Comparative Pharmacokinetic and Pharmacodynamic Study Between 2 Agalsidase Beta Formulations at a Single Dose of 1 mg/kg of Agalsidase (Biosidus) and Fabrazyme (Sanofi Genzyme) as Intravenous Infusion in Male Healthy Volunteers
Phase: PHASE1
Conditions: Fabry Disease
Sponsor: Bio Sidus SA

 
 
PROTOCOL 
 
COMPARATIVE STUDY ON THE PHARMACOKINETICS AND PHARMACODYNAMICS 
BETWEEN TWO AGALSIDASE BETA FORMULATIONS IN A SINGLE DOSE OF 1 
MG/KG AGALSIDASE (BIOSIDUS) AND FABRAZYME® (SANOFI GENZYME) BY 
INTRAVENOUS INFUSION IN HEALTHY MALE VOLUNTEERS 
 
Protocol Number: RD-BIO-AGA-01/03/21 
 
Date of Protocol:  
Version: Amendment 2 – September 2, 2021 
 
 
Sponsor: Biosidus S.A. 
 
 
 
 
 
 
 
 
 
This document contains confidential information, unless otherwise agreed in writing. By 
accepting or reviewing this material, you agree to keep such information secret and not to 
copy and disclose it to any third party (except as required by the applicable law), or use it for 
unauthorized purposes. 
 
 

SIGNATURES AND OTHER PARTICIPANTS 
Protocol Nº RD-BIO-AGA-01/03/21 
 
APPROVAL 
 
Sponsor and Principal Investigator ´s signatures confirm the approval of this protocol and ensure that the 
study will be conducted in accordance with the clinical and administrative sections of the protocol and the CRF 
book, including all confidentiality agreements.   
 
The Principal Investigator is informed of his/her responsibilities and of the Good Clinical Practice and local 
Regulations, and agrees to conduct this study in accordance with these standards and cooperate with the staff 
involved. 
 
 
Sponsor: 
Biosidus S.A. 
Constitución 4232 (1254), C.A.B.A, Argentina 
Office. +541149098000  ext 8404 
Signature:  
 
 
Date: 
 
 
Principal Investigator  
Eduardo Pirotzky, MD 
Lavalle 1537 10 E (1048) 
C.A.B.A., Buenos Aires, Argentina  
Phone: (+54 9 11) 6554-7584 
 
 
 
Signature:  
 
 
Date: 
 
 
Co Investigador 
Hernán Taconelli, MD 
Sanatorio Nuestra Señora del Pilar  
Av. Gaona 3979, (C.P.: 1702) 
Ciudadela, Prov. de Buenos Aires  
Phone: (+54 11) 6846-3670 
 
 
Signature: 
 
 
Date: 

Other Participants 
 
Statistical Analysis: 
K&H Consulting S.A.SL. 
Sarmiento 4508, 8°B, 
Ciudad Autónoma de Buenos Aires - Argentina 
Phone.:4952-1360 
 
Medical Site: 
Sanatorio Nuestra Señora del Pilar 
Av. Gaona 3979 (B1702API) Ciudadela, Provincia 
de Buenos Aires - Argentina 
Phone: 011 4469-6700 
 
Virological Analysis Laboratory:  
Immune Virology Laboratory  
Academia Nacional de Medicina Pacheco de Melo 
3081 3er Piso CABA, Argentina 
Phone +54 9 11 5497-5179 
 
Immunogenicity Laboratory 
 
 
Biosidus Department of Development 
Constitución 4234 - C1254ABX Buenos Aires, 
Argentina –  
Phone.4909 8000 - Fax 4909 8055. 
 
Pharmacokinetics Laboratory: 
Shin Nippon Biomedical Laboratories, Ltd 
2438, Miyanoura, Kagoshima 891-1394, Japan, 
Phone: +81-99-294-2600; fax: +81-99-294-3619 
 
Enzyme Activity Laboratory: 
 
Biosidus Department of Development  
Constitución 4234 - C1254ABX Buenos Aires, 
Argentina –Phone 4909 8000 - Fax 4909 8055. 
 
Ethics Committee Participant: 
Research Ethics Committee at DIM Clínica 
Privada 
Avenida Rivadavia 17.624, Morón, Provincia de 
Buenos Aires Argentina, Phone: 5554-8904/8905 
 
 
 
 

Version History 
 
Number 
of
substantial 
amendment 
Final version 
involved 
Nature of the amendment 
1 
17Aug2021 
Request for changes by the Regulatory Authority –Communication #01 
Dose selection is accounted for 
Changes were made to the window periods for COVID19 vaccines for 
the first and second doses 
The possibility of voluntarily withdrawing from the study is made explicit 
to the volunteer given the immediate possibility of receiving COVID19 
vaccine  
The number of volunteers to be hospitalized is clarified 
2 
02Sept2021 Response to stopclock by ANMAT dated 13 September, 2021 
 
 
Rationale was extended to justify the dose 
The possibility of receiving a COVID vaccine was added as a criterion 
for withdrawal from the study 
The number of volunteers to be admitted was changed 
The window of time in which the volunteer may receive COVID-19 
vaccine was clarified in an inclusion criterion 
 
Amendment Version 1 – 17Aug2021 
Reviews for version dated July 20, 2021 are the following: 
Justification for dose selection: 
Section # 7 DOSING REGIMEN AND ADMINISTRATION, the following statement was added to account 
for the selected dose: 
“A dose of 1mg/kg is the one that did not cause adverse events in preclinical animal studies, conducted 
with the same Biosidus molecule, which defines this dose as the NOAEL (Non observed adverse event 
level).” 
Changes in the exclusion criteria: 
New Exclusion Criterion #7 (Exclusion Criterion #7: Reception of live or dead virus, bacteria 
vaccines, including COVID-19 vaccine (In this case, patients who have received a single dose of 
vaccine in the month prior to recruitment will be excluded.) 
It was divided into two parts to clearly state the window periods for the COVID-19 vaccine): 
Exclusion Criterion #7: Reception of live or dead virus, bacteria vaccines within fifteen days prior to 
visit 1 or 2. 
The following Exclusion Criterion is added: 
Exclusion Criterion #8: In the case of COVID vaccines, a window period longer than 15 days must 
be observed before visits 1 or 2. This period is contemplated based on the recommendations by 
the National Ministry of Health published on 5-29-2021. 

Possibility of withdrawing from the study due to the potential for receiving COVID-19 vaccine: 
In the section STUDY COURSE AND PROCEDURES the following statement was added related to 
COVID-19 Vaccine: 
“Note: The participant may withdraw from the study at any time if they decide to receive the first or 
second dose of the COVID vaccine during the study.” 
Number of volunteers to be hospitalized: 
The following clarification regarding the number of volunteers to be hospitalized was added to the item 
VISIT II (Hospitalization): 
 “The rest of the volunteers will be hospitalized in three groups of 6 volunteers (in compliance with 
the number of volunteers for bioequivalence studies according to authorizing provision, see Annex F) 
as described in the section Study course and Procedure - Visit II (Hospitalization).” 
 
Amendment Version 2 – 02Sept2021 
Revisions for Amendment 1 dated August 17, 2021 are the following: 
Dose justification: 
The rationale was extended to justify the recommended dose of 1 mg/kg /body weight of agalsidase beta. 
Withdrawal from the study due to COVID vaccine: 
The following text was removed: “Note: the participant may withdraw from the study at any time if they 
decide to receive the first or second dose of the COVID vaccine during the study”. The text was modified 
and included as a criterion for study withdrawal: "Participant who has the possibility of receiving the first or 
second dose of the COVID vaccine during the study” 
Number of patients to be hospitalized: 
In relation to the site authorization for the development of phase 1 clinical trials and bioequivalence 
by Provision No. 5344/2019, as it is a phase I trial, the use of up to 2 simultaneous beds is 
authorized. For this reason, the hospitalization of the volunteers was adapted as authorized and a 
staggered hospitalization will be carried out (see Visit II (Hospitalization)). 
Clarification for exclusion criterion: 
The following text was added to exclusion criterion #8: “Volunteer may receive the vaccine 35 days 
after receiving the investigational product.” to make clear the time window in which the 
volunteer may receive the COVID-19 vaccine. 
 
 

PROTOCOL SUMMARY 
TITLE 
COMPARATIVE 
STUDY 
ON 
THE 
PHARMACOKINETICS 
AND 
PHARMACODYNAMICS 
BETWEEN 
TWO 
AGALSIDASE 
BETA 
FORMULATIONS IN A SINGLE DOSE OF 1 MG/KG AGALSIDASE 
(BIOSIDUS) 
AND 
FABRAZYME® 
(SANOFI 
GENZYME) 
BY 
INTRAVENOUS INFUSION IN HEALTHY MALE VOLUNTEERS 
CODE 
RD-BIO-AGA-01/03/21 
SPONSOR 
Biosidus S.A.  
Constitución 4234 (1254) Buenos Aires  
Phone number: 54-11-4909-8049 / 8000  
FAX 54-11-4924-3601 
PRINCIPAL 
INVESTIGATOR 
Eduardo Pirotzky, MD 
Lavalle 1537 10º E, Ciudad Autónoma de Buenos Aires, Argentina 
Phone: 15-6554-7584 
CO – INVESTIGATOR 
Hernán Taconelli, MD 
REGULATORY HEAD 
Pablo Liuboschitz 
SITE  
Sanatorio Nuestra Señora del Pilar 
Av. Gaona 3979 (B1702API) Ciudadela, Provincia de Buenos Aires - 
Argentina 
LABORATORIES 
• 
Enzyme Activity Assessment: 
Biosidus Develpment Department - Constitución 4234 - C1254ABX 
Buenos Aires, Argentina -  Phone.4909 8000 - Fax 4909 8055 
• 
Immunogenicity Assessment: 
Biosidus Develpment Department - Constitución 4234 - C1254ABX 
Buenos Aires, Argentina -  Phone.4909 8000 - Fax 4909 8055 
• 
Pharmacokinetic Assessment: 
Shin Nippon Biomedical Laboratories, Ltd., 2438, Miyanoura, 
Kagoshima 891-1394, Japan, Phone: +81-99-294-2600; fax: +81-99-
294-3619 
PRIMARY OBJECTIVE 
Compare the pharmacokinetic behavior of Biosidus Agalsidase beta and 
Fabrazyme® (Genzyme) after administration by intravenous infusion of 1 
mg/kg over a period of 5 hours of one of the following formulations: 
- Fabrazyme®, by Sanofi Genzyme (Reference Formulation, “R”) 
- Agalsidase beta, biosimilar developed by Biosidus SA (Test 
Formulation, “T”) 

SECONDARY 
OBJECTIVE 
Evaluation of enzyme activity, product safety, incidence of adverse events 
and tolerance. Evaluation of antibody occurrence. 
STUDY DESIGN  
Open, randomized, parallel-arms, balanced, single-dose, and sequential 
DISCONTINUATION 
SCHEDULE 
The sponsor and investigators may independently discontinue the study 
at any time at the sole possibility or occurrence of a serious adverse 
event or situation that could affect the volunteer's health. 
NUMBER 
OF 
VOLUNTEERS 
A total of 20 (twenty) male volunteers will initially be included. As it is a 
sequential design, if the statistical analysis on the first 20 volunteers does 
not demonstrate bioequivalence, an additional number of healthy 
volunteers to be determined will be included to verify the similarity 
criterion with adequate power. 
 
 
 
 
 
 
 
 
 
SCREENING CRITERIA 
 
INCLUSION CRITERIA: 
1) Male subjects aged between 18 and 40 years. 
2) Body mass index (BMI) between 19 and 25 kg/m². 
3) Volunteers whose investigations (ECG, chest X-ray, blood and urine, 
PCR for COVID-19), performed prior to their inclusion and are within 
normal limits and/or without clinical significance at the investigator's 
discretion. 
4) Subjects with systolic blood pressure higher than 110 mm Hg and 
lower than 139 mm Hg, a diastolic pressure higher than 70 mm Hg 
and lower than 89 mm Hg with a heart rate higher than 50 and lower 
than 90 beats per minute after being 5 minutes in a sitting position 
and then in a standing position (outliers included). 
5) Volunteer being willing to participate in the study and who has signed 
the informed consent approved before the study initiation. 
 
EXCLUSION CRITERIA: 
1) History of clinically significant allergies (except asymptomatic 
untreated seasonal allergies). 
2) ≥ 20 mm Hg drop in systolic pressure or ≥ 10 mm Hg in diastolic 
pressure in the first 3 minutes of postural change. 
3) Volunteers who are or have been taking other drugs (prescription or 
over-the-counter) within two weeks prior to the study. 
4) Volunteers with a history of autoimmune diseases. 
5) Organic CNS mental and/or psychiatric conditions: bipolar disorder, 
severe depression, insomnia, changes in personality. 
6) Active or chronic infections 

7) Reception of live or dead virus, bacteria vaccines within fifteen days 
prior to visit 1 or 2. 
8) In the case of COVID vaccines, a window period longer than 15 
days must be observed before visits 1 or 2. This period is 
contemplated based on the recommendations by the National 
Ministry of Health published on 5-29-2021. The volunteer may 
receive the vaccine 35 days after receiving the investigational 
product. 
9) Known allergy to any of the components of the formulations. 
10)  Active smoker of more than 10 cigarettes/day. 
11) Current clinical evidence of severe digestive disorders, digestive 
tract surgeries (except appendectomy) 
12) Current clinical evidence of kidney disease. 
13) Current evidence of liver disorders 
14) Current clinical evidence of respiratory and cardiac diseases. 
15) Presence of diabetes mellitus, thyroid dysfunction or other 
endocrine disorder. 
16) Evidence of active gastroduodenal disease. 
17) History of peripheral thrombotic phenomena. 
18) Underlying neurological disease 
19) Current presence of any progressive chronic disease 
20) History of drug or alcohol abuse or addiction during the last three 
years. 
21) Participation in a clinical study within the last three months. 
22) Use of any drug within fourteen days prior to the study initiation that, 
at the principal investigator´s discretion, may interfere with the drug 
biodistribution. 
23) Subject who donated or experienced blood loss within the last 
twelve weeks prior to the study initiation, or willing to donate blood 
within three months after the end of the study. 
24) Excessive drinker of tea, cocoa, mate, coffee and/or beverages 
containing caffeine (>5 cups/day) or wine (>0.5 L/day) or alcoholic 
drinks (>50 ml/day). 
25) Significant abnormalities in the electrocardiogram 
26) PCR positive for COVID-19. 

27) Serology positive for HIV, hepatitis B or hepatitis C. 
28) Abnormal clinical laboratory results (which, at the principal 
investigator´s discretion, are considered clinically significant) 
29) Non-cooperative volunteer 
STUDY DURATION PER 
VOLUNTEER 
Duration of the clinical study 56 days per volunteer: 21 days (screening) 
and 35 days (follow-up until end-of-study visit) 
STUDY VISITS 
• 
Visit I: Selection of volunteers, obtaining informed consent and 
completion of entrance exams. 
• 
Visit II: It will consist of a 24-h hospitalization for the administration 
of the R or T product, assigned according to the volunteer 
randomization. Sampling times for the investigational product 
pharmacokinetic quantification for each volunteer will be: pre-dose 
(0 h), 1 h, 3 h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h and 12 h. h from the 
beginning of the infusion. For the assessment of enzymatic activity, 
the following assessment times will be taken: pre-dose (0 h), 1 h, 3 
h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h and 12 h. For the assessment of 
immunogenicity, the following evaluation times will be taken: pre-
dose (0 h) and 12 h. 
• 
Visit III, IV, V and VI (telephone contact): The principal 
investigator or the delegated medical staff will contact the 
volunteer by telephone after Visit II to ask about medication 
tolerance and the presence of adverse events to the days 3, 7 and 
14 respectively. 
• 
Visit VII (end of study visit): It will be carried out 35 days after 
Visit I, to evaluate the general clinical and biological status of 
volunteers. The same studies performed at the initial visit will be 
repeated. For the assessment of immunogenicity, samples will be 
taken to be evaluated at 35 days. 
INVESTIGATIONAL 
PRODUCT/S 
DOSAGE 
AND 
ROUTE 
OF 
ADMINISTRATION  
A single dose of 1 mg/kg of agalsidase beta will be administered 
intravenously in an accurate strength to each volunteer for 5 h.  
The following formulations will be used: 
• 
Sanofi Genzyme Fabrazyme® (Reference Formulation, “R”) 
supplied as 35 mg and 5 mg lyophilized powder to be 
reconstituted for infusion. 
• 
Agalsidase beta, a biosimilar developed by Biosidus S.A. (Test 
Formulation, “T”) supplied as 35 mg and 5 mg lyophilized powder 
to be reconstituted for infusion. 
COMPARATIVE 
ASSESSMENT 
OF 
From the results of the serum concentration of agalsidase beta, at the 
different sampling times, the following variables will be estimated: 

PHARMACOKINETICS 
AND 
BIOLOGICAL 
ACTIVITY 
BETWEEN 
FORMULATIONS 
- AUC0-12h: area under the serum concentration curve between time 0 and 
12 hours. 
- AUC0-∞: area under the serum concentration curve, as a result of the 
extrapolation between time 0 and infinite time. 
- Cmax: maximum serum concentration of agalsidase beta. 
- Tmax: time to maximum serum concentration for agalsidase beta. 
The difference between the Tmax of the test formulation and that of the 
reference will be analyzed, and the drug elimination rate constant (Kel) 
and half-life (t1/2) in the body will be calculated. 
Summary measures (arithmetic and geometric means, standard 
deviation, CV% and range) for each formulation, as well as the individual 
results of each volunteer will be presented. 
STATISTICAL 
ANALYSIS:  
 
SAMPLE SIZE CALCULATION: 
The sample size (n: 20) was established based on data from the 
literature. As it is a sequential design, if the statistical analysis on the first 
20 volunteers does not demonstrate bioequivalence, additional volunteers 
will be included for the eventual demonstration of bioequivalence with 
adequate power. 
ANALYSIS PLAN 
COMPARATIVE 
PHARMACOKINETIC 
ANALYSIS 
BETWEEN 
FORMULATIONS 
- ABCT / ABCR: Ratio between the area under the curve for the Test 
formulation and the area under the curve for the Reference formulation. 
For log transformation, the 90% confidence interval of the AUC ratio (for 
AUC0-120h and AUC0-∞) should be within the range of 0.80-1.25. 
- Cmax T / Cmax R: Ratio between the Cmax of the Test formulation and 
Cmax of the Reference formulation. For logarithmic transformation, the 
90% confidence interval of the Cmax ratio should initially be within the 
range of 0.80-1.25. 
 
COMPARATIVE 
ENZYME 
ACTIVITY 
ANALYSIS 
BETWEEN 
FORMULATIONS 
- Variation in the enzymatic activity for agalsidase beta in the sample at 
the end of infusion (5 h) compared to baseline (pre-infusion). For 
logarithmic transformation, the 90% confidence interval of the Cmax ratio 
should initially be within the range of 0.80 -1.25. 
 
For an estimation of the pharmacokinetic parameters, specific 
pharmacokinetic 
computer 
softwares 
will 
be 
used 
(WinNonlin 

Professional Edition, version 5.2.1 (2008). 
Possible inter-subject and inter-treatment differences will be studied using 
ANOVA. 
Outliers: for determination of possible outliers, a selective statistical 
method will be applied. 
 
COMPARATIVE 
IMMUNOGENIC 
ANALYSIS 
BETWEEN 
FORMULATIONS 
The presence of anti-agalsidase beta antibodies, and their eventual titer, 
will be determined in the two groups of volunteers, using the biological 
activity neutralization technique. 
 
 
 
 
 

TABLE OF CONTENTS 
Content 
1 
OVERALL INFORMATION AND BACKGROUND ..................................................... 16 
1.1.1 
Fabry Disease and Enzyme Replacement Therapy .................................... 16 
1.1.2 
Mechanism of Action .................................................................................. 16 
1.1.3 
Pharmacokinetics ....................................................................................... 17 
1.1.4 
Dosing Regimen and Administration ........................................................... 17 
1.1.5 
Contraindications ........................................................................................ 17 
1.1.6 
Warnings .................................................................................................... 17 
1.1.7 
Interactions ................................................................................................. 18 
1.1.8 
Adverse Reactions...................................................................................... 19 
1.1.9 
Pregnancy and Breastfeeding ..................................................................... 19 
1.1.10 
Overdose .................................................................................................... 19 
1.1.11 
Storage Conditions ..................................................................................... 19 
2 
STUDY OBJECTIVES .............................................................................................. 19 
3 
SITE ......................................................................................................................... 20 
4 
STUDY DESIGN ....................................................................................................... 20 
4.1 
RANDOMIZATION PROCEDURE ..................................................................... 21 
4.2 
CODE UNBLINDING IN BLINDED STUDIES .................................................... 21 
4.3 
STUDY DURATION AND DISCONTINUATION SCHEDULE ............................ 21 
4.4 
STUDY COURSE AND PROCEDURES ............................................................ 21 
4.4.1 
Procedures to be made at each Visit .......................................................... 22 
4.4.2 
Blood sampling ........................................................................................... 24 
4.4.3 
Analytical Method ....................................................................................... 26 
4.5 
VOLUNTEER FOLLOW-UP ............................................................................... 27 
4.6 
VOLUNTEER IDENTIFICATION ........................................................................ 27 
4.7 
SAMPLE IDENTIFICATION ............................................................................... 27 
5 
CRITERIA FOR SCREENING VOLUNTEERS.......................................................... 29 
5.1 
POPULATION ................................................................................................... 29 
5.2 
INCLUSION CRITERIA ..................................................................................... 29 
5.3 
EXCLUSION CRITERIA .................................................................................... 29 
5.4 
STUDY WITHDRAWAL CRITERIA .................................................................... 31 
6 
STUDY DRUG .......................................................................................................... 32 

6.1 
DRUG NAME, FORMULATION AND STORAGE .............................................. 32 
6.1.1 
Study Formulations ..................................................................................... 32 
6.1.2 
Content and Potency Uniformity ................................................................. 34 
6.1.3 
Storage ....................................................................................................... 34 
6.2 
MATERIAL PREPARATION AND LABELING .................................................... 34 
6.3 
DOSE AND ROUTE OF ADMINISTRATION ..................................................... 34 
6.4 
TREATMENT DURATION ................................................................................. 34 
7 
ENZYME ACTIVITY ANALYSIS ............................................................................... 35 
7.1 
PHARMACOKINETIC ANALYSIS OF RESULTS............................................... 35 
7.1.1 
Comparative pharmacokinetic analysis between formulations .................... 35 
7.2 
PHARMACODYNAMIC ANALYSIS OF RESULTS ............................................ 37 
8 
IMMUNOGENICITY ANALYSIS ................................................................................ 37 
9 
SAFETY ANALYSIS ................................................................................................. 37 
9.1 
ADVERSE EVENTS .......................................................................................... 37 
9.2 
ADVERSE EVENT REPORTING ....................................................................... 38 
9.3 
ADVERSE EVENT DEFINITION ........................................................................ 40 
9.4 
ABNORMAL FINDINGS IN THE INVESTIGATIONS ......................................... 41 
9.5 
SERIOUS ADVERSE EVENT ............................................................................ 41 
9.6 
HOSPITALIZATION ........................................................................................... 42 
9.7 
SEVERITY ASSESSMENT ................................................................................ 43 
9.8 
CAUSALITY 
ASSESSMENT 
(RELATIONSHIP 
TO 
MEDICATION, 
RELATEDNESS) .......................................................................................................... 43 
10 
STATISTICAL CONSIDERATIONS AND ANALYTICAL PLAN. SAMPLE SIZE 
CALCULATION ................................................................................................................ 44 
11 
ETHICAL ASPECTS ............................................................................................. 44 
11.1 
LOCAL REGULATIONS/ DECLARATION OF HELSINKI .................................. 44 
11.2 
INFORMATION FOR THE SUBJECT AND INFORMED CONSENT.................. 44 
11.3 
INDEPENDENT ETHICS COMMITTEE (IEC) .................................................... 44 
12 
CONDITIONS FOR AMENDING THE PROTOCOL .............................................. 45 
13 
CONDITIONS FOR COMPLETING THE STUDY .................................................. 45 
14 
STUDY DOCUMENTATION, CRF and RECORDS ............................................... 45 
14.1 
INVESTIGATOR FILES / DOCUMENT RETENTION ........................................ 45 
14.2 
AUDITS AND INSPECTIONS ............................................................................ 46 
14.3 
CASE REPORT FORMS (CRF) ......................................................................... 46 

14.4 
CONTRACT RESEARCH ORGANIZATIONS .................................................... 46 
15 
STUDY MONITORING .......................................................................................... 47 
16 
CONFIDENCIALIDAD 
DE 
LOS 
DOCUMENTOS 
DEL 
ENSAYO 
Y 
LOS 
REGISTROS DE LOS PACIENTES ................................................................................. 47 
17 
QUALITY ASSURANCE ........................................................................................ 48 
18 
FINAL REPORT AND PUBLICATION ................................................................... 49 
19 
CALENDAR PROVIDED ....................................................................................... 50 
20 
FUNDING AND INSURANCE ............................................................................... 50 
21 
SIGNATURE PAGE .............................................................................................. 51 
22 
REFERENCES ...................................................................................................... 52 
 
 
 
 

ABBREVIATIONS 
 
Abbreviation 
Meaning 
AE 
Adverse Event 
ANMAT 
Administración Nacional de Medicamentos, Alimentos y Tecnología 
Médica (National Administration of Drugs, Food and Medical Technology) 
AUC 
Area Under the Curve 
BMI 
Body Mass Index 
CHO 
Chinese Hamster Ovary 
CRF 
Case Report Form 
CRO 
Contract Research Organization 
DNA 
deoxyribonucleic acid 
ECG 
Electrocardiogram 
FDA 
Food and Drug Administration 
Gamma GT 
gamma glutamyl transpeptidase 
GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
HIV 
Human Immunodeficiency Virus  
ICH 
International Council for Harmonization  
INAME 
Instituto Nacional del Medicamento (National Drug Institute) 
IRR 
Infusion-related reactions 
KPTT 
Partial thromboplastin time with kaolin 
r.p.m 
Revolutions per minute 
RAD 
Adverse Drug Reaction  
r-hαGAL 
Recombinant human α-galactosidase A  
RIPA 
Radioimmune precipitation 
SAE 
Serious Adverse Event 
SNBL 
Shin Nippon Biomedical Laboratories 
SNSDP 
Sanatorio Nuestra Señora del Pilar 
USP 
U.S. Pharmacopeia  
 
 
 

1 
OVERALL INFORMATION AND BACKGROUND  
Agalsidase beta is a recombinant form of human α-galactosidase A and is obtained by 
recombinant DNA technology from a cell line culture from Chinese hamster ovary (CHO). 
The amino acid sequence of the recombinant form, as well as the nucleotide sequence 
that encoded it, are identical to those of the natural form of α-galactosidase A. ATC Code: 
A16ABO4. 
 
1.1.1 Fabry Disease and Enzyme Replacement Therapy  
Fabry disease is a rare lysosomal disease, associated with the X chromosome and caused 
by mutations in the gene encoding the α-galactosidase A enzyme. Decreased enzyme 
activity results in a progressive lysosomal accumulation of glycosphingolipids, mainly 
globotriaosylceramide (GL-3) in several tissues, including vascular endothelial cells, 
neurons, myocytes, and renal cells. This accumulation leads to cellular dysfunction, tissue 
remodeling, fibrosis, and ultimately, organ failure such as heart failure or stroke. 
The severity of manifestations varies depending on the remaining enzyme activity of α-
galactosidase A. 
Fabry disease is diagnosed by decreased α-galactosidase A activity in plasma leukocytes 
or dermal fibroblast culture. Urine or plasma GL-3 concentrations are used as a biomarker 
for diagnosis and monitoring. 
Plasma globotriaosylsphingosine (lyso-GL-3) concentrations have also been shown to be 
used as a diagnostic marker. This is because lyso-GL-3 level has shown to be increased 
in patients who have not shown increased GL-3 level. 
Treatment with agalsidase beta is indicated as long-term enzyme replacement therapy in 
patients with a confirmed diagnosis of Fabry disease and may be used in adults, 
adolescents, and children ≥ 8 years of age. 
Currently, three therapies are approved worldwide: agalsidase α (Replagal by Shire and 
Agalsidase Beta BS by Amicus) and agalsidase β (Fabrazyme® by Sanofi-Genzyme or 
JR-051 by JCR Pharmaceuticals). Treatment with these drugs decreases GL-3 
accumulation in the kidneys, heart, and epithelium. It also reduces pain and maintains 
renal function. 
 
1.1.2 Mechanism of Action 
The rationale for the enzyme replacement therapy is to restore enzyme activity to a level 
sufficient to remove accumulated substrate from organ tissues, thereby preventing, 
stabilizing, or reversing the progressive decline in organ function before irreversible 
damage occurs. 
Following intravenous infusion, agalsidase beta is rapidly cleared from bloodstream and is 
taken up by the lysosomes of parenchymal and endothelial cells, which they probably 
enter via mannose-6 phosphate, mannose, and asialoglycoprotein receptors. 

 
1.1.3 Pharmacokinetics  
Following intravenous administration of agalsidase beta to adults dosed at 0.3 mg, 1 mg, 
and 3 mg/kg of body weight, AUC values increased more than dose-proportionally due to 
decreased clearance, which indicated saturated clearance. The elimination half-life was 
not dose-dependent and ranged from 45 to 100 minutes. 
Following intravenous administration of agalsidase beta to adults with an infusion time of 
approximately 300 minutes and dosed at 1 mg/kg/body weight, every two weeks, mean 
plasma Cmax ranged from 2,000 to 3,500 ng/mL, while that the AUC0-∞ ranged from 370 to 
780 mcg.min/mL. The mean volume of distribution at steady state ranged from 8.3-40.8 L, 
plasma clearance from 119-345 mL/min. and the mean elimination half-life from 80 to 120 
minutes. 
Beta agalsidase is a protein and is predicted to be metabolically degraded by peptic 
hydrolysis. Therefore, hepatic impairment is not expected to impact on the 
pharmacokinetics of agalsidase beta in a clinically significant manner. Renal clearance of 
agalsidase beta is considered a secondary clearance pathway. 
 
1.1.4 Dosing Regimen and Administration  
The dose to be used in this study on agalsidase beta is 1 mg/kg/body weight, administered 
as an intravenous infusion. 
The rationale is as follows: 
• 
The recommended dose of agalsidase beta comparator is 1 mg/kg of body weight, 
administered once every 2 weeks by intravenous infusion. 
• 
It is the dose that did not cause adverse events in preclinical studies in animals, 
conducted on the same Biosidus S.A. molecule, which defines this dose as the 
NOAEL. 
• 
It is the dose used in the biosimilar study conducted in Japan15. 
The initial infusion rate should not exceed 0.25 mg/min (15 mg/h) to minimize the potential 
for infusion-related reactions. Once the volunteer's tolerance has been established, the 
infusion rate can be gradually increased for subsequent infusions. 
 
1.1.5 Contraindications 
Life-threatening hypersensitivity (anaphylactic reaction) to the drug substance or to any of 
the inactive ingredients. 
 
1.1.6 Warnings 
Immunogenicity: 

As agalsidase beta (recombinant human α-galactosidase A or r-hαGAL) is a recombinant 
protein, development of IgG antibodies is expected to occur in patients with little or no 
residual enzyme activity. In studies that resulted in the approval of the reference medicine, 
most patients developed IgG antibodies to r-hαGAL, mainly within 3 months of the first 
drug infusion. Over time, most seropositive patients in clinical trials showed a decreasing 
trend in titers (based on a ≥ 4-fold reduction in titers from peak measurement to the last 
measurement) (40% patients) or they showed tolerance (no detectable antibody was 
confirmed by 2 consecutive radioimmune precipitation assays (RIPA)) (14% patients) or 
showed a plateau (35% patients). Healthy volunteers participating in this study, whose 
enzyme activity is normal, are expected to have little or no antibody response to 
agalsidase beta administration. 
 
Infusion-Related Reactions: 
Patients with antibodies to recombinant agalsidase beta (recombinant human α-
galactosidase A or r-hαGAL) are more likely to experience infusion-related reactions 
(IRRs) defined as any related adverse effect occurring on the day of the infusion. These 
patients should be treated with caution when re-administering agalsidase beta. Antibody 
status in patients with Fabry disease and on treatment should be monitored regularly. 
 
Hypersensitivity 
Allergic hypersensitivity reactions may occur, as with any other protein medication for 
intravenous administration. 
If serious allergic or anaphylactic reactions occur, agalsidase beta administration should 
be discontinued immediately and appropriate treatment should be initiated. Current 
standards for emergency medical treatment of this situation will be followed. 
 
Patients with Advanced Kidney Disease 
The renal effect of agalsidase beta treatment may be limited in patients with advanced 
kidney disease. 
1.1.7 Interactions 
Biosidus has not conducted any in vitro metabolism or drug interaction studies. A literature 
search did not provide data on this type of study either. Given its metabolism, cytochrome 
P450-mediated drug interactions are unlikely to occur with agalsidase beta. It should not 
be administered with chloroquine, hydroxychloroquine, amiodarone, or aminoglycosides 
(amikacin, streptomycin, gentamicin, or tobramycin), due to the theoretical risk of inhibition 
of α-galactosidase A activity. 
 

1.1.8 Adverse Reactions 
Safety Profile Summary 
As agalsidase beta (r-hαGAL) is a recombinant protein, development of IgG antibodies is 
expected to occur in patients with little or no residual enzyme activity. 
Patients with antibodies to r-hαGAL are more likely to experience infusion-related 
reactions. Cases of reactions suggestive of immediate hypersensitivity (type I) have been 
reported in a small number of patients. 
Chills, pyrexia, feeling cold, nausea, vomiting, headache and paraesthesia are included as 
very common adverse reactions. 67% of patients experienced at least one infusion-related 
reaction. In some cases anaphylactoid reactions have been reported. Treatment 
considered 
for 
anaphylactic 
reactions 
is 
adrenaline, 
corticosteroids 
(dexamethasone/hydrocortisone) and antihistamine (diphenhydramine). 
 
1.1.9 Pregnancy and Breastfeeding 
There are no sufficient data from the use of agalsidase beta in pregnant women. 
Studies in animals do not indicate direct or indirect harmful effects on embryonic/fetal 
development. 
Agalsidase beta should not be used during pregnancy unless clearly necessary. 
Agalsidase beta may be excreted in breast milk. As there are no data on the effects on 
newborns exposed to the drug through breast milk, breastfeeding discontinuation is 
recommended when initiating treatment. 
 
1.1.10 Overdose 
Clinical studies with agalsidase beta have used up to 3 mg/kg of body weight. 
 
1.1.11 Storage Conditions 
Store in the refrigerator at a temperature between 2 °C and 8 °C in its original container.  
Reconstituted solution should not be stored and should be diluted quickly. Diluted solution 
may be kept for up to 24 hours at a temperature between 2 °C and 8 °C. 
 
2 
STUDY OBJECTIVES 
Primary Objective 
The objective of this trial is to compare the pharmacokinetic behavior after administration 
by intravenous infusion over a period of 5 hours of a single administration of 1 mg/kg 

Biosidus Agalsidase beta (Test Formulation, “T”) versus an intravenous infusion of Sanofi-
Genzyme Fabrazyme® at the same strength (Reference Formulation, “R”). 
 
Secondary Objectives 
Evaluate and compare the plasma enzyme activity of agalsidase beta, the eventual 
induction of anti-agalsidase beta antibodies and the occurrence and frequency of adverse 
effects and tolerance to the formulations studied. 
 
3 
SITE 
Volunteer recruitment, evaluation, control, clinical follow-up, hospitalization and processing 
of the samples to obtain the serum and plasma aliquots applicable to each time point will 
be conducted at Sanatorio Nuestra Señora del Pilar Research Center (SNSDP). It was 
authorized by ANMAT to conduct Phase I studies. The authorizing provision is attached in 
Annex 6 (DI-2019-5344-APN-ANMAT#MSYDS). 
Samples will be sent with a numerical code (see section 6.2 MATERIAL PREPARATION 
AND LABELING) to be able to later identify the patient and the formulation received, with 
the submission of results. In this way, not only is the anonymity of the volunteer ensured, 
but also the work done in a blinded manner. 
Volunteers will be hospitalized at SNSDP and the serum and plasma samples will be sent 
to Biosidus S.A. Code described in section 6.2 using a dry ice transport system for 
biological substances. 
All members in the research team agree to adhere to the ethical, legal and administrative 
standards that govern research on human beings. 
 
4 
STUDY DESIGN 
Pharmacokinetics, biological activity and immunogenicity will be studied in 20 healthy 
volunteers starting with a single dose of 1 mg/kg/body weight after administration via 
intravenous infusion of one (1) dose of: 
• 
Sanofi-Genzyme Fabrazyme®. (Reference Formulation, “R”), or 
• 
Biosidus SA Agalsidase beta (Test Formulation, “T”) 
This study was conducted under an open, randomized, parallel-arm, balanced, and 
sequential design. As it is a sequential design, if the statistical analysis of the first 20 
volunteers does not show bioequivalence, additional volunteers will be included until the 
similarity criterion is verified with adequate power. 
For volunteers and clinical staff, the study is open. Laboratory determinations will be 
completed in a blinded fashion, with coded samples. 
Below (Figure 1), a study visit scheme is presented: 

 
Figure 1 Study Visit Scheme  
 
4.1 
RANDOMIZATION PROCEDURE  
Randomization to the study will be performed by chance, using a table of random 
numbers, using the RANDOM software of Randomness and Integrity Services Ltd. 
Premier Business Centres, 8 Dawson Street, Dublin 2, D02 N767, Ireland. It will be carried 
out in a balanced way in the anthropometric variables between both arms. 
Annex 4 contains the randomization table, which indicates the treatment (R or T) assigned 
to each patient. 
 
4.2 
CODE UNBLINDING IN BLINDED STUDIES 
This study is open to clinical staff, and is only blind for measurements to be made in the 
laboratory, which will be further carried out, when all the study samples are collected. 
Therefore, both the sponsor and the principal investigator will know the treatment received 
by each volunteer. The pharmacokinetic, enzyme activity and immunogenicity samples will 
be blind during dosing, and will be sent to the relevant laboratories presenting only the 
identification code of each subject. 
 
4.3 
STUDY DURATION AND DISCONTINUATION SCHEDULE  
The study duration is approximately 6 months since the approval by the ANMAT, for the 
recruitment of volunteers, study conduction, analysis of results and final report preparation. 
Times for completion are subject to the COVID-19 Pandemic progress in Argentina, 
especially in the Province of Buenos Aires, and the availability of beds at SNSP. 
The sponsor and the investigators independently may discontinue the study at any time at 
the possibility or occurrence of a serious adverse event or situation that could impact on 
the volunteer´s health. 
 
4.4 
STUDY COURSE AND PROCEDURES  
The study will be conducted by staff trained in the Good Clinical Practice (GCP) Standards 
by the International Council for Harmonization (formerly International Conference on 
Harmonization), in compliance with ANMAT Regulation 6677/10.  
Principal Investigator, Dr. Eduardo Pirotzky will be responsible for conducting this study. 
SCREENING 
VISIT I 
 
HOSPITALIZATION 
AND TREATMENT 
VISIT II 
FOLLOW-UP 
VISITS III, IV, V and 
VI 
END OF STUDY 
VISIT VII 
HEALTHY 
VOLUNTEERS 
PHONE 
CONTACT 
FINAL ON-SITE 
VISIT 
R or T 

 
4.4.1 Procedures to be made at each Visit  
 
VISIT I (Screening) 
After signing the informed consent, each volunteer will undergo a physical examination 
that will include: medical history, neurological semiology, cardiovascular, (ECG, blood 
pressure, chest x-ray). Volunteers weight will be measured, taking this body weight to 
adjust the dose should they enter the study. Lab tests will also be done within 30 days, 
including: 
- 
CBC 
- 
Erythrocyte sedimentation 
- 
Uremia 
- 
Blood glucose 
- 
Uricemia 
- 
Gamma GT 
- 
LFT 
- 
Quick time 
- 
KPTT 
- 
Platelet count 
- 
Hepatitis B and C and HIV screening tests 
- 
Urinalysis for screening drugs of abuse (cannabinoids and cocaine) 
- 
Serum electrolyte panel 
- 
Complete urinalysis 
- 
PCR for COVID-19 
- 
Serum creatinine 
- 
Glomerular filtration rate (GFR) 
- 
Urine protein 
 
In the event that outliers are detected, volunteer will be contacted by phone, email or by 
mail to arrange a new visit where he/she will be informed about the reasons why he/she 
cannot participate and will also be instructed to see an applicable medical specialist. 
 
VISIT II (Hospitalization) 
During this visit, volunteer will receive the formulation randomly assigned. Volunteers must 
come to the clinic the day before, at 8:00 p.m. and the infusion will begin at 8:00 a.m. the 
following day, with the volunteer in fasting state. 
In this instance, the Principal Investigator will ensure whether volunteers meet the 
conditions to proceed with the drug infusion (interrogation and physical examination). 
There will be a staggered hospitalization of two volunteers at a time. 

After the hospitalization of the first two volunteers (1 Reference volunteer and 1 Test 
volunteer), a week will be allowed to admit the next two volunteers. At the end of each 
hospitalization, volunteers will be evaluated for AE occurrence. If no AEs that jeopardize 
the participant´s safety occur, the next volunteers will be hospitalized according to the 
randomization table at the time intervals deemed appropriate by the Principal Investigator. 
To draw blood samples and avoid multiple punctures, an abbocath-type intravenous line 
with a three-way stopcock will be placed in the arm contralateral to the one used for 
agalsidase infusion. Through it, all blood samples will be drawn during hospitalization. 
At each time point, 30 mL of blood will be drawn, which will be placed in adequate tubes to 
obtain serum or plasma, as appropriate (for example, up to 500 mL of blood is drawn from 
a volunteer blood donor at a time and without consequences for him/her). 
Samples to be obtained from each volunteer will be drawn from the contralateral arm to 
which the infusion of the experimental drug is being made. 
After verifying its correct operation, the pre-dose sample (hour 0) will be drawn. Next, the 
intravenous administration of the corresponding infusion will begin according to the order 
of randomization. 
 
Sampling times during Visit II will be the following: 
1. For pharmacokinetic analysis: predose (0 hour), 1 h, 3 h, 5 h, 5,5 h, 6 h, 7 h, 
8 h, 10 h, 12 h from the beginning of infusion. 
2. For enzyme activity analysis: predose (0 hour), 1 h, 3 h, 5 h, 5,5 h, 6 h, 7 h, 8 
h, 10 h, 12 h from the beginning of infusion. 
3. For immunogenicity analysis: predose (0 h), 12 h and day 35. 
 
On the study day, volunteers will be allowed to drink fluids freely. 
Alcoholic drinks or xanthines will not be allowed (since 48 hours before the test). 
No medication (neither allopathic nor homeopathic) will be allowed during the two weeks 
prior to the study. 
 
Snacks during Visit II: 
During hospitalization, volunteers may consume only standardized food, provided by the 
clinic. No other type of food will be allowed other than that which complies with the 
guidelines that are developed below: 
 
 
 

Meal 
Time Schedule 
Menu 
 
 
Lunch 
 
4 hrs after the beginning of 
the infusion (approximately 
between 12 and 12:30 p.m.) 
A piece of chicken (breast or leg and thigh), 
with no skin  
A portion of white rice  
500 mL of mineral water 1 sachet of salt  
1 small package of crackers 
1 apple. 
 
Afternoon 
snack 
8 hrs after the beginning of 
infusion (approx. 4:00 p.m.) 
1 cup of coffee or tea (decaffeinated with 
100 mL of whole milk), 2 butter croissants 
each with 2 cooked ham slices and 2 
cheese slices. 
 
 
Dinner 
 
 
12 hrs after the beginning of 
the infusion (approx. 8:00 
p.m.) 
A piece of chicken (breast or leg and thigh), 
with no skin  
A portion of white rice  
500 mL of mineral water 1 sachet of salt  
1 small package of crackers 
1 apple. 
 
VISIT III, IV, V y VI (phone contact): 
The principal investigator or the delegated medical staff will contact the volunteer by 
telephone after visit II to ask about tolerance to medication and the occurrence of adverse 
events on days 3, 7, 14 and 21 after visit II respectively. 
VISIT VII (on-site) 
Volunteers will come to SNSDP after 35 days, to complete the end-of-study visit on an 
outpatient basis in order to assess the patient overall clinical status. The same studies 
carried out at the initial visit will be repeated. 
For the immunogenicity analysis, a serum sample will be collected. 
 
4.4.2 Blood sampling 
Sterile, disposable tubes with a stopper will be used for the collection of blood samples for 
complete blood count. A 30-mL sample of venous blood should be drawn before the 
beginning of the infusion (time point 0, pre-dose).  
Once the infusion begins, venous blood collection will continue at the following time points: 
• 
1 h, 3 h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h, 12 h from the beginning of the infusion 
for the pharmacokinetic study. 

• 
Day 35 from the beginning of infusion for immunogenicity study. 
• 
1 h, 3 h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h, 12 h from the beginning of the infusion 
for the enzymatic activity study  
 
Once the infusion finishes (visit II), volunteers will be observed until 20:00 p.m, the same 
day. Then, they will be discharged to return to their homes. 
 
For immunogenicity sample collection: At visit II, racks with 2 2-mL polypropylene tubes 
with stoppers will be prepared to separate serum from each volunteer in two samples 
(sample and counter sample) from each collection time point for anti-agalsidase beta 
antibody measurement, immunogenicity measurement. 
After collecting the blood sample, tubes will allowed to reach room temperature for 30 
minutes to separate the clot and serum. Then they will be centrifuged at 2000g for 15 
minutes to separate the serum. 
To receive the serum, polypropylene tubes will be used. Each serum sample will be 
separated into 2 aliquots (sample and counter sample) to measure immunogenicity. 
 
For pharmacokinetic sample collection: At visit II, racks with 10 2-mL polypropylene tubes 
with stoppers (heparinized tubes) will be prepared for each volunteer for the separation of 
plasma into two samples (sample and counter sample) applicable to each collection 
timepoint for the quantification of agalsidase beta, pharmacokinetics measurement. After 
collecting the blood sample, tubes will be kept cold and then centrifuged at 4 °C at 8000 
rpm for 10 min for plasma separation. 
Sample quantification will be carried out in an independent laboratory. The mass of 
agalsidase present in each sample will be quantified. 
 
For sample collection for enzyme activity: At visit II, the same tubes used for PK 
measurement will be used for each volunteer. An aliquot of the plasma separated at each 
time point will be used for measuring enzyme activity. 
After collecting the blood sample, tubes will be kept cold and then centrifuged at 4 °C at 
8000 rpm for 10 min to separate the plasma. 
Biological activity will be measured by the fluorimetry method in plasma samples, obtained 
in heparin tubes. 
To receive the serum, polypropylene tubes will be used. Each serum sample will be 
separated into 2 aliquots (sample and counter sample) for agalsidase beta, enzyme 
activity or immunogenicity measurements. Tubes will be labeled with the study code, 
volunteer number and collection time points. 

The complete process from the blood sample collection to obtaining the 2 (two) aliquots of 
serum will take 60 minutes. Tubes will be frozen at -20 °C at the Site's laboratory and will 
be taken after each hospitalization by Biosidus for their storage until measurements are 
made. 
Criteria for rejecting a sample: pronounced hemolysis, accidental thawing. 
 
All tubes for blood collections, and polypropylene tubes for serum collection will be labeled 
with a self-adhesive label where the study code, volunteer number and collection timepoint 
will be indicated. 
 
Storage of Plasma and Serum Samples  
Serum or plasma obtained at each time point will be placed in duplicate in polypropylene 
tubes properly labeled and stored at -20 ºC in a secured freezer with temperature control, 
at the Site laboratory, with restricted access to the staff until its transfer to Biosidus. 
 
4.4.3 Analytical Method  
Samples 
All serum or plasma samples will be stored in duplicate at -20°C. Before the assay, the 
samples should be allowed to reach room temperature (15 °C - 28 °C) and mixed by 
gentle stirring. Aliquots from the samples will be made to prevent freezing - repeated 
thawing. The duplicate shall be kept at -20 °C. 
 
Quantification Method  
Pharmacokinetic Quantification: 
Samples will be shipped at -20°C to Shin Nippon Biomedical Laboratories (SNBL), Ltd. 
The agalsidase content will be quantified by the electrochemiluminescence method. The 
SNBL laboratory will validate the quantification method before the study is initiated. 
 
Enzyme Activity Quantification 
For the measurement of the biological enzyme activity in the plasma samples, the 
fluorimetric method of hydrolysis of the synthetic substrate 4-MU-Gal will be used. 
 
Immunogenicity Measurement 
Serum samples will be incubated with 1 ng beta agalsidase for 15 minutes at room 
temperature. Then, the sample enzymatic activity will be measured using the 4-MU-Gal 
substrate. 

The inhibition percentage of the samples will be determined by comparing the activity 
value of 1 ng agalsidase beta with the treated samples. Those samples with percentage 
inhibition > 50% will be designated as positive. 
 
4.5 
VOLUNTEER FOLLOW-UP 
The study will be conducted under medical supervision and all the information will be 
transcribed to the Medical Record and the CRF. 
During the hospitalization period, the volunteer will undergo a permanent clinical 
evaluation by the principal investigator and his group of collaborators. 
Before each blood draw, blood pressure, heart rate and respiratory rate will be measured. 
In the pre-dose and final draw, hematocrit will also be recorded. These data together with 
the theoretical and real times of collection will be recorded in the Medical Record and will 
be transcribed to the Case Report Form (CRF). 
During the study, signs and symptoms of intolerance to the medication must be observed, 
which will be documented in the Medical Record and the CRF. 
 
4.6 
VOLUNTEER IDENTIFICATION 
The system for assigning numbers to volunteers (after signing the ICF) is based on the 
obligation to keep anonymity. Volunteers will be assigned a number according to their 
order of inclusion in the study (according to the table in Annex 4 - Randomization Table). 
They will be identified by the number assigned from 01 to 20 consecutively (in case new 
volunteers need to be added because it is a sequential study, they will be assigned 
numbers from 21), a two-letter code (first letter of the last name and first letter of the first 
name) and their date of birth (day-month-year): 
• 
Assignment Number: based on the randomization table and according to the order 
of inclusion: NN 
• 
First letter of the last name and first letter of the first name. XX 
• 
Date of birth (day-month-year): xxxxxx 
NNXXxxxxxx 
EXAMPLE: Juan Perez, Assigned number 004, Date of birth May 27, 1960 Patient ID: 
04PJ270560 
4.7 
SAMPLE IDENTIFICATION  
Collection tubes: 
Tubes must be identified with the study protocol code (RD-BIO-AGA-01/03/21) at the top, 
then the patient identification code (NNXXxxxxx) must be placed, then two letters that will 
identify the type of quantification to be made on the sample (pharmacokinetics: PK, 
enzyme activity: AE or Immunogenicity: IM), then a two-digit number that corresponds to 
the sample number. 

EXAMPLE: 
RD-BIO-AGA-01/03/21 
04PJ270560: PK 10 
In this case, this sample applies to a pharmacokinetic quantification, to the tenth sample 
from volunteer number 04. 
 
Aliquot tubes: 
The aliquot number will be added to the above information (sample: A1 or counter sample: 
A2) with a digit followed by the letter A. 
EXAMPLE: 
RD-BIO-AGA-01/03/21 
04PJ270560: PC 10 A1 
In this case, this sample is to perform the pharmacokinetic quantification of Aliquot 1 of the 
tenth sample from volunteer number 04. 
 
The types of sample obtained in this study are detailed below: 
Table 1 PK Samples (PK) 
Timepoint 
(h) 
0 
1 
3 
5 
5.5 
6 
7 
8 
10 
12 
Sample 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
Aliquot 1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
Aliquot 2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
 
Table 2 Samples for Enzymatic Activity (EA) 
Timepoint 
(h) 
0 
1 
3 
5 
5.5 
6 
7 
8 
10 
12 
Sample 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
Aliquot 1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
A1 
Aliquot 2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
A2 
 
 
 
 

Table 3 Immunogenicity Samples (IM) 
Timepoint 
(h 
or 
Days) 
0 h 
 
 
 
 
 
 
 
12 h 
Day 30 
Sample 
1 
 
 
 
 
 
 
 
10 
3 
Aliquot 1 
A1 
 
 
 
 
 
 
 
A1 
A1 
Aliquot 2 
A2 
 
 
 
 
 
 
 
A2 
A2 
 
 
5 
CRITERIA FOR SCREENING VOLUNTEERS  
5.1 
POPULATION 
In order to be selected and included in the study, volunteers must meet all the screening 
and inclusion criteria and none of the exclusion criteria. 
The number of subjects to be included in the study will be a total of 20 male volunteers. As 
a sequential study, additional volunteers may be included. 
Those volunteers who meet all the inclusion criteria and none of the exclusion criteria will 
be incorporated into the study, after having explained the purpose of the clinical trial and 
having signed the informed consent for participation. 
Presence of the following inclusion criteria and absence of exclusion criteria must be 
verified by the Principal Investigator in the medical record and in the applicable form in the 
CRF book of each subject. 
 
5.2 
INCLUSION CRITERIA 
1) Male subjects aged between 18 and 40 years. 
2) Body mass index (BMI) between 19 and 25 kg/m² 
3) Volunteers whose investigations (ECG, chest X-ray, blood and urine, PCR for 
COVID-19), performed prior to their inclusion and are within normal limits 
and/or without clinical significance at the investigator's discretion. 
4) Subjects with systolic blood pressure higher than 110 mm Hg and lower than 
139 mm Hg, a diastolic pressure higher than 70 mm Hg and lower than 89 
mm Hg with a heart rate higher than 50 and lower than 90 beats per minute 
after being 5 minutes in a sitting position and then in a standing position 
(outliers included). 
5) Volunteer being willing to participate in the study and who has signed the 
informed consent approved before the study initiation. 
 
5.3 
EXCLUSION CRITERIA 

1) History of clinically significant allergies (except asymptomatic untreated 
seasonal allergies), 
2) ≥ 20 mm Hg drop in systolic pressure or ≥ 10 mm Hg in diastolic pressure in 
the first 3 minutes of postural change. 
3) Volunteers who are or have been taking other drugs (prescription or over-the-
counter) within two weeks prior to the study. 
4) Volunteers with a history of autoimmune diseases. 
5) Organic CNS mental and/or psychiatric conditions: bipolar disorder, severe 
depression, insomnia, changes in personality. 
6) Active or chronic infections 
7) Reception of live or dead virus, bacteria vaccines within fifteen days prior to 
visit 1 or 2. 
8) In the case of COVID vaccines, a window period longer than 15 days must be 
observed before visits 1 or 2. This period is contemplated based on the 
recommendations by the National Ministry of Health published on 5-29-2021. 
The volunteer may receive the vaccine 35 days after receiving the 
investigational product. 
9) Known allergy to any of the components of the formulations. 
10) Active smoker of more than 10 cigarettes/day. 
11) Current clinical evidence of severe digestive disorders, digestive tract 
surgeries (except appendectomy) 
12) Current clinical evidence of kidney disease. 
13) Current evidence of liver disorders 
14) Current clinical evidence of respiratory and cardiac diseases.  
15) Presence of diabetes mellitus, thyroid dysfunction or other endocrine disorder. 
16) Evidence of active gastroduodenal disease. 
17) History of peripheral thrombotic phenomena 
18) Underlying neurological disease 
19) Current presence of any progressive chronic disease  
20) History of drug or alcohol abuse or addiction during the last three years. 
21) Participation in a clinical study within the last three months. 
22) Use of any drug within fourteen days prior to the study initiation that, at the 
principal investigator´s discretion, may interfere with the drug biodistribution. 

23) Subject who donated or experienced blood loss within the last twelve weeks 
prior to the study initiation, or willing to donate blood within three months after 
the end of the study. 
24) Excessive drinker of tea, cocoa, mate, coffee and/or beverages containing 
caffeine (>5 cups/day) or wine (>0.5 L/day) or alcoholic drinks (>50 ml/day). 
25) Significant abnormalities in the electrocardiogram 
26) PCR positive for COVID-19 
27) Serology positive for HIV, hepatitis B or hepatitis C. 
28) Abnormal clinical laboratory results (which, at the principal investigator´s 
discretion, are considered clinically significant) 
29) Non-cooperative volunteer 
 
5.4 
STUDY WITHDRAWAL CRITERIA  
Subjects may freely withdraw from the study at any time, and without giving reasons. For 
safety, behavior or administrative reasons, they may also be withdrawn from the study, at 
the Principal Investigator´s discretion. If a subject does not come for the next scheduled 
visit, every effort should be made to contact the subject, document the reasons for his 
withdrawal, and ask for any adverse events. 
If the subject decides to withdraw during the study, the safety telephone follow-up will be 
made on days 3, 7, 14 and 21. The investigator may retain and continue using any data 
collected prior to the withdrawal of consent. 
A patient will be withdrawn from the study due to any of the following causes: 
1. If you have study drug related- adverse events considered unacceptable. 
2. Free decision by the volunteer who no longer wishes to participate in the study 
without any type of reason 
3. Serious adverse event 
4. Decision by ANMAT 
5. Decision by the Sponsor, or Ethics Committee at DIM CLINICA PRIVADA Site. 
6. Risk to the volunteer 
7. Decision by the principal investigator, who considers the volunteer is at undue risk 
8. Participant who has the possibility of receiving the first or second dose of the 
COVID vaccine during the study. 
9. If the study doctor thinks that continuing in the study may not be at patient's 
health best interest. 
10. If the Sponsor, the Argentine Public Health authority (A.N.M.A.T.) or other 
applicable regulatory authorities decide to discontinue the study. 

11. Non-compliance by the volunteer 
12. Not meeting the inclusion criteria or meeting some exclusion or withdrawal criteria 
at the time of hospitalization. 
 
6 
STUDY DRUG 
 
6.1 
DRUG NAME, FORMULATION AND STORAGE  
 
6.1.1 Study Formulations 
Test Formulation (T) 
Biosidus beta agalsidase. Powder for concentrate for solution for infusion. It is supplied as 
both 35 mg and 5 mg per vial to adjust for the indicated dose 
The following data will be specified on the labels of the infusion vials: Lot, Study and 
expiration date. 
 
Qualitative-quantitative formula: 
Biosidus beta agalsidase 35 mg: Each vial contains lyophilized powder to reconstitute: 
 
Drug Substance 
Agalsidase beta 
35 mg 
Inactive 
ingredients 
Sodium phosphate 
monobasic 
anhydrous 
42 mg 
Sodium Hydroxide 
q.s. pH 6.8 
Mannitol 
210 mg 
Once reconstituted with 7.2 mL WFI, each vial contains 5 mg/mL (35 mg/7 mL) agalsidase 
beta. Reconstituted solution must then be diluted.  
 
Biousidus agalsidase beta 5 mg: each vial contains lyophilized powder to reconstitute:  
Drug Substance 
Agalsidase beta 
5.5 mg 
Inactive 
ingredients 
Sodium phosphate 
monobasic 
anhydrous 
6 mg 
Sodium Hydroxide 
q.s. pH 6.8 

Mannitol 
30 mg 
Once reconstituted with 1.1 mL WFI, each vial contains 5 mg/mL agalsidase beta. 
Reconstituted solution must then be diluted. 
 
Reference Formulation (R) 
Sanofi-Genzyme Fabrazyme®. Powder for concentrate for solution for infusion. It is 
supplied as both 35 mg and 5 mg per vial to adjust for the indicated dose.  
The following information shall be specified on the labels of infusion bottles: Lot, Study and 
expiration date. 
Qualitative-Quantitative Formula: 
Fabrazyme® 35 mg: each vial contains lyophilized powder to reconstitute:  
Drug Substance 
Agalsidase beta 
37 mg 
Inactive 
ingredients 
Sodium phosphate 
monobasic 
monohydrate 
20.4 mg 
sodium phosphate 
dibasic heptahydrate  
59.2 mg 
Mannitol 
222 mg 
Once reconstituted with 7.2 mL WFI, each vial contains 5 mg/mL (35 mg/7 mL) 
Fabrazyme® agalsidase beta. Reconstituted solution must then be diluted. 
 
Fabrazyme® 5 mg: each vial contains lyophilized powder to reconstitute:  
Drug Substance 
Agalsidase beta 
5.5 mg 
Inactive 
ingredients 
sodium phosphate 
monobasic 
monohydrate 
3 mg 
sodium phosphate 
dibasic heptahydrate  
8.8 mg 
Mannitol 
30 mg 
Once reconstituted with 1.1 ml WFI, each vial contains 5 mg/mL Fabrazyme agalsidase 
beta. Reconstituted solution must then be diluted. 
 
Justification for the difference in drug substance Agalsidase Biosidus and Frabrazyme® 
strengths is found in Annex 7. 

 
6.1.2 Content and Potency Uniformity 
Prior to the clinical phase of this study, the Biosidus Development Department will 
evaluate the comparative enzyme activity of all batches of the Test and Reference 
products to be used in the clinical study. Content uniformity of the formulations will also be 
determined by reverse phase HPLC. 
Justification for the difference in drug substance Agalsidase Biosidus and Frabrazyme® 
strengths is found in Annex 7. 
 
6.1.3 Storage 
All the medication for this study will be stored in the refrigerator at 2 °C – 8 °C, not in the 
freezer, under adequate conditions, indicated by the sponsor in writing, with restricted 
access to the staff in charge. 
 
6.2 
MATERIAL PREPARATION AND LABELING 
Medication used in the study will be provided by Biosidus S.A., in accordance with current 
regulations. Preparation for the medication used for each volunteer will be made by the 
principal investigator, or his designee, as follows. 
Each volunteer will receive one (1) intravenous infusion in the arm. Vials will be labeled 
with a self-adhesive label where the study code and other information necessary for the 
label of the investigational product will be mentioned, in accordance with ANMAT Provision 
6677/2010, as indicated in Annex 3. Reconstitution and dilution for each volunteer will be 
made based on his body weight. 
 
6.3 
DOSE AND ROUTE OF ADMINISTRATION 
Each volunteer will receive a single 1 mg/kg agalsidase beta dose. 
To adjust the required dose based on the volunteer´s weight on the hospitalization day, the 
following formulations will be available: 
1. Fabrazyme® 35 mg lyophilized powder to be reconstituted for infusion 
2. Fabrazyme® 5 mg lyophilized powder to be reconstituted for infusion 
3. Biosidus Agalsidase 35 mg lyophilized powder to be reconstituted for infusion 
4. Biosidus Agalsidase 5 mg lyophilized powder to be reconstituted for infusion 
 
6.4 
TREATMENT DURATION 

Study duration will be approximately 56 days for each volunteer, assuming: 21 days for the 
screening period (Visit 1), 1 day for hospitalization and treatment (Visit 2), 4 telephone 
contacts (days 3, 7, 14 and 21 - Visit 3, 4, 5 and 6) and 1 day for the Final Visit (day 35 – 
End of study). 
Each volunteer will receive only one study formulation (T or R) at visit II (Hospitalization). 
7 
ENZYME ACTIVITY ANALYSIS 
For measurement of the biological enzyme activity in plasma samples, the fluorimetric 
method of hydrolysis of the synthetic substrate 4MU-Gal will be used. 
7.1 
PHARMACOKINETIC ANALYSIS OF RESULTS 
From the results of plasma concentration of agalsidase beta at the different sampling 
times, the following variables will be calculated: 
ABC0-t: Area under the plasma concentration curve between time 0 and time t (in this case 
12 hours). 
ABC0-∞: Area under the plasma concentration curve, result of extrapolation between time 0 
and time ∞. 
Cmax: Peak plasma concentration of agalsidase beta. 
Tmax: Time to reach the peak plasma concentration of agalsidase beta. 
Cmax / ABC0-∞: ratio between the peak plasma concentration and the area under the 
plasma concentration curve between time 0 to ∞, as an absorption rate index of less 
variability than the Tmax. 
 
The drug elimination constant and elimination half-life in the body will be calculated. The 
summary measures (arithmetic and geometric means, standard deviation, CV% and 
range) for each infusion will be presented, as well as the individual results of each 
volunteer. 
 
7.1.1 Comparative pharmacokinetic analysis between formulations 
ABCT/ABCR: Ratio between the area under the curve of each test formulation and the area 
under the curve of the reference formulation. 
For logarithmic transformation, the 90% confidence interval of the AUC ratio (for ABC0- 120 h 
and ABC0-∞) should be within the range of 0.80-1.25. 
 
Cmax T/Cmax R: Ratio between the Cmax of each Test formulation and Cmax of the Reference 
formulation. For logarithmic transformation, the 90% confidence interval of the Cmax ratio 
should be first within the range 0.80-1.25 (80% to 125%). Considering the high intra-
individual variability of this pharmacokinetic parameter, if a variability > 30% is confirmed, 
this interval for the Cmax may be extended to 0.75-1.33 (75-133%). 

 
The FDA assumes that the test product is considered bioequivalent to the reference 
product if the 90% confidence interval (CI) of the geometric mean ratio of AUC and Cmax 
between the test product and the reference product fall within the range 80-125%, even 
though the application of such criterion to drugs with high variability is controversial and 
regulatory agencies as strict as the FDA use different criteria. In Argentina, renowned 
specialists such as R. Bolaños, MD and S. Giarcovich, MD have analyzed the current 
status of the bioequivalence in the world proposing the most appropriate study designs. 
Globally, drugs with high intra-individual variability are considered to be those with a 
coefficient of variation (CV) greater than 30% in pharmacokinetic parameters such as Cmax 
and AUC. Different strategies have been tested given the high variability issue and in order 
to meet the conventional bioequivalence principles (90% CI 80-125). One has been to 
significantly increase the number of volunteers, sometimes to an unfeasible number, 
although this procedure does not guarantee that the results will fall within that equivalence 
interval. In this regard, there are reference products with high variability that are not 
capable of demonstrating bioequivalence with themselves. Additionally, the increase in 
volunteers to minimize variability poses an ethical problem, since we expose a greater 
number of healthy individuals to drugs that involve a risk in order to meet the 
bioequivalence criteria, which have been arbitrarily established. Other approaches have 
additional problems. 
Given this difficulty, most regulatory agencies consider expanding the classical limits of 
bioequivalence (80-125%) in bioequivalence studies with drugs with high intra-individual 
variability. These considerations are made on the Cmax and not on the AUC, where there is 
a consensus to keep the range 80 to 125% in all products. The reason for the greatest 
Cmax variability compared to the AUC is the fact that this parameter arises from the 
measurement of an individual point, which makes it highly dependent on time, sampling 
frequency, and drug elimination rate. 
Several proposals are available in the literature to modify the bioequivalence criteria for 
drugs with high variability. In general, the proposals are based on reducing the level of the 
confidence interval, on widening the equivalence limits, or on both. 
A reduction in the level of the confidence interval, for example, from 90 to 85%, would 
imply a possible increase in the risk of patients, which is usually not acceptable. In 
contrast, the extension of the equivalence limits, which is the generally accepted criterion 
and the one used here for Cmax, represents what limits are admissible and tolerable for the 
ratios (or differences) of the means between the compared products. 
Any adjustment of these limits should be based on considerations of the statistical data 
properties, as well as the clinical characteristics of the individual drug. Statistically, the 
extension of the limits can be done by directly modifying the tolerated value of the 
bioequivalence limits, which is what some agencies such as the EMEA, Japan and South 
Africa have adopted. In countries like Canada, confidence limits for Cmax are not required, 
but rather that the geometric mean ratios for T/R be within 80 -125%. 

Using simulation techniques, Tothfalusi L et al.8 compared the gradual limit escalation for 
high variability drugs and the direct expansion of the limits around Cmax. Escalation 
involves extending the limits of the confidence interval based on the variability of the 
reference product. Authors conclude that escalation would significantly reduce the sample 
size needed for bioequivalence studies of high variability drugs. They also conclude that 
the same results are obtained simply by extending the regulatory limits to 75-133% or 
even 70-143% for Cmax. 
To calculate the pharmacokinetic/pharmacodynamic parameters, specific pharmacokinetic 
software will be used (EquivTest version 2.0 (Equivalence Testing Software) from 
Statistical Solutions Ltd.) 
 
Outliers: for measuring potential outliers, a selective statistical method will be applied. K&H 
Consulting S.A. will be responsible for statistical processing. 
 
7.2 
PHARMACODYNAMIC ANALYSIS OF RESULTS 
As an estimator of agalsidase beta biological activity, blood samples will be drawn to 
determine the variation in the agalsidase beta enzyme activity of the end-of-infusion 
sample (5 h) compared to baseline activity (pre-infusion).  
Ratio of activity variation in both groups to be close to the unit and that the 90% CI is 
within the range 0.8 to 1.25 will be taken as bioequivalence criterion. 
8 
IMMUNOGENICITY ANALYSIS 
The incidence of anti-agalsidase beta antibodies will be determined both at the start of 
treatment, at the end of visit II and 31 days after the end of the infusion, for both treatment 
groups. 
Serum samples will be incubated with 1 ng agalsidase beta for 15 minutes at room 
temperature. Then, the enzyme activity for the samples will be determined using the 4-MU-
Gal substrate. 
The inhibition percentage of the samples will be determined by comparing the activity 
value of 1 ng agalsidase beta with the treated samples. Those samples with percentage 
inhibition > 50% will be reported as positive. 
 
9 
SAFETY ANALYSIS 
9.1 
ADVERSE EVENTS 
All adverse events that occurred in the subjects during their participation in the study will 
be recorded. The record will be documented in the subject's Medical Record and on the 
applicable pages of the Case Report Form. 

All solicited and unsolicited reported adverse events, regardless of treatment group or 
presumed causal relationship to the investigational product(s), will be reported as 
described in the following sections. 
For all adverse events, the investigator must search and obtain adequate information, both 
to determine the outcome of the adverse event, and to assess whether or not it meets the 
criteria for its classification as a serious adverse event, which require immediate reporting 
to the ANMAT.  For all adverse events, the investigator must obtain sufficient information 
to determine the adverse event causality. The investigator is required to assess such 
causality. For adverse events with a causal relationship to the investigational product, 
follow-up is required by the investigator until the event or its sequelae resolve or stabilize 
at a level acceptable to the investigator. 
 
9.2 
ADVERSE EVENT REPORTING 
All adverse events detected by the investigator or his collaborators, or reported 
spontaneously by the subjects at each visit, must be reported in a special section in the 
Case Report Form. Adverse events will be described preferably using WHO terminology. 
The study monitor will check the completion in the CRF. 
The following information must be recorded for each adverse event, whether it is related to 
the study drug or not: 
• 
description of the adverse event 
• 
start date / end date 
• 
duration of the event 
• 
characteristics of the event (severity, expectedness, seriousness) 
• 
action taken (report any required treatment or dose adjustments in the CRF) 
• 
progress 
• 
relationship to study drug (causality) 
All adverse events must be documented and followed until their resolution or adequate 
justification, even when the subject has completed the study. Any change in the dose of 
the drug administered will be reported on the Case Report Form. 
All non-serious adverse events (expected and unexpected) will be described in the final 
study report. 
The Serious Adverse Event report will be prepared in compliance with ANMAT 6677/2010 
provision (Section B, item 7). 
The investigators will report Serious Adverse Events (SAE) immediately, within 24 hours, 
to the sponsor by phone 49098049/ fax no. 49243601 (Biosidus Development Department) 
and/or email farmacovigilancia@biosidus.com.ar, except those events that the protocol 
identifies as not requiring immediate reporting. 

According to provision 6677/2010, item 7.1, the Sponsor must inform ANMAT of all 
Serious and Unexpected Adverse Drug Reactions (RAMSI – by its Spanish acronym) 
related to the product under study within 10 administrative business days of becoming 
aware of it. The RAMSI caused by a comparator product already registered in ANMAT for 
being marketed in the country or those related to placebo must be reported only to the 
ANMAT Pharmacovigilance System. The Investigator must report Serious and Unexpected 
Adverse Events to the Teaching and Research and Ethics Committee(s) in accordance 
with their requirements, in a timely manner. 
 
Based on provision 6677/2010, item 7.3, RAMSI reporting must include the following 
information: 
• 
name of the adverse reaction; 
• 
investigational product: code, INN name and/or trade name; 
• 
type of report: initial, update (update number) or final; 
• 
sponsor name; 
• 
investigational site and investigator name; 
• 
number of ANMAT Provision authorizing the study; 
• 
study name or title; 
• 
participant code, age and sex; 
• 
dates of RAMSI occurrence, investigator's report and date it is received by the 
sponsor; 
• 
RAMSI seriousness criterion; 
• 
brief description for the RAMSI; 
• 
investigational product information: daily dose, route of administration, start and 
stop dates, duration of therapy, and indication; 
• 
specify if the reaction went away when treatment was discontinued; 
• 
specify if the reaction reoccurred with treatment rechallange; 
• 
concomitant medications: dosage and start and end dates; 
• 
Participant history relevant to the RAMSI. 
A single RAMSI Report per drug will be accepted, referring to all files and approval 
provisions by A.N.M.A.T. of studies that are being conducted in the country with that drug. 
The Sponsor must immediately notify the Investigator(s) and the Ethics Committee at DIM 
CLINICA PRIVADA site that approved the study of all Adverse Drug Reactions that are 
both Serious and Unexpected that occur in other national investigational sites. 
The Sponsor must inform the Investigator(s) and the Ethics Committee at DIM CLINICA 
PRIVADA that approved the study of all other relevant information regarding the research 

product within a period not exceeding 10 business days from receiving initial information 
by the Sponsor. 
Adverse Reactions attributable to placebo or to the comparator will not be subject to this 
individualized reporting system. Reports submitted to ANMAT will comply with the item 
7.1, provision 6677/2010. 
According to section E: Glossary, provision 6677/2010 establishes the following definitions 
of Adverse Drug Reaction (ADR) and Serious and Unexpected Adverse Drug Reaction 
(RAMSI). 
 
Adverse Drug Reaction (ADR): noxious and unintentional response to a medicinal 
product related to any dose. In clinical experience prior to the approval of a new medicinal 
product or its new uses, particularly when the therapeutic dose cannot be established, an 
adverse drug reaction should be considered any reaction that involves a causal 
relationship between a medicinal product and an adverse event such as a reasonable 
possibility, that means, that the relationship cannot be ruled out. 
 
Serious and Unexpected Adverse Drug Reaction (RAMSI): is the adverse reaction that 
results in death, is life threatening, requires hospitalization or prolongs an existing 
hospitalization or produces persistent or significant disability or incapacity, and their nature 
or severity is not consistent with the product information described in the IB or other 
documentation. 
 
9.3 
ADVERSE EVENT DEFINITION  
An adverse event is any adverse medical event, occurring in a clinical research subject 
who has been administered a product or who uses a medical device. The event does not 
necessarily have to have a causal relationship with the treatment or use. 
Examples of adverse events include, but are not limited to, the following: 
• 
Abnormal findings in investigations. 
• 
Clinically significant symptoms and signs; 
• 
Changes in physical examination findings; 
• 
Hypersensitivity; 
• 
Progression/worsening of an underlying disease; 
 
Additionally, they may include signs and symptoms resulting from: 
• 
Drug overdose; 
• 
Drug withdrawal; 
• 
Drug abuse; 
• 
Inappropriate drug use; 

• 
Drug interactions; 
• 
Drug dependency; 
• 
Extravasation 
 
9.4 
ABNORMAL FINDINGS IN THE INVESTIGATIONS 
Criteria for determining whether an abnormal finding on investigations should be reported 
as an adverse event are as follows: 
• 
The result of the investigations is associated with accompanying symptoms, 
and/or 
• 
The test result requires additional diagnostic testing and/or medical/surgical 
intervention, and/or 
• 
The investigator considers the test result as an adverse event. 
• 
A repeated abnormal investigation, in the absence of any of the conditions 
mentioned above, does not constitute an adverse event. Any result of an 
abnormal investigation determined as an error does not require being reported 
as an adverse event. 
 
9.5 
SERIOUS ADVERSE EVENT 
A serious adverse event is any unfavorable medical event that at any dose: 
• 
results in death; 
• 
is life-threatening; (immediate risk of death) 
• 
requires subject hospitalization or prolongation of existing hospitalization; 
• 
results in a persistent or significant disability/incapacity; 
• 
results in a congenital abnormality/birth defect. 
• 
Important / significant medical events. 
In determining whether an event is a significant medical event, medical and scientific 
judgment must be exercised. An important medical event may not be immediately life-
threatening and/or result in death or hospitalization. However, if it is determined that the 
event may jeopardize the subject and require intervention to prevent one of the other 
outcomes listed in the definition above, the important medical event should be reported as 
serious. 
Examples of such events are: intensive treatments in an emergency room or at home for 
allergic bronchospasms; blood dyscrasias or seizures that do not result in hospitalization; 
or the development of drug dependence or abuse 
 

9.6 
HOSPITALIZATION 
Reported adverse events from clinical trials associated with hospitalizations, or 
prolongation of hospitalizations, are considered serious. Any initial admission (even for 
less than 24 hours) to a health care institution meets such criteria. Admission also includes 
transfer within the hospital to an acute/intensive care unit (e.g., from a psychiatric ward to 
a clinical ward, clinical ward to coronary unit, neurology ward, etc.). 
Hospitalizations do not include the following: 
- 
rehabilitation institutions; 
- 
hospices; 
- 
qualified nursing facilities; 
- 
nursing sites; 
- 
routine admission to an emergency room; 
- 
same-day surgeries (outpatient/same day procedures). 
- 
any hospitalization or prolongation of an existing hospitalization, in the 
absence of a clinical adverse event that precipitates it, does not constitute, in 
itself, a Serious Adverse Event. Examples include the following: 
- 
admission for treatment of a pre-existing condition, not associated with the 
development of a new adverse event, or with the worsening of a pre-existing 
condition (for example, to treat a persistent laboratory abnormality prior to 
treatment); 
- 
admission for social reasons (for example, the subject has nowhere to sleep); 
- 
administrative admission (for example, for an annual physical examination); 
- 
protocol-specified admission during a clinical trial (eg, for a procedure 
required by the trial protocol); 
- 
optional admission not associated with a precipitating adverse event (for 
example, for elective cosmetic surgery); 
- 
previously planned surgical treatments or procedures must be recorded in the 
baseline documentation for the entire protocol and/or for the individual 
subject. 
- 
non-invasive and invasive diagnostic and therapeutic procedures, such as 
surgery, should not be reported as adverse events. However, the medical 
condition for which the procedure is performed must be reported, if it meets 
the definition of an adverse event. For example, acute appendicitis beginning 
during the adverse event reporting period should be reported as an adverse 
event, and the resulting appendectomy recorded as treatment of the adverse 
event. 
 

 
9.7 
SEVERITY ASSESSMENT 
If required by the adverse event case report forms, the investigator will use the adjectives 
MILD, MODERATE, or SEVERE to describe the maximum intensity of the adverse event. 
For purposes of consistency of information, these degrees of intensity are defined as 
follows: 
 
MILD 
Does not interfere with the subject's usual activities   
MODERATE 
Somehow interferes with the subject's usual activities 
SEVERITY 
Significantly interferes with the subject's usual activities 
 
Please note the differentiation between severity and seriousness of an adverse event. A 
severe event is not necessarily a serious event. For example, a headache may be severe 
(significantly interferes with the performance of the subject's usual functions), but would 
not be classified as serious, unless it meets one of the criteria considered for serious 
adverse events, detailed above. 
 
9.8 
CAUSALITY ASSESSMENT (RELATIONSHIP TO MEDICATION, RELATEDNESS) 
The investigator's causality assessment must be provided for all Adverse Events (serious 
and non-serious). The investigator's causality assessment consists of determining whether 
or not there is a reasonable possibility that the product under investigation caused or 
contributed to an adverse event. 
Causality may be assigned with one of the following two possibilities: 
• 
No: Definitely not drug related. 
• 
Yes: remotely, possibly, probably, or definitely related to the drug 
If the investigator's final determination of causality is unknown, and the investigator does 
not know whether the event was caused by the investigational product, then the event will 
be considered “related to the investigational product” for the purposes of event reporting. If 
the investigator's assessment of causality is “unknown, but clearly not related to the 
investigational product,” that fact must be clearly documented in the study records. In 
addition, if the investigator determines that a serious adverse event is associated with the 
trial procedures, the investigator must record such causal relationship in the source 
documents and Case Report Forms, as applicable, and report such assessment in 
accordance with the requirements for reporting serious adverse events, if applicable. 
All Adverse Events judged by the investigator or sponsor to be "drug related" qualify as an 
Adverse Drug Reaction (ADR). 
 

 
10 STATISTICAL CONSIDERATIONS AND ANALYTICAL PLAN. SAMPLE 
SIZE CALCULATION 
A recent study with JR-051, the first agalsidase beta biosimilar to be approved in Japan, 
was conducted in 20 volunteers (10 per treatment group), based on the fact that previous 
studies with this molecule assumed the geometric mean of Cmax and ABC0–24 with a value 
of 1.0 and a coefficient of variation of 0.15. 
Therefore, with a power = 0.80, significance level α = 0.05, and a limit for accepting 
bioequivalence where the 90% confidence interval of the AUC ratio (for AUC0-120h and 
AUC0-∞) should be within the range of 0.80-1.25 9 volunteers per group are required for 
the study, estimating the possibility of 1 volunteer lost to follow-up per group, the sample 
size was calculated in 20 volunteers. 
11 ETHICAL ASPECTS 
11.1 LOCAL REGULATIONS/ DECLARATION OF HELSINKI 
The Principal Investigator, the Co-Investigators and the members of the Research team 
will ensure that this study is conducted in full compliance with the principles of the 
"Declaration of Helsinki 1964" (latest version 64th General Assembly, Fortaleza, Brazil de 
October 2013) and with the Argentine laws and regulations, in accordance with the ones 
that most protects the individual. The study must strictly adhere to the principles 
enunciated in the "Guidelines for Good Clinical Practice" (ICH Tripartite Guideline, January 
1997) or local laws, if they provide greater protection for the patient. 
 
11.2 INFORMATION FOR THE SUBJECT AND INFORMED CONSENT 
The informed consent form must be approved by DIM CLINICA PRIVADA Ethics 
Committee and local regulatory agencies (ANMAT) and must be in accordance with the 
the International Conference on the Harmonization Good Clinical Practice guidelines and 
local legal and regulatory requirements. 
The investigator or the investigator's designee (if permitted by local regulations) is 
responsible for obtaining the written informed consent from each individual participating in 
this study or undergoing any examination to assess inclusion in the study, after the 
objectives, methods, and potential risks of the study have been adequately explained. It is 
also necessary to explain to the volunteers that they are completely free to refuse to 
participate in the study or to withdraw from it at any time and for any reason and they will 
be provided with the corresponding forms for the written informed consent. 
 
11.3 INDEPENDENT ETHICS COMMITTEE (IEC) 

It is the investigator's responsibility to obtain advance approval of the Study protocol, 
protocol amendments, Informed Consent forms and other relevant documents, notices, if 
applicable, by the DIM CLINICA PRIVADA Ethics Committee 
The Principal Investigator will submit to DIM CLINICA PRIVADA Ethics Committee, this 
protocol, any protocol amendment, as well as all accompanying material. Before starting 
the study, approval by this Committee must be obtained, which must be documented in a 
letter addressed to the investigator, specifying the date on which the Committee met and 
granted its approval. 
All correspondence and emails with the DIM CLINICA PRIVADA Ethics Committee must 
be kept at the Investigator's file. 
12 CONDITIONS FOR AMENDING THE PROTOCOL 
All amendments to the protocol must be submitted to DIM CLINICA PRIVADA Ethics 
Committee and ANMAT. No changes may be implemented until such approval is obtained, 
except if they are necessary to eliminate an immediate risk to trial patients or if the 
change(s) involve(s) logistical or administrative aspects of the trial (e.g., change of 
monitor(s), change of telephone number(s)). 
13 CONDITIONS FOR COMPLETING THE STUDY 
Study early termination may occur upon Principal Investigator, Sponsor, ANMAT 
Regulatory Authorities, the Ethics Committee decision or due to drug safety problems. 
The investigator has the right to terminate the study at any time. When terminating a study, 
the investigator should ensure that special consideration has been given to patient 
protection. 
Following notification, the investigator must contact all participating subjects within one 
month of becoming aware of trial termination or discontinuation. 
 
14 STUDY DOCUMENTATION, CRF and RECORDS 
14.1 INVESTIGATOR FILES / DOCUMENT RETENTION 
The investigator, the clinical test laboratory and the plasma measurement laboratory must 
keep accurate records so that the study is adequately documented and its data can be 
verified. These documents should be classified into two different categories: (1) 
Investigator file and (2) patient source clinical documents. 
Patient source clinical documents will include hospital/clinic generated patient records, 
physician and nursing notes, visit schedule, original laboratory reports, ECG, pathology 
and special assessment reports, signed informed consent forms, mailing and patient 
screening and recruitment logs. 
Principal Investigator will file copies of all the information and documentation submitted to 
the ANMAT regulatory authorities, treatment sequence randomization, monitoring reports 

conducted at the hospitalization site, the original copies of receipt and return of materials 
and leftover drugs that is carried out in each hospitalization cycle. 
Medical records, clinical laboratory reports, anteroposterior chest X-ray and admission 
electrocardiogram, Case Report Forms, copies of receipt and return of materials, will be 
available at the hospitalization sit. 
The data and all information from the agalsidase beta enzyme activity method and its 
results will be available at Biosidus Analytical Department - Constitución 4234 - 
C1254ABX Buenos Aires, Argentina – Phone 4909 8000 - Fax 4909 8055. 
The investigator must retain these documents in his file for at least 10 years from the study 
termination or discontinuation. In case of not being able to retain the documentation during 
that period, it will be kept by the sponsor. After that period, documents may be destroyed, 
in accordance with local regulations. (GCP - Version July 1991). 
 
14.2 AUDITS AND INSPECTIONS 
Principal Investigator and his team will accept that the source documents for this trial are 
available to the study monitors, previously agreed sponsor audits, members of DIM 
CLINICA PRIVADA Ethics Committee and inspections by the ANMAT or other regulatory 
authorities with prior notification and agreement of inspection date and time. Verification of 
the data contained in the CRFs will be carried out by direct inspection of the source 
documents. In no case may the inspectors remove original documents, photocopies or 
data from the study volunteers from Sanatorio Nuestra Señora del Pilar site. 
 
14.3 CASE REPORT FORMS (CRF) 
Principal investigator or authorized study staff delegated must complete and sign a CRF 
for each enrolled patient. This will also apply to the records of those patients who do not 
complete the study (even when this occurs during the screening period prior to 
randomization, if a CRF has already been initiated). If a patient withdraws from the study, 
the reason should be recorded in the CRF. If a patient is withdrawn from the study due to 
an adverse treatment effect, every effort should be made to clearly document the course. 
Forms must be completed in printing or with indelible black ink and must be legible. Errors 
must be crossed out with a transversal line —never erased—, the correction must be 
inserted, and the change must be signed and dated by the investigator or its authorized 
designee. Investigator must ensure that data reported to the sponsor in the CRFs and in 
all requested reports are complete, accurate, legible, and timely. 
 
14.4 CONTRACT RESEARCH ORGANIZATIONS 
The study protocol has been prepared by Dr. Eduardo Pirotzky, who assumes the duties of 
Principal Investigator for this study, as agreed with the sponsor in the agreement signed 
between both parties. 

15 STUDY MONITORING 
The monitor will contact and meet regularly with the investigator and upon request, will be 
allowed to inspect the various trial records (CRF and other relevant data) as long as 
patient confidentiality is kept, in accordance with the local requirements. 
The monitor will be responsible for inspection of CRFs at regular intervals during the study 
to verify the degree of compliance with the protocol and that the data entered are 
complete, consistent and accurate. 
The monitor will be able to access laboratory test reports and other patient records needed 
to verify CRF entries. 
Investigator (or his representative) will agree to cooperate with the monitor to ensure the 
any problems detected during these visits are resolved. 
Monitors should review source documents to confirm that the data recorded on the CRFs 
is accurate. 
Investigator and the Hospitalization Center must allow monitors, regulatory authorities and 
members of the DIM Clínica Privada Ethics Committee who require it, direct access to the 
source documents to comply with this verification. 
In no case will it be authorized to withdraw from the hospitalization center: originals, 
photocopies or written data of the volunteers of the source documents. 
It is important that both the respondent and relevant personnel are available during 
monitoring visits, and possible audits or inspections, and that they are able to devote 
sufficient time to this process. 
 
16 CONFIDENTIALITY OF THE STUDY DOCUMENTS AND PATIENT 
RECORDS  
The Principal Investigator and his team, Sanatorio Nuestra Señora del Pilar site must 
ensure the anonymity of the participating individuals and it will not be authorized to pick up 
the original, photocopy or annotation that violates the data confidentiality, violating the 
confidentiality and anonymity of the volunteer. 
In the CRF or other documents, patients will not be identified by their names but by their 
identification code. 
The investigator must keep a patient recruitment log in which the codes, names and 
addresses appear. 
All information resulting from this study may be seen by monitors, auditors, members of 
the DIM CLINICA PRIVADA Ethics Committee and ANMAT regulatory authorities, after 
arranging the date and time of the visit with the investigator, without violating confidentiality 
and secrecy about the identity of the volunteer to the extent permitted by applicable laws 
and regulations. 

If the results of the study are published, the volunteer identity will be kept confidential. 
According to Law 25,326 on Personal Data Protection: the information collected "...cannot 
be used for purposes other than or incompatible with those for which it was obtained...", 
"...must be destroyed when it is no longer necessary or pertinent to the purposes for which 
it was collected..." and "...the processing of personal data is unlawful when the owner has 
not given his free, express and informed consent, which must be in writing, or by another 
equal means, according to the circumstances.” 
According to the same law, the personal data bearer is allowed to exercise the right of 
access to them free of charge at intervals no shorter than six months, unless a legitimate 
interest is accredited to that effect, in accordance with the provisions of section 14, 
paragraph 3 of Law No. 25326. 
The National Directorate for the Personal Data Protection, controlling Body of Law No. 
25326, has the power to deal with complaints and claims that are filed in relation to non-
compliance with the rules on personal data protection. 
All data generated during the study (Registration Forms, patient identification list, 
laboratory findings) will be kept for 10 years. In the event that the principal investigator 
could not retain the documentation during this time, the sponsor will be informed, who will 
be in charge of safeguarding it. 
If the results of the study are published, the volunteer identity will be kept confidential. 
Data related to this study and its results are the property of the sponsor and may not be 
made public, including presentations at congresses or scientific publications, without the 
sponsor written permission. 
 
17 QUALITY ASSURANCE 
Good Clinical Practice 
The study will be conducted adhering to the GCPs, following the international 
recommendations (ICH) or, otherwise, the European (EMA) and national (ANMAT) 
Guidelines. 
The study quality control and the GCP audits may be carried out by the Sponsor and its 
quality assurance service or by an inspector from ANMAT. In particular, this protocol will 
be submitted for the Independent Ethics Committee (IEC) approval. Consistency between 
study development and the protocol will be controlled. No amendment to the protocol will 
be made without approval by the CEI that has agreed to the study conduct, and without 
documented authorization from the Sponsor. Sponsor will be notified of any minor changes 
to the protocol. In case of important and written modification (modification that could 
compromise the integrity of the people participating in the study and modifications that 
could change the scientific validity of the study) the protocol will be submitted again for IEC 
approval. 

Each patient will receive detailed written information, in the information letter, including 
active name under study, a summary of its properties and potential side effects, doses 
administered, administration times, number of visits, nature and number of previous 
exams, previously sent to the IEC. 
The patient will be informed that she can withdraw from the study at any time without 
justification. 
The system for assigning numbers to patients is based on the obligation to maintain 
anonymity. Patients will be numbered in the order of inclusion in the study. They will be 
identified by the Sponsor by the assigned number, a two-letter code (first letter of the last 
name and first letter of the first name), their date of birth (day-month-year). 
Compliance with the study development will be controlled from the patient screening and 
throughout the trial. To control the study development, the Principal Investigator will make 
available to the monitor or to any other person designated, the necessary documents for 
the control of the data reported in the observation book (medical records, investigations 
results, etc.). 
The Principal Investigator will be available by phone for each patient he has included. The 
CRFs will be completed legibly with a black pen without deletions or overwriting. Deletions 
and overwriting, modifications, will be corrected by the Principal Investigator by signing 
and dating them. The CRF and any other document considered source document that may 
be filed (biological tests, written informed consent, etc.). 
 
Quality Control 
Control actions will be taken within the frame of the Quality Assurance system to verify that 
the quality requirements of the trial are met. 
The original documents, produced during the study, will be controlled by both the Sponsor 
representative of the Sponsor, the person responsible for the study, and the Principal 
Investigator at each stage of the study in order to guarantee the reliability of the data 
analyzed. 
Internal audits may be carried out during the course of the study by the representative of 
the Sponsor or an independent body. These would make it possible to verify that the study 
is conducted in accordance with the protocol and the regulations and recommendations in 
force. 
 
18 FINAL REPORT AND PUBLICATION 
The Principal Investigator or the Person responsible for conducting the Study agrees to:  
• 
Keep the Sponsor informed in writing of the results of the study. 
• 
Write a study results report for the Sponsor. 
 

19 CALENDAR PROVIDED 
The clinical study will begin immediately after the approval of the protocol by the DIM 
CLINICA PRIVADA Ethics Committee and obtaining the authorizing provision by ANMAT. 
The duration of the study for a patient will be 56 days. 
 
20 FUNDING AND INSURANCE 
The Sponsor will regulate all expenses related to the study. A commercial agreement has 
been signed between the Sponsor and Research & Development RA. S.A. (CRO 
Contracted) and with Dr. Eduardo Pirotzky and/or all the doctors participating in the study 
who are under his control.  
The Sponsor assures full and total responsibility for any damage that may result from the 
experimentation described in this protocol, subject to the fact that the terms and conditions 
of the study have been thoroughly respected by the Principal Investigator. 
 
 
 

21 SIGNATURE PAGE 
I have carefully read and reviewed the protocol. Having read and understood the 
requirements and conditions of the protocol, I agree to conduct this study following 
international guidelines for good clinical practice and local regulatory requirements, for 
verification of source documents, study audits and inspections. 
I agree to use the study material, including medication, only as stated in this protocol. 
I understand that any change to the protocol must be made in the form of an amendment 
that must be previously approved by the sponsor, DIM CLINICA PRIVADA Ethics 
Committee and the ANMAT. 
I understand that any protocol violation may lead to the study early termination. I agree 
with the following study schedule. It will begin after the ANMAT approval and will end in a 
period not exceeding five months later. 
I agree to report to Biosidus S.A. within 24 hours of any serious adverse event, whether 
considered related or not to the drug administered. 
I understand that any protocol violation may lead to early study termination. 
 
Principal Investigator 
 
___________________ 
___________________ 
___________________ 
Date 
 
 
 
 
Signature 
 
Name 
 
Biosidus S.A. 
 
___________________ 
___________________ 
___________________ 
Date 
 
 
 
 
Signature 
 
Name 
 
 

22 REFERENCES 
1. H. Blume, K.K. Midha. Report of Consensus Meeting: Bio-international ’92. 
Conference on Bioavailability, Bioequivalence and Pharmacokinetic Studies. 
Bad Homburg, Germany, 20-22 May, 1992. Eur. J. Pharm. Sci. 1:165-171 
(1993). Bio-international ’94, Munich Germany. 14-18 June, 1994. 
2. Giarcovich SS, Bolaños R. El “ser o no ser “de la Bioequivalencia: Rangos 
one size fits all vs. case by case y otras opciones. Revista de la SAFYBI 
(Sociedad Argentina de Farmacia y Bioquímica Industrial) 44(108):6-25 
(2004). 
3. Health Canada, Therapeutic Products Directorate (TPD). Discussion paper 
on 'Bioequivalence requirements - highly variable drugs and highly variable 
drug products: issues and options'. Expert Advisory Committee on 
Bioavailability and Bioequivalence (EAC-BB) Meeting, June 26-27, 2003. 
4. A.W. Boddy, F.C. Snikeris, R.O. Kringle, G.C.G. Wei, J.A. Oppermann, and 
K.K. Midha. An approach for widening the bioequivalence acceptance limits 
in the case of highly variable drugs. Pharm. Res. 12:1865-1868 (1995). 
5. L. Tothfalusi, L. Endrenyi, K.K. Midha, M. J. Rawson, and J.W. Hubbard. 
Evaluation of the bioequivalence of highly variable drugs and drug products. 
Pharm. Res. 18:728-733 (2001). 
6. Committee for Proprietary Medicinal Products (CPMP), the European 
Agency for the Evaluation of Medicinal Products (EMEA). Note for Guidance 
on the Investigation of Bioavailability and Bioequivalence. 2001. 
7. Japan National Institute of Health, Division of Drugs. Guideline for 
Bioequivalence Studies of Generic Drug Products. 1997. 
8. L. Tothfalusi, L. Endrenyi, and K.K. Midha. Scaling or wider bioequivalence 
limits for highly variable drugs and for the special case of Cmax. Int. J. Clin. 
Pharmacol. Ther. 41:217-225 (2003). 
9. Shin, et al. A Liquid Chromatography-Quadrupole-Time-of-Flight Mass 
Spectrometric Assay for the Quantification of Fabry Disease Biomarker 
Globotriaosylceramide (GB3) in Fabry Model Mouse; Pharmaceutics 2018, 
10(2), 69; https://doi.org/10.3390/pharmaceutics10020069 
10. Karen Lee, X. J.-M. (Abril de 2003). A biochemical and pharmacological 
comparison of enzyme replacement therapies for the glycolipid storage 
disorder 
Fabry 
disease. 
Glycobiology, 
13(4), 
305-313. 
doi:https://doi.org/10.1093/glycob/cwg034 
11. Hideto Morimoto, Y. I. (Septiembre de 2018). Non-clinical evaluation of JR-
051 as a biosimilar to agalsidase beta for the. Molecular Genetics and 
Metabolism, 125, 153-160. 

12. Scott C.Garman, D. N. (19 de Marzo de 2004). The Molecular Defect 
Leading to Fabry Disease: Structure of Human α-Galactosidase. Journal of 
Molecular 
Biology, 
337(2), 
319-335. 
doi:https://doi.org/10.1016/j.jmb.2004.01.035 
13. Tian, W. Y. (2019). The glycosylation design space for recombinant 
lysosomal 
replacement 
enzymes 
produced 
in 
CHO 
cells. 
Nature 
Communications, 10. doi:https://doi.org/10.1038/s41467-019-09809-3 
14. Yiannis A. Ioannou, K. M. (Enero de 2001). Fabry Disease: Preclinical 
Studies Demonstrate the Effectiveness of α-Galactosidase A Replacement in 
Enzyme-Deficient 
Mice. 
AJHG, 
68(1), 
14-25. 
doi:https://doi.org/10.1086/316953 
15. Nakamura K, Kawashima S, Tozawa H, Yamaoka M, Yamamoto T, Tanaka 
N, 
Yamamoto 
R, 
Okuyama 
T, 
Eto 
Y. 
Pharmacokinetics 
and 
pharmacodynamics of JR-051, a biosimilar of agalsidase beta, in healthy 
adults and patients with Fabry disease: Phase I and II/III clinical studies. Mol 
Genet Metab. 2020 Jul;130(3):215-224. doi: 10.1016/j.ymgme.2020.04.003. 
Epub 2020 May 1. PMID: 32389574 
 
 
 
 

Annex 1: SCHEDULE OF ACTIVITIES 
 
Protocol activity 
Visit 1 
Visit 2 
Visit 3 to 6 Visit 7 
Screening Hospitalizati
on 
Phone 
Follow-up 
Final Visit 
21 days 
1 
3, 7, 14 and
21 
35 
Informed Consent 
X 
Preparation of Medical Record/Progress 
Form  
X 
X 
Physical Examination 
X 
ECG 
X 
Laboratory tests 
X 
Evaluation of inclusion/exclusion criteria X 
Investigational Product Administration 
X 
BP, HR and RR measurement prior to 
blood 
sampling 
for 
evaluation 
of 
pharmacokinetic 
parameters 
and 
immunogenicity 
X 
Sampling 
for 
the 
evaluation 
of 
pharmacokinetic, pharmacodynamic and 
immunogenicity parameters 
X 
* Adverse event evaluation * 
X 
X 
X 
Clinical status evaluation 
X 
X 
X 
PCR COVID-19 
X 
COVID-19 Rapid Test 
X 
BP: blood pressure, HR: heart rate and RR: Respiratory Rate 
*.- Please note that the Serious Adverse Event reporting must be made from the time the 
volunteer signs the Informed Consent and in the time and manner indicated in the 
protocol. 
 
 

Annex 2: Declaration of Helsinki of the World Medical Association Ethical Principles 
for Medical Research Involving Human Subjects 
Adopted by the  
18th  General Assembly, Helsinki, Finland, June 1964 and amended by the: 
29th  General Assembly, Tokyo, Japan, October 1975 
35th  General Assembly, Venice, Italy, October 1983 
41st  General Assembly, Hong Kong, September 1989 
48th  General Assembly, Somerset West, South Africa, October 1996 
52nd  General Assembly, Edinburgh, Scotland, October 2000 
Note of Clarification added by the WMA General Assembly, Washington 2002 
Note of Clarification added by the WMA General Assembly, Tokyo 2004 
59th  General Assembly, Seoul, Korea, October 2008 
64th  General Assembly, Fortaleza, Brazil, October 2013 
• 
Introduction 
• 
Basic Principles for all Medical Research  
• 
Principles Applicable When Medical Research Is Combined with Healthcare 
 
DECLARATION OF HELSINKI - Introduction 
1. 
The World Medical Association (WMA) has developed the Declaration of Helsinki as 
a statement of ethical principles for medical research involving human subjects, 
including research on identifiable human material and data. 
The Declaration is intended to be read as a whole and each of its constituent paragraphs 
should be applied with consideration of all other relevant paragraphs. 
2. 
Consistent with the mandate of the WMA, the Declaration is addressed primarily to 
physicians. The WMA encourages others who are involved in medical research 
involving human subjects to adopt these principles. 
 
General Principles 
3. 
The Declaration of Geneva of the WMA binds the physician with the words, “The 
health of my patient will be my first consideration,” and the International Code of 
Medical Ethics declares that, “A physician shall act in the patient’s best interest 
when providing medical care.” 
4. 
It is the duty of the physician to promote and safeguard the health, well-being and 
rights of patients, including those who are involved in medical research. The 
physician’s knowledge and conscience are dedicated to the fulfilment of this duty. 

5. 
Medical progress is based on research that ultimately must include studies involving 
human subjects. 
6. 
The primary purpose of medical research involving human subjects is to understand 
the causes, development and effects of diseases and improve preventive, 
diagnostic and therapeutic interventions (methods, procedures and treatments). 
Even the best proven interventions must be evaluated continually through research 
for their safety, effectiveness, efficiency, accessibility and quality. 
7. 
Medical research is subject to ethical standards that promote and ensure respect 
for all human subjects and protect their health and rights. 
8. 
While the primary purpose of medical research is to generate new knowledge, this 
goal can never take precedence over the rights and interests of individual research 
subjects. 
9. 
It is the duty of physicians who are involved in medical research to protect the life, 
health, dignity, integrity, right to self-determination, privacy, and confidentiality of 
personal information of research subjects. The responsibility for the protection of 
research subjects must always rest with the physician or other health care 
professionals and never with the research subjects, even though they have given 
consent. 
10. 
Physicians must consider the ethical, legal and regulatory norms and standards for 
research involving human subjects in their own countries as well as applicable 
international norms and standards. No national or international ethical, legal or 
regulatory requirement should reduce or eliminate any of the protections for 
research subjects set forth in this Declaration.  
11. 
Medical research should be conducted in a manner that minimises possible harm to 
the environment. 
12. 
Medical research involving human subjects must be conducted only by individuals 
with the appropriate ethics and scientific education, training and qualifications. 
Research on patients or healthy volunteers requires the supervision of a competent 
and appropriately qualified physician or other health care professional.  
13. 
Groups that are underrepresented in medical research should be provided 
appropriate access to participation in research.  
14. 
Physicians who combine medical research with medical care should involve their 
patients in research only to the extent that this is justified by its potential preventive, 
diagnostic or therapeutic value and if the physician has good reason to believe that 
participation in the research study will not adversely affect the health of the patients 
who serve as research subjects.  
15. 
Appropriate compensation and treatment for subjects who are harmed as a result of 
participating in research must be ensured.  
 

Risks, Burdens and Benefits  
16. 
 In medical practice and in medical research, most interventions involve risks and 
burdens.  
Medical research involving human subjects may only be conducted if the importance of the 
objective outweighs the risks and burdens to the research subjects.  
17. 
All medical research involving human subjects must be preceded by careful 
assessment of predictable risks and burdens to the individuals and groups involved 
in the research in comparison with foreseeable benefits to them and to other 
individuals or groups affected by the condition under investigation.  
Measures to minimize the risks must be implemented. The risks must be continuously 
monitored, assessed and documented by the researcher. 
18. 
 Physicians may not be involved in a research study involving human subjects 
unless they are confident that the risks have been adequately assessed and can be 
satisfactorily managed. 
When the risks are found to outweigh the potential benefits or when there is conclusive 
proof of definitive outcomes, physicians must assess whether to continue, modify or 
immediately stop the study. 
 
Vulnerable Groups and Individuals 
19. 
Some groups and individuals are particularly vulnerable and may have an increased 
likelihood of being wronged or of incurring additional harm. 
All vulnerable groups and individuals should receive specifically considered protection. 
20. 
Medical research with a vulnerable group is only justified if the research is 
responsive to the health needs or priorities of this group and the research cannot be 
carried out in a non-vulnerable group. In addition, this group should stand to benefit 
from the knowledge, practices or interventions that result from the research. 
 
Scientific Requirements and Research Protocols  
21. 
Medical research involving human subjects must conform to generally accepted 
scientific principles, be based on a thorough knowledge of the scientific literature, 
other relevant sources of information, and adequate laboratory and, as appropriate, 
animal experimentation. The welfare of animals used for research must be 
respected. 
22. 
The design and performance of each research study involving human subjects must 
be clearly described and justified in a research protocol.  
The protocol should contain a statement of the ethical considerations involved and should 
indicate how the principles in this Declaration have been addressed. The protocol should 
include information regarding funding, sponsors, institutional affiliations, potential conflicts 

of interest, incentives for subjects and information regarding provisions for treating and/or 
compensating subjects who are harmed as a consequence of participation in the research 
study. 
In clinical trials, the protocol must also describe appropriate arrangements for post-trial 
provisions. 
 
Research Ethics Committees  
23. 
The research protocol must be submitted for consideration, comment, guidance and 
approval to the concerned research ethics committee before the study begins. This 
committee must be transparent in its functioning, must be independent of the 
researcher, the sponsor and any other undue influence and must be duly qualified. 
It must take into consideration the laws and regulations of the country or countries 
in which the research is to be performed as well as applicable international norms 
and standards but these must not be allowed to reduce or eliminate any of the 
protections for research subjects set forth in this Declaration. 
The committee must have the right to monitor ongoing studies. The researcher must 
provide monitoring information to the committee, especially information about any serious 
adverse events. No amendment to the protocol may be made without consideration and 
approval by the committee. After the end of the study, the researchers must submit a final 
report to the committee containing a summary of the study’s findings and conclusions. 
 
Privacy and Confidentiality 
24. 
Every precaution must be taken to protect the privacy of research subjects and the 
confidentiality of their personal information. 
 
Informed Consent  
25. 
Participation by individuals capable of giving informed consent as subjects in 
medical research must be voluntary. Although it may be appropriate to consult 
family members or community leaders, no individual capable of giving informed 
consent may be enrolled in a research study unless he or she freely agrees. 
26. 
In medical research involving human subjects capable of giving informed consent, 
each potential subject must be adequately informed of the aims, methods, sources 
of funding, any possible conflicts of interest, institutional affiliations of the 
researcher, the anticipated benefits and potential risks of the study and the 
discomfort it may entail, post-study provisions and any other relevant aspects of the 
study. The potential subject must be informed of the right to refuse to participate in 
the study or to withdraw consent to participate at any time without reprisal. Special 
attention should be given to the specific information needs of individual potential 
subjects as well as to the methods used to deliver the information. 

After ensuring that the potential subject has understood the information, the physician or 
another appropriately qualified individual must then seek the potential subject’s freely-
given informed consent, preferably in writing. If the consent cannot be expressed in 
writing, the non-written consent must be formally documented and witnessed. 
All medical research subjects should be given the option of being informed about the 
general outcome and results of the study. 
27. 
When seeking informed consent for participation in a research study the physician 
must be particularly cautious if the potential subject is in a dependent relationship 
with the physician or may consent under duress. In such situations the informed 
consent must be sought by an appropriately qualified individual who is completely 
independent of this relationship.  
28. 
For a potential research subject who is incapable of giving informed consent, the 
physician must seek informed consent from the legally authorised representative. 
These individuals must not be included in a research study that has no likelihood of 
benefit for them unless it is intended to promote the health of the group represented 
by the potential subject, the research cannot instead be performed with persons 
capable of providing informed consent, and the research entails only minimal risk 
and minimal burden. 
29. 
When a potential research subject who is deemed incapable of giving informed 
consent is able to give assent to decisions about participation in research, the 
physician must seek that assent in addition to the consent of the legally authorised 
representative. The potential subject’s dissent should be respected. 
30. 
Research involving subjects who are physically or mentally incapable of giving 
consent, for example, unconscious patients, may be done only if the physical or 
mental condition that prevents giving informed consent is a necessary characteristic 
of the research  group. In such circumstances the physician must seek informed 
consent from the legally authorised representative. If no such representative is 
available and if the research cannot be delayed, the study may proceed without 
informed consent provided that the specific reasons for involving subjects with a 
condition that renders them unable to give informed consent have been stated in 
the research protocol and the study has been approved by a research ethics 
committee. Consent to remain in the research must be obtained as soon as 
possible from the subject or a legally authorised representative.   
31. 
The physician must fully inform the patient which aspects of their care are related to 
the research. The refusal of a patient to participate in a study or the patient’s 
decision to withdraw from the study must never adversely affect the patient-
physician relationship. 
32. 
For medical research using identifiable human material or data, such as research 
on material or data contained in biobanks or similar repositories, physicians must 
seek informed consent for its collection, storage and/or reuse. There may be 
exceptional situations where consent would be impossible or impracticable to obtain 

for such research. In such situations the research may be done only after 
consideration and approval of a research ethics committee. 
 
 
Use of placebo 
33. 
The benefits, risks, burdens and effectiveness of a new intervention must be tested 
against those of the best proven intervention(s), except in the following 
circumstances: 
Where no proven intervention exists, the use of placebo, or no intervention, is acceptable; 
or where for compelling and scientifically sound methodological reasons the use of any 
intervention less effective than the best proven one, the use of placebo, or no intervention 
is necessary to determine the efficacy or safety of an intervention. 
The patients who receive any intervention less effective than the best proven one, 
placebo, or no intervention will not be subject to additional risks of serious or irreversible 
harm as a result of not receiving the best proven intervention. 
Extreme care must be taken to avoid abuse of this option. 
 
 
Post-Trial Provisions 
34. 
 In advance of a clinical trial, sponsors, researchers and host country governments 
should make provisions for post-trial access for all participants who still need an 
intervention identified as beneficial in the trial. This information must also be 
disclosed to participants during the informed consent process. 
 
Research Registration and Publication and Disemination of Results 
35. 
Every research study involving human subjects must be registered in a publicly 
accessible database before recruitment of the first subject. 
36. 
Researchers, authors, sponsors, editors and publishers all have ethical obligations 
with regard to the publication and dissemination of the results of research. 
Researchers have a duty to make publicly available the results of their research on 
human subjects and are accountable for the completeness and accuracy of their 
reports. All parties should adhere to accepted guidelines for ethical reporting. 
Negative and inconclusive as well as positive results must be published or 
otherwise made publicly available. Sources of funding, institutional affiliations and 
conflicts of interest must be declared in the publication. Reports of research not in 
accordance with the principles of this Declaration should not be accepted for 
publication. 
 

Unproven Interventions in Clinical Practice  
37. 
In the treatment of an individual patient, where proven interventions do not exist or 
other known interventions have been ineffective, the physician, after seeking expert 
advice, with informed consent from the patient or a legally authorised 
representative, may use an unproven intervention if in the physician’s judgement it 
offers hope of saving life, re-establishing health or alleviating suffering. This 
intervention should subsequently be made the object of research, designed to 
evaluate its safety and efficacy. In all cases, new information must be recorded and, 
where appropriate, made publicly available. 

RD-BIO-AGA-01/03/21 
 
Confidential Information –BIOSIDUS S.A. 
Page 62 
 
 
Annex 3: Storage and labeling 
Each treatment unit corresponds to one bottle for intravenous administration of Biosidus 
Agalsidase or Sanofi Genzyme Fabrazyme® for a period of 5 hours.  
Bottle label for each patient contains the following information: Protocol No: RD-BIO—
AGA-01/03/21 
Bottle  N°: 
Patient Initials:  
 
Pacient N°:  
 
 
Investigator´s Name: Eduardo Pirotzky, MD 
Date of provision:  
 
 
Lot N°: 
Date of Expiration: November 30, 2021 
Store between 2°C to 8°C, protect from heat, light and excessive moisture. 
For exclusive use in clinical research. Keep out of reach of children. 
Biosidus Cazadores de Coquimbo 2841 - Building 4 – 1st Floor (B1605AZE) Munro - 
Buenos Aires, Argentina 
 
Note: Bottle N° 1 with 35 mg or Bottle N° 2 with 5 mg. 

RD-BIO-AGA-01/03/21 
 
Confidential Information –BIOSIDUS S.A. 
Page 63 
 
 
Annex 4: Randomization Table 
 
Patient 
number 
Treatment 
assigned 
1 
R 
2 
T 
3 
T 
4 
R 
5 
T 
6 
R 
7 
R 
8 
T 
9 
T 
10 
R 
11 
R 
12 
R 
13 
R 
14 
T 
15 
T 
16 
R 
17 
T 
18 
R 
19 
T 
20 
T 
 
Table generated using RANDOM software by Randomness and Integrity Services Ltd. Premier 
Business Centres, 8 Dawson Street, Dublin 2, D02 N767, Ireland. 

RD-BIO-AGA-01/03/21 
Confidential Information –BIOSIDUS S.A. 
Page 64 
 
 
Annex 5: Adverse reactions organized by system and frequency – Reference 
Product  
 
Classification 
System 
Very Common 
Common 
Uncommon 
Unknown 
frequency 
infections and 
infestations  
Nasopharyngitis 
Rhinitis 
Immune system 
Anaphylactoid 
reaction 
Nervous system 
Headache, 
paraesthesia 
Dizziness, 
drowsiness, 
hypoesthesia, 
burning, lethargy, 
syncope 
Hyperesthesia, 
tremor 
Eye 
Increased lacrimation pruritus, ocular 
hyperaemia 
Ear and labyrinth 
Tinnitus, vertigo 
Auricular 
inflammation 
earache 
Cardiac 
Tachycardia, 
bradycardia, 
palpitations 
Sinus bradycardia 
Vascular 
Flushing, 
hypertension, 
paleness, 
hypotension, hot 
flushes 
Limb coldness 
Respiratory, 
thoracic and 
mediastinal 
Dyspnoea, nasal 
congestion, throat 
tightness, wheezing, 
cough, exacerbated 
dyspnea 
Bronchospasm, 
pharyngolaryngeal 
pain, rhinorrhea, 
tachypnoea, upper 
respiratory tract 
congestion 
hypoxia 
Gastro-intestinal 
Nausea, vomiting Abdominal pain, 
abdominal discomfort, 
upset stomach, oral 
hypoaesthesia, 
diarrhea 
Dyspepsia, 
dysphagia 
Skin and 
subcutaneous tissue 
Pruritus, hives, rash, 
erythema, generalized 
pruritus, angioneurotic 
edema, facial 
swelling, 
Livedo reticularis, 
erythematous, rash, 
pruritic rash, skin 
dispigmentation, 
Classical 
leukocytic 
vasculitis 

RD-BIO-AGA-01/03/21 
Confidential Information –BIOSIDUS S.A. 
Page 65 
 
 
maculopapular rash 
skin discomfort 
Musculoskeletal and 
connective tissue 
Limb pain, myalgia, 
back pain, muscle 
spasms, arthralgia, 
muscle tightness, 
musculoskeletal 
stiffness 
Musculoskeletal 
pain 
Administration site 
and general 
disorders 
Chills, pyrexia, 
feeling cold 
Fatigue, chest 
discomfort, feeling 
hot, peripheral 
edema, pain, 
asthenia, chest pain, 
face edema, 
hyperthermia 
Feeling hot and 
cold, flu-like 
syndrome, pain at 
the infusion 
administration site, 
reaction at infusion 
site, thrombosis at 
injection site, 
general discomfort, 
edema 
Investigations 
Decreased 
oxygen 
saturation 
 
 
 

RD-BIO-AGA-01/03/21 
Confidential Information –BIOSIDUS S.A. 
Page 66 
 
 
Annex 6: Authorization to conduct Phase I Clinical Pharmacology Studies  
 
Let the company Sanatorio Nuestra Señora del Pilar S.A. be authorized to conduct 
PHASE I CLINICAL PHARMACOLOGY STUDIES IN THE ADULT POPULATION and 
BIOEQUIVALENCE STUDIES, at Sanatorio Nuestra Señora del Pilar. 
 
[Argentina’s coat of arms] 
Argentine Republic – National Executive Branch 
2019 – Export Year  
 
Provision 
 
 
Number: DI-2019-5344-APN-ANMAT#MSYDS 
 
 
 
 
 
 
         CITY OF BUENOS AIRES 
Wednesday, July 3, 2019 
 
Reference: EX-2019-00632005-APN-DGA#ANMAT 
 
IN VIEW OF ANMAT Provisions N° 6677/10, 4008/17 and 4009/17 and File N° EX-2019-00632005-
APN-DGA#ANMAT of the “Registro de la Administración Nacional de Medicamentos, Alimentos y 
Tecnología Médica” (Registry of the National Administration of Drugs, Food and Medical 
Technology) and, 
 
WHEREAS, 
By means of these proceedings the company Sanatorio Nuestra Señora del Pilar S.A., requests 
authorization to conduct Phase I Clinical Pharmacology Studies and Bioequivalence Studies, under 
the terms of ANMAT Provisions N° 4008/17 and 4009/17 at Sanatorio Nuestra Señora del Pilar. 
ANMAT Provision No. 4008/17 in section 11 states the Specific Considerations for Phase I Clinical 
Pharmacology Studies, indicating in its sections 11.2 and 11.4 the specifications for the 
investigational site and for the approval and authorizations, respectively. 
 

RD-BIO-AGA-01/03/21 
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Page 67 
 
 
ANMAT Provision No. 4009/17 approves the requirements and conditions that Healthcare Sites 
must meet in order to be authorized to conduct Phase I Clinical Pharmacology Studies and/or 
Bioequivalence studies. 
Sanatorio Nuestra Señora del Pilar is duly authorized and registered in Registro Federal de 
Establecimientos de Salud (REFES) (Federal Registry of Healthcare Sites) of the National Ministry 
of Health. 
Dirección de Evaluación y Registro de Medicamentos (DERM) (Directorate of Drug Evaluation and 
Registration), through Inspection Order 2019/838-INAME-116, carried out the verification procedure 
provided for in ANMAT Provision No. 4009/17. 
Final technical report by DERM is added, considering site is acceptable for conducting Phase I 
Clinical Pharmacology Studies in the adult population and Bioequivalence studies. 
Directorate of Drug Evaluation and Registration of the National Drug Institute and the General 
Directorate of Legal Affairs have intervened accordingly. 
Action is taken under the powers conferred by Decree No. 1490/92 and its amendments. 
 
Therefore, 
 
THE 
NATIONAL 
ADMINISTRATOR 
OF 
THE 
ADMINISTRACION 
NACIONAL 
DE 
MEDICAMENTOS, ALIMENTOS Y TECNOLOGÍA MÉDICA  
STATES THAT: 
 
1° SECTION: Let the company Sanatorio Nuestra Señora del Pilar S.A. be authorized to 
conduct PHASE I CLINICAL PHARMACOLOGY STUDIES IN THE ADULT POPULATION 
and BIOEQUIVALENCE STUDIES, at Sanatorio Nuestra Señora del Pilar, located at Av. 
Gaona 3979 PB, Ciudadela, Province of Buenos Aires – CP1702, in accordance with data 
and conditions described in Annex I herein. 
 
2° SECTION: Let it be stated that the authorization granted by virtue of 1° section will be valid for 5 
(five) years, being the bearer obliged to notify this National Administration of any modification in the 
conditions of health authorization or the conditions of approval in accordance with this provision and 
ANMAT Provision No. 4009/17. 
 
3° SECTION: - Let it be stated that the authorization granted in accordance with 1° section will be 
without effect, by operation of law, in the event the Site does not conduct any Phase I Clinical 
Pharmacology or Bioequivalence research study during the 2 (two) years following the date of 
granting such authorization. 
 
4° SECTION.- Let it be registered. Let the Admission Desk Department notify the interested party, 
delivering this provision. Let the Directorate of Drug Evaluation and Registration be notified. Let it 
be filed once completed these proceedings. 

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Page 68 
 
 
 
File: EX-2019-00632005-APN-DGA#ANMAT 
ANNEX I  
DATA AND CONDITIONS FOR THE HEALTHCARE SITE AUTHORIZATION TO CONDUCT 
PHASE I CLINICAL PHARMACOLOGY STUDIES IN THE ADULT POPULATION AND 
BIOEQUIVALENCE STUDIES 
 
Information on Investigational Site for Phase I Clinical Pharmacology studies in the adult 
population and Bioequivalence studies 
Site Name 
Sanatorio Nuestra Señora del Pilar 
Site address 
Av. Gaona 3979 PB, Ciudadela, Province of Buenos Aires- 
CP1702  
Name or Corporate Name 
of the bearer of the Site 
Authorization 
Sanatorio Nuestra Señora del Pilar 
Name of Site Director 
Juan Domingo Pugliese, MD (Medical Chief Officer) 
Authorized activity 
INVESTIGATIONAL 
SITE 
FOR 
PHASE 
I 
CLINICAL 
PHARMACOLOGY STUDIES IN ADULT POPULATION AND 
BIOEQUIVALENCE STUDIES 
Location of area authorized 
destined to studies 
Ground Floor 
- 
Recovery room: 8 beds for bioequivalence. When phase 1 
studies are conducted in the area, the use of up to 2 
simultaneous beds is authorized. 
- 
ICU 
- 
Rehabilitation Site 
- 
Kitchen 
- 
Pharmacy 
- 
Doctor offices on Ground Floor, 1° and 2° Floors 
3° Floor 
- 
Meeting room, Filing room, administrative office 
Registration 
in 
Registro 
Federal 
de 
Establecimientos de Salud 
(REFES) 
Disposition 1070/09 
Code: 10068402300461 
 
Healthcare Facility with general hospitalization.  
Category: High risk with intensive therapy 
Phone/Fax 
4469-6700/6767 
Contact 
Medical Manager: Néstor Zungri, MD 
nestor@sanatoriodelpilar.com.ar 
Principal Investigator: Eduardo Pirotzky MD 

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Page 69 
 
 
epirotzky@yahoo.com 
15 6554-7584  Eduardo Pirotzky MD 
 
File: EX-2019-00632005-APN-DGA#ANMAT 
[There follows a digital logo and a statement that reads:] Carlos Alberto Chiale – Administrator – 
National Administration of Drugs, Food and Medical Technology 

RD-BIO-AGA-01/03/21 – Amendment 2 02SEPT2021 
 
 
Confidential Information- BIOSIDUS S.A. 
  
Page 70 of 71 
 
Annex 7: Justification for the difference in drug substance strengths of both 
Agalsidase Biosidus and Frabrazyme®. 
 
Fabrazyme and AGA Biosidus content 
To determine the beta agalsidase content in the 35 mg and 5 mg presentations according 
to our methodology, volume and mass measurements were performed on Fabrazyme. 
 
To that end, several vial batches of each of the Fabrazyme® presentations were 
reconstituted according to the instructions on the package insert (7.2 mL for the 35 mg 
presentation and 1.1 mL for the 5 mg presentation). The extractable volume was 
measured according to RTD-AGA-091 and the agalsidase strength by reverse phase 
HPLC (RTD-AGA-022). 
 
The RF-HPLC method uses a standard with a quantification value that is previously 
assigned by optical density (OD). The extinction coefficient to determine the agalsidase 
beta content in the standard was 2.55 based on the experimental results carried out in 
Protagen (Germany) using time resolved analysis of said standard (RTD AGA117). 
 
The Fabrazyme results were: 
 
Fabrazyme® 35 mg Date 
cc 
(mg/mL) 
Volume 
(mL) 
Content (mg) 
Fabrazyme® 
8W0716 
February 
21, 2019 
5.4 
7.2 
39 
Fabrazyme® 
8W1890 
February 
21, 2019 
5.4 
7.2 
39 
 
Fabrazyme® 5 mg Date 
cc 
(mg/mL) 
Volume 
(mL) 
Content (mg) 
Fabrazyme® 
7HV1176 
November 
18, 2018 
5.4 
1.0 
5.4 
Fabrazyme® 
8HV1569 
February 
21, 2019 
5.3 
1.0 
5.3 
Fabrazyme® 
November 5.3 
1.0 
5.3 

RD-BIO-AGA-01/03/21 – Amendment 2 02SEPT2021 
 
 
Confidential Information- BIOSIDUS S.A. 
  
Page 71 of 71 
 
9W1763 
15, 2019 
 
In other words, although Fabrazyme declares a total of 37 mg (in 7.2 ml), with our 
methodology the content was 39 mg. 
 
In order to have a target concentration in our product and to be biosimilar to Fabrazyme 
(not to differ more than 3% in content), the value of 5.3 mg/mL was taken as the target 
concentration of agalsidase beta in the reconstituted product. Therefore, our product is 
formulated with approximately 6% overage compared to the declared theoretical 
concentration. 
 
In other words, each vial of Biosidus Agalsidase labeled 35 mg will contain 5.3 mg/mL 
when reconstituted in 7.2 mL, total: 38 mg. 
 
With these values, we ensure biosimilarity in content with respect to the reference 
Fabrazyme, which according to our methodology has a total of 38-39 mg.