology

8. General Toxicology
1. Analytical toxicology
2. Applied toxicology
3. Clinical toxicology
4. Veterinary toxicology
5. Forensic toxicology
6. Environment toxicology
7. Industrial toxicology

9. 1. Analytical toxicology:
It is the branch of toxicology which deals with the study of detection and assay of
poisonous chemicals including their metabolites that could affect the biological
system.
2. Applied toxicology:
It is the application of new and modern methods or technologies for early detection of
toxicants in the field setting or practice area.
3.Clinical toxicology:
It is mainly involved in the study of diagnosis and treatment of poisoning that can
occur in humans.
4.Veterinary toxicology:
Veterinary toxicology focus in the study of Diagnosis and treatment of animal
poisoning including the transmission of toxin from animals to humans via milk, meat,
fish, food stuffe etc.

10. 5. Forensic toxicology:
Forensic toxicology is a multidisciplinary field involving the detection
and interpretation of the presence of drugs and other potentially toxic
compounds in bodily tissues and fluids.
6. Environmental toxicology:
It is the branch of toxicology in which study of presence of different
toxicants including their metabolites and degradation products in the
environment and their effects on humans and animals.
7. Industrial toxicology:
It is the study of selective and specific area of environmental toxicology.

11. Mechanistic Toxicology
■ It is a branch of toxicology that focuses on how the cellular,
biochemical, and molecular mechanisms of chemicals exert toxic
effects on living organisms and how the biological system protects
themselves against these adverse effects.
■ It aims at identifying the molecular events that lead from initial
exposure to the chemical to the ultimate manifestation of toxic injury
in an organism.

12. Regulatory Toxicology
■ It consists of collecting, processing and evaluating incidents,
distribution, and control of diseases towards the protection of health
against harmful toxicants.
■ It supports the development of standard protocols and new testing
methods.
■ Its aim is to control production and use of dangerous materials to
prevent adverse effects on human health and the environment.

13. National & International Collaboration In
Rregulatory Toxicology
■ These standards are implemented worldwide for sustainable
development with the goal of improving the quality of life for all
people.
■ A number of international bodies and authorities promote the sound
management of chemicals at national and international level. They
are:
• ICH
• WHO
• FDA
• OECD

14. Importance of Guidelines in Regulatory
Toxicity Studies
■ Prevent duplication of clinical trials in humans .
■ Ensure SAFETY, EFFICACY and QUALITY of medicines .
■ Minimize the use of animal testing.
■ Provides the definite parameters of evaluation.
■ Provides a roadmap to prepare a study protocol.
■ Increase international harmonization of technical requirements toensure
that safe, effective, and high quality medicines are developed.
■ Supports economic growth, raise living standards, maintain financial
stability and contribute to growth in world trade

15. Descriptive Toxicology
■ It is concerned directly with toxicity testing, which provides
information for safety evaluation and regulatory requirements.
■ Focuses on toxicity testing of chemicals, usually on animals and then
correlated to human conditions.
■ It provides dose-response information upon exposure to a harmful
toxic agent.
■ The results from the toxicity testing are typically applied to approval
of product use and regulating allowable concentrations in the
environment.

16. Regulatory Guidelines For
Conducting Toxicity Studies

17. OECD GUIDELINES

18. OECD Guidelines
■ OECD guidelines for the testing of chemicals are a set of
internationally accepted specifications for testing of chemicals
decided on by the Organization for Economic Co-operation &
development (OECD).
■ They are split into five sections:
Section 1: physical chemical properties
Section 2: effects on biotic system
Section 3: degradation and accumulation
Section 4: health effects
Section 5: other test guidelines

19. Continue…
■ Here our main focus is to study the regulatory guidelines for conducting toxicity
studies, therefore we would focus mainly on
■ Guidelines are under constant review, with guidelines being periodically updated,
new guidelines being adopted.
■ Following is the list of guidelines included in health effects.

26. ICH GUIDELINES

27. ICH Guidelines
■ ICH is the “International Conference on Harmonization of Technical
Requirements for Registration of Pharmaceuticals for Human Use”
■ ICH is a joint initiative involving both regulators and research-based
industry representatives of the EU, Japan and the USA in scientific and
technical discussions of the testing procedures required to assess and
ensure the safety, quality and efficacy of medicines
■ The ICH Secretariat is based in Geneva. The biennial meetings and
conferences of the ICH Steering Committee rotate between the EU, Japan,
and the USA.

28. Objectives of ICH
■ To increase international harmonization of technical requirements to
ensure that safe, effective and high quality medicines are developed.
■ To harmonize technical requirements for registration or marketing
approval.
■ To develop and register pharmaceuticals in the most efficient and
cost effective manner.
■ To promote public health.
■ To prevent unnecessary duplication of clinical trials on humans.
■ To minimize the use of animal testing without compromising safety
and effectiveness of drug.

29. Guidelines

30. ■ “Quality” Topics i.e., those relating to chemical and pharmaceutical
Quality Assurance (Stability Testing, Impurity Testing, etc.)
■ “Efficacy” Topics, i.e., those relating to clinical studies in human
subject (Dose Response Studies, Good Clinical Practices, etc.)
■ “Safety” Topics, i.e., those relating to in vitro and in vivo pre-clinical
studies (Carcinogenicity Testing, Genotoxicity Testing, etc.)
■ “Multidisciplinary” Topics, i.e., cross-cutting Topics which do not fit
uniquely into one of the above categories.

32. Carcinogenicity studies(S1A-S1C)
S1A: Guideline on the Need for Carcinogenicity Studies of
Pharmaceuticals
■ Carcinogenicity studies should be performed for any pharmaceutical whose expected
clinical use is continuous for at least 6 months.
■ This document provides a consistent definition of the circumstances under which it is
necessary to undertake carcinogenicity studies on new drugs. These recommendations
take into account the known risk factors as well as the intended indications and
duration of exposure.
■ The objectives of carcinogenicity studies are to identify a tumorigenic potential in
animals and to assess the relevant risk in humans.

33. S1B: Testing for Carcinogenicity of Pharmaceuticals
■ This document provides guidance on the need to carry out
carcinogenicity studies in both mice and rats, and guidance is also
given on alternative testing procedures which may be applied without
jeopardizing safety.
S1C(R2): Dose Selection for Carcinogenicity Studies of
Pharmaceuticals
■ This document addresses the criteria for the selection of the high
dose to be used in carcinogenicity studies on new therapeutic agents
to harmonize current practices and improve the design of studies.

34. S2– Genotoxicity
S2(R1): Guidance on Genotoxicity Testing and Data
Interpretation for Pharmaceuticals Intended for Human Use
■ This guidance is a combination of ICH S2A and S2B guidelines:
S2A: Guidance on Specific Aspects of Regulatory Genotoxicity
Tests for Pharmaceuticals;
■ This document provided specific guidance and recommendations for in
vitro and in vivo tests and on the evaluation of test results. It includes terms
related to genotoxicity tests to improve consistency in applications.

35. S2B: A Standard Battery for Genotoxicity Testing for
Pharmaceuticals :
■ This document addresses two fundamental areas of genotoxicity testing:
1.the identification of a standard set of assays to be conducted for registration,
and the extent of confirmatory experimentation in any particular genotoxicity
assay in the standard battery.
2.Registration of pharmaceuticals requires a comprehensive assessment of
their genotoxic potential. It is clear that no single test is capable of detecting all
relevant genotoxic agents. Therefore, the usual approach should be to carry out
a battery of in vitro and in vivo tests for genotoxicity
■ The purpose of this guideline is to optimize the standard genetic toxicology
battery for prediction of potential human risks, and for interpretation of
results, with the ultimate goal of improving risk characterization for
carcinogenic effects that have their basis in changes in the genetic
material.out

36. S3A-S3B Toxicokinetics andPharmacokinetics:
S3A: Note for Guidance on Toxicokinetics: The Assessment of
Systemic Exposure in Toxicity Studies
■ ICH guidelines do not require toxicokinetic studies to be conducted, except in
pregnant, lactating animals, before initiating reproductive studies.
■ In this context, toxicokinetics is defined as the generation of pharmacokinetic
data, either as an integral component in the conduct of non-clinical toxicity
studies or in specially designed supportive studies, in order to assess systemic
exposure.
■ This document gives guidance on developing test strategies in toxicokinetics and
the need to integrate these pharmacokinetics into toxicity testing, in order to aid
in the interpretation of the toxicology findings and and their relevance to clinical
safety issues
■ The primary objective of toxicokinetics is: to describe the systemic exposure
achieved in animals and its relationship to dose level and the time course of the
toxicity study.

37. S3B: Pharmacokinetics: Guidance for Repeated Dose Tissue
Distribution Studies
■ Tissue distribution studies are essential in providing information on
distribution and accumulation of the compound and/or metabolites,
especially in relation to potential sites of action; this information may
be useful for designing toxicology and pharmacology studies and for
interpreting the results of these experiments.
■ This document gives guidance, when the repeated dose tissue
distribution studies should be considered (i.e., when appropriate data
cannot be derived from other sources). It also gives
recommendations on the conduct of such studies

38. S4: Duration of Chronic Toxicity Testing in Animals
(Rodent and Non-Rodent Toxicity Testing)
■ The objective of this guidance is to set out the considerations that
apply to chronic toxicity testing in rodents and non rodents as part of
the safety evaluation of a medicinal product
• Rodents (a study of 6 months duration)
• Non-rodents (a study of nine months duration).

39. S5: Detection of Toxicity to Reproduction for Medicinal
Products & Toxicity to Male Fertility
■ This document provides guidance on tests for reproductive toxicity.
■ It defines the periods of treatment to be used in animals to better
reflect human exposure to medical products and allow more specific
identification of stages at risk.
■ It should encourage the full assessment on the safety of chemicals
on the development of the offspring.

40. S6: Preclinical Safety Evaluation of Biotechnology-
Derived Pharmaceuticals
■ This document covers the pre-clinical safety testing requirements for
biotechnological products. It addresses the use of animal models of
disease, determination of when genotoxicity assays and
carcinogenicity studies should be performed, and the impact of
antibody formation on duration of toxicology studies.
■ The primary goals of preclinical safety evaluation are:
1) to identify an initial safe dose and subsequent dose in humans;
2) to identify potential target organs for toxicity and for the study of
whether such toxicity is reversible;
3) to identify safety parameters for clinical monitoring.

41. S7A: Safety Pharmacology Studies for Human
Pharmaceuticals
■ This guideline was developed to protect clinical trial participants and
patients receiving marketed products from potential adverse effects
of pharmaceuticals
■ This document addresses the definition, objectives and scope of
safety pharmacology studies.
■ It also addresses which studies are needed before initiation of Phase
1 clinical studies as well as information needed for marketing.

42. S7B : The Nonclinical Evaluation of the Potential for
Delayed Ventricular Repolarization (QT Interval
Prolongation) By Human Pharmaceuticals
■ This guideline describes a non-clinical testing strategy for assessing
the potential of a test substance to delay ventricular repolarization.
■ This guideline includes information concerning non-clinical assays
and integrated risk assessments.

43. S8 : Immunotoxicity Studies for Human
Pharmaceuticals
■ This guideline addresses the recommendations on nonclinical testing
for immunosuppressant.
■ The guideline might also apply to drugs in which clinical signs of
immunosuppressant are observed during clinical trials and following
approval to market.
■ The term immunotoxicity in this guideline will primarily refer to
immunosuppressant, i.e. a state of increased susceptibility to
infections or the development of tumors.

44. S9: Nonclinical Evaluation for Anticancer
Pharmaceuticals
■ This guideline provides information for pharmaceuticals that are only
intended to treat cancer in patients with late stage or advanced disease
regardless of the route of administration, including both small molecule
and biotechnology-derived pharmaceuticals.
■ It describes the type and timing of nonclinical studies in relation to the
development of anticancer pharmaceuticals and references other
guidance as appropriate.
■ This guideline aims to facilitate and accelerate the development of
anticancer pharmaceuticals and to protect patients from unnecessary
adverse effects, while avoiding unnecessary use of animals and other
resources

45. S10 guidelines-Photo safety evaluation of
pharmaceuticals
■ These guideline applies to new APIs . New excipients clinical formulations
for dermal application and photodynamic therapy products.
■ It is an integrated process that can involve an evaluation of photochemical
characteristics, data from non clinical studies and human safety
information.
■ The photo safety assessment aims to determine weather risk
minimization measures are warranted to prevent adverse events in
humans.
■ In-vitro assay for photo-toxicity is the 3T3 neutral red uptake assay.
■ in vivo for species selection irradiation sensitivity, heat tolerance,
performance of reference substance should be considered.

46. S11-FOR NON CLINICAL SAFETY TESTING IN SUPPORT
OF DEVELOPMENT OF PRDIATRIC MEDICINES
■ This guideline is needed to recommended standards for the conditions
under which non clinical juvenile animal testing is considered informative
and support pediatric clinical trails .
■ The expert working group (EWG) Will consist of two nonclinical experts
nominated by EU,EFPIA,FDA, MHLW, JPMA, HEALTH Canada and Swiss
medic. One member can also be nominated by WHO Observer, as well as
RHIs, DRAs/doH
■ The ICH M3 (R2) Guideline state, the conduct of any juvenile animal toxicity
studies should be considered when pervious animal data and human safety
data ,including effects from other drugs of pharmacological class, are
judged to be sufficient to support pediatric studies

47. EPA GUIDELINES

48. EPA Guidelines
■ The Environmental Protection Agency (EPA) is an agency of the
federal government of the United States charged with protecting
human health & with safeguarding the natural environment , air ,
water , & land
■ The EPA began operation on December 2 , 1970 , when it was
established by President Richard Nixon
■ EPA recommends the following means to reduce the number of
animals used to evaluate acute effects of chemical exposure while
pre serving its ability to make reasonable judgements about safety:

49. Continue…
1. Use of data from structurally related substances or mixtures. In
order to minimize the need for animal testing for acute effects, the
Agency encourages the review of existing acute toxicity information
on chemical substances that are structurally related to the agent
under investigation. In certain cases, it may be possible to obtain
enough information to make preliminary hazard evaluations that
may reduce the need for further animal testing for acute effects.
Similarly, mixtures or formulated products that are substantially
similar to well-characterized mixtures or products may not need
additional testing if there are sufficient bridging data available for
meaningfu extrapolation. In those cases, classification would be
extrapolated from the mixture already tested.onal

50. 2. EPA recommends the Up-and-Down Procedure (UDP), as detailed in this
guideline and adopted by OECD as test Guideline 425 to access acute oral
toxicity. This method provides a point estimate of lethality and confidence
interval. A dedicated program (AOT425StatPgm) has been developed by EPA to
assist laboratories in the conduct of this protocol. Acute oral toxicity testing
may also be performed using the Fixed Dose Method of OECD Guideline 420 or
the Acute Toxic Class Method of OECD Guideline 423 . These methods assess
lethality within a dose range.

51. Schedule Y

52. Schedule Y
REQUIREMENTS AND GUIDELINES FOR PERMISSION TO
IMPORT AND / OR MANUFACTURE OF NEW DRUGS FOR SALE
OR TO UNDERTAKE CLINICAL TRIALS

53. 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

54. Appendix III:-Animal Toxicology
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

56. Reproductive Toxicity
1. Male Fertility Study
2. Female Reproduction and Developmental Toxicity Studies
3. Teratogenicity Study
4. Perinatal Study

57. Teratogenicity Study Perinatal 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:
1. signs of intoxication,
2. effect on body weight,
3. effect on food intake,
4. examination of uterus, ovaries and uterine
5. contents,
6. number of corpora lutea,
7. implantation sites, resorptions (if any);
8. the fetuses, the total number, gender, body length,
weight and gross/ visceral/ skeletal abnormalities, if
any.
• 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
• Animals should be sacrificed at the end of the
study and the observation parameters should
include
1. body weight,
2. food intake,
3. general signs of intoxication,
4. progress of gestation/ parturition periods and
gross pathology (if any);
5. 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.

58. 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

59. 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.

60. 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.

61. Clinical Toxicology Study
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

62. 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

63. 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)

64. 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

65. References
■ http://www.srmuniv.ac.in/sites/default/files/downloads/Acute_Sub
acute_And_Chronic_ToxicityAnimals.pdf
■ https://www.slideshare.net/ShitalMagar2/oecd-guidline-on-acute-
and-chronic-toxicity
■ https://outage.niehs.nih.gov/iccvam/suppdocs/feddocs/oecd/oecd_
gl420.pdf
■ http://www.cdsco.nic.in/
■ http://www.csir.res.in/
■ http://www.cdriindia.org/