The document discusses guidelines for clinical trial protocols according to various regulatory agencies. It provides definitions of a protocol, describes common components of protocols such as objectives, study design, and safety assessments. It also outlines regulatory requirements for bioequivalence studies from agencies like FDA, EMA, and CDSCO regarding issues like study design, sample size, acceptance criteria, product handling and more. Requirements for conducting fed and fasting studies are also covered.

1. PROTOCOL PREPARATION
ACCORDING TO VARIOUS
REGULATORIES
Mr. Ajit Thorat
Accutest Research laboratories

2. OBJECTIVES
 Definition
 Components of Protocol
 Introduction to various regulatories

3. DEFINITION
 A document that states the background,
objectives, rationale, design, methodology
(including the methods for dealing with AEs,
withdrawals etc.) and statistical
considerations of the study. It also states the
conditions under which the study shall be
performed and managed.

4. COMPONENTS OF PROTOCOL
A well designed study relies predominantly
on a thoroughly considered, well-structured
and complete protocol
 General information
 Objectives and Justification

5. COMPONENTS OF PROTOCOL
 Ethical Considerations
 Study design
 Inclusion, Exclusion and Withdrawal of
Subjects

6. COMPONENTS OF PROTOCOL
 Handling of the Product(s)
 Assessment of Efficacy
 Assessment of Safety
 Statistics

7. COMPONENTS OF PROTOCOL
 Data handling and management
 Quality control and quality assurance
 Finance and insurance
 Publication policy

8. COMPONENTS OF PROTOCOL
 Suppementaries and appendices

9. VARIOUS REGULATORIES
FDA – Food & Drug Administration
EMA – European Medicines Agency
MCC – Medicines Control Council
ANVISA – National Health Surveillance Agency
Brazil
TGA – Therapeutics Good’s Administration
CDSCO – Central Drugs Standard Control
Organization
WHO – World Health Organization
Canada – Health Products and Food Branch

10. SEMI REGULATORIES
 Asia Pacific
 Asean Countries
 Middle- East
 African Countries
 Latin America

11. STUDY DESIGN
REGULATORY STUDY DESIGN
CDSCO Two studies fast & fed, If the effect of food on the reference
product is not known, two separate, two-way cross-over
studies.
USFDA Non-replicate crossover study designs are recommended for
BE studies of IR and MR dosage forms.
BRAZIL Open randomized crossed type studies.
EUROPE A two-period, two-sequence single dose crossover design &
steady state studies.
CANADA A two-period cross-over design.
SOUTH AFRICA A balanced two-period, two-sequence crossover design.

12. SAMPLE SIZE
Regulatory
Agency
Sample size
CDSCO Minimum number of subjects should not be less than 16 unless
justified for ethical reasons.
USFDA NLT 12.
ANVISA NLT 24.
TGA NLT 12.
MCC NLT 12 to provide at least 80% power of meeting the criteria and 20
subject for modified release tablet.
EMA NLT 12.
CANADA NLT 12.
WHO A minimum of 12 subjects is required.

13. DEMOGRAPHIC REQUIREMENT: AGE
Regulatory Agency Age
CDSCO Healthy adult volunteers, for elderly- 60 yrs & Above
USFDA 18 yrs of age or older and capable of giving informed consent.
For Elderly- 60 yrs & Above.
Brazil Subjects aged anywhere between 18 and 50 capable of
expressing their free and informed consent, of the male or
female gender or both.
Australia 18-55 within Normal Range as per Body mass Index
South Africa 18-55 within ± 15 % of Ideal body mass
Europe 18 years or older having BMI between 18.5 and 30 kg/m2
Canada Age range of 18 to 55 years, inclusive.
WHO Age range of 18 to 55 years and their weight should be within
the normal range.

14. DEMOGRAPHIC REQUIREMENT: SEX
Regulatory Agency Sex
CDSCO If the drug product is intended for use in both sexes attempt
should be made to include similar proportion of males and
females
USFDA If the drug product is intended for use in both sexes, the
sponsor attempt to include similar proportions of males and
females in the study
EMA Subjects could belong to either sex
South Africa Subjects may be selected from either sex
WHO If the pharmaceutical product is intended for use in both
sexes the sponsor may wish to include both males and
females in the study.
ANVISA Number of men and women are equally distributed between
the sequences.

15. DEMOGRAPHIC REQUIREMENT:
Regulatory Agency Demographic Requirements
USFDA Within normal range as per BMI 18.5 to 24.9 kg/m2
EMA Weight within the normal range according to accepted
normal values for the BMI between 18.5 and 30 kg/m2.
South Africa Within normal range as per BMI or within 15 % of the ideal
body mass, or any other recognized reference
ANVISA The weight of the subjects shall be within a limit of ±15% of
the weight considered normal for men and women, taking
into account height and physical structure
CANADA Subjects should preferably have a Body Mass Index within
18.5 and 30 kg/m2.
CDSCO Not Specified

16. DEMOGRAPHIC REQUIREMENT: ALCOHOL / SMOKING
RESTRICTIONS
Regulatory Agency Alcohol / smoking restrictions
USFDA Abstain from alcohol for 24 hours before each study period
and until after the last sample from each period is collected
CDSCO Implementation of these restriction from 48 hrs prior to study
initiation through end of study
Europe Without history of alcohol. (non-smoker, no drug abuse
history).
MCC Subjects should preferably be non-smokers and without a
history of alcohol or drug abuse
Canada Non smokers, but stipulate that subject who smokes may
included if they identified. alcohol use should also be
standardized
WHO Should have no history of alcohol.

17. FOOD RESTRICTIONS
Regulatory Agency Food Restrictions
USFDA 10 Hrs Before and 4 Hrs after drug administration
ANVISA Fasting for 8 Hrs before drug Administration.
CDSCO Same as USFDA
Europe Subjects should fast for at least 8 hours prior to
administration of the products, unless otherwise justified
MCC In fasted studies the period of fasting prior to dosing should
be standardised and supervised. All meals and fluids taken
after dosing should also be standardised in regard to
composition and time of administration and in accordance
with any specific requirements for each study
Canada & WHO Same as USFDA

18. WATER RESTRICTION
Regulatory Agency Water Restriction
USFDA Pre-Dose- 1 hr. Post Dose- 1 hr
Europe Water is allowed at desired except for 1 hr before after
drug administration
CDSCO As per USA .
Canada Two hours before and after drug administration
WHO Pre-Dose- 1 hr. Post Dose -2 hrs

19. WATER CONTENT
Regulatory Agency Water content
USFDA Subject should be administered drug product with 240 ml
water.
ANVISA Usually 200ml.
WHO Standard volume of water (150-250 ml).
TGA & EMA At least 150 ml
CDSCO Standard quantity of water (one glass : 200 ml) should be
allowed for proper ingestion of the dosage form
CANADA Standard volume (e.g., 150 ml).
SOUTH AFRICA Should be constant (e.g. 200 ml).

20. WASH OUT PERIOD
Regulatory Agency Wash Out Period
CDSCO >5 half lives of Analyte .
USA >5 half lives of Analyte .
ANVISA At least 7 times the studied drug half-life
EU At least 5 times the terminal half-life
TGA Adequate Duration
South Africa adequate wash-out periods
Canada NLT 10 times the mean terminal half-life of the drug.
WHO An adequate wash-out period should follow the
administration of each product

21. CONDITION FOR FED & FASTING STUDIES
Regulatory
Agency
Recommendation
CDSCO Generally, a single dose fasting study should be conducted after an
overnight fast (at least 10 hr), with subsequent fast of 4 hr of
dosing. For multiple dose fasting state studies, when an evening
dose must be given, 2 hr of fasting before & after the dose is
considered acceptable. For Fed sate studies require the
consumption of a high-fat breakfast (950-1000 KCals) before dosing
USFDA In fasting studies unless the studies is food effect. In fed study
high fat breakfast consisting 800-1000 Kcals.
ANVISA For fed studies reference product contain specific recommendation
about food effect.
CANADA For fed study high-fat, high-calorie meal (800 to 1000 kilocalories).
For Fasting study subjects should fast for eight hours before drug
administration.

22. CONDITION FOR FED & FASTING STUDIES
Regulatory
Agency
Recommendation
EUROPE The study should be conducted during fasting conditions unless
the SPC (Summary of Product Characteristics) recommends intake
of the originator product only in the fed state.
If the recommendation of food intake in the SPC is based on
pharmacokinetic properties such as higher BA, the BE study
should be conducted in the fed state.
Also if recommendation of food intake is intended to decrease
adverse events or to improve tolerability, it is recommended to
conduct the BE study in fed state, although a bioequivalence study
under fasting conditions could be acceptable if this has been
adequately justified.
In fed conditions, the composition of the meal should be
according to recommendations in the SPC of the reference
product. If no recommendation on the composition of the meal is
given in the reference product SPC, the meal should be a
"standardized non high-fat meal"(about 650 kcal with about 30% of
calories derived from fat).

23. CONDITION FOR FED & FASTING STUDIES
Regulatory
Agency
Recommendation
SOUTH AFRICA IR dosage forms should be done under fasting conditions. If
the reference product dosage directions state either “with or
without food” or make no statement with respect to food, the
study needs only be done under fasting conditions. for MR
dosage forms should demonstrate any influence of food in
order to exclude any possibility of dose dumping; hence, both
fed and fasted studies are required.

24. BLOOD SAMPLE COLLECTION
Regulatory
Agency
Blood sample collection
CDSCO For at Least 3 elimination Half Life, 3 sample During absorption, 3-
4 during T max, 4 during elimination.
USFDA 12 to 18 samples including a pre-dose sample.
BRAZIL Schedule must observe a length of time equal or greater than 3-5
times the half-life of elimination of the drug.
SOUTH
AFRICA
12 to 18 samples including a pre-dose sample .
CANADA A minimum of 12 samples should be collected per subject per dose.

25. BLOOD SAMPLE COLLECTION
Regulatory
Agency
Blood sample collection
EUROPE AUC- 80% of AUCinf, at least 3,4 samples during terminal log-linear
phases. In Multiple dose studies - The pre dose sample should be
taken immediately before (within 5 Min.) dosing and last sample
taken within 10 min of nominal time of dosing interval.
WHO Sampling points should include a pre-dose sample, at least 1–2
points before Cmax, 2 points around Cmax and 3–4 points during
the elimination phase. Consequently at least 7 sampling points-
necessary for estimation of the required pharmacokinetic
parameters.

26. USE OF DRUG METABOLITES
Regulatory
Agency
Use of Drug metabolite
USFDA Parent drug not measurable, significant active
and pre-systemic metabolism (present as
supportive data) They recommend that both the
parent drug and its major active metabolites be
measured, if analytically feasible.
EU Metabolite can be estimated if concentration of
active substance is too low, difficulty in analytical
method, product is unstable or has short half life.
The use of metabolite should be justified in each
case.
CDSCO Active substance if not possible then active
metabolite.

27. BIOEQUIVALENCE ACCEPTANCE CRITERIA
Regulatory
agency
90 % confidence interval on Log transformed data
Cmax % AUC0-t % AUC0-∞ %
CDSCO 80-125 80-125 80-125
USFDA 80-125 80-125 80-125
BRAZIL 80-125 80-125 NA
SOUTH
AFRICA
75-133 80-125 NA
CANADA 80-125 80-125 NA
CDSCO 80-125 80-125 NA
EUROPE 80-125 80-125 NA
AUSTRALIA 80-125 80-125 NA

28. HANDLING OF INVESTIGATIONAL PRODUCT
Regulatory
Agency
Retention period Retention quantity
USFDA Retain IP for 5 years. 5 Times or 300 Tablet each for test
and reference.
EUROPE Retain IP for 1 yrs in excess of
acceptable shelf life or 2 yrs
after completion of Trial or
approval whichever is greater.
Sufficient numbers to allow
retesting.
CANADA Retention period is not
applicable for Canada.
As specified and provided by
sponsor
CDSCO 3 Yrs after the conduct of study
or 1 yr after the expiry of the
drug.
As specified and provided by
sponsor

29. HANDLING OF INVESTIGATIONAL PRODUCT
Regulatory Agency Retention period Retention quantity
AUSTRALIA Retain IP for 1 yrs in excess of
acceptable shelf life or 2 yrs
after completion of Trial or
approval whichever is greater.
As specified and provided
by sponsor
SOUTH AFRICA one year in excess of the
accepted shelf-life, or two
years after completion of the
trial or until approval,
whichever is longer.
As specified and provided
by sponsor
BRAZIL 2 years Quantity performed to
required new study.