This presentation provides a knowledge about Safety Pharmacology, It's aim & objectives, issues, consideration in selection and design of study and test study, duration of study, various studies involved in safety pharmacology, its guidelines, preclinical safety pharmacology. An assignment for the subject, Clinical Research and Pharmacovigilance, 1st year M.Pharm, 2nd semester.

1. Clinical Research And Pharmacovigilance
“Safety Pharmacology”
by
Chetan A., M.Pharm 1st Year (Pharmacology)
K.K. College of Pharmacy
Chennai, TamilNadu

2. Learning Objective
• Introduction
• Safety Pharmacology
• General Considerations in Selection & Design of Study
• General Consideration on Test Systems:
• Safety Pharmacology Studies
• Guidelines
• Conditions under which Safety Pharmacology Studies are not necessary
• Pre-clinical Safety Pharmacology
• Conclusion
• Recent Research
• Reference

3. Pharmacology
Pharmacology is the branch of pharmaceutical sciences which is concerned with
the study of drug action on body.
• Types of Pharmacology
• Primary Pharmacology - Effects of a substance in relation to its desired therapeutic
target (Mechanism of Action; MOA).
• Secondary Pharmacology - Identifying/understanding “off-target effects”. Effects
of a substance not related to its desired therapeutic target
• Safety Pharmacology - Investigates the potential undesirable pharmacodynamic
effects of a substance on physiological functions in relation to exposure in the
therapeutic range and above.

4. Safety
Pharmacology

5. Safety Pharmacology
• Safety pharmacology is a branch of pharmacology specialising in
detecting and investigating potential undesirable pharmacodynamic
effects of new chemical entities (NCEs) on physiological functions in
relation to exposure in the therapeutic range and above.
• Safety pharmacology studies are required to be completed prior to
human exposure (i.e., Phase I clinical trials), and regulatory guidance is
provided in ICH S7A and other documents.
• These test are conducted at doses not too much in excess of the intended
clinical dose.

6. General Principle
• Rational approach in Design and Conduct based on
Pharmaceutical’s Properties and uses.
• Scientifically valid methods to be used.
• Use of new technologies and methodologies is encouraged.
• Safety pharmacology end points can be incorporate in design of
Toxicology, kinetics, clinical studies etc.

7. Key Aim of Safety Pharmacology
• The aims of nonclinical safety pharmacology evaluations are three-fold:
• To protect Phase I clinical trial volunteers from acute adverse effects
of drugs
• To protect patients (including patients participating in Phase II and III
clinical trials)
• To minimize risks of failure during drug development and post-
marketing phases due to undesirable pharmacodynamic effects

8. Objectives of Safety Pharmacology
• HAZARD IDENTIFICATION
To identify undesirable pharmacodynamic properties of a
substance that may have relevance to its human safety
• RISKASSESSMENT
To evaluate adverse pharmacodynamic or pathophysiological
effects of a substance observed in toxicology studies
• RISK MANAGEMENT/MITIGATION
To investigate the mechanism of the adverse pharmacodynamic
effects observed

9. Key Issues
• The following key issues have to be considered within safety
pharmacology:
• The detection of adverse effects liability (i.e. hazard identification)
• Investigation of the mechanism of effect (risk assessment)
• Calculating a projected safety margin
• Implications for clinical safety monitoring
• Mitigation strategies (risk management)

10. General Considerations in Selection & Design of Study
• The following factors should be considered (the list is not comprehensive):-
• * Effects related to the therapeutic class of the test substance, since the
mechanism of action may suggest specific adverse effects.
• e.g., proarrhythmia is a common feature of antiarrhythmic agents
• * Adverse effects associated with members of the chemical or therapeutic
class, but independent of the primary pharmacodynamics effects.
• e.g., anti-psychotics and QT prolongation
• * Ligand binding or enzyme assay data suggesting a potential for adverse
effects .

11. Continuation.,,
• Results from previous safety pharmacology studies, from secondary
pharmacodynamics studies, from toxicology studies, or from human use that warrant
further investigation to establish and characterize the relevance of these findings to
potential adverse effects in humans .
• * During early development, sufficient information (e.g., comparative metabolism)
may not always be available to rationally select or design the studies in accordance
with the points stated above; in such circumstances, a more general approach in
safety pharmacology investigations can be applied.
• • A hierarchy of organ systems can be developed according to their importance with
respect to life-supporting functions.
• * Vital organs or systems, the functions of which are acutely critical for life, such as
the cardiovascular, respiratory and central nervous systems, are considered to be the
most important ones to assess in safety pharmacology studies.

12. Continuation..,
• Other organ systems, such as the renal or gastrointestinal system, the
functions of which can be transiently disrupted by adverse
pharmacodynamics effects without causing irreversible harm, are of less
immediate investigative concern.
• Safety pharmacology evaluation of effects on these other systems may be
of particular importance when considering factors such as the likely
clinical trial or patient population .

13. Test System
• General Consideration on Test Systems:
• Consideration should be given to the selection of relevant animal models
or other test systems so that scientifically valid information can be
derived.
• Data from humans (e.g., in vitro metabolism), when available, should
also be considered.
• The time points for the measurements should be based on
pharmacodynamic and pharmacokinetic consideration
• Justification should be provided for the selection of the particular animal
model or test system.

14. Use of In Vivo and In Vitro Studies
• Animal Models as well as ex vivo and in vivo preparations can be used as test
systems.
• Ex vivo and in vivo systems can include:
a) Isolated organs & tissues
b) Cell Culture
c) Cell Organelles
d) Subculture Organelles etc
• In vivo animal study include,
a) Use of unanaesthetized animals
b) Data from unrestrained animals used for telemetry
c) For unanesthetized animals, the avoidance of discomfort or pain is a foremost
consideration

15. Experimental Design
• Sample size and use of controls
a) Size of the group should be sufficient to allow meaningful scientific interpretation.
b) No. of animals should be adequate
c) Negative and Positive control groups should be included.
d) In vivo System - positive group may not be necessary.
e) Exclusion of controls from subject should be justified.
• Route of Administration:
a) Clinical route of administration should be used
b) Exposure to the parent substance and its metabolites should be similar to that
achieved in humans
c) If clinical use involves multiple routes, consider more than one route.

16. Dose levels or Concentrations of Test Substances
• In Vivo Studies:
a) It should be designed to define the dose-response relationship of the adverse effects
observed.
b) The dose elicting the adverse effect should be compared to the doses elicting the
primary pharmacodynamic effect in the test species or the proposed therapeutic
effect in humans, feasible.
c) It is recognized that there are species differences in pharmacodynamic sensitivity.
Therefore, should include and exceed the primary pharmacodynamic or therapeutic
range.
d) In the absence of an adverse effect, the highest tested dose should be a dose that
produces moderate adverse effects in this or in other studies of similar route &
duration. These adverse effects can include dose-limiting pharmacodynamics effects
or toxicity.

17. Continuation..,
• Testing of a single group at the limiting dose as described above may be
sufficient in the absense of an adverse effect on safety pharmacology
endpoints in the test species.
• In Vitro Studies
a) It should be deesigned to establish a concentration-effect relationship.
b) The range of concentration used should be selected to increase the likelihood
of detecting an effect on the test system.
c) The upper limit of this range may be influenced by physio-chemical
properties of the test substance and other assay specific factors.
d) In the absence of an effect, the range of concentration selected should be
justified.

18. Timing of Safety Pharmacology Studies in Relation to
Clinical Development
• Conducted prior to Administration in Humans (First in Huaman
[FIH];Investigational New Drug [IND] application, “IND-enabling”)
• During Clinical Development (Clinical Trials Phases 1-3)
• Before Approval (NDA application)
• Post-approval safety pharmacology investigations

19. Drug Discovery Cycle

20. Duration of Studies
• Are generally performed in Single dose administration
• The duration of the safety pharmacology studeis to address the following
effects should be rationally based on,
1. When pharmacodynamic effects occur only after a certain duration of
treatment
2. When results from repeat dose non-clinical studies
3. Results from use in humans give rise to concerns about safety
pharmacological effects.

21. Safety Pharmacology Studies

22. Safety Pharmacology Studies
I. Safety Pharmacology Core Battery
• Safety pharmacology core battery is to investigate the effects of the test
substance on vital functions.
a) Follow-up Studies For Safety Pharmacology Core Battery
These are meant to provide a greater depth of understanding than, or
additional knowledge to, that provided by the core battery on vital functions for
potential adverse pharmacodynamic effects
II. Supplemental Safety Pharmacology Studies
• To evaluate potential adverse pharmacodynamic effects on organ system
functions not addressed by the core battery or repeated dose toxicity studies

23. I. Safety Pharmacology Core Battery
• Tier 1
1. Central Nervous System
2. Cardiovascular System
3. Respiratory System
1. Central Nervous System
In Core battery
• Motor activity
• Behavioral changes
• Coordination
• Sensory/motor reflex response
• Body Temperature
In Follow-up studies
• Learning and memory
• Ligand-specific binding
• Neurochemistry
• Visual & auditory examination

24. 1. Central Nervous System
• Evaluation Methods
1. Functional Observation Battery (FOB)
2. Irwin’s test
FOB method of evaluation involves Neurological and
Neuropathological investigations

25. Irwin’s Test
• This method is to evaluate the qualitative effect of test substance on behavioural and
physiological functions and also duration of action
• The parameters observed are,
AUTONOMIC
EFFECTS
SENSORIMOTOR
EFFECTS
NEUROMUSCULAR
EFFECTS
BEHAVIOURAL
EFFECTS
Salivation Touch response Posture Arousal
Lacrimisation Palpebral reflex Grip strength Vocalisation
Pilorection Startle reflex Tremor Aggressiveness
Rectal temperature Pinna reflex Traction response Sniffing
Abnormal urination,
defecation, respiration
Writhing reflex Twitches Grooming

26. Continuation..,
Conclusion
• This test provide a rapid detection of test substances toxicity, active
dosage range Effects on behavioural and physiological function
Modification:
• Functional Observation Battery(FOB)

27. 1. Central Nervous System
• Other evaluation techniques
1. Rotarod
2. Hot plate test, Tail flick, paw pressure
3. Photoelectric beam interruption techniques
4. Passive avoidance tests
5. Pentylenetetrazol (PTZ) seizure tests
6. Electroencephalography (EEG)
• Emerging Techniques
1. Automated video systems
2. Integrated video and EEG systems

28. 2. Cardiovascular Systems (CVS)
Core Battery
• Blood Pressure
• Heart rate
Follow up Studies
• Cardiac output
• Ventricular contractility
• Vascular resistance
• Endogeneous & Exogeneous
substances on the Cardiovascular response

29. 2. Cardiovascular Systems (CVS)
• Evaluation Methods
1. Electrocardiogram
2. hERG asssay
3. Manual patch clamp
4. Automated high-throughput patch clamp
5. Isolated organ preparation
6. Whole heart preparation
7. Isolated purkinje fibres

30. hERG assay (human Ether-a-go-go Related Gene)
• The alpha subunit of a potassium ion channels in the heart that codes for
a protein known as Kv11.1
• Ion channel proteins (the ‘rapid’ delayed rectifier current (IKr)) that
conducts potassium (K+) ions out of the muscle cells of the heart.
• Inhibition of the hERG current causes QT interval prolongation resulting
in potentially fatal ventricular tachyarrythmia called Torsade de Pointes

31. 3. Respiratory System
Core Battery
1. Respiratory rate
2. Tidal Volume
3. Haemoglobin
oxygen
saturation
Follow up studies
1. Airway resistance
2. compliance
3. pulmonary arterial pressure
4. blood gases
5. blood pH

32. 3. Respiratory System
1. Plethysmography
2. Head out Plethysmography
3. Whole body Plethysmography
• Respiratory parameters:
1. Insoiratoey Time (Ti, ms)
2. Expiratory Time (Te, ms)
3. Peak Inspiratiory Flow (PIF, ml/s)
4. Tidal Volume (TV, ml)
5. Respiratory Rate (ResR, breaths/min)
6. Relaxation Time (Tr, ms)

33. II. Supplement Safety Pharmacology Studies
• Tier 2
1. Renal/Urinary System
2. Gastrointestinal System
3. Other organs - Skeletal System, Immune & Endocrine functions
1. Renal/Urinary System
Renal parameters should be assessed are
a) Urinary volume
b) Specific gravity
c) Osmolarity
d) pH, fluid/electrolyte balance
e) Proteins, cytology &
f) Blood chemistry determinations such as blood urea nitrogen, Na+, Cl-, K+, creatinine
and plasma proteins

35. 2. Gastrointestinal System
• Gastric secretion
• Gastrointestinal injury potential
• Bile secretion
• Transit time in vivo
• Ileal contraction in vivo
• Gastric pH measurement
• Gastric emptying
• Intestinal motility
• Emesis induction

36. 2. Gastrointestinal System
• Evaluaion Methods
a) Barium sulphate (BaSO4) or a charcoal test meal
b) Pylorus ligation test
• Emerging techniques
a) Endoscopy
b) Biomarkers
c) EMG Citrulline
d) miR-194
e) Calprotectin

37. Alternative Methods
• Zebrafish model: Anticonvulsant activity, locomotor activity,
behavioural paradigms such as addiction, memory and anxiety.
• human Embryonic Stemcell derived Cardiomyocytes (hESC-CM)
and human inducible Pluriopotent Stemcell derived Cardiomyocytes
(hiPS-CM) as models of in vitro high throughput drug screening and
CVS safety assessment.

38. Guidelines

39. Objectives of the Guideline
• 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

40. Guidlines
• Safety pharmacology studies are described in ICH guidances
• ICH S7A outlines the core battery studies and discusses the supplemental
studies
• ICH S7B details the procedures for conducting an in vitro evaluation of
delayed ventricular prolongation (QT interval prolongation)
• All safety pharmacology must be conducted in accordance with these
guidances

41. Conditions under which Safety Pharmacology Studies are
not necessary
• Safety pharmacology studies are usually not required for locally
applied agents eg, Dermal or Ocular, in cases when the
pharmacology of the investigational drug is well known, and/or
when systemic absorption from the site of application is low.
• Safety pharmacology testing is also not necessary, in the case of
a new derivative having similar pharmacokinetics and
pharmacodynamics.

42. Country-Specific Highlights: INDIA
• Agency: Central Drugs Standard Control Organization
• Website: http://www.cdsco.nic.in/
• Has specific guidelines for drug/vaccine development
• Drugs and Cosmetics (IIND Amendment) Rules “Schedule Y”
(2005) – Treats vaccine like a small molecule and requires
studies in two species, and requires both single-dose and repeat-
dose toxicity studies

43. Preclinical safety pharmacology
• Preclinical safety pharmacology integrates in silico, in vitro and in vivo
approaches.
• In vitro safety pharmacology studies are focused on early hazard
identification and subsequent compound profiling in order to guide
preclinical in vivo safety and toxicity studies.
• Early compound profiling can flag for receptor-, enzyme-, transporter-,
and ion channel-related liabilities of NCEs (e.g., inhibition of the human
ether-a-go-go related gene protein (hERG)). Classically in vivo
investigations comprise the use of young adult conscious animals.

44. Conclusion
• In summary, safety pharmacology is a discipline that plays an important
role in protecting human volunteers and patients from a new compound’s
potential for undesirable pharmacodynamic effects.
• Safety pharmacology studies evaluate nervous system, cardiovascular
and respiratory functionover a range of doses in the therapeutic range
and above that do not cause overt toxicity.
• Safety pharmacology is a diverse and exciting field that offers careers in
a number of different areas such as academia, industry and regulatory
agencies.

45. Recent Research
Precise safety pharmacology studies of lapatinib for onco-cardiology assessed using in vivocanine models
- Kentaro Ando et,al. (2020)
Cancer chemotherapies have improved prognosis in cancer patients, resulting in a large and rapidly
increasing number of cancer survivors. “onco-cardiology” or “cardio-oncology” is a new discipline for
addressing the unanticipated cardiac side effects of newly developed cancer drugs. Lapatinib, a tyrosine kinase
inhibitor suppressing the epidermal growth factor receptor and ErbB2, has been used in advanced or metastatic
breast cancer treatment. Reportedly, lapatinib has induced cardiovascular adverse events including Qt-interval
prolongation and heart failure. However, they have not been predicted by preclinical studies. Hence, a new
method to assess the tyrosine kinase inhibitor-induced adverse effects needs to be established. Here, they
intravenously administered lapatinib to halothane-anaesthetised dogs, evaluating cardiohemodynamic,
electrophysiological, and echocardiographic profiles for pharmacological safety assessments. They
intravenously administered lapatinib to chronic atrioventricular block beagle dogs to assess its proarrhythmic
potential. the therapeutic concentration of lapatinib significantly increased total peripheral vascular resistance,
QT, QTc, monophasic action potential (MAp)90(sinus), MAp90(CL400), effective refractory period, and
plasma concentration of cardiac troponin i (ctni), suggesting that lapatinib prolonged the ventricular
repolarization without inducing lethal ventricular arrhythmia. careful monitoring of plasma ctni concentration
and an electrocardiogram could be supportive biomarkers, predicting the onset of lapatinib-induced
cardiovascular adverse events.

46. Reference
• Wikipedia.org
• Slideshare.net
• Google search
• ProQuest LLC. (www.nature.com/scientificreports) -
DOI:10.1038/s41598-020-57601-x

47. Quote
~“Peace comes from within. Do not seek it without.”
-Gautama Buddha

48. Thank You