29/11/2007KLE University, Belgaum CURRENT PHARMACOLOGY & TOXICOLOGY GUIDLINES FOR PHARMACEUTICAL INDUSTRY Dr. Basavaraj K. Nanjwade M.Pharm., Ph.D Associate Professor Department of Pharmaceutics KLE University BELGAUM - 10 By

29/11/2007 KLE University, Belgaum Objective of the Guidance Guidance was developed to help protect clinical trial participants and patients receiving marketed products from potential adverse effects of pharmaceuticals, Avoiding unnecessary use of animals and other resources. Guidance provides a definition, general principles and recommendations for safety pharmacology studies.

29/11/2007 KLE University, Belgaum Background The term safety pharmacology studies first appeared in ICH M3 Timing of Non-clinical Safety Studies for the Conduct of Human Clinical Trials for Pharmaceuticals and S6 Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals as studies that should be conducted to support use of therapeutics in humans

29/11/2007 KLE University, Belgaum Scope of Guidance Guidance generally applies to new chemical entities and biotechnology-derived products for human use. Guidance can be applied to marketed pharmaceuticals when appropriate

29/11/2007 KLE University, Belgaum Safety Pharmacology Safety pharmacology studies are defined as those studies that investigate the potential undesirable Pharmacodynamic effects of a substance on physiological functions in relation to exposure in the therapeutic range and above.

29/11/2007 KLE University, Belgaum Pharmacology Guidance Objectives of Studies General Considerations in Selection and Design of Safety Pharmacology Studies Test Systems Dose Levels or Concentration of Test Substance Duration of Studies Studies on Metabolites, Isomers and Finished Products

29/11/2007 KLE University, Belgaum Pharmacology Guidance Safety Pharmacology Core Battery Follow-up and Supplemental Safety Pharmacology Studies Conditions Under Which Studies Are Not Necessary Timing of Safety Pharmacology Studies in Relation to Clinical Development Application of Good Laboratory Practice

29/11/2007 KLE University, Belgaum Objective of Studies 1.To identify undesirable Pharmacodynamic properties of a substance that may relevance to its human safety 2.To evaluate adverse Pharmacodynamic and/or pathophysiological effects of a substance observed in toxicology and /or clinical studies 3.To investigate the mechanism of the adverse Pharmacodynamic effects observed and/or suspected.

29/11/2007 KLE University, Belgaum General Considerations in Selection and Design of Safety Pharmacology Studies 1.Effects related to the therapeutic class of the test substance. (e.g. Proarrhythmia is a common feature of antiarrhythmic agents) 2.Adverse effects associated with members of the chemical or therapeutic class. (e.g. Antipsychotics and QT prolongation)

29/11/2007 KLE University, Belgaum General Considerations in Selection and Design of Safety Pharmacology Studies 3. Ligand binding or enzyme assay data suggesting a potential for adverse effects 4.Results from previous safety pharmacology studies from secondary Pharmacodynamic studies from toxicology studies

29/11/2007 KLE University, Belgaum Test Systems 1. General considerations on test systems 2. Use of In Vivo and In Vitro Studies 3. Experimental Design a.Sample size and use of controls b.Route of administration

29/11/2007 KLE University, Belgaum Dose Levels or Concentrations of Test Substance 1.In Vivo Studies In vivo safety pharmacology studies should be designed to define the dose-response relationship of the adverse effect observed The time course of the adverse effect should be investigated e.g. onset and duration of response 2.In Vitro studies In vitro studies should be designed to establish a concentration-effect relationship

29/11/2007 KLE University, Belgaum Duration of Studies Pharmacology studies are generally performed by single-dose administration. Pharmacodynamic effects occur only after a certain duration of treatment OR Results from repeat dose nonclinical studies or results from use in humans give rise to concerns about safety pharmacology effects. The duration of the safety pharmacology studies to address effects should be rationally based.

29/11/2007 KLE University, Belgaum Studies on Metabolites, Isomers and Finished Products Metabolites from humans are known to substantially contribute to the pharmacological actions of the therapeutic agent, it could be important to test such active metabolites. In vitro or in vivo testing of the individual isomers should also be considered when the product contains an isomeric mixture. The finished product formulations should be conducted only for formulations that substantially alter the pharmacokinetics and/or Pharmacodynamic of the active substance in comparison to formulations previously tested.

29/11/2007 KLE University, Belgaum Safety Pharmacology Core Battery 1.Central nervous system 2.Cardiovascular system 3.Respiratory system

29/11/2007 KLE University, Belgaum Follow-up and Supplemental Safety Pharmacology Studies 1.Follow-up studies for safety pharmacology core battery a.Central nervous system b.Cardiovascular system c.Respiratory system 2. Supplemental safety pharmacology studies a.Renal/Urinary system b.Autonomic nervous system c.Gastrointestinal system d.Other organ systems (e.g. Skeletal Muscle, immune and endocrine functions)

29/11/2007 KLE University, Belgaum Conditions Under Which Studies Are Not Necessary Safety pharmacology studies may not be needed for locally applied agent. Safety pharmacology studies prior to the first administration in humans may not be needed for cytotoxic agents for treatment of end-stage cancer patients.

29/11/2007 KLE University, Belgaum Conditions Under Which Studies Are Not Necessary Biotechnology-derived products that achieve highly specific receptor targeting. Biotechnology-derived products that represent a novel therapeutic class and/or those products that do not achieve highly specific receptor targeting. Additional exceptions where safety pharmacology testing is not needed.

29/11/2007 KLE University, Belgaum Timing of Safety Pharmacology Studies in Relation to Clinical Development 1.Studies prior to first administration in humans 2. Studies during clinical development 3. Studies before approval

29/11/2007 KLE University, Belgaum Application of Good Laboratory Practice It’s important to ensure the quality and reliability of nonclinical safety studies. It has to be emphasized that data quality and integrity in safety pharmacology studies should be ensure even in the absence of formal adherence to the principles of GLP. Safety pharmacology investigations can be part of toxicology studies in such case, these studies would be conducted in compliance with GLP. Primary Pharmacodynamic studies do not need to be conducted in compliance with GLP.

29/11/2007 KLE University, Belgaum TOXICOLOGY “The science of poisons". It is the study of the opposing effects of physical agents or chemicals on living organisms.

29/11/2007 KLE University, Belgaum Recommended Toxicology Testing Genetic toxicity tests Short-term toxicity studies with rodents(14 to 21 days) Subchronic toxicity studies with rodents(90 days) Subchronic toxicity studies with non-rodents One-year toxicity studies with non-rodents

29/11/2007 KLE University, Belgaum Recommended Toxicology Testing Chronic toxicity or combined chronic toxicity/Carcinogenicity ( 2 years) Carcinogenicity studies with rodents (2 years) Reproductive studies  Developmental toxicity studies  Metabolism and Pharmacokinetic studies  Human studies 

29/11/2007 KLE University, Belgaum Genetic toxicity tests Test for gene mutations in bacteria In vitro test with cytogenetic evaluation of chromosomal damage using mammalian cells or In vitro mouse lymphoma thymidine kinase +/- gene mutation assay In vivo test for chromosomal damage using mammalian hematopoietic cells

29/11/2007 KLE University, Belgaum Short-term toxicity studies with rodents I. Good laboratory practice II. Test animals a.Care, maintenance and housing b.Selection of rodent species, strains and sex c.Age d.Number and sex e.Infected animals f.Animal identifications

29/11/2007 KLE University, Belgaum Short-term toxicity studies with rodents g. Caging h. Diet i. Assignment of control and compound treated animals j. Mortality k. Autolysis l. Necropsy

29/11/2007 KLE University, Belgaum Short-term toxicity studies with rodents III. Test substance a.Identity b.Composition/Purity c.Conditions of storage d.Expiration date IV. Experimental Design a.Duration of testing b.Route of administration c.Dose groups 1. Selection of treatment doses 2. Controls d. Computerized systems

29/11/2007 KLE University, Belgaum Short-term toxicity studies with rodents V. Observations and Clinical Tests a.Observations of test animals b.Body weight and feed intake data c.Clinical testing 1. Ophthalmic examination 2. Hematology 3. Clinical chemistry 4. Urinalyses 5. Neurotoxicity screening/testing 6. Immunotoxicity

29/11/2007 KLE University, Belgaum Short-term toxicity studies with rodents VI. Necropsy and microscopic examination a.Gross necropsy b.Organ weight c.Preparation of tissues for microscopic examination d.Microscopic evaluation. e.Histopathology of Lymphoid Organs

29/11/2007 KLE University, Belgaum Chronic toxicity or Combined chronic toxicity/carcinogenicity studies with rodents A. Experimental animals i. Age ii. Species and strains iii. Number and sex B. Administration of the test substance i. Duration of testing ii. Dosed groups a. Assessment of the carcinogenicity of the test substance High dose Low dose Intermediate dose Optional fourth dose b. Assessment of the chronic toxicity of the test substance

29/11/2007 KLE University, Belgaum Chronic toxicity or Combined chronic toxicity/carcinogenicity studies with rodents C. Observations and Clinical Tests i. Observation of test animals ii. Clinical testing Ophthalmological examination Hematology Clinical chemistry Urinalyses D. Necropsy and histopathology examination

29/11/2007 KLE University, Belgaum Reproduction studies A.General Recommendations B.Dose Range-Finding Study C.Main Study 1. Experimental Animals, Species and Strain Selection and Housing 2. Number, Sex, and Age 3. Assignment to Dose Groups 4. Dose Selection 5. Control Group's 6. Duration of Test

29/11/2007 KLE University, Belgaum Reproduction studies 7. Substance Administration 8. Mating Procedures 9. Standardizing the Number of Pups per Litter 10. Selection of Parental Animals for Next Generation 11. Optional Third Generation 12. Optional Second Mating 13. Optional Teratology Phase 14. Clinical Observation

29/11/2007 KLE University, Belgaum Reproduction studies 15. Growth of Offspring 16. Optional Neurotoxicity Screening 17. Optional Immunotoxicity Screening 18. Gross Necropsy and Microscopic Examination a.Necropsy of Weanlings b.Necropsy of Parental Animals i.Fixation of Tissues and Organs ii.General Histopathology iii.Histopathology of Female Reproductive Organs iv.Histopathology of Male Reproductive Organs

29/11/2007 KLE University, Belgaum Reproduction studies D. End Points of Female Reproductive Toxicity 1.Female Fertility Index 2.Gestation Index 3.Live-born Index 4.Weaning Index 5.Sex Ratio and Percentage by Sex 6.Viability Indices

29/11/2007 KLE University, Belgaum Reproduction studies E. End Points of Male Reproductive Toxicity 1.Evaluation of Testicular Spermatid Numbers 2.Sperm Evaluation for Motility, Morphology and Numbers F. Analysis of Data G. Reporting the Results of Reproduction Studies

29/11/2007 KLE University, Belgaum Development toxicity studies A.General Recommendations B.Dose Range-Finding Study C.Main Study 1. Experimental Animals, Species and Strain Selection 2. Animal Husbandry 3. Number, Sex, and Age 4. Duration of Testing 5. Route of Administration 6. Mating Procedures

29/11/2007 KLE University, Belgaum Development toxicity studies 7. Control and Dosed Groups 8. Maternal Toxicity and its Significance 9. Clinical Observation and Examination of Dams and Fetuses 10. Histopathology 11. End Points Measured 12. Analysis of Data D. Reporting the Results of Developmental Toxicity Studies

29/11/2007 KLE University, Belgaum Metabolism and Pharmacokinetic studies 1.Considerations in the design of analysis of and use of data from metabolic and pharmacokinetic studies a.Design and analysis of metabolic and pharmacokinetic studies i. Test compound ii. Animals iii. Route of administration iv. Dosage regimen v. Sampling vi. In vitro studies vii. Analysis of data

29/11/2007 KLE University, Belgaum Metabolism and Pharmacokinetic studies b. Use of data from metabolism and pharmacokinetic studies Design of toxicity studies Setting dose levels Determining mechanisms of toxicity Improving the risk assessment process 2. Recommended metabolism and pharmacokinetic studies 3. Additional studies

29/11/2007 KLE University, Belgaum Human studies 1.General considerations for clinical studies 2.Specific considerations for clinical studies a.Protocol design b.The study population c.Statistical analyses 3. Sequence of clinical studies a.Early clinical studies b.Further clinical studies

29/11/2007 KLE University, Belgaum Human studies 4. Submitting reports of clinical studies to CFSAN (Center for Food Safety and Applied Nutrition) 5. Appendix A a. Principles of institutional review b. Principles of informed consent.

29/11/2007 KLE University, Belgaum THANK YOU