A Double-blind, Randomized, Controlled Clinical Study of the Pharmacokinetics, Pharmacodynamics, Tolerability, and Safety of Multiple Intravenous Injections of BCD-066 and Aranesp® in Healthy Volunteers
Phase: PHASE1
Conditions: Healthy
Sponsor: Biocad

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
Page 1 of 10 
 
 
CLINICAL STUDY PROTOCOL 
 
Protocol Title: 
 
A Double-blind, Randomized, Controlled Clinical Study of the 
Pharmacokinetics, Pharmacodynamics, Tolerability, and Safety 
of Multiple Intravenous Injections of BCD-066 (JSC BIOCAD) 
and Aranesp® (Amgen Europe B.V., the Netherlands) in 
Healthy Volunteers) 
Protocol ID: 
BCD-066-3, NCT03693950 
Protocol Date: 
October 02, 2016 
Protocol Amendment Number: 
Not applicable. 
Protocol Amendment Date: 
Not applicable. 
Protocol Version: 
1.0 
Study Sponsor/Monitor: 
 
JSC BIOCAD, Russia   
Legal address: 34-A, Ul. Svyazi, Strelna, Petrodvortsoviy District, 
Saint Petersburg, Russian Federation, 198515 
Postal address: Petrovo-Dalnee, Krasnogorskiy District, Moscow 
Region, Russian Federation, 143422  
Tel.: +7 (495) 992-66-28, Fax: +7 (495) 992-82-98 
Sponsor’s Contact Person 
Responsible for Signing the 
Protocol and Protocol 
Amendments: 
 
Roman Alexeevich Ivanov, Vice President for Research & 
Development 
Address: JSC BIOCAD, Petrovo Dalnee, Krasnogorskiy District, 
Moscow Region,  
Russian Federation, 143422 
e-mail: ivanov@biocad.ru 
Tel.: + 7 (495) 992-66-28 (ext. 154) 
Sponsor’s Responsible Medical 
Expert: 
Bolsun D.D., Medical Expert, Category II 
JSC BIOCAD, 34 A, Ulitsa Svyazi, Strelna, Petrodvortsoviy 
District, Saint Petersburg, Russian Federation, 198515 
Email: bolsun@biocad.ru 
Tel.: +7 (812) 380 49 33 (ext. 745); 7 (911) 137-89-29 
e-mail: bolsun@biocad.ru 
The information contained in this document is confidential and intended to be used solely by 
investigators, ethics committee members and health authorities. This information may not be 
transferred to any third party without the prior written permission of JSC BIOCAD, except when 
necessary for obtaining the patient’s consent to participate in the study. The above-mentioned 
requirements become effective upon the signing of this Protocol. 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
Page 2 of 10 
 
SYNOPSIS 
Protocol ID 
BCD-066-3, NCT03693950 
Study Title 
A Double-blind, Randomized, Controlled Clinical Study of the 
Pharmacokinetics, Pharmacodynamics, Tolerability, and Safety of 
Multiple Intravenous Injections of BCD-066 (JSC BIOCAD) and 
Aranesp® (Amgen Europe B.V., the Netherlands) in Healthy 
Volunteers 
Phase 
Phase I 
Study Sponsor 
JSC BIOCAD, Russia 
Postal address: Petrovo Dalneye, Krasnogorskiy District, Moscow 
Region, Russian Federation, 143422 
Legal address: 34 A, Ul. Svyazi, Strelna, Petrodvortsoviy District, 
Strelna, St. Petersburg, 198515 
Test Drug 
BCD-066 (INN: darbepoetin alfa, JSC BIOCAD, Russia), solution 
for injection 
Reference Drug 
Aranesp® (INN: darbepoetin alfa, Amgen Europe B.V., the 
Netherlands), solution for injection 
Study Purpose and 
Objectives 
The purpose of the study is to confirm the equivalent 
pharmacokinetics, pharmacodynamics, safety, and tolerability of 
multiple IV injections of BCD-066 and Aranesp® in healthy 
volunteers. 
Objectives: 
1. To evaluate and compare the key pharmacokinetic 
parameters of darbepoetin alfa in the serum after repeated-
dose intravenous injections of 1 μg/kg BCD-066 (JSC 
BIOCAD, Russia) and 1 μg/kg Aranesp® (Amgen Europe 
B.V., the Netherlands) in healthy volunteers. 
2. To evaluate and compare the key hemoglobin-based 
pharmacodynamic 
parameters 
after 
repeated-dose 
intravenous injections of 1 μg/kg BCD-066 (JSC BIOCAD, 
Russia) and 1 μg/kg Aranesp® (Amgen Europe B.V., the 
Netherlands) in healthy volunteers. 
3. To evaluate and compare the safety and tolerability 
characteristics after repeated-dose intravenous injections of 
1 μg/kg BCD-066 (JSC BIOCAD, Russia) and 1 μg/kg 
Aranesp® (Amgen Europe B.V., the Netherlands) in healthy 
volunteers. 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
Page 3 of 10 
 
Study Design 
Clinical study BCD-066-3 is a double-blind controlled, 
randomized, parallel-group study of the pharmacokinetics, 
pharmacodynamics, tolerability, and safety of repeated-dose 
intravenous BCD-066 and Aranesp®. 
The study is planned to include 62 healthy male volunteers 
(56 active participants and 6 back up subjects to replace dropouts). 
Before inclusion in the study, all potential participants are given full 
information about the study. This information is presented in the 
Participant Information Sheet. If the potential participant gives his 
consent to be in the study, he has to sign the Informed Consent Form 
for participation in a clinical study and then undergo a 14-day 
screening examination to confirm that he is eligible for the study.  
After passing the screening and being considered eligible for 
the study, subjects are centrally randomized at a 1:1 ratio to one of 
two treatment groups (#1 and #2). Subjects in Group #1 will receive 
IV injections of 1.0 μg/kg BCD-066 on Day 1, Day 8, Day 15, and 
Day 22. Subjects in Group #2 will receive IV injections 1 μg/kg 
Aranesp® on Day 1, Day 8, Day 15, and Day 22. This is a double-
blind study, which means that neither the investigator nor the 
subject will know what drug is used in each group. 
Blood samples for evaluation of darbepoetin alfa 
concentration will be collected 30 min, 20 min, and 10 min before 
the injection, immediately before the injection (not more than 5 min 
before drug administration), and then 5 min, 10 min, 15 min, 20 
min, 30 min, 45 min, 1 h, 2 h, 4 h, 8 h, 16 h, 24 h, 48 h, and 72 h 
after the first and the fourth IV injection. There will be 4 blood 
collections before and 14 blood collections after the first and the 
fourth IV injection of darbepoetin alfa. The serum concentrations 
will be used to calculate the key PK parameters of darbepoetin alfa. 
Before the second and the third injection, blood samples will be 
collected only once. During the entire study, there will be 38 blood 
collections for PK evaluation. 
Blood samples for evaluation of hemoglobin (the primary PD 
endpoint) and secondary PD parameters (reticulocytes, RBC, 
hematocrit) will be collected before each injection of the test 
drug/reference drug, then 1 day, 3 days, and 5 days after the first, 
second, and third injection, and 1 day, 3 days, and 7 days after the 
fourth injection. Two more blood collections will be performed for 
immunogenicity assessment (at screening and on Day 29). 
This study includes a screening examination (not more than 
14 days) and 16 study visits. After the first and fourth injection of 
the test/reference drug, all volunteers will be required to stay in the 
clinic for at least 24 h for monitoring purposes. The duration of the 
study from the first drug injection to the final visit is 29 days. 
Special requirements to volunteers 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
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Study subjects are required not to drink any alcohol for 24 h 
before and 72 h after each injection of BCD-066/Aranesp®; not to 
drink more than 10 units of alcohol per week (1 unit of alcohol is 
equivalent to 0.5 L of beer, 200 mL of wine, or 50 mL of a strong 
alcohol beverage); and not to smoke for 1 h before and 3 h after 
each injection of BCD-066/Aranesp® and for 1 h before each blood 
pressure measurement.  
Study Population 
Healthy male volunteers from 18 to 45 years old (inclusive). 
Planned Sample Size 
Sixty-two healthy male volunteers (56 volunteers in the main 
sample plus 6 backup volunteers to replace the dropouts). 
Inclusion Criteria 
1. Signed informed consent form  
2. Men from 18 to 45 years old (inclusive) 
3. BMI within the normal limits (18.5 to 30 kg/m2) 
4. Hemoglobin from 120 g/L to 150 g/L and hematocrit 41% to 
49% at screening (before the first injection) 
5. Serum transferring 2.15 g/L to 3.6 g/L; serum ferritin from 
20 μg/L to 250 μg/L 
6. Vitamin B12 from 187 pcg/mL to 883 pcg/mL; folic acid 
from 3.1 ng/mL to 20.5 ng/mL 
7. Endogenous serum erythropoietin < 30 mIU/mL at screening 
8. The subject is verified as “Healthy” according to results of 
standard clinical, laboratory and instrumental tests 
9. Subject’s ability (in the investigator’s opinion) to follow the 
protocol procedures 
10. The subject and his sexual partner with retained childbearing 
potential consent to implement reliable contraceptive 
methods starting 2 weeks before inclusion in the study and 
up to 2 weeks after the last dose of the test/reference drug. 
This requirement does not apply to surgically sterile subjects. 
Reliable contraception methods mean one barrier method in 
combination with one of the following: spermicides, 
intrauterine device and/or oral contraceptives used by the 
subject's partner. 
11. The subject agrees not to drink alcohol for 24 h prior to each 
injection of the test/reference drug and for 72 h after the 
injection. 
Exclusion Criteria 
1. Psychiatric disorders or other conditions that can affect the 
ability of the subject to follow the study protocol 
2. Acute infections within 4 weeks before the study start 
3. Results of laboratory and/or instrumental tests are outside 
the normal range 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
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4. Chronic cardiovascular, bronchial and/or pulmonary, 
neuroendocrine GI, liver, kidney, and blood diseases, 
including CAD, arterial hypertension, peripheral vascular 
and/or cerebral vascular disorders, and thrombocytosis  
5. A history of chronic hemorrhages 
6. Any malignancy (or a prior history of malignancy) 
7. Prior treatment with any erythropoietin/darbepoetin product 
or other products promoting erythropoiesis received at any 
time before enrollment 
8. Intravenous iron therapy within two years prior to 
enrollment 
9. Treatment with any drugs (including over-the-counter 
drugs, herbal drugs, or nutritional supplements) within 14 
days prior to the first administration of the investigational 
product. The restriction does not apply to the use of 
paracetamol (less than 3 g a day) or ibuprofen (less than 1 g 
a day) 
10. Epileptic seizures within six months prior to the first 
administration of the investigational product 
11. Extensive surgery within one month before enrollment 
12. Impossibility to insert an intravenous catheter for blood 
sampling (e.g., because of a skin condition at the 
venepuncture site) 
13. Hypersensitivity to any components of BCD-066 (JSC 
BIOCAD), 
Aranesp® 
(Amgen 
Europe 
B.V., 
the 
Netherlands) or drug products of the same therapeutic 
category; intolerance to erythropoietins and/or other 
recombinant human proteins; intolerance to iron (III) 
products 
14. Episodes of thrombosis and/or thromboembolism in the past 
(myocardial infarction, stroke, transient ischemic attacks, 
deep vein thrombosis, pulmonary embolism within 6 
months before enrollment); increased risk of deep vein 
thrombosis 
15. Known severe allergies (anaphylaxis or multiple drug 
allergy) 
16. Antibodies to darbepoetin alfa at screening 
17. Smoking of more than 10 cigarettes a day 
18. The subject consumes more than 10 units of alcohol per 
week (1 unit equals to 0.5 L of beer, 200 mL of wine or 50 
mL of a strong alcohol beverage) or has a history of alcohol, 
recreational drug or medication abuse 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
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Version 1.0 dated 02-Oct-2016 
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19. Acute hemorrhage or blood/plasma donation or blood 
transfusion within 2 months before enrollment 
20. Participation in other clinical studies within 30 calendar 
days before enrollment in this study 
21. Prior participation in this study. 
Study Therapy 
The study subjects will receive 1 μg/kg darbepoetin alfa (BCD-066 
or Aranesp®) as weekly IV injections on Day 1, Day 8, Day 15, and 
Day 22. Blood samples will be collected for investigation of the PK 
and PD. 
Study Procedures 
In this study, blood samples will be collected according to the 
schedule to measure the concentration of darbepoetin alfa, evaluate 
the hemoglobin-based PD parameters, and to evaluate additional 
characteristics of the red blood (reticulocytes, RBC, hematocrit). 
The study also involves procedures to monitor the safety. 
Blood samples for darbepoetin alfa concentration will be collected 
before and after each IV injection of the test/reference drug. 
Collections will be performed as follows: 
• Visit 1 (Day 1 – Day 2): 30 min, 20 min, 10 min and 
immediately before the first injection of darbepoetin alfa and 
then 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 1 h, 2 h, 
4 h, 8 h, 16 h, and 24 h after the first injection. 
• Visit 2 (Day 3): 48 h after the first injection.  
• Visit 3 (Day 4): 72 h after the first injection.  
• Visit 5 (Day 8): immediately before the second injection. 
• Visit 9 (Day 15): immediately before the third injection. 
• Visit 13, (Day 22 – Day 23): 30 min, 20 min, 10 min and 
immediately before the fourth injection of darbepoetin alfa 
and then 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 1 h, 
2 h, 4 h, 8 h, 16 h, and 24 h after the fourth injection. 
• Visit 14 (Day 24): 48 h after the fourth injection. 
• Visit 15 (Day 25): 72 h after the fourth injection. 
Blood samples for PD study will be collected as follows: 
• Visit 1 (Day 1 – Day 2): before the injection and 24 h (one 
day) after the first injection. 
• Visit 3 (Day 4): 72 h (3 days) after the first injection. 
• Visit 4 (Day 6): 120 h (5 days) after the first injection. 
• Visit 5 (Day 8): 168 h after the first injection (immediately 
before the second injection). 
• Visit 6 (Day 9): 192 h after the first injection (24 h after the 
second injection). 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
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• Visit 7 (Day 11): 240 h after the first injection (3 days after 
the second injection). 
• Visit 8 (Day 13): 288 h after the first injection (5 days after 
the second injection). 
• Visit 9 (Day 15): 336 h after the first injection (immediately 
before the third injection). 
• Visit 10 (Day 16): 360 h after the first injection (one day after 
the third injection). 
• Visit 11 (Day 18): 408 h after the first injection (3 days after 
the third injection). 
• Visit 12 (Day 20): 432 h after the first injection (5 days after 
the third injection). 
• Visit 13 (Day 22 – Day 23): 504 h after the first injection 
(immediately before the fourth injection); 528 h after the first 
injection (one day after the fourth injection). 
• Visit 15 (Day 25): 576 h after the first injection (3 days after 
the fourth injection). 
• Visit 16 (Day 29): 672 h after the first injection (7 days after 
the fourth injection). 
Safety monitoring procedures: 
For safety evaluation, the study team will monitor vital signs (BP, 
pulse, respiration rate, and body temperature), perform regular 
physical examinations including assessment of the injection site, 
and run hematology and chemistry tests, blood tests for anti-
darbepoetin alfa binding and neutralizing antibodies, urinalysis, and 
ECG. 
Total Study Duration 
Expected study duration is 2 years including up to 18 months of 
enrollment and 6 months of data collection and statistical 
processing. 
Each study subject is expected to be in the study for 64 days, which 
includes the screening (14 days), administration of the 
test/reference drug and investigation of the PK and PD (29 days), 
and the follow-up period (Day 30 to Day 50).  
PK Endpoints 
• Endpoints: 
− The area under the Time vs. Concentration curve for 
darbepoetin alfa from injection to 72 h (AUC(0-72)) after the 
first and the fourth IV injection of BCD-066 or Aranesp® 
− The maximum concentration of darbepoetin alfa in the 
serum after the first and the fourth IV injection of BCD-066 
or Aranesp® (Cmax) 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
CONFIDENTIAL 
Version 1.0 dated 02-Oct-2016 
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− The elimination half-life (T½)  
− The total area under the Concentration vs. Time curve from 
0 to infinity (AUC0-168) 
− 
The time to maximum drug concentration in the serum 
(Tmax) 
− 
The residual area 
− 
The elimination constant (kel) 
− 
The total clearance (CL). 
 
Methods used for PK assessment: 
The PK analysis will include all subjects who have received 
all doses of the test/reference drug as per protocol. 
Not more than 2 PK blood collections after the first injection 
and not more than 2 PK blood collections after the fourth injection 
can be missed. Subjects who have two consecutive blood samples 
missed will be not included in the analysis.  
The PK parameters will be analyzed based on darbepoetin 
alfa concentrations in the subject’s serum. 
 
PD Endpoints 
Endpoints: 
• Primary endpoint: 
⎯ The area under the baseline-adjusted Effect vs. Time 
curve from injection to Day 29 after the repeated-
dose IV administration of the test/reference drug 
(AUEC (1-29)). 
• Secondary endpoints: 
⎯ The maximum increase in hemoglobin from 
baseline, estimated from injection to Day 29 after 
the repeated-dose IV administration of the 
test/reference drug (AC-Emax). 
Supplementary endpoints: 
⎯ The time to the absolute maximum increase in 
hemoglobin from injection to Day 29 after the 
repeated-dose IV administration of the test/reference 
drug (Tmax). 
In addition, PD parameters (AUEC, AC-Emax, and Tmax) will be 
estimated based on the reticulocyte and RBC counts and on 
hematocrit values. 
 
Methods used for pharmacodynamic assessment: 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
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Version 1.0 dated 02-Oct-2016 
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The PD analysis will include all subjects who have received 
all doses of the test/reference drug as per protocol. 
Not more than 3 PD blood collections can be missed over 
the entire study. Subjects who have two consecutive blood samples 
missed will be not included in the analysis. 
The PD analysis will be based on the hemoglobin 
concentrations, reticulocyte and RBC counts, and hematocrit. 
The limiting hemoglobin level in this study is 180 g/L. If the subject 
reaches this level before the last injection of darbepoetin alfa, he 
will be withdrawn from the study. If the subject reaches the limiting 
Hb concentration after he receives the last (fourth) injection of 
darbepoetin alfa, he will not be withdrawn from the study. 
Safety Assessment 
Endpoints: 
• The proportion of subjects who developed AE(s) after 
receiving at one injection of darbepoetin alfa 
• The rate of SAEs 
• The rate of  CTCAE grade 3/4 AEs 
• The rate of withdrawals due to AEs 
• The proportion of subjects who had injection site reactions 
• The proportion of subjects who had antibodies to 
darbepoetin alfa on Day 29. 
Methods used for safety assessment: 
The safety analysis will include all subjects who have 
received at least one dose of the test/reference drug. 
The safety will be analyzed based on the information about: 
− The reported AEs/SAEs, 
− Results of physical examinations with an assessment of the 
injection site, vital signs, and instrumental and laboratory 
test results (hematology and chemistry, blood testing for 
binding and neutralizing anti-darbepoetin alfa antibodies, 
and urinalysis). 
Statistical Analysis 
Determination of sample size 
The sample size was determined using the variation coefficients for 
hemoglobin increase (Hb AUEC) obtained by Cheung, 20011. The 
estimated Type I error was 5% (α=0.05) and Type II error was 20% 
(β=0.2), with the test power of 80%.  
The sample size was calculated with the “sampleN.TOST” function 
of the “PowerTOST” package for R. The calculation used the CV 
values and the ratios of the mean Hb increases (AUEC). The study 
                                                          
1 W.Cheung, N. Minton, K.Gunawardena. Pharmacokinetics and pharmacodynamics of epoetin alfa once 
weekly an three times weekly, 2001 

Clinical Study Protocol 
Protocol ID: BCD-066-3 
NCT03693950 
 
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Version 1.0 dated 02-Oct-2016 
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should include 28 subjects per group and 6 subjects as backups to 
replace potential early withdrawals (62 volunteers total). 
Methods used to analyze PK and PD parameters 
Descriptive statistics for the data on drug concentration in 
the serum, effect values, and protocol-specified PK and PD 
parameters will be performed using the following: mean values, 
geometric mean values, standard deviations, medians, upper and 
lower quartiles, min and max, and the coefficient of variation.  
The statistical comparison involves the calculation of two-
sided parametric 90% confidence intervals (CIs) for the ratios of the 
AUEC(1-29) means for hemoglobin and darbepoetin alfa following 
the IV injection of BCD-066 and Aranesp®, and comparison of 
these CIs with the  PD equivalence interval: 
AUEC(1-29),:  [0.8; 1.25] (80-125%). 
The study drugs will be considered equivalent if the 
estimated 90% CIs for the mean AUEC(1-29) ratio for darbepoetin in 
BCD-066 and darbepoetin in Aranesp® will fall in the said range. 
Methods used to analyze the safety and demographic data 
The interval (quantitative) data will be presented as mean 
values, standard deviations, medians, upper and lower quartiles, 
min and max. The normality of the interval data will be tested with 
the Shapiro-Wilk test. 
The categorical (qualitative) data will be presented as the 
absolute and relative (proportions or percentages) number of 
observations.  
The normally distributed interval data will be compared 
using the ANOVA and t-test for dependent and independent 
samples. Non-normally distributed interval data will be analyzed 
using non-parametric tests: Friedman’s ANOVA, Kruskal-Wallis H 
test, Wilcoxon T test, and Mann-Whitney test. Categorical data will 
be compared using the χ2 test or the Fisher’s exact test. 
Correction for multiple comparisons will be performed with 
the Benjamini-Yekutieli procedure.  
Statistical methods will be chosen based on the type and distribution 
of raw data. Applicability of certain statistical tests will be 
evaluated after all the data are collected because it is impossible to 
predict the distribution pattern, data homogeneity and other data 
characteristics in advance.