BC Diltiazem Stada

MINISTRY OF HEALTH
NATIONAL INSTITUTE OF DRUG QUALITY CONTROL

----O----
BIOEQUIVALENCE REPORT
IN VIVO BIOEQUIVALENCE STUDY OF

Diltiazem STADA® 60 mg (Diltiazem hydrochloride 60 mg
extended-release tablets) versus TILDIEM® 60 mg
under fed condition
Sponsor: STADA - VN JOINT VENTURE CO., LTD.

Address: 40 Tu Do Avenue, Vietnam-Singapore Industrial park, Binh Duong,
Vietnam
Contract Research organization: National Institute of Drug Quality Control
Address: 48 – Hai Ba Trung, Hanoi, VN
Study No.: 45BE 26
HA NOI 2016
BE Center-NIDQC BE report – Diltiazem STADA® 60 mg
GENERAL INFORMATION
Study Title: In vivo bioequivalence study of Diltiazem STADA® 60 mg (Diltiazem
hydrochloride 60 mg extended-release tablets) versus TILDIEM® 60 mg under fed
condition.
1. Study Number: 45BE 26
2. Name and address of Sponsor
STADA - VN JOINT VENTURE CO., LTD.
Address: 40 Tu Do Avenue, Vietnam-Singapore Industrial park, Binh Duong, Vietnam
Tel: 84.650.3767470 Fax: 84.650.3767469
3. Name and address of Institution
BE Center, National Institute of Drug Quality Control
Address: 48 – Hai Ba Trung, Hanoi, Vietnam
Tel: 04.39365601 Fax: 04.3825.6911
4. Name and address of Principal Investigator
Pharm. Ta Manh Hung, MSc., BE Center, NIDQC
Address: 48 - Hai Ba Trung, Hanoi, VN, Email: tahungvkn@gmail.com
5. Name and address, person in charge of Clinical Investigator
Pharm. Ha Thi Tuyen, BE Center, NIDQC
Address: 48 - Hai Ba Trung, Hanoi, VN, Email: tuyenvkn@gmail.com
6. Name and address, person in charge and address of clinical laboratory
MD. Nguyen Nghiem Luat; Medical Laboratory Technology Company
Address: 42 – Nghia Dung Street, Hanoi, VN
7. Name and address, person in charge and address of analytical laboratory
Pharm. Tran Hoang, MSc., BE Center, NIDQC
Address: 48 – Hai Ba Trung, Hanoi, VN. Email: tranhoangds@yahoo.com
8. Name and address, person in charge and address for pharmacokinetics and statistical
analysis
Pharm. Phan Thi Nghia, MSc., BE Center, NIDQC
Address: 48 – Hai Ba Trung, Hanoi, VN. Email: nghiavkn@gmail.com
9. Name and address, person in charge of QA
Pharm. Le Thi Thu Huyen, MSc., BE Center, NIDQC
Address: 48 – Hai Ba Trung, Hanoi, VN; Email: thuhuyenvkn@gmail.com
10. Study products
- Test Product: Diltiazem STADA® 60 mg
(Diltiazem hydrochloride 60 mg extended-release tablets)
Producer: STADA - VN JOINT VENTURE CO., LTD.
- Reference Product: TILDIEM® 60 mg
(Diltiazem hydrochloride 60 mg tablets)
Manufactured by: Sanofi Winthrop Industrie – France
BE/FM/REP.01.01 i Study number: 45BE 26

11. Start and end date of clinical and analytical study
- Clinical period: 16/12/2015 – 27/12/2015
- Analytical period: 06/01/2016 – 12/01/2016
- Statistical period: 17/02/2016 – 18/02/2016
12. Signature and date of investigators
This study was performed in compliance with the guidelines for Good Clinical Practice,
Good Laboratory Practice and the Declaration of Helsinki
We, the undersigned, have read this report and confirm that the information and data in
the report are correct.
Position Name Signature/ Date

Date:
Clinical Investigator Pharm. Ha Thi Tuyen Signature:
Date:
Clinical Doctor MD. Pham Viet Tuan, MSc. Signature:
Date:
Analytical Investigator Pharm. Tran Hoang, MSc. Signature:
Date:
Statistical Investigator Pharm. Phan Thi Nghia, MSc. Signature:
Date:
Quality assurance Pharm. Le Thi Thu Huyen, MSc. Signature:
Date:
Principal Investigator Pharm. Ta Manh Hung, MSc. Signature:
Date: ................................... 2016

Contract Research Organization
BE/FM/REP.01.01 ii Study number: 45BE 26

STUDY SYNOPSIS
In vivo bioequivalence study of Diltiazem STADA® 60 mg (Diltiazem
Study Title hydrochloride 60 mg extended-release tablets) versus TILDIEM® 60
mg under fed condition.
Contract
National Institute of Drug Quality Control
Research
Address: 48 – Hai Ba Trung, Hanoi, Vietnam.
Organization
Principal
Pharm. Ta Manh Hung, MSc., BE Center, NIDQC
Investigator
Duration of
study 12/2015 – 02/2016
This study will assess the bioequivalence of Diltiazem STADA® 60 mg

(Diltiazem hydrochloride 60 mg extended-release tablets) manufactured
by STADA - VN Joint Venture Co., Ltd. versus TILDIEM® 60 mg
Objective (Diltiazem hydrochloride 60 mg tablets) manufactured by Sanofi
Winthrop Industrie - France in healthy adult volunteers after single-
dose administration (1 tablet x 60 mg) under fed condition.
Diltiazem STADA® 60 mg
(Diltiazem hydrochloride 60 mg extended-release tablets
Lot No.: 020915; Mfg. date: 170915; Exp. date: 170917
Test product
Reg. No: VD-14553-11
Producer: STADA - VN Joint Venture Co., Ltd.
Dose: 1 tablet x 60 mg
TILDIEM ® 60 mg
(Diltiazem hydrochloride 60 mg tablets)
Reference Lot No.: 4T014; Mfg. date: 04/2014; Exp. date: 03/2017;
product Reg. No: VN-17695-14
Manufactured by: Sanofi Winthrop Industrie - France
Dose: 1 tablet x 60 mg
Randomized, single-dose, two-treatment, two-sequence, two-period,
crossover study. Take drug 30 minutes after consuming the standard test
Study design
meal (high fat, high energy). Wash-out period is 7 days. The study is
conducted on 24 subjects.
Healthy male or female from 18 to 55 years of age at the time of dosing.
The BMI is in the range 18–25 kg/m2. All the subjects comply with the
Inclusion
inclusion criteria mentioned in protocol, including general examination,
Criteria
screening clinical laboratory, HIV Antibody and Hepatitis B Surface
Antigen, urine test to detect pregnancy (for women).
BE/FM/REP.01.01 iii Study number: 45BE 26

- Dosing and sampling: 24
Subjects - Sample analysis: 23
- Statistical analysis: 23
Bioanalytical Determine Diltiazem concentration in plasma by validated LCMS
method method.
- Analyze the variance for C max and AUC, determine the 90%
Statistical confidence interval for the ratio of Test and Reference products
analysis (calculated on log-transformed data).
- Comparison Tmax by non-parametric method (Wilcoxon signed-rank test)
For Cmax and AUC, 90% confidence interval for the ratio of the Test and
Assessment
Reference products must be in the range 80.00 – 125.00%.
criteria
Tmax: No statistically significant difference
Cmax: 149.09%-199.78%; AUC0-t: 98.12%-120.73%;
Summary of AUC0-: 91.75%-110.10%;
results Tmax: No statistically significant difference at 90% confidence level
(p>0.05)
BE/FM/REP.01.01 iv Study number: 45BE 26

TABLE OF CONTENTS
1. INTRODUCTION ABOUT DILTIAZEM ...................................................................... 1
2. OBJECTIVE ..................................................................................................................... 1
3. INVESTIGATIONAL PRODUCTS ................................................................................ 2
3.1. Test Product Information ........................................................................................... 2
3.2. Reference Product Information.................................................................................. 3
3.3. Pharmaceutical Equivalence Data ............................................................................. 3
3.4. Comparison of Dissolution Profiles........................................................................... 4
3.5. Test product and dossier submitted for marketing authorization .............................. 4
4. METHODOLOGY OF CLINICAL STUDY ................................................................... 4
4.1. Study design............................................................................................................... 4
4.2. Approved protocol ..................................................................................................... 4
4.3. Subjects ...................................................................................................................... 4
4.3.1. Number of subjects ............................................................................................. 4
4.3.2. Inclusion Criteria ................................................................................................ 4
4.3.3. Exclusion Criteria ............................................................................................... 5
4.4. Implementation procedures........................................................................................ 5
4.4.1. Subject screening ................................................................................................ 5
4.4.2. Subject confinement............................................................................................ 6
4.4.3. Drug administration ............................................................................................ 6
4.4.4. Sampling ............................................................................................................. 6
4.4.5. Sample processing and storage ........................................................................... 7
4.4.6. Food, drink regimen and activities of subjects ................................................... 7
4.4.7. Concomitant medication ..................................................................................... 8
4.4.8. Health monitoring ............................................................................................... 8
4.4.9. Adverse events and treatment ............................................................................. 8
4.5. Withdrawal criteria .................................................................................................... 9
4.6. Replacement of withdrawal subjects ......................................................................... 9
5. BIOANALYTICAL AND STATISTICAL METHODOLOGY ..................................... 9
5.1. Method validation ...................................................................................................... 9
5.2. Analysis of subject samples ....................................................................................... 9
5.3. Statistical analysis .................................................................................................... 12
5.3.1. Pharmacokinetic Parameters ............................................................................. 12
5.3.2. Assessment of bioequivalence .......................................................................... 13
6. DATA QUALITY ASSURANCE.................................................................................. 13
7. RESULTS AND DISCUSSION .................................................................................... 13
7.1. Clinical results ......................................................................................................... 13
7.1.1. Subject screening results ................................................................................... 14
7.1.2. Randomization and dosing schedule................................................................. 17
7.1.3. Dosing and sampling process............................................................................ 17
BE/FM/REP.01.01 v Study number: 45BE 26

7.1.4. Health monitoring ............................................................................................. 18
7.1.5. Adverse events .................................................................................................. 18
7.1.6. Concomitant medication ................................................................................... 18
7.1.7. Protocol deviation ............................................................................................. 19
7.2. Summary of analytical results.................................................................................. 19
7.3. Pharmacokinetic analyses ........................................................................................ 34
7.4. Statistical analyses ................................................................................................... 37
7.4.1. Statistical summary of pharmacokinetic parameters ......................................... 37
7.4.2. Comparison of pharmacokinetic parameters of the Test versus the Reference
product ........................................................................................................................ 38
7.4.3. Analysis of variance (ANOVA) ........................................................................ 41
7.4.4. Comparison of Tmax ........................................................................................... 42
8. CONCLUSIONS ........................................................................................................... 44
BE/FM/REP.01.01 vi Study number: 45BE 26

LIST OF TABLES
Table 1: Summary of bioanalytical method validation results
Table 2: Results of general examination and screening tests of 24 official subjects
Table 3: Randomization and dosing schedule
Table 4: Summary of analysis process
Table 5: Concentration of Diltiazem in plasma after taking the Test product
Table 6: Concentration of Diltiazem in plasma after taking the Reference product
Table 7: Individual pharmacokinetic parameters after taking the Test product
Table 8: Individual pharmacokinetic parameters after taking the Reference product
Table 9: Statistical summary of pharmacokinetic parameters
Table 10: Individual comparison of Cmax of the Test versus the Reference product
Table 11: Individual comparison of AUC0-t of the Test versus the Reference product
Table 12: Individual comparison of AUC0-∞ of the Test versus the Reference product
Table 13: ANOVA for Cmax
Table 14: ANOVA for AUC0-t
Table 15: ANOVA for AUC0-∞
Table 16: Summary of comparative bioavailability data
Table 17: Comparison of Tmax
LIST OF FIGURES
Figure 1: Mean plasma concentration - time curve of 23 subjects
Figure 1: Mean semi-logarithmic plasma concentration - time curve of 23 subjects
LIST OF APPENDICES
- Appendix 1: Clinical data.
o Protocol, Informed Consent form.
o Letter of approval, list of members of Ethical Committee.
o Report of clinical data.
o Case Report Form of 5 Subjects.
- Appendix 2: Information of the investigational products.
o Certificates of analysis of the investigational products.
o Certificates of analysis of standards
o Formulation and manufacturing process of the Test product
o Comparison of Dissolution profiles
- Appendix 3: Bioanalytical method validation report and 20% chromatograms.
- Appendix 4: Analytical report and 20% chromatograms.
- Appendix 5: Statistical results
o Statistical report
o Individual plasma concentration – time curves
- Appendix 6: Certificates of facilities and information of investigators
BE/FM/REP.01.01 vii Study number: 45BE 26

o Certificates of facilities
o Information of investigators
O List of some SOPs used in study
ABBREVIATION
Abbreviation Meaning
ADR Adverse drug reaction
AE Adverse event
AUC Area under the curve
BE Bioequivalence
Cmax Maximum concentration
CRF Case Report Form
CV Coefficient of variation
EC Ethics committee
Exp. Date Expiration date
GCP Good Clinical Practice
GLP Good Laboratory Practice
HQC High quality control
ISR Incurred Sample Reanalysis
LLOQ Lower limit of quantification
LQC Low quality control
MQC Medium quality control
Mfg. date Manufacturing date
NIDQC National Institute of Drug Quality Control
QC Quality control
R The Reference product
RA Re analysis
Reg. No. Register number
SD Standard deviation
Subj. Subject
T The Test product
Tmax Time to maximum concentration
ULOQ Upper limit of quantification
BE/FM/REP.01.01 viii Study number: 45BE 26

1. INTRODUCTION ABOUT DILTIAZEM
Diltiazem is a calcium-channel blocker, used for treatment of angina pectoris and
hypertension.
Pharmacokinetics:
Diltiazem is absorbed well following oral administration, bioavailability is about 40%
due to extensive first-past hepatic metabolism. The drug is about 70% – 80% bound to
plasma proteins. The drug is mainly metabolised in liver and excreted in urine and feces.
Indications:
- Treatment of mild to moderate hypertension.
- Treatment and prevention of angina pectoris.
Contraindications:
- Sinus node operation dysfunction, pre-existing second or third-degree
atriventricular block.
- Hypersensitivity to Diltiazem.
- Left ventricular failure with pulmonary congestion.
- Heartbeat less than 50 beats/minute
Adverse events and treatments:
- Common, ADR>1/100:
+ Body: Ankle oedema, headache, dizziness, somnolence.
+ Circulation: First-degree atriventricular block.
+ Gastrointestinal: Nausea, constipation.
+ Skin: Itching, rash.
- Less common, 1/1000<ADR<1/100: Bradycardia, urticaria
- Rare, ADR,1/1000:
+ Body: Board with facial flushing feeling.
+ Circulation: Second or third-degree atriventricular block, sinus arrest, angina
increased, palpitation, hypotension, tachycardia, ventricular beats.
+ Gastrointestinal: Pypertrophic gingivitis, hepatitis.
+ Skin: Erythema multiforme, edema Quincke.
+ Musculoskeletal: myalgia, arthralgia.
+ Neurological: Confusion or insomnia.
+ Rash due to hypersensitivity, are normally mild and transient but in a few cases
erythema multiforme or exfoliative dermatitis has developed.
+ Transient elevations in liver enzyme values and hepatitis.
+ Diltiazem can cause congestive heart failure, requiring care of patients with the
left ventricular dysfunction.
- Treatments: In a few cases, it can be seen rising liver enzymes (GOT, GPT; Grama GT;
LDH) and alkaline phosphatase with symptoms similar to acute hepatitis. So, it is
necessary evaluate the parameters of liver regularly. Specially when used with high
doses and/or a history of heart disease, bradycardia, conduction disturbances (sinoatrial or
BE/FM/REP.01.01 1/44 Study number: 45BE 26

atrioventricular block), hypotension, palpitations and heart failure may occur. Physicians
should monitor regularly and treat symptoms.
Dosage and Administration:
- The usual dose is 60 mg, three times daily before meal.
- Dose for treatment of angina pectoris is 60 mg, three times daily. For patients with
unstable angina, may use sustained-release tablets with dose from 360 – 480 mg daily.
- For treatment of hypertension: Use sustained-release tablets with initial dose of 60-120
mg, two times daily. Maximum dose is 360 mg daily.
Drug interaction:
- Antiarrhythmics: Diltiazem should not be used in combination with other
antiarrhythmic drugs because they increase the adverse effects on the heart.
- Carbamazepine, Cyclosporine, Theophyline: Concomitant use of diltiazem and these
drug may result in increased plasma concentration of these drug. Therefore, adjust dosage
in combination time and after combination.
- Digoxin: Concomitant use of diltiazem and Digoxin may result in increased plasma
concentration of Digoxin.
- H2 - receptor antagonists: Concomitant use of Diltiazem and Cimetidin or Ranididin
may result in increased plasma concentration of Diltiazem.
- Antiepilepic drugs: Diltiazem can result in increased toxicity of carbamazepine.
2. OBJECTIVE
This study will assess the bioequivalence of Diltiazem STADA® 60 mg (Diltiazem
hydrochloride 60 mg extended-release tablets) manufactured by STADA - VN Joint
Venture Co., Ltd. versus TILDIEM® 60 mg (Diltiazem hydrochloride 60 mg tablets)
manufactured by Sanofi Winthrop Industrie - France in healthy adult volunteers after
single-dose administration (1 tablet x 60 mg) under fed condition.
3. INVESTIGATIONAL PRODUCTS
3.1. Test Product Information
(Diltiazem hydrochloride 60 mg extended-release tablets)
Lot No.: 020915, Mfg. date: 170915, Exp. date: 170917; Reg.No: VD-14553-11
Manufactured by: STADA - VN Joint Venture Co., Ltd.
Address: 40 Tu Do Avenue, Vietnam-Singapore Industrial park, Binh Duong,
Vietnam
Description: White round tablets, engraved with “STADA” on one side and with “60” on
the other side. Package of 3 blisters x 10 tablets per carton. The label on the blisters and
the carton are printed with the product name, strength and information about
manufacturer, Lot No., Mfg. date, Exp. Date and Reg. No.
Batch size: 1.000.000 tablets
Product formulation:

Ser. Ingredients Quantity/1 tablet Quantity/1 batch
(mg) (kg)
1. Diltiazem hydrocloride 60.0 60.0
2. Dicalci phosphate dihydrate 112.0 112.0
Glycerol dibehenate
3. 15.0 15.0
(Compritol 888 ATO)
4. Povidon K30 5.0 5.0
5. Magnesi stearate 4.0 4.0
6. Talc 4.0 4.0
7. Ethanol 96%* (10.0) (10.0)
8. Purified water* (10.0) (10.0)
Total 200 (mg) 200 (kg)
(*): Solvent was evaporated during the process.
Finished Product Specifications: In-house specification, enclosed in Appendix 2.
3.2. Reference Product Information
TILDIEM® 60 mg (Diltiazem hydrochloride 60 mg tablets)
Lot.No.: 4T014; Mfg.date: 04/2014; Exp.date: 03/2017;
Reg.No.: VN-17695-14
Manufactured by: Sanofi Winthrop Industrie - France
Address: 30-36 avenue Gustave Eiffel, 37100 Tours, France
Description: White round tablets, engraved with “TILDIEM 60” on one side and plain on
the other side. Package of 3 blisters x 10 tablets per carton. The label on the blisters and
the carton are printed with the product name, strength, information about manufacturer,
Lot No., Mfg. date, Exp. date and Reg. No.
The investigational drugs were provided by STADA - VN Joint Venture Co., Ltd. and
sent to BE center by post.
3.3. Pharmaceutical Equivalence Data
Test products and Reference products are tablets containing 60 mg of Diltiazem
hydrochloride. Test products complies with in-house specification. Certificate of Analysis
is enclosed in Appendix 2.
The investigational products were tested for some characteristics (identification, assay
and uniformity of weight) according to in-house specification of the Test product. The
contents of Diltiazem hydrochloride in two products are listed below:
Test products: 102.1%
Reference products: 100.5%
The difference of quantitative contents of Diltiazem hydrochloride between two products
are less than 5% and both meet the uniformity of content requirement.

The copies of Certificates of Analysis for tested characteristics of Test and Reference
products are enclosed in Appendix 2.
3.4. Comparison of Dissolution Profiles
Directly provided by sponsor, enclosed in Appendix 2.
3.5. Test product and dossier submitted for marketing authorization
Applicant/sponsor will directly provide formula, manufacturing process of Test product
in comparison with the product submitted for marketing authorization.
4. METHODOLOGY OF CLINICAL STUDY
4.1. Study design
Randomized, single-dose, two-treatment, two-sequence, two-period, crossover study.
Take drug 30 minutes after consuming the standard test meal (high fat, high energy).
Wash-out period is at least 7 days. The study is conducted on 24 subjects.
4.2. Approved protocol
Protocol was approved by NIDQC’s Ethical Committee (Letter for approval
No.23/2015/HĐ-ĐĐ dated on December 15th, 2015).
Approved protocol, letter for approval and list of Ethical Committee members are
enclosed in Appendix 1.
4.3. Subjects
4.3.1. Number of subjects
Number of subjects participating in study (sample size) is estimated based on:
+ The intra-subject coefficient of variation of pharmacokinetic parameters.
+ The significance level desired (α = 0.05).
+ The expected ratio of the geometric mean of pharmacokinetic parameters
between the Test and the Reference product.
+ The required statistical power of study.
Sample size will be agreed between BE Center and the sponsor but not less than 12
subjects (according to ASEAN Guidelines).
After calculation and agreement, BE Center and the sponsor decided to conduct this study
on 24 subjects.
4.3.2. Inclusion Criteria
- Healthy men or women, 18 – 55 years of age and with normal civil behavior.
- The BMI will be in the range of 18 – 25 kg/m2 for body frames as per Desirable
Weights for Adults –1983 Metropolitan Height and Weight Table.
- Negative screening test for HIV and HBsAg (by Elisa test or ECLIA test).
- Negative pregnancy test for female subjects (by quick stick).
- Laboratory parameters (haematology and clinical chemistry) are in acceptable
range.
- Agree to implement all requirements of the protocol and sign Informed Consent
Form voluntarily.

4.3.3. Exclusion Criteria
- Volunteers with abnormal civil behavior.
- Volunteers don’t meet inclusion criteria.
- Volunteers with a recent history of drug abuse, alcoholic addiction.
- Volunteers with presence of significant smoking (more than 5 cigarettes/day).
- Volunteers with the presence of a clinically significant disorder involving the
cardiovascular, respiratory, renal, gastrointestinal, immunological, hematological,
endocrine, or neurological system(s) or psychiatric disease (as determined by the
clinical investigators).
- Volunteers with a positive hepatitis B surface antigen screen or a reactive HIV
antibody screen (by Elisa test or ECLIA test).
- Volunteers with history of allergic response(s) to Diltiazem or other in the same
group.
- Volunteers with a clinically significant illness during the 2 weeks prior to Period 1
dosing.
- Volunteers who have taken any drug known to induce or inhibit hepatic drug
metabolism (such as MAOIs group, Cimetidin, Amiodaron …) in the 28 days prior
to Period 1 dosing.
- Volunteers who have participated in blood donation or any other clinical study
within 90 days prior to Period 1 dosing.
- Volunteers who have received any investigational or prescription drug within 14
days prior to Period 1 dosing.
- Volunteers who have a history of difficulty in swallowing, or any gastrointestinal
disease which could affect the drug absorption
- Women who are pregnant (positive with pregnancy test-stick), or breastfeeding, or
intend to be pregnant in the next 2 months.
- Volunteers with history of orthostatic hypotension, fasting hypoglycemia or fear of
blood.
4.4. Implementation procedures
4.4.1. Subject screening
After study protocol is approved by Ethics Committee and within 4 weeks prior to period
1 dosing, 24 official and about 02 stand-by subjects will be recruited.
The subjects will be informed full of the study contents and sign the Informed Consent
Form voluntarily before screening.
Determine body weight (kg), height (m), BMI (Kg/m2) and review medical history… of
subjects. Then, each subject will undergo a physical examination and laboratory tests
including:
 Haematology: haematocrite, haemoglobin, WBC count, RBC count, platelet
count…
 Clinical Chemistry: AST (GOT), ALT (GPT), serum creatinine, glucose, urea.

 HIV Antibody and Hepatitis B Surface Antigen.
 Urine test to detect pregnancy (for women).
Physician, principal investigator will review and compare all information of each subject
with inclusion and exclusion criteria to conclude that he or she is eligible or ineligible to
participate in the study.
4.4.2. Subject confinement
In each period, subjects will be confined at clinical area from at least 10 hours before
drug administration to 12 hours post dose, then they will come back to their residence and
return to clinical area for the ambulatory sample (24 hours after dosing) as schedule.
During confinement, each subject will be worn an identified card containing the
following information: subject code, subject name, and the study protocol number. This
card will identify subjects in every study-related activities such as blood sampling, health
monitoring or AE treatment…
4.4.3. Drug administration
Using Excel software to randomize the dosing sequences of each subject. Divide into two
groups and take drugs as:
Period
Group 1 2
1 T R
2 R T
- Dose: Single dose, 1 tablet x 60 mg of test or reference drug per period.
- Administration: Take an intact tablet with 240 mL of warm water. After dosing, subjects
should remain in an upright position for at least 2 hours. Oral cavity check will be carried
out immediately after drug administration to confirm complete medication.
- The randomization code will only be informed to the physician in case of AE treatment
and will be only sent to statistical staff with the aim of statistical analysis and final report
preparation. The staffs who are involved in subject management, sampling and sample
analysis will be blinded to the randomization code.
- Before dosing day of each period, the drug will be packed into jars for each subject. Jars
are labeled with the study number, subject code and study period.
4.4.4. Sampling
- Sampling points in each period (hour):
0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 8.0 10 12 24
- Pre-dose sample (zero point) will be collected within one hour prior to dosing. The post
dose samples collected within 1 minutes for the first 4 hours, within 3 minutes for 5-hour
and 6-hour samples, within 5 minutes for samples from 8 – 12 hours and within 30
minutes for 24-hour sample deviated from the scheduled sampling time will not be
considered as protocol deviations. Any deviations greater than the mention above will be
reported as protocol deviations and will be corrected when statistical analysis.

- Blood volume: For each subject, at each sampling time-point, 6 mL of blood will be
collected. Thus total 30 blood samples (180 mL) of each subject will be collected for both
periods.
- Sample collection:
+ In the day of dosing, samples will be collected through an indwelling
cannula placed in a forearm-vein of the subject. Each post dose sample, about 5.5 mL of
blood will be collected after discarding the first 0.5 mL of heparinized blood (0.9% NaCl
solution containing heparin 5UI/mL) from cannula. In case the cannula is blocked,
samples will be collected directly at forearm vein.
+ For ambulatory samples (24 hours): about 5.5 mL of blood will be collected
directly at forearm vein.
4.4.5. Sample processing and storage
After being collected, blood samples are transferred to the tubes containing Na2EDTA as
anticoagulant. Collection tubes will be inverted 3 to 4 times immediately, centrifuged at
approximately 4000 rpm for 10 minutes. The plasma will be pipetted into labeled
polyethylene tubes, frozen at approximately -350C ± 50C and stored until analysis. The
labels for all biological sample collection and storage tubes will contain information of
the subject’s code, period, sampling time-point and study number.
4.4.6. Food, drink regimen and activities of subjects
- Drinks:
+ In the dosing day, drinking water will not be allowed within 1 hour prior to and
after dosing, except that given with drug administration. At other time, the subjects can
drink water as their demand.
+ Consuming of products containing alcohol or caffeine is prohibited from at least
48 hours prior to dosing until the last blood sample in each period. Consuming of these
products should be limited in wash-out period.
- Food: In the dosing day, subjects take drug 30 minutes after consuming the standard test
meal (high fat, high energy). Standardized meals will be served to each subject
approximately 4 hours and 11 hours after dosing as scheduled. Standard meals are
homogenous for both periods.
- Activities:
+ Subjects should remain in an upright position for the first 2 hours after dosing.
+ During confinement, subjects should avoid severe physical activities after
dosing.
+ In the dosing day, subjects are only in clinical area within the first 4 hours after
dosing. After that, subjects can relax in campus of NIDQC but must be agreed by
investigators.
+ Severe physical activities should be limited during wash-out period.

4.4.7. Concomitant medication
- No medication is allowed from two weeks prior to dosing until the study is completed
unless required to treat any adverse events. Medication for treatment of adverse events
will be indicated by clinical doctor depending on symptom and severity.
- If the subjects used any non-study medication prior to dosing or during the study or in
the washout period, the principal investigator will decide to remove the subject or not,
based on the following informations:
 The pharmacology and pharmacokinetic of the non-study medication.
 The likelihood of drug-drug interactions.
 The time and duration of administration of the non-study medication.
4.4.8. Health monitoring
- During the study, vital signs (blood pressure, pulse rate, respirator rate, body
temperature) will be measured at the following time- point:
+ Prior to drug administration
+ 2.0, 4.0, 8.0 hours post dose.
+ At the time before exit.
+ At each ambulatory sample (24 hours after dosing).
+ At any time when the subjects feel abnormal.
- Subjects will undergo physical examination before drug administration and at the time
before exit.
- Subjects only exit the clinical area if having agreement of investigator. Entry time and
exit time will be recorded in CRF.
4.4.9. Adverse events and treatment
- Monitor adverse events during dosing and blood sampling, in washout period and 1
week after finishing of sampling.
- The adverse events can be detecsted by following means:
+ When reported by the subjects (report directly in the sampling day or by phone in
washout period or after finishing study).
+ At the time of the vital sign measurements.
- Treatments:
+ Rescue and treat AEs, if occur, according to standard therapeutic regimen (if
necessary):
 Shock: Use standard therapeutic regimens issued by the Ministry of Health
(enclosed with the circular No. 08/1999-TT-BYT dated 4th May, 1999).
 Hypotension: Relax in bed, take about 200 mL of ginger tea mixed with
glucose.
 Hypoglycemia: Relax in bed, drink about 200mL of Glucose 10% or Glucose
20% (w/v) solution or administered slow intravenously of Glucose 10% (w/v)
solution, depending on the severity.
 Adverse events that related to specific investigational products will be treated

following the guidances of the Vietnam National Drug Formulary.
+ For severe cases, after initial treatments, the subjects will be transferred to the intensive
care unit of Vietnam-Cuba hospital (37 Hai Ba Trung, Hanoi) or the intensive care unit of
Dong Da hospital (192- Nguyen Luong Bang, Hanoi).
+ Subjects who experience adverse events will be cared and followed up until they
become well being.
+ All SAEs will be reported to the Sponsor and EC promptly by telephone within 24
hours and in writing within 7 calendar days (for fatal or life-threatening events) or 15
calendar days (for non-fatal or life-threatening events).
+ Discontinue study in case ≥ 20% of subjects experience serious adverse event during
study period and report to related parties as soon as possible according to the current
regulations.
+ All adverse events should be recorded in Case Report Form, including: date and time,
severity, relationship with investigational product, treatments and drug used to treat (if
any) and results after treatment.
4.5. Withdrawal criteria
Subjects can withdraw from the study at any time for any reason. In addition, Principal
Investigators may consider and decide to remove a subject from the study for any of the
following:
- The subject is found to be non compliant to protocol requirements which can affect
on study results.
- The subject experiences adverse events.
- The subject suffers from significant illness or has to undergo surgery during the
study.
- The subject vomits within the first 2 hours after drug administration.
4.6. Replacement of withdrawal subjects
- In case there are subjects withdrawing from the study, Principal Investigator will
discuss with the Sponsor to make a decision of subject replacement for each specific
circumstance.
- If necessary, the withdrawal subjects will be replaced by the stand-by ones in order to
suffice the intended size.
- In case the number of withdrawal subjects is higher than the reserved quantity, CRO
will discuss with the Sponsor to find out the solution. Report to EC before conducting the
dosing and sampling process for additional subjects.
5. BIOANALYTICAL AND STATISTICAL METHODOLOGY
5.1. Method validation
The analytical method by LCMS was validated according to the current guidelines for
bioanalytical method validation with criteria: specificity/selectivity, linearity range, lower
limit of quantification, intra-day and inter-day accuracy and precision, stability of plasma

samples, autosampler stability…
Full validation report (BE/DIL.01F/2010); partial validation report
(BE/LCMS/DIL.01.01P/2015) and 20% chromatograms of validation process are enclosed
in Appendix 3.
Sample extraction procedure: Plasma sample were thawed at room temperature. Pipette
0.5 mL of plasma sample, add the internal standard working solution and 0.1M NH4OH
solution. Mix and then add mixture of diethylether : chloroform (suitable ratio). Shake
and centrifuge to get the upper solvent layer, evaporate to completed dryness under a
nitrogen stream. The residue was dissolved in mobile phase for direct injection into the
LC-MS/MS system.
Chromatographic conditions:
- Column: C18; 100 x 2.1 mm; 1.9 µm.
- Guard column C18; 4 x 3 mm
- Column temperature: 40oC
- Mobile phase: MeCN : MeOH : IPA : 2 mM ammonium acetate (suitable ratio).
- Flow rate: 300 µL/min
- Injection volume: 5 µL.
- Detector: TSQ Quantum Ultra
- Autosampler temperature: 20oC
Mass spectrometric parameters:
Analytes
Diltiazem Felodipine (IS)
Parameters
Ionization mode ESI (+) ESI (+)
Spray voltage (V) 3200 3200
o
Spray temperature ( C) 100 100
Sheath gas pressure (psi) 20 20
Auxilary gas pressure (psi) 5 5
Ion sweep gas pressure (psi) 2 2
o
Capillary temperature ( C) 360 360
Tube lens offset (V) 108 108
Parent ion (Dalton) m/z = 415.4 m/z = 384.1
Collision energy (V) 22 14
Product ion (Dalton) m/z = 177.9 m/z = 338.1
Standards:
Content Water
Standard Purpose Manufacturer Lot No.
(%) (%)
Diltiazem
Standard NIDQC WS.0108218 100.85 0.3
hydrocloride
Felodipine IS NIDQC WS.0107222 99.30 0.06

Certification of standards are inclosed in Appendix 2.
Bioanalytical method validation results are summarized in table 1.
Table 1: Summary of bioanalytical method validation results
Validation criteria Result
Specificity/selectivity Specific with Diltiazem and Felodipine
Carry over No carry-over
Matrix effect No matrix-effect
LLOQ 5 ng/mL
Linearity range 5 – 500 ng/mL
Calibration curve Y = ax + b (r  0,95); Weight = 1/x2
LQC concentration 15 ng/mL
SQC concentration 90 ng/mL
MQC concentration 250 ng/mL
HQC concentration 350 ng/mL
LLOQ : 90.6% CV% : 9.5 %
LQC : 98.8 % CV% : 5.4 %
Intra-day accuracy and precision SQC : 99.6% CV% : 1.3%
MQC : 100.3 % CV% : 2.2 %
HQC : 95.3% CV% : 3.1 %
LLOQ : 96.6% CV% : 10.6%
LQC : 103.0% CV% : 6.1%
Inter-day accuracy and precision
MQC : 100.9% CV% : 4.3%
HQC : 101.2% CV% : 4.8%
Recovery of IS 107.0%
Recovery of Diltiazem LQC: 88.8%; MQC: 91.8%; HQC: 89.0%
Short-term stability of plasma
3.5 hours/ room temperature
samples
Long term stability of plasma
47 days/-350C ± 50C
samples
Freeze and thaw stability of
3 cycles
plasma samples
Auto-sampler stability 21 hours/200C
5.2. Analysis of subject samples
- Check the suitability of the analytical method prior to sample analysis. The analytical
method can be modified but then must be validated partially following the guidelines
before using, if necessary.

- Only analyze the plasma samples of those subjects who participate completely in both
periods.
- Analysis will be performed as soon as possible after finishing the sampling of period 2
and the analytical duration must be within the stability period of plasma sample.
- All the samples of one subject were analyzed in one day and under the same analytical
conditions.
- Samples of subject will not be used for analysis if:
+ The subject withdraws from the study and does not finish both periods.
+ During the study or after finishing the study, the subject is found to be non
compliant to protocol requirements which can affect on study results (such as
subjects use non-study medicine or drinks affecting pharmacokinetics of the study
medicine…).
+ Subjects with four or more missing samples in any period.
+ Subjects with three or more consecutive missing samples in any period.
5.3. Statistical analysis
5.3.1. Pharmacokinetic Parameters
Determine pharmacokinetic parameters of each subject based on drug concentration in
plasma, including:
Maximum concentration (Cmax): directly on measurement
Time to maximum concentration (Tmax): directly on measurement.
Elimination constant (kel): the rate at which drugs are removed from plasma, determined
as slope of linear log-transformed concentration–time curve in elimination phase with at
least 3 concentration values.
Plasma elimination half-life (t1/2): The time required for the plasma concentration of the
drug to eliminate half of its original value, calculated by linearity pharmacokinetic, non-
compartmental model, as formula (1):
0.693
(1) t1/2 =
/ kel /
Area under the curve:
AUC0–t: Using the trapezoidal method from time 0 to last quantifiable concentration time-
point, by formula (2):
(2)
AUC t0   C i  C i 1  t i 1  t i 
n 1
i0 2
in which: Ci is the drug concentration at ti point.
AUC0–: extrapolated from AUC0-t, estimated to inifinity by linearity pharmacokinetic,
non-compartmental model, as formula (3):
(3) Cn
AUC0-∞ = AUC0-t +
/kel/

in which: Cn is the drug concentration at the last quantifiable sampling point
kel is the elimination constant.
Time adjustment: If the actual sampling time deviates from the scheduled time more than
criteria specified in part 4.4.4, adjust measured concentration by the time difference.
5.3.2. Assessment of bioequivalence
Statistical analysis and bioequivalence assessment following ASEAN guidelines (2004).
- Comparison of Cmax and AUC: Analyze the variance (ANOVA), determine the 90%
confidence interval (CI) for the ratio of the Test and Reference products, calculated on
log-transformed data using WinNonlin 5.2 software.
- Comparison of Tmax: using the non – parametric method (Wilcoxon signed – rank test),
based on determining sum of positive/negative rank or p-value according to Z value as
below:
R  N ( N  1) / 4
Z
N ( N  1/ 2)(N  1) / 12
In which: R: Either larger or smaller rank sum
N: Sample size (number of paired Tmax value of the Test and Reference
product that are different)
From Z value, check in table: Cumulative normal distribution to determine the value
Area and the p-value = 1- Area.
- Acceptance criteria for bioequivalence:
+ For Cmax and AUC, 90% confidence interval for the ratio of the Test and
Reference products must be in the range of 80.00% – 125.00%.
+ The difference of T max between the Test and Reference product is not
statistically significant (when sum of positive/negative rank is greater than the theoretical
value with size of n (n is number of paired Tmax value of the Test and Reference product
that are different) or p-value > 0.05).
6. DATA QUALITY ASSURANCE

This study complies with the requirements of Good Laboratory Practice (GLP) and
management of laboratory according to ISO/IEC 17025 concretized on Quality
Management System of National Institute of Drug Quality Control. This also complies
with the ethical principle in the guidelines of WHO’s Good Clinical Practice. Facilities
have been evaluated and inspected periodically. The equipments used in study were
validated and investigators were trained and provided with the knowledge of GLP, GCP
and general methodology for bioequivalence assessment.
7. RESULTS AND DISCUSSION
7.1. Clinical results
Perform subject recruitment, dosing and sampling after study protocol was approved by
Ethics Committee. Duration of clinical phase is from 16/12/2015 to 27/12/2015.

Number of subjects taking part in recruitment : 33
Number of subjects meeting criteria to take part in study (official + standby) : 27
Number of subjects withdrawing from study before dosing of period 1 :0
Number of subjects discontinuing the study after dosing of period 1 : 01
Number of subjects replaced :0
Number of subjects completing the study : 23
Number of subjects used for analysis : 23
Number of subjects used for statistical analysis : 23
Number of subjects had adverse event : 01
Number of subjects used concomitant medication : 01
Time of recruitment : 16/12/15
Time of dosing and sampling in period 1 : 19-20/12/2015
Time of dosing and sampling in period 2 : 26-27/12/2015
Washout period : 07 days
Detailed results are shown in the report of clinical data which is enclosed in Appendix 1.
7.1.1. Subject screening results
24 official and 03 stand-by subjects were recruited to participate in the study. Results of
general examination and screening tests of 24 official subjects are shown in table 2.

Table 2: Results of general examination and screening tests of 24 official subjects
BE/FM/REP.01.01
BE Center-NIDQC
Subj.code
D01 D02 D03 D04 D05 D07 D08 D10 D12 D16 D17 D19
Index
Age (18 - 55) 23 22 23 24 22 23 23 21 19 22 33 20
Heigh (m) 1.61 1.65 1.68 1.65 1.45 1.58 1.63 1.63 1.74 1.65 1.59 1.66
Weight (Kg) 58 59 58 65 43 51 69 57 63 57 53 52
2
BMI (18.0 – 25.0 kg/m ) 22.4 21.7 20.5 23.9 20.5 20.4 25.5 21.4 20.8 20.9 20.9 18.8
Blood pressure
(140/90-90/60 mmHg) 120/60 120/70 110/60 120/60 120/70 120/60 120/70 130/80 100/60 137/80 110/70 140/70
Heart rate
(60-80 rhythm/min) 70 70 70 68 70 79 68 72 72 75 70 79
Breathing rhythm
15/44
(14-20 rhythm/min) 16 18 17 17 18 18 18 18 17 17 17 17
Temperature (36.0-37.5 oC) 36.7 36.5 36.7 36.7 36.5 36.7 36.6 36.6 36.7 36.7 36.7 36.7
9
WBC(4-10x10 /L) 6.3 4.2 6.6 4.5 6.2 7.7 5.4 6.1 6.1 7.3 9.3 4.2
12
RBC (3.8-5.8x10 /L) 5.77 4.93 5.33 5.56 4.24 4.17 4.51 4.77 5.47 5.21 3.92 4.89
BE report – Diltiazem STADA® 60 mg

Hb (12 -16.5g/dL) 16.5 14.9 16.3 16.0 13.4 13.1 13.3 15.3 15.6 16.2 12.5 14.8
HCT (35 - 48%) 50.5 44.8 49.7 47.7 39.0 39.4 37.7 43.7 45.7 48.1 36.6 44.1
9
PLT (150-450x10 /L) 201 240 198 193 376 208 221 230 212 292 194 176
AST (< 40 u/L) 24.6 23.2 17.6 24.4 15.6 31.6 17.1 18.5 17.9 25.0 13.5 21.1
Study number: 45BE 26
ALT (< 41 u/L) 39.9 25.2 15.3 17.2 8.8 34.4 11.3 9.0 15.8 23.5 7.0 6.7
Glucose (4.2-6.4 mmol/L) 4.85 4.30 4.32 4.95 4.93 4.73 4.52 4.33 4.53 4.20 4.74 4.28
Urea (2.5 – 8.07 mmol/L) 4.7 3.6 5.2 5.9 3.5 3.5 7.0 4.6 3.5 4.7 4.0 3.9
Creatinin (44 -106 µmol/L) 76.2 88.2 77.5 78.2 48.2 56.4 63.9 66.0 75.6 82.2 52.3 71.4
HbsAg (<1) 0.227 0.221 0.213 0.251 0.254 0.200 0.178 0.232 0.590 0.246 0.206 0.213
HIV (<1) 0.140 0.163 0.105 0.275 0.240 0.260 0.248 0.271 0.119 0.178 0.311 0.277
BE/FM/REP.01.01
Table 2: Results of general examination and screening tests of 24 official subjects (cont.)
BE Center-NIDQC
Subj.code
D20 D21 D22 D23 D24 D26 D28 D29 D30 D31 D32 D33
Index
Age (18 - 55) 23 23 23 21 21 21 23 21 25 35 21 22
Heigh (m) 1.68 1.59 1.75 1.55 1.49 1.71 1.66 1.65 1.60 1.66 1.62 1.55
Weight (Kg) 58 57 59 48 44 59 57 64 58 63 59 52
2
BMI (18.0 – 25.0 kg/m ) 20.5 22.5 19.2 19.9 19.8 20.1 20.6 23.5 22.6 22.8 22.4 21.6
Blood pressure
(140/90-90/60 mmHg) 130/80 120/70 110/60 105/60 110/60 120/70 120/70 100/60 100/60 110/60 110/60 110/60
Heart rate
(60-80 rhythm/min) 70 70 70 69 67 70 70 72 67 73 69 70
16/44
Breathing rhythm
(14-20 rhythm/min) 18 18 18 18 17 18 18 17 17 17 17 18
Temperature (36.0-37.5 oC) 36.5 36.5 36.5 36.7 36.7 36.6 36.6 36.7 36.7 36.7 36.7 36.5
9
WBC(4-10x10 /L) 5.0 7.1 8.6 5.2 7.8 7.0 6.0 9.5 6.0 7.3 4.6 5.6
12
RBC (3.8-5.8x10 /L) 4.99 5.13 5.09 4.47 4.25 4.77 5.09 4.72 4.47 4.41 4.96 4.62
Hb (12 -16.5g/dL)

14.9 15.3 15.9 14.1 12.6 14.7 15.3 14.7 13.4 13.9 15.9 12.9
HCT (35 - 48%) 44.7 45.3 45.9 42.6 37.7 43.6 44.3 42.9 39.8 41.1 46.8 39.6
9
PLT (150-450x10 /L) 242 255 238 233 261 263 271 252 277 207 247 296
AST (< 40 u/L) 15.1 19.3 23.2 18.0 16.1 22.7 18.1 21.4 25.3 14.1 19.9 24.9
ALT (< 41 u/L) 7.0 25.2 26.2 11.7 5.8 12.1 21.3 16.8 20.4 2.3 11.5 21.3
Glucose (4.2-6.4 mmol/L) 4.68 4.75 4.32 4.67 4.95 4.73 4.56 4.60 4.81 4.91 5.06 4.33
Urea (2.5 – 8.07 mmol/L) 5.7 2.7 4.5 5.3 5.4 4.6 4.8 6.2 3.3 5.9 5.1 4.5
Creatinin (44 -106 µmol/L) 76.2 56.4 81.5 66.9 47.5 84.8 74.9 73.6 50.9 62.5 72.8 69.6
HbsAg (<1) 0.217 0.233 0.211 0.188 0.239 0.197 0.189 0.195 0.213 0.227 0.260 0.206
HIV (<1) 0.172 0.278 0.321 0.315 0.259 0.257 0.100 0.338 0.124 0.193 0.341 0.158
7.1.2. Randomization and dosing schedule
The results of randomization process are shown in Table 3.
Table 3: Randomization and dosing schedule
Drug
No. Subj.Code Sequences
Period 1 Period 2
1 D01 RT R T
2 D02 RT R T
3 D03 TR T R
4 D04 TR T R
5 D05 RT R T
6 D07 TR T R
7 D08 RT R T
8 D10 TR T R
9 D12 TR T R
10 D16 RT R T
11 D17 RT R T
12 D19 TR T R
13 D20 RT R T
14 D21 RT R T
15 D22 TR T R
16 D23 TR T R
17 D24 TR T R
18 D26 RT R T
19 D28 TR T R
20 D29 RT R T
21 D30 TR T R
22 D31 RT R T
23 D32 TR T R
24 D33 RT R T
In which:
T - Test product (1 tablet of Diltiazem STADA® 60 mg )
R - Reference product (1 tablet of TILDIEM® 60 mg )
7.1.3. Dosing and sampling process
- Subject took drugs following sequences in table 3. Take intact drug 30 minutes after
consuming the standard test meal with 240 mL of warm water.
- Bood sampling was performed according to the approved protocol and standard

operation procedure. Subject D23 was removed from the study before period 2 due to
adverse event, so there were only 15 blood samples collected in period 1. Thus, total 705
blood samples of 24 subjects were collected. Actual sampling time of all samples was not
deviated from the scheduled time.
- Food, drink regimen and activities of subjects during dosing and sampling process was
performed according to approved protocol. These main meals were served in NIDQC’s
canteen. The food was controlled and homogenous among subjects for both periods with
following menu:
Contents Menu
(Period 1 + Period 2)
Bread……………….………. 2 slices x 35g
Fried eggs……………………….…..2 eggs
Breakfast
Potato chips……………………………80g
(30 minutes before dosing) Butter………………………………… 10g
Fresh milk……………………………110mL
Vietnamese pork pie ………….……..50g
Sweet and sour pork ribs....................... 100g
Boiled pork meat..................... ............... 100g
Lunch Roasted Peanuts . ....................................100g
(about 4 hours after dosing) Stir-fried bok choy with garlic................150g
Green pumpkin soup with pork rib......200ml
Steamed rice (equivalence to rice) . ........250g
Fried chicken ............................................. 100g
Braised pork ................................................ 100g
Dinner Fried tofu in tomato sauce .......................... 100g
(about 11 hours after dosing) Stir-fried bok choy with garlic.................... 150g
Shrinkage soup with minced pork ...........200ml
Steamed rice (equivalence to rice) ............. 250g
7.1.4. Health monitoring

Subjects were monitored throughout the dosing and sampling duration as recommended in
the protocol.
7.1.5. Adverse events
Subject D23 had dengue before period 2. The subject was removed from the study and
hospotalized to treat adverse event. This adverse event was considerd to be moderate and
unrelate to study medicine
7.1.6. Concomitant medication
Subject D23 took Panadol (paracetamol 500 mg): 2 tablets x 2 times/day x 4 days (from
22/12/2015 – 25/12/2015) because of having fever. This subjects was removed from the
study before period 2.

7.1.7. Protocol deviation
Subject D23 was removed from the study before period 2 due to dengue, so there were 23
subjects completing the study.
7.2. Summary of analytical results
Diltiazem concentration in plasma samples of subjects were determined by the validated
bioanalytical method. Bioanalytical method validation results is summarized in section
5.1.
Analytical procedure started after finishing the second sampling period and last from
06/01/2016 to 12/01/2016 at BE center - NIDQC. All the samples of one subject were
analyzed in one day and under the same analytical conditions. Total 690 samples of 23
subjects were analyzed within 07 days. The period from the first sampling point to the last
day of analysis are 27 days, within the stability period of plasma samples (47 days).
Summary of analytical process is presented in table 4.
Diltiazem concentration in plasma of each subject are presented in Table 5, 6.
The mean plasma concentration versus time curve is presented in Figure 1.
The semi-logarithmic of mean plasma concentration - time curve is presented in Figure 2.
Analytical report and 20% analytical chromatograms are enclosed in Appendix 4.
The individual plasma concentration - time curves are enclosed in Appendix 5.
Table 4: Summary of analytical process
Total analytical samples
Analytical
No. Subject code CC QC Subj.
date
samples samples samples
D01 08 08 30
D02 08 08 30
D03 08 08 30
01 06/01/2016
D04 08 08 30
D05 08 08 30
D07 08 08 30
D16 08 08 30
D08 08 08 30
D10 08 08 30
02 07/01/2016 D12 08 08 30
D17 08 08 30
D19 08 08 30
D20 08 08 30

D21 08 08 30
D22 08 08 30
D24 08 08 30
03 08/01/2016
D26 08 08 30
D28 08 08 30
D29 08 08 30
D30 08 08 30
04 11/01/2016 D31 08 08 30
D32 08 08 30
D33 08 08 30
05 12/01/2016 ISR 08 08 53

Mean concentration - time curve

(n =23)
200.0
Concentration (ng/mL)
TILDIEM® 60 mg
150.0
100.0
50.0
0.0
0.0 6.0 12.0 18.0 24.0
Time (hour)
Figure 1: Mean plasma concentration - time curve of 23 subjects
Semi logarithmic of mean concentration - time curve

(n = 23)
100000.0
Diltiazem STADA® 60 m g
TILDIEM® 60 mg
1000.0
10.0
0.1
0.0 6.0 12.0 18.0 24.0
Time (hour)
Figure 2: Semi-logarithmic of mean plasma concentration - time curve of 23 subjects

Individual plasma concentration - time Individual semi-logarithmic of plasma
curve concentration - time curve
Subject D01 Subject D01

300.0 10000.0
Diltiazem STADA® 60 mg Diltiazem STADA® 60 mg
TILDIEM® 60 mg TILDIEM® 60 mg
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0
Time (hour) Time (hour)

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
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0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
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100.0
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0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
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100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
TILDIEM® 60 mg 1000.0 TILDIEM® 60 mg

200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
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100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0


300.0 10000.0
TILDIEM® 60 mg
TILDIEM® 60 mg 1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

300.0 10000.0
1000.0
200.0
100.0
100.0
10.0
0.0 1.0
0.0 6.0 12.0 18.0 24.0 0.0 6.0 12.0 18.0 24.0

BE/FM/REP.01.01
Table 5: Concentration of Diltiazem in plasma after taking the Test product (ng/mL)
BE Center-NIDQC
Code Sampling time-point (hour)
of
Subj. 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 8.0 10.0 12.0 24.0
D01 0.0 59.7 155.4 170.5 158.7 128.6 104.1 90.0 91.7 64.7 52.0 37.4 25.6 18.4 7.3
D02 0.0 6.9 42.2 82.5 102.1 107.0 92.0 92.6 79.6 59.3 52.1 35.8 23.2 22.0 0.0
D03 0.0 40.2 102.1 222.4 187.1 139.3 130.8 107.8 103.5 75.6 58.9 36.9 23.1 17.4 5.8
D04 0.0 12.2 99.6 138.6 134.0 137.2 96.9 79.6 60.2 48.1 38.4 25.4 17.3 12.2 0.0
D05 0.0 26.6 69.5 95.6 135.2 86.8 68.6 51.0 59.1 40.5 19.4 14.1 13.2 8.7 0.0
30/44
D07 0.0 0.0 9.8 39.8 72.8 85.6 114.6 96.8 77.0 57.5 42.5 32.3 26.2 19.7 0.0
D08 0.0 48.1 115.7 88.1 68.7 50.9 39.8 30.1 24.0 18.0 13.6 10.1 8.0 0.0 0.0

D10 0.0 0.0 7.0 46.6 65.9 78.3 75.7 59.7 51.6 32.3 22.9 16.9 9.4 0.0 0.0
D12 0.0 0.0 49.6 87.7 100.6 105.0 77.1 56.7 51.7 32.9 30.5 24.8 20.7 12.6 0.0
D16 0.0 18.1 70.0 124.6 133.1 116.2 98.0 75.8 65.6 45.7 35.0 26.2 16.9 9.3 0.0
D17 0.0 33.6 82.4 107.1 109.7 155.0 104.6 82.5 80.1 54.3 43.4 36.6 25.7 17.1 0.0
D19 0.0 0.0 0.0 40.9 41.0 42.0 56.4 60.7 74.3 52.4 38.6 30.1 20.3 15.1 0.0
D20 0.0 0.0 20.2 80.6 92.6 102.2 85.2 64.8 51.3 39.1 36.1 27.1 15.9 9.8 0.0
BE/FM/REP.01.01
Table 5: Concentration of Diltiazem in plasma after taking the Test product (ng/mL) (cont.)
BE Center-NIDQC
of
Subj. 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 8.0 10.0 12.0 24.0
D21 0.0 18.7 95.0 163.6 197.8 158.5 129.5 97.5 95.3 64.9 54.9 26.9 17.1 12.6 0.0
D22 0.0 7.7 43.0 66.8 110.9 109.5 75.3 59.5 58.6 42.3 35.4 28.8 21.2 15.1 0.0
D24 0.0 16.9 23.0 44.3 44.2 95.7 70.8 61.9 65.8 65.7 40.8 32.4 22.3 12.9 0.0
D26 0.0 45.0 116.5 137.0 112.3 86.1 62.3 63.9 48.6 34.1 30.3 26.6 14.2 8.3 0.0
D28 0.0 6.4 48.7 129.0 175.2 152.0 123.5 94.2 78.3 62.0 48.4 42.3 31.3 21.9 0.0
31/44
D29 0.0 35.4 48.2 102.6 164.9 124.2 96.2 75.3 64.0 49.2 29.7 21.3 16.1 10.7 0.0
D30 0.0 0.0 64.2 75.0 88.1 72.7 51.2 38.9 34.9 21.3 15.9 12.6 9.0 5.1 0.0
BE report - Diltiazem STADA® 60 mg

D31 0.0 10.2 64.1 66.6 90.2 91.9 78.7 67.1 56.5 32.0 26.4 16.0 11.9 7.5 0.0
D32 0.0 0.0 0.0 0.0 11.8 51.3 55.5 65.0 70.5 50.6 36.7 18.6 12.5 10.6 0.0
D33 0.0 15.2 33.3 53.9 73.0 109.2 91.5 68.2 53.6 37.0 30.9 20.6 15.4 13.3 0.0
Mean 0.0 17.4 59.1 94.1 107.4 103.7 86.0 71.3 65.0 46.9 36.2 26.1 18.1 12.2 0.6
SD 0.0 18.1 41.5 50.7 48.3 32.8 25.1 19.3 18.7 15.0 12.2 8.8 6.1 5.9 1.9
CV
+ 103.9 70.2 53.9 45.0 31.7 29.2 27.1 28.8 31.9 33.6 33.6 34.0 48.6 333.7
(%)
(+): unidentified
BE Center-NIDQC
BE/FM/REP.01.01
Table 6: Concentration of Diltiazem in plasma after taking the Reference product (ng/mL)
of
Subj. 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 8.0 10.0 12.0 24.0
D01 0.0 0.0 66.0 117.4 95.8 83.6 70.8 64.6 59.4 42.9 35.2 25.8 24.0 24.7 9.5
D02 0.0 0.0 0.0 0.0 0.0 8.0 15.0 36.8 48.9 103.7 78.9 56.4 36.0 31.2 7.6
D03 0.0 30.3 83.2 87.2 91.7 87.5 78.2 80.9 67.3 51.0 42.0 32.0 26.8 22.8 7.8
D04 0.0 5.2 60.6 96.7 77.3 62.9 55.2 45.6 36.8 32.0 28.9 29.3 19.7 16.0 0.0
D05 0.0 31.5 46.0 44.1 46.9 38.1 34.4 27.8 22.8 19.1 18.5 14.0 13.3 7.9 0.0
32/44
D07 0.0 0.0 17.9 56.5 64.3 74.6 67.7 67.2 59.2 44.0 38.1 27.2 29.6 21.5 5.5
D08 0.0 11.2 19.5 24.3 30.6 33.2 36.2 36.0 29.5 25.9 28.1 13.7 10.0 6.4 0.0

D10 0.0 0.0 32.8 41.4 36.9 40.0 41.4 40.3 40.5 39.2 29.3 23.4 16.3 9.0 0.0
D12 0.0 29.8 53.1 74.1 72.4 60.5 55.2 52.1 51.0 49.0 42.0 36.1 24.1 17.5 0.0
D16 0.0 8.1 113.8 114.6 106.0 85.9 98.8 83.8 79.7 60.3 44.5 36.5 40.9 34.6 8.9
D17 0.0 0.0 75.0 92.9 88.9 80.9 79.1 62.9 54.9 56.1 44.2 39.0 24.7 17.5 0.0
D19 0.0 0.0 0.0 13.4 31.6 85.8 66.2 54.6 48.5 39.9 30.5 27.6 24.2 19.8 0.0
D20 0.0 9.7 26.6 48.5 45.2 44.2 46.3 40.7 38.9 27.5 29.4 28.9 20.0 15.7 0.0
BE Center-NIDQC
BE/FM/REP.01.01
Table 6: Concentration of Diltiazem in plasma after taking the Reference product (ng/mL) (cont.)
of
Subj. 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 8.0 10.0 12.0 24.0
D21 0.0 8.4 53.4 84.7 88.2 99.3 95.7 85.8 82.7 65.6 51.0 32.0 21.2 16.1 0.0
D22 0.0 0.0 29.7 33.0 40.0 40.7 36.0 38.4 38.8 49.3 41.1 29.9 26.3 21.5 0.0
D24 0.0 5.7 13.6 38.8 50.2 75.2 50.0 59.1 61.9 79.2 60.3 48.1 31.8 26.1 0.0
D26 0.0 20.5 66.5 63.8 54.8 44.7 43.3 40.4 34.2 25.4 17.1 13.1 11.0 9.2 0.0
D28 0.0 14.3 34.2 71.8 131.7 104.4 90.4 90.5 94.3 141.5 114.3 82.0 73.4 50.1 13.8
33/44
D29 0.0 23.5 27.1 31.0 25.8 29.6 29.0 23.6 22.9 23.8 21.9 21.4 23.6 21.3 6.2
D30 0.0 20.7 50.7 50.9 61.6 56.2 47.2 40.8 35.6 25.8 17.6 14.7 14.8 12.3 0.0

D31 0.0 0.0 24.6 56.2 78.6 63.4 70.4 61.2 52.4 48.0 37.5 25.3 16.6 11.5 0.0
D32 0.0 0.0 13.8 23.8 54.4 51.3 45.8 40.3 40.3 46.2 33.0 24.1 19.8 14.5 0.0
D33 0.0 0.0 30.4 33.6 37.0 32.4 29.9 25.1 21.3 19.0 16.4 13.0 10.9 6.6 6.2
Mean 0.0 9.5 40.8 56.5 61.3 60.1 55.8 52.1 48.8 48.4 39.1 30.2 24.3 18.9 2.8
SD 0.0 11.1 27.9 31.6 30.5 25.2 22.7 19.8 19.4 28.6 22.0 15.7 13.3 10.1 4.3
CV
+ 117.2 68.5 56.0 49.7 42.0 40.7 38.0 39.8 59.0 56.2 52.1 54.7 53.7 149.6
(%)
(+): unidentified
7.3. Pharmacokinetic analyses
Parameter Definition Unit
Cmax Maximum observed concentration ng/mL
Tmax Sampling time at which Cmax occured hour
AUC0-t Area under the concentration versus time curve ng.hour/mL

calculated by the trapezoidal rule from time 0 to t*
AUC0-∞ Area to infinity, extrapolated from AUC0-t and ng.hour/mL
plasma elimination half-life
F=AUC0-t/AUC0-∞ Percentage of AUC0-t relative to AUC0-∞ %
F# (%AUCextra) (a) Percentage of extrapolated AUC relative to AUC0-∞ %
t* Last quantifiable concentration time-point Hour
Kel Elimination constant 1/hour
t1/2 Plasma elimination half-life Hour
Elimination phase Points included for calculation of kel, t1/2 and Hour
extrapolation of AUCt-∞ value
(a) F# (%AUCextra ) = (AUC0-∞ - AUC0-t)/AUC0-∞ x 100%

Table 7: Individual pharmacokinetic parameters after taking the Test product
Elimi-
Subj. Sequ- #
Cmax Tmax AUC0-t AUC0-∞ F F t* Kel T1/2 nation
Code ences
phase
D01 RT 170.5 1.5 944.1 994.7 94.9 5.1 24.0 0.144 4.8 1.5-24.0
D02 RT 107.0 2.5 599.8 723.2 82.9 17.1 12.0 0.178 3.9 2.5-12.0
D03 TR 222.4 1.5 983.0 1018.0 96.6 3.4 24.0 0.166 4.2 1.5-24.0
D04 TR 138.6 1.5 597.6 647.5 92.3 7.7 12.0 0.245 2.8 1.5-12.0
D05 RT 135.2 2.0 444.0 477.3 93.0 7.0 12.0 0.260 2.7 2.0-12.0
D07 TR 114.6 3.0 525.4 629.6 83.4 16.6 12.0 0.189 3.7 3.0-12.0
D08 RT 115.7 1.0 305.4 332.0 92.0 8.0 10.0 0.301 2.3 1.0-10.0
D10 TR 78.3 2.5 314.9 347.2 90.7 9.3 10.0 0.290 2.4 2.5-10.0
D12 TR 105.0 2.5 459.5 525.1 87.5 12.5 12.0 0.193 3.6 2.5-12.0
D16 RT 133.1 2.0 560.8 597.1 93.9 6.1 12.0 0.256 2.7 2.0-12.0
D17 RT 155.0 2.5 658.8 742.2 88.8 11.2 12.0 0.205 3.4 2.5-12.0
D19 TR 74.3 4.0 402.6 482.6 83.4 16.6 12.0 0.189 3.7 4.0-12.0
D20 RT 102.2 2.5 450.4 493.6 91.2 8.8 12.0 0.227 3.0 2.5-12.0
D21 RT 197.8 2.0 749.7 795.0 94.3 5.7 12.0 0.278 2.5 2.0-12.0
D22 TR 110.9 2.0 490.8 569.6 86.2 13.8 12.0 0.192 3.6 2.0-12.0
D24 TR 95.7 2.5 476.9 544.1 87.7 12.3 12.0 0.191 3.6 2.5-12.0
D26 RT 137.0 1.5 517.3 551.3 93.8 6.2 12.0 0.245 2.8 1.5-12.0
D28 TR 175.2 2.0 726.9 838.3 86.7 13.3 12.0 0.197 3.5 2.0-12.0
D29 RT 164.9 2.0 550.9 591.5 93.1 6.9 12.0 0.264 2.6 2.0-12.0
D30 TR 88.1 2.0 314.7 333.7 94.3 5.7 12.0 0.268 2.6 2.0-12.0
D31 RT 91.9 2.5 411.7 439.8 93.6 6.4 12.0 0.265 2.6 2.5-12.0
D32 TR 70.5 4.0 323.0 366.3 88.2 11.8 12.0 0.246 2.8 4.0-12.0
D33 RT 109.2 2.5 431.1 491.1 87.8 12.2 12.0 0.221 3.1 2.5-12.0
Mean 125.8 2.3 532.1 588.3 90.3 9.7 12.9 0.227 3.2
SD 40.0 0.7 183.4 192.1 4.0 4.0 3.6 0.043 0.6
CV
31.8 31.9 34.5 32.7 4.5 41.5 27.7 18.9 20.2
(%)

Table 8: Individual pharmacokinetic parameters after taking the Reference product
Elimi-
Subj. Sequ-
Cmax Tmax AUC0-t AUC0-∞ F F# t* Kel T1/2 nation
Code ences
phase
1.5-4.0;
D01 RT 117.4 1.5 718.6 805.1 89.2 10.8 24.0 0.109 6.3
8.0-24.0
D02 RT 103.7 5.0 737.6 795.0 92.8 7.2 24.0 0.132 5.2 5.0-24.0
D03 TR 91.7 2.0 758.2 826.3 91.8 8.2 24.0 0.115 6.0 2.0-24.0
1.5-3.5;
D04 TR 96.7 1.5 418.6 520.5 80.4 19.6 12.0 0.157 4.4
5.0-12.0
D05 RT 46.9 2.0 260.8 311.2 83.8 16.2 12.0 0.156 4.4 2.0-12.0
D07 TR 74.6 2.5 616.9 662.9 93.1 6.9 24.0 0.119 5.8 2.5-24.0
D08 RT 36.2 3.0 239.3 271.7 88.1 11.9 12.0 0.196 3.5 3.0-12.0
D10 TR 41.4 3.0 318.4 373.3 85.3 14.7 12.0 0.165 4.2 3.0-12.0
D12 TR 74.1 1.5 486.5 623.1 78.1 21.9 12.0 0.128 5.4 1.5-12.0
D16 RT 114.6 1.5 943.4 1024.5 92.1 7.9 24.0 0.110 6.3 1.5-24.0
D17 RT 92.9 1.5 548.3 661.6 82.9 17.1 12.0 0.155 4.5 1.5-12.0
2.5-5.0;
D19 TR 85.8 2.5 371.0 513.4 72.3 27.7 12.0 0.139 5.0
8.0-12.0
D20 RT 48.5 1.5 344.7 494.9 69.7 30.3 12.0 0.105 6.6 1.5-12.0
D21 RT 99.3 2.5 584.4 663.1 88.1 11.9 12.0 0.204 3.4 2.5-12.0
D22 TR 49.3 5.0 383.0 568.9 67.3 32.7 12.0 0.115 6.0 5.0-12.0
D24 TR 79.2 5.0 548.4 713.8 76.8 23.2 12.0 0.158 4.4 5.0-12.0
D26 RT 66.5 1.0 301.1 348.5 86.4 13.6 12.0 0.195 3.5 1.0-12.0
D28 TR 141.5 5.0 1396.6 1512.6 92.3 7.7 24.0 0.119 5.8 5.0-24.0
1.5; 2.5-3.0;
D29 RT 31.0 1.5 444.9 537.9 82.7 17.3 24.0 0.067 10.4
5.0-24.0
2.0-5.0;
D30 TR 61.6 2.0 314.4 388.4 81.0 19.0 12.0 0.166 4.2
8.0-12.0
D31 RT 78.6 2.0 415.9 476.1 87.4 12.6 12.0 0.192 3.6 2.0-12.0
D32 TR 54.4 2.0 342.8 455.1 75.3 24.7 12.0 0.129 5.4 2.0-12.0
D33 RT 37.0 2.0 285.3 324.9 87.8 12.2 24.0 0.157 4.4 2.0-12.0
Mean 74.9 2.5 512.1 603.2 83.7 16.3 16.2 0.143 5.2
SD 29.6 1.3 267.0 274.8 7.6 7.6 5.8 0.035 1.5
CV
39.5 51.8 52.1 45.6 9.0 46.3 36.1 24.2 29.2
(%)

7.4. Statistical analyses
7.4.1. Statistical summary of pharmacokinetic parameters
Statistical results of pharmacokinetic parameters are summarized in Table 9.
Table 9: Statistical summary of pharmacokinetic parameters
Cmax Tmax AUC0-t AUC0-
T R T R T R T R
Number of
23 23 23 23 23 23 23 23
subjects
Arithmetic
125.8 74.9 2.26 2.48 532.1 512.1 588.3 603.2
mean
Geometric
120.1 69.1 2.16 2.22 505.1 462.1 560.1 554.5
mean
SD 40.0 29.6 0.72 1.28 183.4 267.0 192.1 274.8
CV (%) 31.8 39.5 31.9 51.8 34.5 52.1 32.7 45.6
Minimum 70.5 31.0 1.00 1.00 305.4 239.3 332.0 271.7
1st quartile 99.0 48.9 2.00 1.50 421.4 330.6 479.9 421.7
Median 114.6 74.6 2.00 2.00 490.8 418.6 551.3 537.9
3 rd quartile 146.8 94.8 2.50 2.75 598.7 600.6 685.4 688.5
Maximum 222.4 141.5 4.00 5.00 983.0 1396.6 1018.0 1512.6

7.4.2. Comparison of pharmacokinetic parameters of the Test versus the Reference
product
The results for comparison of pharmacokinetic parameters of the Test produsct and
Reference product such as Cmax, AUC0-t and AUC0-∞ are summarized in table 10, 11, 12.
Table 10: Individual comparison of Cmax of the Test versus the Reference product
Subj. Cmax Ln(Cmax) Difference Ratio
Ln(T/R)
code T R T R T-R T/R
D01 170.5 117.4 5.139 4.766 53.1 1.452 0.373
D02 107.0 103.7 4.673 4.641 3.3 1.032 0.032
D03 222.4 91.7 5.404 4.518 130.7 2.426 0.886
D04 138.6 96.7 4.932 4.571 42.0 1.434 0.361
D05 135.2 46.9 4.907 3.849 88.3 2.881 1.058
D07 114.6 74.6 4.742 4.312 40.0 1.536 0.429
D08 115.7 36.2 4.751 3.590 79.5 3.195 1.161
D10 78.3 41.4 4.360 3.723 36.9 1.892 0.638
D12 105.0 74.1 4.654 4.305 31.0 1.418 0.349
D16 133.1 114.6 4.891 4.742 18.4 1.161 0.149
D17 155.0 92.9 5.043 4.531 62.1 1.669 0.512
D19 74.3 85.8 4.309 4.452 -11.4 0.867 -0.143
D20 102.2 48.5 4.627 3.881 53.8 2.109 0.746
D21 197.8 99.3 5.287 4.598 98.6 1.993 0.690
D22 110.9 49.3 4.708 3.897 61.6 2.251 0.811
D24 95.7 79.2 4.562 4.372 16.6 1.209 0.190
D26 137.0 66.5 4.920 4.197 70.5 2.060 0.723
D28 175.2 141.5 5.166 4.952 33.8 1.239 0.214
D29 164.9 31.0 5.105 3.432 134.0 5.329 1.673
D30 88.1 61.6 4.478 4.120 26.5 1.430 0.358
D31 91.9 78.6 4.521 4.364 13.4 1.170 0.157
D32 70.5 54.4 4.256 3.997 16.1 1.296 0.259
D33 109.2 37.0 4.693 3.612 72.1 2.948 1.081
Mean 125.8 74.9 4.788 4.236 50.9 1.913 0.553
SD 40.0 29.6 0.310 0.421 37.9 0.982 0.427

Table 11: Individual comparison of AUC0-t of the Test versus the Reference product
Subj. AUC0-t Ln(AUC0-t) Difference Ratio

Ln(T/R)
D01 944.1 718.6 6.850 6.577 225.5 1.314 0.273

D02 599.8 737.6 6.397 6.603 -137.8 0.813 -0.207
D03 983.0 758.2 6.891 6.631 224.8 1.296 0.260
D04 597.6 418.6 6.393 6.037 179.0 1.428 0.356
D05 444.0 260.8 6.096 5.564 183.2 1.702 0.532
D07 525.4 616.9 6.264 6.425 -91.5 0.852 -0.161
D08 305.4 239.3 5.722 5.478 66.1 1.276 0.244
D10 314.9 318.4 5.752 5.763 -3.4 0.989 -0.011
D12 459.5 486.5 6.130 6.187 -27.1 0.944 -0.057
D16 560.8 943.4 6.329 6.850 -382.6 0.594 -0.520
D17 658.8 548.3 6.490 6.307 110.5 1.202 0.184
D19 402.6 371.0 5.998 5.916 31.5 1.085 0.082
D20 450.4 344.7 6.110 5.843 105.6 1.306 0.267
D21 749.7 584.4 6.620 6.371 165.3 1.283 0.249
D22 490.8 383.0 6.196 5.948 107.8 1.282 0.248
D24 476.9 548.4 6.167 6.307 -71.5 0.870 -0.140
D26 517.3 301.1 6.249 5.708 216.2 1.718 0.541
D28 726.9 1396.6 6.589 7.242 -669.7 0.520 -0.653
D29 550.9 444.9 6.311 6.098 106.0 1.238 0.214
D30 314.7 314.4 5.751 5.751 0.3 1.001 0.001
D31 411.7 415.9 6.020 6.030 -4.2 0.990 -0.010
D32 323.0 342.8 5.778 5.837 -19.7 0.942 -0.059
D33 431.1 285.3 6.066 5.654 145.8 1.511 0.413
Mean 532.1 512.1 6.225 6.136 20.0 1.137 0.089
SD 183.4 267.0 0.326 0.444 206.0 0.310 0.299

Table 12: Individual comparison of AUC0-∞ of the Test versus the Reference product
Subj. AUC0- Ln(AUC0-) Difference Ratio

Ln(T/R)
D01 994.7 805.1 6.902 6.691 189.6 1.236 0.212

D02 723.2 795.0 6.584 6.678 -71.8 0.910 -0.095
D03 1018.0 826.3 6.926 6.717 191.7 1.232 0.209
D04 647.5 520.5 6.473 6.255 127.1 1.244 0.218
D05 477.3 311.2 6.168 5.740 166.1 1.534 0.428
D07 629.6 662.9 6.445 6.497 -33.2 0.950 -0.051
D08 332.0 271.7 5.805 5.605 60.3 1.222 0.201
D10 347.2 373.3 5.850 5.922 -26.1 0.930 -0.073
D12 525.1 623.1 6.264 6.435 -98.0 0.843 -0.171
D16 597.1 1024.5 6.392 6.932 -427.4 0.583 -0.540
D17 742.2 661.6 6.610 6.495 80.7 1.122 0.115
D19 482.6 513.4 6.179 6.241 -30.8 0.940 -0.062
D20 493.6 494.9 6.202 6.204 -1.3 0.997 -0.003
D21 795.0 663.1 6.678 6.497 131.9 1.199 0.181
D22 569.6 568.9 6.345 6.344 0.7 1.001 0.001
D24 544.1 713.8 6.299 6.571 -169.7 0.762 -0.271
D26 551.3 348.5 6.312 5.854 202.8 1.582 0.459
D28 838.3 1512.6 6.731 7.322 -674.2 0.554 -0.590
D29 591.5 537.9 6.383 6.288 53.6 1.100 0.095
D30 333.7 388.4 5.810 5.962 -54.7 0.859 -0.152
D31 439.8 476.1 6.086 6.166 -36.2 0.924 -0.079
D32 366.3 455.1 5.903 6.121 -88.9 0.805 -0.217
D33 491.1 324.9 6.197 5.783 166.2 1.512 0.413
Mean 588.3 603.2 6.328 6.318 -14.9 1.045 0.010
SD 192.1 274.8 0.320 0.410 203.0 0.275 0.272

7.4.3. Analysis of variance (ANOVA)
Results of ANOVA for Cmax, AUC0-t, AUC0-∞ are showed in tables 13-16.
Table 13: ANOVA for Cmax
Sum Mean
Variable source Df F P
square square
Total 45 9.5168
Sequence 1 0.0194 0.0194 0.1000 0.7523
Subject 21 3.9881 0.1899 2.2900 0.0322
Formulation 1 3.5072 3.5072 42.270 0.0000
Period 1 0.2598 0.2598 3.1300 0.0913
Error 21 1.7423 0.0830
Root mean square error 0.2881 CV (%) 6.4
Intra-subject variability (%) 29.4% Power (%) 81.19
90% confidence interval for the ratio of the Test and Reference products calculated on
log- transformed data:
Lower limit: 149.09% Upper limit: 199.78%
Table 14: ANOVA for AUC0-t
Sum Mean
square square
Total 45 6.7598
Sequence 1 0.0000 0.0000 0.0000 0.9938
Subject 21 5.6859 0.2708 6.5000 0.0000
Formulation 1 0.0909 0.0909 2.1800 0.1545
Period 1 0.1079 0.1079 2.5900 0.1226
Error 21 0.8751 0.0417
90% confidence interval for the ratio of the Test and Reference products calculated on

Table 15: ANOVA for AUC0-∞
Sum Mean
square square
Total 45 5.9536
Sequence 1 0.0220 0.0220 0.0900 0.7667
Subject 21 5.1141 0.2435 7.5600 0.0000
Formulation 1 0.0011 0.0011 0.0300 0.8539
Period 1 0.1400 0.1400 4.3500 0.0494
Error 21 0.6764 0.0322
90% confidence interval for the ratio of the Test and Reference products, calculated on
Table 16: Summary of comparative bioavailability data

Geometric
Geometric Ratio of
mean of 90% confidence
Parameters mean of Test geometric mean
Reference interval
product (%)
product
Cmax 120.1 69.1 173.8 149.09%-199.78%
AUC0-t 505.1 462.1 109.3 98.12%-120.73%
AUC0- 560.1 554.5 101.0 91.75%-110.10%
7.4.4. Comparison of Tmax

Using the non – parametric method (Wilcoxon signed – rank test) for comparison of T max.
The results are showed in Table 17.

Table 17: Comparison of Tmax by the non-parametric statistical method
Subj. Tmax (hour) Ratio Difference Rank
code T R T/R (+) (-) (+) (-)
D01 1.5 1.5 1.0
D02 2.5 5.0 0.5 2.5 16.5
D03 1.5 2.0 0.8 0.5 5.0
D04 1.5 1.5 1.0
D05 2.0 2.0 1.0
D07 3.0 2.5 1.2 0.5 5.0
D08 1.0 3.0 0.3 2.0 14.5
D10 2.5 3.0 0.8 0.5 5.0
D12 2.5 1.5 1.7 1.0 11.0
D16 2.0 1.5 1.3 0.5 5.0
D17 2.5 1.5 1.7 1.0 11.0
D19 4.0 2.5 1.6 1.5 13.0
D20 2.5 1.5 1.7 1.0 11.0
D21 2.0 2.5 0.8 0.5 5.0
D22 2.0 5.0 0.4 3.0 18.5
D24 2.5 5.0 0.5 2.5 16.5
D26 1.5 1.0 1.5 0.5 5.0
D28 2.0 5.0 0.4 3.0 18.5
D29 2.0 1.5 1.3 0.5 5.0
D30 2.0 2.0 1.0
D31 2.5 2.0 1.3 0.5 5.0
D32 4.0 2.0 2.0 2.0 14.5
D33 2.5 2.0 1.3 0.5 5.0
Mean 2.3 2.5 1.1
SD 0.7 1.3 0.5 Sum 90.5 99.5
The theoretical value with size of n = 19 (n is number of paired Tmax value of the test and
reference product that are different) is 46. Sum of positive rank and sum of negative rank
is more than the theoretical value.
R  N ( N  1) / 4 99.5  19(19  1) / 4
Z   0.18
N ( N  1 / 2)( N  1) / 12 19(19  1 / 2)(19  1) / 12
Area = 0.5714 (base on: Cumulative Normal Distribution)
p-value = 1-0.5714 = 0.4286; more than 0.05
Tmax of test and reference products is no statistically significant difference with 95%
confidence level.

8. CONCLUSIONS
In vivo bioequivalence study for Diltiazem STADA® 60 mg (Diltiazem hydrochloride 60
mg extended-release tablets, Lot No.: 020915, Mfg. date: 170915, Exp. date: 170917;
Reg. No: VD-14553-11; manufactured by STADA - VN Joint Venture Co., Ltd.. versus
TILDIEM® 60 mg (Diltiazem hydrochloride 60 mg tablets), Lot No: 4T014, Mfg. date:
04/2014, Exp. date: 03/2017, Reg. No: VN-17695-14, manufactured by Sanofi Winthrop
Industrie - France, after single-dose administration (1 tablet x 60 mg) in 23 healthy
Vietnamese volunteers under fed condition has the following results:
Mean pharmacokinetic (n=23):

Parameters Cmax  SD AUC0–t  SD AUC0-  SD Tmax 
Products (ng/mL) (ng.hour/mL) (ng.hour/mL) SD (hour)
Diltiazem
125.8  40.0 532.1  183.4 588.3  192.1 2.3  0.7
STADA® 60 mg
TILDIEM® 60 mg 74.9  29.6 512.1  267.0 603.2  274.8 2.5  1.3
Bioequivalence comparison:
- 90% confidence interval for the ratio of the Test and Reference products, calculated on
log-transformed data:
Parameter Acceptance criteria Results
Cmax 80.00% - 125.00% 149.09%-199.78%
AUC0-t 80.00% - 125.00% 98.12%-120.73%
AUC0- 80.00% - 125.00% 91.75%-110.10%
- Tmax: No statistically significant difference with 95% confidence level.
CONCLUSION:
Two products Diltiazem STADA® 60 mg and TILDIEM® 60 mg
are not in vivo bioequivalent under fed condition
according to ASEAN guidelines.

BC Diltiazem Stada

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MINISTRY OF HEALTH

NATIONAL INSTITUTE OF DRUG QUALITY CONTROL

IN VIVO BIOEQUIVALENCE STUDY OF

Sponsor: STADA - VN JOINT VENTURE CO., LTD.

BE/FM/REP.01.01 i Study number: 45BE 26

Position Name Signature/ Date

Date: ................................... 2016

BE/FM/REP.01.01 ii Study number: 45BE 26

This study will assess the bioequivalence of Diltiazem STADA® 60 mg

BE/FM/REP.01.01 iii Study number: 45BE 26

BE/FM/REP.01.01 iv Study number: 45BE 26

BE/FM/REP.01.01 v Study number: 45BE 26

BE/FM/REP.01.01 vi Study number: 45BE 26

BE/FM/REP.01.01 vii Study number: 45BE 26

BE/FM/REP.01.01 viii Study number: 45BE 26

BE/FM/REP.01.01 1/44 Study number: 45BE 26

BE/FM/REP.01.01 2/44 Study number: 45BE 26

BE/FM/REP.01.01 3/44 Study number: 45BE 26

BE/FM/REP.01.01 4/44 Study number: 45BE 26

BE/FM/REP.01.01 5/44 Study number: 45BE 26

BE/FM/REP.01.01 6/44 Study number: 45BE 26

BE/FM/REP.01.01 7/44 Study number: 45BE 26

BE/FM/REP.01.01 8/44 Study number: 45BE 26

BE/FM/REP.01.01 9/44 Study number: 45BE 26

BE/FM/REP.01.01 10/44 Study number: 45BE 26

BE/FM/REP.01.01 11/44 Study number: 45BE 26

BE/FM/REP.01.01 12/44 Study number: 45BE 26

6. DATA QUALITY ASSURANCE

BE/FM/REP.01.01 13/44 Study number: 45BE 26

BE/FM/REP.01.01 14/44 Study number: 45BE 26

BE report – Diltiazem STADA® 60 mg

BE report – Diltiazem STADA® 60 mg

BE/FM/REP.01.01 17/44 Study number: 45BE 26

7.1.4. Health monitoring

BE/FM/REP.01.01 18/44 Study number: 45BE 26

BE/FM/REP.01.01 19/44 Study number: 45BE 26

BE/FM/REP.01.01 20/44 Study number: 45BE 26

Mean concentration - time curve

Figure 1: Mean plasma concentration - time curve of 23 subjects

Semi logarithmic of mean concentration - time curve

Figure 2: Semi-logarithmic of mean plasma concentration - time curve of 23 subjects

BE/FM/REP.01.01 21/44 Study number: 45BE 26

Subject D01 Subject D01

Subject D02 Subject D02

Subject D03 Subject D03

BE/FM/REP.01.01 22/44 Study number: 45BE 26

Subject D04 Subject D04

Subject D05 Subject D05

Subject D07 Subject D07

BE/FM/REP.01.01 23/44 Study number: 45BE 26

Subject D08 Subject D08

Time (hour) Time (hour)

Subject D10 Subject D10

Subject D12 Subject D12

Time (hour) Time (hour)

BE/FM/REP.01.01 24/44 Study number: 45BE 26

Subject D16 Subject D16

Subject D17 Subject D17

Subject D19 Subject D19

BE/FM/REP.01.01 25/44 Study number: 45BE 26

Subject D20 Subject D20

Subject D21 Subject D21

Subject D22 Subject D22