A Phase I Study to Evaluate the Tolerance, Safety and Pharmacokinetics of ALT-BB4 in Healthy Volunteer
Phase: PHASE1
Conditions: Health, Subjective
Sponsor: Alteogen, Inc.

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
 
 
 
 
Clinical Study Protocol 
 
 
 
A Phase I Study to Evaluate the Tolerance,  
Safety and Pharmacokinetics of ALT-BB4  
in Healthy Volunteers 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
CONFIDENTIAL 
All information contained in this Protocol is provided for the Principal Investigator (PI), 
subinvestigator, Institutional Review Board and health authorities, and cannot be disclosed 
to a third party without prior written consent of Alteogen, Inc. unless it is necessary to obtain 
written informed consent for the study participation from subjects who will receive the 
products used in the study. 
 
Investigational Product (IP) 
ALT-BB4 (Recombinant Hyaluronidase) 
Protocol No. 
ALT-BB4-01 
Protocol Version 
3.2 
Study Phase 
Phase 1 
Sponsor 
Alteogen, Inc. 
Protocol Preparation Date 
2022-06-10 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
 Term Definition and Abbreviation 
ADR 
Adverse drug reaction 
AE 
Adverse event 
ALP 
Alkaline phosphatase 
ALT 
Alanine aminotransferase 
ANOVA 
Analysis of variance 
AST 
Aspartate aminotransferase 
AUC 
Area under the concentration-time curve 
AVG 
Average 
BLQ 
Below the limit of quantification 
BPM 
Beats per minute 
BUN 
Blood urea nitrogen 
CFR 
Code of federal regulations 
CIOMS 
Council for international organizations of medical sciences 
Cl 
Clearance 
Co-mix 
Concomitant 
CS 
Clinically significant 
DB 
Database 
ECM 
Extracellular matrix 
ECG 
Electrocardiogram 
EDC 
Electronic data capture 
eCRF 
Electronic case report form 
FDA 
Food and drug administration 
GCP 
Good clinical practice 
HA 
Hyaluronic acid 
HBV 
Hepatitis B virus 
hCG 
human Chorionic gonadotrophin 
HCV 
Hepatitis C virus 
HIV 
Human immunodeficiency virus 
HYALP1 
Hyaluronoglucosaminidase pseudogene 1  
IB 
Investigator's brochure 
IgE 
Immunoglobulin E 
IRB 
Institutional review board 
Kcat 
Catalytic rate constant 
KGCP 
Korean good clinical practice 
MedDRA 
Medical dictionary for regulatory activities 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
NCS 
Not clinically significant 
NOAEL 
No observed adverse effect level 
NRS 
Numeric rating scale 
NSAIDs 
Non-steroidal anti-inflammatory drugs 
RBC 
Red blood cell 
RPM 
Revolutions per minute 
SADR 
Serious adverse drug reaction 
SAE 
Serious adverse event 
SEM 
Standard error of the mean 
Seq 
Sequential 
SI 
Stimulation Index 
SOC 
System organ class 
SPAM 1 
Sperm adhesion molecule 1 
SUSAR 
Suspected unexpected serious adverse reaction 
OTC 
Over-the-counter 
PK 
Pharmacokinetics 
PT 
Preferred term 
USV 
Unscheduled visit 
VDRL 
Venereal disease research laboratory 
WBC 
White blood cell 
WOCBP 
Women of childbearing potential 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
 Table of Contents 
 TERM DEFINITION AND ABBREVIATION ............................................................................. 2 
1. 
STUDY TITLE, PHASE, PROTOCOL IDENTIFICATION NO. AND CREATION/REVISION 
HISTORY ....................................................................................................................................... 8 
1.1. Title .............................................................................................................................. 8 
1.2. Phase ........................................................................................................................... 8 
1.3. Protocol Identification No. and Creation/Revision History ........................................... 8 
2. 
SYNOPSIS, TABLE OF SCHEDULE, AND DIAGRAM ................................................... 10 
2.2 Table of Study Schedule .............................................................................................. 19 
2.2.1 
Part I (Allergy assessment) ......................................................................... 19 
2.2.2 
Part II-A (PK assessment) .......................................................................... 21 
2.2.3 
Part II-B (Safety assessment) ..................................................................... 23 
2.3 Diagram ........................................................................................................................ 25 
3. 
INTRODUCTION ............................................................................................................... 26 
3.1. Study Background ..................................................................................................... 26 
3.2. Study Rationale ......................................................................................................... 27 
3.3. Risk and Benefit Assessment .................................................................................... 27 
3.3.1. 
In Vitro Pharmacology ................................................................................ 27 
3.3.2. 
In Vivo Pharmacology ................................................................................. 29 
3.3.3. 
Nonclinical Pharmacokinetics ..................................................................... 30 
3.3.4. 
Toxicology ................................................................................................... 34 
3.4. Rationale for Dose Selection ..................................................................................... 36 
4. 
STUDY OBJECTIVES ....................................................................................................... 38 
4.1. Primary Objective ...................................................................................................... 38 
4.2. Secondary Objectives ................................................................................................ 38 
4.3. Exploratory Objectives ............................................................................................... 38 
5. 
STUDY POPULATION (SAMPLE SIZE AND SUBJECT SELECTION) .......................... 39 
5.1. Sample Size ............................................................................................................... 39 
5.2. Rationale for Sample Size Determination ................................................................. 40 
5.3. Subject Selection ....................................................................................................... 42 
5.3.1. 
Inclusion Criteria ......................................................................................... 42 
5.3.2. 
Exclusion Criteria ........................................................................................ 42 
5.3.3. 
Withdrawal Criteria ..................................................................................... 43 
5.3.4. 
Protocol Deviation/Violation ........................................................................ 44 
6. 
STUDY DESIGN ................................................................................................................ 45 
6.1. Study Period .............................................................................................................. 45 
6.2. Study Group/Control Group ....................................................................................... 45 
6.3. Randomization ........................................................................................................... 46 
6.4. Blinding ...................................................................................................................... 46 
6.4.1. 
Double-blinding ........................................................................................... 46 
6.4.2. 
Unblinding ................................................................................................... 47 
7. 
CRITERIA FOR STUDY TERMINATION AND EARLY TERMINATION ......................... 48 
7.1. Scheduled Termination .............................................................................................. 48 
7.2. Early Termination ....................................................................................................... 48 
8. 
INVESTIGATIONAL PRODUCT INFORMATION AND MANAGEMENT ........................ 49 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
8.1. Investigational Product (IP) ....................................................................................... 49 
8.1.1. 
Study Drug .................................................................................................. 49 
8.1.2. 
Comparator ................................................................................................. 49 
8.2. Production/Packaging and Labeling .......................................................................... 49 
8.3. Investigational Product Management ........................................................................ 49 
9. 
METHODS AND ADMINISTRATION PLAN ..................................................................... 50 
9.1. Overall Methods ......................................................................................................... 50 
9.2. Investigational Product Dosage and Administration Method ..................................... 50 
9.3. Concomitant Medications and Therapies .................................................................. 51 
9.3.1. 
Permitted Concomitant Medications and Therapies ................................... 51 
9.3.2. 
Forbidden Concomitant Medications and Therapies .................................. 51 
9.4. Treatment Compliance .............................................................................................. 52 
10. 
STUDY PROCEDURES AND ASSESSMENT ................................................................. 53 
10.1. Observations .............................................................................................................. 53 
10.1.1. 
Written Informed Consent Form and Assignment of Screening Number ... 53 
10.1.2. 
Demographic Data ...................................................................................... 53 
10.1.3. 
Investigation of Medication History and Medical History ............................ 53 
10.1.4. 
Vital Sign ..................................................................................................... 53 
10.1.5. 
Electrocardiogram (ECG) ........................................................................... 53 
10.1.6. 
Physical Examination .................................................................................. 53 
10.1.7. 
Laboratory Test ........................................................................................... 54 
10.1.8. 
Immunogenicity Test ................................................................................... 54 
10.1.9. 
Pregnancy Test ........................................................................................... 54 
10.1.10. Randomization ............................................................................................ 54 
10.1.11. PK Assessment........................................................................................... 55 
10.1.12. Investigational Product Administration ....................................................... 55 
10.1.13. Monitoring of Adverse Event ....................................................................... 55 
10.1.14. Photographing of Investigational Product administration Site .................... 56 
10.2. Efficacy Assessment.................................................................................................. 57 
10.2.1. 
Efficacy Endpoints ...................................................................................... 57 
10.3. Safety Assessment .................................................................................................... 57 
10.3.1. 
Safety Endpoints ......................................................................................... 57 
10.3.1.1. Primary Endpoints .............................................................................. 57 
10.3.1.2. Secondary Endpoints ......................................................................... 57 
10.3.1.3. Other Safety Endpoints ...................................................................... 57 
10.3.2. 
Safety Assessment Method ........................................................................ 57 
10.3.2.1. Primary Endpoints .............................................................................. 58 
10.3.2.2. Secondary Endpoints ......................................................................... 58 
10.3.2.3. Other Safety Endpoints ...................................................................... 59 
10.4. Pharmacokinetic Assessment ................................................................................... 60 
10.4.1. 
PK Endpoints .............................................................................................. 60 
10.4.2. 
PK Assessment Method ............................................................................. 60 
10.5. Evaluation Criteria and Report of Adverse Events .................................................... 60 
10.5.1. 
Definition of Safety-related Terms .............................................................. 60 
10.5.2. 
Evaluation Criteria of Adverse Events ........................................................ 61 
10.5.2.1. Severity............................................................................................... 61 

 
 
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Protocol No.: ALT-BB4-01 
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10.5.2.2. Causality to the IP .............................................................................. 61 
10.5.2.3. Action taken and Outcome ................................................................. 62 
10.5.3. 
Reporting Method for Adverse Events ........................................................ 62 
10.5.3.1. Reporting of AE .................................................................................. 62 
10.5.3.2. Expedited Reporting ........................................................................... 63 
10.5.3.3. Responsibilities of Staff in case of Serious Adverse Drug Reactions 63 
10.5.3.4. Pregnancy .......................................................................................... 64 
11. 
DATA ANALYSIS AND STATISTICAL CONSIDERATIONS .......................................... 65 
11.1. Analysis Sets ............................................................................................................. 65 
11.1.1. 
Safety Set ................................................................................................... 65 
11.1.2. 
Full Analysis Set ......................................................................................... 65 
11.1.3. 
PK Analysis Set .......................................................................................... 65 
11.2. Statistical Analysis Method ........................................................................................ 65 
11.2.1. 
General Principle of Result Analysis........................................................... 65 
11.2.2. 
Demographic Data and Medical History ..................................................... 65 
11.2.3. 
Efficacy Endpoints ...................................................................................... 65 
11.2.4. 
Safety Endpoints ......................................................................................... 65 
11.2.4.1. Primary Endpoints .............................................................................. 65 
11.2.4.2. Secondary Endpoints ......................................................................... 66 
11.2.4.3. Other Safety Endpoints ...................................................................... 67 
11.2.5. 
PK Endpoints .............................................................................................. 67 
11.3. Planned Analysis Time .............................................................................................. 67 
12. 
DATA MANAGEMENT ..................................................................................................... 68 
12.1. Record and Access.................................................................................................... 68 
12.2. Data Collection .......................................................................................................... 68 
12.3. Record Protection and Storage ................................................................................. 68 
13. 
ETHICAL CONSIDERATIONS AND ADMINISTRATIVE PROCEDURES ...................... 69 
13.1. Compliance with Regulations, including Korean Good Clinical Practice, and Ethical 
Standards .................................................................................................................. 69 
13.2. Institutional Review Board (IRB) ................................................................................ 69 
13.3. Informed Consent Process ........................................................................................ 69 
13.4. Confidentiality ............................................................................................................ 69 
13.5. Subject Safeguards ................................................................................................... 69 
13.5.1. 
Actions Taken for Adverse Events ............................................................. 69 
13.5.2. 
Post-Clinical-Study Treatment and Therapy for Subjects .......................... 69 
13.5.3. 
Patient Compensation Rule ........................................................................ 70 
13.6. Quality Control and Reliability Assurance ................................................................. 70 
13.6.1. 
Monitoring of Institutions ............................................................................. 70 
13.6.2. 
Audit ............................................................................................................ 70 
13.6.3. 
Inspection .................................................................................................... 70 
14. 
SPONSOR’S INFORMATION, AND PRINCIPAL INVESTIGATOR’S NAME AND 
POSITION ................................................................................................................................... 71 
14.1. Sponsor Information .................................................................................................. 71 
14.2. Institution, and Principal Investigator’s Name and Position....................................... 71 
15. 
OTHER MATTERS REQUIRED FOR SAFE AND SCIENTIFIC CONDUCT OF STUDY 72 
15.1. Protocol Amendment ................................................................................................. 72 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
16. 
REFERENCES .................................................................................................................. 73 
17. 
APPENDIX ........................................................................................................................ 75 
18. 
ANNEX .............................................................................................................................. 76 
 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
1. Study Title, Phase, Protocol Identification No. and 
Creation/Revision History 
1.1. Title 
A Phase I Study to Evaluate the Safety, Tolerability and Pharmacokinetics of ALT-BB4 
(Recombinant Hyaluronidase) in Healthy Volunteers 
 
1.2. Phase 
 
Part I (Allergy assessment): Multicenter, 2-arm (study drug and comparator), randomized 
(randomization of dosing site of the study drug and comparator), double-blinded, placebo-
controlled, intradermal injection, Phase I 
 
Part II-A (PK assessment): Single center, open-labeled, subcutaneous injection, Phase I 
 
Part II-B (Safety assessment): Multicenter, 2-arm (study group and control group), 
randomized, double-blinded, placebo-controlled, subcutaneous injection, Phase I 
 
1.3. Protocol Identification No. and Creation/Revision History 
 
Protocol Identification No.: ALT-BB4-01 
 
Creation/Revision History 
Version 
Date 
Major Changes 
1.0 
2020-12-18 
Created. 
2.0 
2021-07-16 
- 
Design changed (2 arms → 3 arms). 
- 
Sample size changed. 
- 
Institutions/Principal Investigators added. 
2.1 
2021-07-30 
- 
Section 3.4: Rationale for dose selection added. 
- 
Section 5.3.1: Change of the inclusion criteria (in the 
contraception, exclude ‘concomitant use of male condom 
and female condom’ from double barrier methods) 
- 
Section 5.3.2: Change of the exclusion criteria (‘Past 
history of drug abuse’ added) 
- 
Section 10.3.1: Photographing of administration site at 
Visit 4 (2D) in Part I becomes mandatory. 
- 
Section 10.3.3: Photographing of administration site at 
Visit 4 (0D) and Visit 5 (2D) in Part II-B becomes 
mandatory. A measure to collect AEs is added. 
- 
Section 10.1.13: A measure to document rationale for 
photographing of the administration site is added. 
- 
Section 11.1.1: Definition of safety set is modified. 
- 
Section 11.2.1: Imputation method of missing data is 
added 
- 
Section 11.2.4.1: Statistical analysis method of primary 
endpoints is added 
- 
Others: Terms modified, description clarification, etc. 
3.0 
2021-12-28 
- 
Section 2.2.1: Timing of vital signs and laboratory tests at 
baseline is clarified before the ID injection of the IP). 
- 
Section 2.2.2 and Section 2.2.3: Corrections are made to 
vital signs and laboratory tests at Visit 4: they are to be 
performed before the ID injection of the IP and may be 
replaced with the results obtained at Visit 4 of Part I. 
- 
Section 5.3.2: Change of the exclusion criteria 
 
1) Common 1. Prohibited concomitant medications before 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
Version 
Date 
Major Changes 
study participation should be checked relative to baseline, 
and the timeframe for NSAIDs is changed to within 14 days 
from baseline since 14 days are enough to wash out for 
NSAIDs. 
2) The criteria is clarified to exclude current clinically 
significant diseases. 
3) Common 4. The criteria is clarified to exclude subjects 
who are diagnosed with hypertension/hypotension or 
subjects who have clinically significant blood pressure. 
4) Part II-A 13, 14, 15, and 16. The exclusion criteria should 
be checked relative to Visit 4 (baseline of Part II-A). 
- 
Section 9.3.2: It is stated that whole blood donation, 
apheresis, and transfusion are prohibited only for subjects 
in Part II-A. 
- 
Section 10.1.2: Height, weight, and BMI are deleted since 
they are measured in the physical examination, and history 
of whole blood donation, apheresis, and transfusion are to 
be investigated only for subjects considering participating 
in Part II-A. 
- 
Section 10.1.5: ECG parameters are modified. 
- 
Section 10.1.7: It is allowed to replace test results for 
baseline (Visit 2) with the results obtained within 7 days, if 
available. 
- 
Section 10.1.13: The time window for observation of 
immediate allergic reactions is changed. 
- 
 Section 10.3.1.2 and Section 10.3.2.2: The statement that 
elevation considered due to an injection is not considered 
wheal is added. 
- 
 Section 11.2.4.1: The statistical analysis method is 
corrected to one that is appropriate for result analysis of 
the primary and secondary endpoints.  
- 
 Section 11.2.4.2: Details description of the comparison 
time points within a group and comparative analysis 
between the groups at each time point after IP 
administration are added. 
- 
Other: Errors corrected, description clarification, etc. 
3.1 
2022-02-22 
- 
Section 11.2.4.1: Corrected the statistical analysis method 
to the method suitable for the analysis of primary endpoint 
results 
- 
Section 11.2.4.2: Corrected the analysis for comparison 
between the groups by dividing into Part 1 and Part 2 for 
the analysis of secondary endpoint results 
3.2 
2022-06-10 
- 
Section 5: Changed the total number of subjects 
- 
Section 6.4.2: Corrected the unblinding method  
 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
2. Synopsis, Table of Schedule, and Diagram  
2.1 Synopsis 
Study Title 
A Phase I Study to Evaluate the Safety, Tolerability and Pharmacokinetics 
of ALT-BB4 (Recombinant Hyaluronidase) in Healthy Volunteers 
Phase and 
Design 
 
Part I (Allergy assessment): Multicenter, 2-arm (study drug and 
comparator), randomized (randomization of dosing site of the study 
drug and comparator), double-blinded, placebo-controlled, intradermal 
(ID) injection, Phase I 
 
Part II-A (PK assessment): Single center, single arm, open-labeled, 
subcutaneous (SC) injection, Phase I 
 
Part II-B (Safety assessment): Multicenter, 2-arm (study group and 
control group), randomized, double-blinded, placebo-controlled, SC 
injection, Phase I 
Institutions and 
Principal 
Investigators 
 
See Annex 1. 
Coordinating 
Investigator 
 
Professor Dong Hoon Lee, Dept. of Dermatology, Seoul National 
University Hospital 
Sponsor 
Alteogen, Inc. 
Study 
Objective 
Primary Objective 
To evaluate the allergic reactivity, safety and tolerability of ALT-BB4 
(recombinant hyaluronidase) by checking for drug allergy reactivity 
following single ID injection of ALT-BB4 and monitoring systemic reactions 
and adverse events following its single SC injection 
 
Secondary Objective 
To evaluate the PK profile of single dose of ALT-BB4 
 
Exploratory Objective 
To evaluate the immunogenicity of ALT-BB4 (Visit 1 and End of Study visit) 
Study 
Population 
Healthy volunteers aged ≥19 years 
Inclusion 
Criteria 
Common 
1. 
Healthy volunteers aged ≥19 years at the time of Screening (Visit 1) 
2. 
Female subjects or male subjects’ female partner must be menopausal 
or should have received a sterilization procedure or have agreed to use 
contraceptive methods during the study period, as defined below: 
 
Post-menopausal female subjects or male subjects’ female 
partners (non-drug induced amenorrhea for at least 12 months or 
confirmed diagnosis with menopause) 
 
Female subjects or male subjects’ female partners who have 
received a sterilization procedure (removal of ovary and/or uterus) 
 
Subjects who have agreed to practice total abstinence during the 
study period [For female subjects, periodic abstinence (e.g., 
ovulation, symptothermal, or post-ovulation methods) and 
withdrawal are not acceptable methods of contraception.] 
 
When female subjects or male subjects’ female partners are 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
women of childbearing potential (WOCBP) who have not received 
a sterilization procedure, they must agree to use of following 
contraceptive methods: 
- 
Hormones (implantable, patch, and oral) 
- 
Intrauterine device (IUD) 
- 
Double barrier methods (concomitant use of two of following 
contraceptive methods: male condom, female condom, 
cervical cap, diaphragm, sponge, spermicide) (However, 
concomitant use of male condom and female condom is 
excluded from double barrier methods.) 
3. 
WOCBP or females who have the last menstrual period within 12 
months must have a negative serum or urine pregnancy test at 
Screening (Visit 1). 
4. 
Subjects who have voluntarily decided to take part in the study and able 
to comply with the study protocol 
5. 
Subjects who have no tattoo, acne, dermatitis, pigmentation or lesion 
on the administration site and who have no damage in the skin, so that 
they can receive the investigational product (IP) and allergy test 
6. 
Subjects determined eligible for the study through Screening tests (vital 
signs, physical examination, medical history and surgery history, 
electrocardiogram (ECG), and laboratory tests) 
 
Part II-A 
7. 
Subjects with BMI no less than 18.5 kg/m2, no greater than 24.9 kg/m2 
Exclusion 
Criteria 
Below are the study exclusion criteria: 
 
Common 
1. 
Subjects who have received or treated with following medications 
within the specified timeframe prior to Baseline (Visit 2) or who are 
expected to receive them during the study period: 
 
Within 1 month: Hyaluronidase, Chemotherapeutic agent, 
penicillins 
antibiotics 
(e.g.: 
Amoxicillin, 
Ampicillin, 
etc.), 
cephalosporins antibiotics (e.g.: Cefaclor, Cefadroxil, Cefixime, 
etc.), 
sulfonamides 
antibiotics 
(e.g.: 
Sulfadiazine, 
Sulfamethoxazole, 
etc.), 
quinolones 
antibiotics 
(e.g.: 
Ciprofloxacin, 
Levofloxacin, 
etc.), 
Glucocorticosteroid, 
Immunosuppressive agent 
 
Within 
14 
days: 
Antihistamine 
(e.g.: 
Chlorpheniramine, 
Hydroxyzine, Ketotifen, etc.), non-steroidal anti-inflammatory 
drugs (NSAIDs; e.g.: Aspirin, Aceclofenac, etc.) 
2. 
Subjects who have current or prior history of clinically significant liver, 
kidney, 
gastrointestinal, 
cardiovascular, 
respiratory, 
endocrine, 
immune system, psychiatric/nerve system, and blood/oncological 
disorders 
3. 
Subjects with acute fever > 37.5℃ within 7 days from the expected IP 
administration or showing symptoms suspecting acute diseases within 
14 days from the expected IP administration 
4. 
Subjects who are diagnosed with hypertension/hypotension or subjects 
who are with clinically significant blood pressure* 
*Hypertension=systolic blood pressure ≥ 140 mmHg and/or diastolic blood 
pressure ≥ 90 mmHg 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
*Hypotension=systolic blood pressure ≤ 90 mmHg and/or diastolic blood 
pressure ≤ 60 mmHg 
5. 
Subjects who persistently drink more than the weekly recommended 
alcohol units* 
* 14 g of alcohol content per unit is applied, which corresponds to 1 can (small 
bottle) of beer (5%), 350 mL of draft beer (5%), 1 cup (300 mL) of makgeoli, 1 
glass (150 mL) of wine (12%), 1/4 bottle (90 mL) of soju (20%), and 1 shot (45 
mL) of liquor (40%). Moderate amount of alcohol by age and sex is provided 
below: 
Age 
Recommended unit per week 
Adults (≥ 19 and < 65 years) 
Male: 8 units/week 
Female: 4 units/week 
Elderly (≥ 65 years) 
Male: 4 units/week 
Female: 2 units/week 
 
6. 
Subjects of usual smoker (exceeding 10 cigarettes per day) 
7. 
Subjects who have a past history of autoimmune diseases (e.g.: 
rheumatoid arthritis, etc.) or active immune diseases that may affect 
the immune system [e.g.: flu, cancer, Human immunodeficiency virus 
(HIV), Hepatitis B virus (HBV), Hepatitis C virus (HCV) etc.], diabetes 
mellitus, heart diseases, asthma, sinusitis, chronic urticaria, 
dermographism, or any skin conditions that may affect post-IP 
administration assessments (e.g.: Dermatitis, dermatomycosis and 
other skin diseases or tattoo) 
8. 
Subjects with known allergic reactions or hypersensitivity to bee sting 
or 
other 
common 
allergens 
or 
known 
contraindications 
to 
hyaluronidase, thimerosal (disinfectants) and EDTA 
9. 
Subjects with hypersensitivity to the IP or its ingredients or a history of 
anaphylaxis 
10. Past history of drug abuse 
11. Subjects who have participated in other clinical trials within 6 months 
prior to the expected IP administration 
12. Others determined ineligible for the study participation in the opinion of 
investigator 
 
Part II-A 
13. Subjects who have been treated with a drug-metabolizing enzyme 
inducers and inhibitors* within 30 days prior to Visit 4 
* Example: Phenytoin, Carbamazepine, Barbiturates, Rifampicin, Griseofulvin, 
Cimetidine, 
Disulfiram, 
Erythromycin, 
Ketoconazole, 
Fluconazole, 
Itraconazole, 
Valproic 
acid, 
Isoniazid, 
Ciprofloxacin, 
Omeprazole, 
Clarithromycin, Quinidine, Sulfonamides, etc. 
14. Subjects who have significant bleeding or blood loss within 60 days 
prior to Visit 4 
15. Subjects who have donated whole blood within 60 days or donated 
blood by apheresis within 14 days or received transfusion within 14 
days prior to Visit 4 
16. Subjects who have consumed grapefruit juice within 7 days prior to Visit 
4 
Target Number 
of Subjects 
1. Total number of subjects: At least 231 subjects (Maximum of 290 
including subjects with allergic reactions, dropped out and excluded 
from analysis) 
2. Number of subjects for each Part 

 
 
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Protocol No.: ALT-BB4-01 
Version: 3.2 (2022-06-10) 
 
Part I (Allergy assessment): At least 231 subjects 
 
Part II-A (PK assessment): Total 23 subjects (including 10% 
drop-out rate) 
 
Part II-B (Safety assessment): At least 208 subjects  
(Study group- at least 139 subjects, Control group - at least 69 
subjects; randomized in a 2:1 ratio) 
Study Period 
1. 
Entire Study Period 
Approximately 18 months from approval of the Institutional Review 
Board (IRB) (however, it may be changed depending on the subject 
enrollment rate.) 
 
2. 
Study Period for Individual Subjects 
 
Part I (Allergy assessment):  
- 
Screening period: Up to 2 weeks 
- 
Treatment with IP: 1 day (ID single dose) 
- 
Follow-up period: 2 days (48 hours) 
 
 
Part II-A (Pharmacokinetic (PK) assessment): 
- 
Treatment with IP: 1 day (SC single dose) 
- 
PK Assessment: Total 15 times 
◼ 
Pre-dose: -2h, -1h, and 0h  
◼ 
Post-dose: 10 min (0.17h), 20 min (0.3h), 30 min (0.5h), 
40 min (0.7h), 50 min (0.8h), 1h, 1h 15 min (1.25h), 1h 30 
min (1.5h), 2h, 2 h 30 min (2.5h), 3h and 24h 
- 
Follow-up period: 4 weeks 
 
 
Part II-B (Safety assessment): 
- 
Treatment with IP: 1 day (SC single dose) 
- 
Follow-up period: 4 weeks 
Investigational 
Product (IP) 
1. 
Study Drug: ALT-BB4 (Recombinant Hyaluronidase) 
 
Active ingredient and Content: ALT-BB4 (recombinant 
hyaluronidase) 1,500 IU/mL 
 
Formulation and Appearance: Colorless, clear liquid injection  
 
Manufacturer: Alteogen, Inc. 
2. 
Comparator: Normal saline (0.9% NaCl)  
 
Active ingredient and Content: 0.9% NaCl 
 
Formulation and Appearance: Colorless, clear liquid injection  
 
Manufacturer: JW Pharmaceutical Corp.  
Administration Method 
(Assessment) 
Study drug (ALT-BB4,  
1,500 IU/mL) 
Comparator (0.9% 
NaCl) 
ID injection (Allergy 
assessment) 
30 IU (0.02 mL) 
0.02 mL 
SC injection (PK 
assessment) 
1,500 IU (1 mL) 
N/A 
SC injection (Safety 
assessment) 
1,500 IU (1 mL) 
1 mL 
Dosage and 
Administration 
1. 
Part I (Allergy assessment) 
Subjects will be randomized into the study group or control group at 
Baseline visit (Visit 2), and receive single dose of the study drug or 

 
 
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comparator intradermally into the right or left forearm.  
Treatment group 
Study group 
Control group 
IP 
Study drug  
(ALT-BB4, 1,500 IU/mL) 
Comparator (0.9% NaCl) 
ID dose 
30 IU (0.02 mL) 
0.9% NaCl (0.02 mL) 
Administration site* 
Right or left forearm 
Forearm in the opposite 
side of the previous dose 
* For example, when the study drug is intradermally injected into the left forearm, 
the 
comparator 
will 
be 
intradermally 
injected 
into 
the 
right 
forearm. 
 
2. 
Part II-A (PK assessment) 
Part II-A PK assessment will be performed only in subjects with drug allergy 
test negative at Visit 4, 2 days (48 hours) after the ID injection in Part I. All 
subjects participating in Part II-A will receive single dose of the study drug 
subcutaneously into the right or left upper arm. 
Treatment group 
Study group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
SC dose 
1,500 IU (1 mL) 
Administration site 
Right or left upper arm 
 
3. 
Part II-B (Safety assessment) 
Part II-B safety assessment will be performed only in subjects with drug 
allergy test negative at Visit 4, 2 days (48 hours) after the ID injection in Part 
I, randomized into the study group and control group in a 2:1 ratio. Subjects 
assigned in each group of Part II-B will receive single dose of the study drug 
or comparator subcutaneously into the right or left upper arm.  
Treatment group 
Study group 
Control group 
IP 
Study drug  
(ALT-BB4, 1,500 IU/mL) 
Comparator (0.9% NaCl) 
SC dose 
1,500 IU (1 mL) 
1 mL 
Administration site 
Right or left upper arm 
Right or left upper arm 
Methods 
This is a Phase 1 study to perform allergy assessment and PK profiling as 
well as to evaluate the safety and tolerability following administration of 
ALT-BB4 (recombinant hyaluronidase) in healthy volunteers. This study 
consists of Part I (Allergy assessment), Part II-A (PK assessment) and Part 
II-B (Safety assessment). In Part I, drug allergy assessment will be 
performed first. Then only in eligible subjects, PK of ALT-BB4 will be 
assessed in Part II-A, and the safety and tolerability assessment will be 
performed in Part II-B. 
 
Part I (Allergy assessment) 
For subjects who have agreed to participate by giving a written informed 
consent, Screening tests will be performed. By confirming the Screening 
test results and the inclusion/exclusion criteria, eligible subjects for 
participating in Part I of the study for the drug allergy assessment will be 
enrolled. Enrolled subjects will visit the institution and receive the IP 
intradermally. For up to 30 minutes after the IP administration, subjects will 
be observed for cutaneous hypersensitivity of immediate allergic reaction 
and subjects without any adverse event will be discharged. Later at Visit 3, 
subjects will be contacted via phone to monitor concomitant medications 

 
 
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Protocol No.: ALT-BB4-01 
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and adverse events. At Visit 4, subjects will visit the institution for monitoring 
of cutaneous hypersensitivity of delayed allergic reaction occurring within 
48 hours after the ID injection, blood collection, change of concomitant 
medications and adverse events, performing scheduled tests. ALT-BB4 
allergy assessment will be conducted through scheduled tests and 
evaluations for each visit. 
 
Part II-A (PK assessment) 
Eligible subjects for Part II-A of the study will be enrolled only from subjects 
with drug allergy test negative in Part I. Enrolled subjects will receive the IP 
subcutaneously at the institution and perform scheduled tests and blood 
collection at designated time points. At 3 hours after the IP administration, 
scheduled tests will be completed and subjects without any adverse event 
will be discharged. In a case of adverse events, the subjects will be treated 
by investigator as required before discharge. Later at Visit 5, subjects will 
visit the institution for blood collection, review of any change in concomitant 
medications and adverse events, and scheduled tests, and at Visit 6 (End 
of Study visit), subjects will visit the institution for review of any change in 
concomitant medications and adverse events and scheduled tests. ALT-
BB4 PK assessment will be conducted through scheduled tests and blood 
collection for PK assessment for each visit. 
 
Part II-B (Safety assessment) 
Eligible subjects for Part II-B of the study will be enrolled only from subjects 
with drug allergy test negative in Part I. Subjects enrolled in Part II-B study 
will be randomized into the study group or control group at the institution 
and receive single dose of the IP subcutaneously into the right or left upper 
arm. For up to 30 minutes after the IP administration, subjects will be 
discharged if there is no adverse event when monitoring systemic and 
administration site adverse events. In a case of adverse events, the 
subjects will be treated by investigator as required before discharge. Later 
at Visit 5, subjects will visit the institution for blood collection, review of any 
change in concomitant medications and adverse events, and scheduled 
tests. Visit 6 is a phone visit, where concomitant medications and adverse 
events including systemic and administration site events will be monitored. 
At Visit 7, the End of Study visit, scheduled tests and safety assessment 
will be performed to evaluate the safety and tolerability of ALT-BB4. 
Endpoints and 
Assessment 
Method 
Efficacy Endpoints 
The study aims to evaluate allergy, PK, safety and tolerability and efficacy 
analysis will not be separately performed. 
 
Safety Endpoints 
Part I study will evaluate the drug allergy and adverse events occurring 
following ID injection of the IP. Part II-A study will evaluate adverse events 
occurring following SC injection of the IP. Part II-B study will evaluate the 
safety and tolerability following SC injection of the IP.  
 
1. 
Primary Endpoints 
1) Incidence rate of drug allergy following ID injection of the IP* in 
Part I 
*Subjects developing either immediate or delayed allergic reaction are 
considered to have drug allergy.  
The number and percentage (%) of subjects showing any change of 
drug allergy from the IP administration to the end of study or early 

 
 
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termination in the control group and study group will be provided. In 
addition, as the primary analysis, the upper limit of binomial exact 95% 
two-sided confidence interval (97.5% one-sided confidence interval) < 
10% is considered statistically significant. In addition, the analysis will 
be performed using the McNemar’s test to compare the incidence rate 
of drug allergy between the study group and the control group. 
 
2) Safety and tolerability assessment following SC injection of the 
IP** in Part II 
** Incidence rate of adverse events, including systemic and administration 
site-related adverse events 
To evaluate the safety and tolerability, systemic and administration 
site-related adverse events will be assessed following SC injection of 
the IP. 
In addition to the routine safety tests of physical examination, vital 
signs, ECG and laboratory tests, comprehensive data about clinical 
adverse events will be evaluated to determine relationship of the 
adverse drug reactions, including the nature, signs and type of adverse 
events, severity, frequency and duration, occurrence site and types of 
disease, and symptoms of AEs by system organ class (SOC). 
The number and percentage (%) of subjects developing adverse 
events will be provided and the analysis will be performed using the 
Pearson’s chi-square test or Fisher’s exact test to compare the 
incidence rate of AEs between the study group and the control group.. 
 
2. 
Secondary Endpoints 
1) Incidence rate of immediate allergic reactions† occurring within 
30 minutes after the ID injection of the IP 
†defined as drug allergy associated with wheal ≥ 8 mm with or without 
pseudopod (elevation considered due to an injection is not considered 
wheal), erythema or localized itching occurring in the administration site 
within 30 minutes after IP administration 
 
The number and percentage (%) of subjects showing any change of 
immediate allergic reaction occurring within 30 minutes after the IP 
administration in the control group and study group will be provided and 
analyzed using the McNemar’s test to compare the incidence of 
immediate allergic reaction between the study group and the control 
group. 
 
2) Incidence rate of delayed allergic reactions‡ occurring within 48 
hours after the ID injection of the IP 
‡drug allergy occurring from 30 minutes to 48 hours after the IP 
administration, including but not limited to redness, rash, urticaria, edema, 
and erythema in the administration site as well as acute generalized 
exanthematous pustulosis and erythema multiforme. Investigator will decide 
whether an adverse event is a delayed allergic reaction. 
 
The number and percentage (%) of subjects showing any change of 
delayed allergic reaction occurring from 30 minutes to 48 hours after 
the IP administration in the control group and study group will be 
provided and analyzed using the McNemar’s test to compare the 
incidence of delayed allergic reaction between the study group and the 
control group. 

 
 
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Protocol No.: ALT-BB4-01 
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3) Incidence rate of administration site events* at 30 min. and 48 
hours after the IP administration 
* classified into wheal, erythema, localized itching and other adverse events, 
occurring after the IP administration, at each time point 
 
For wheal, erythema, localized itching and other adverse events at 30 
minutes and 48 hours after the IP administration in the control group 
and study group, the number and percentage (%) of subjects showing 
any change of each event will be provided and analyzed using the 
McNermar’s test in Part 1 and Pearson’s chi-square test or Fisher’s 
exact test in Part 2 to compare the incidence of allergic reaction 
between the study group and control group by administration site-
related AE. 
 
4) Size of wheal and erythema occurring at 30 min. and 48 hours after 
the IP administration 
For the size of wheal and erythema occurring at 30 minutes and 48 
hours and change in the size of wheal and erythema occurring at 48 
hours after the IP administration compared to 30 minutes after the IP 
administration in the control group and study group, descriptive 
statistics (mean, standard deviations, median, min and max) will be 
provided. The analysis will be performed using the Paired t-test or 
Wilcoxon signed-rank sum test to compare the change in the size of 
wheal and erythema occurring at 48 hours after the IP administration 
compared to 30 minutes, and for the comparison between the study 
group and control group, an analysis of covariance (ANCOVA) with the 
baseline as a covariate will be performed. If the assumption of normality 
is not satisfied, a rank ANCOVA will be performed. 
 
3. 
Other Safety Endpoints 
1) Adverse Events 
All adverse events will be coded by using the most recent version of 
Medical dictionary for regulatory activities (MedDRA). The number of 
subjects with adverse events and adverse drug reactions, incidence 
rate (%) and frequency by treatment group will be provided, along with 
its 95% confidence interval. 
In addition, the number of subjects with adverse events and adverse 
drug reactions, incidence rate and frequency by SOC and PT will be 
provided. 
 
2) Physical Examination, Vital Signs, ECG and Laboratory Tests 
For continuous data, descriptive statistics (mean, standard deviation, 
median, min and max) for absolute values and changes from baseline 
at each time point of visits and Paired t-test or Wilcoxon signed-rank 
test will be performed. For laboratory tests and ECG values, the 
frequency and proportion of subjects shifting from normal/clinically 
insignificant abnormal value before the IP administration (Screening) 
to clinically significant abnormal value after the IP administration will be 
provided and McNemar’s test will be performed. 
 
PK Endpoints 

 
 
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1. 
Primary endpoints: Cmax, AUClast, t1/2, and Tmax of ALT-BB4 
2. 
Secondary endpoints: Baseline-adjusted Cmax, AUClast, t1/2, and Tmax of 
ALT-BB4 
 
For PK endpoints, statistical test is not separately performed. PK 
assessment results can be provided in a separate report. 
 
 

 
 
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2.2 Table of Study Schedule 
After allergy assessment in Part I, Part II-A and Part II-B will be performed as provided in the 
schedule below. 
 
2.2.1 
Part I (Allergy assessment) 
 
1. 
Screening visit and baseline visit can be performed on the same day. 
2. 
Vital signs and laboratory tests will be performed before the ID injection of the IP. 
3. 
Visit 4 will be the last visit of Part I for allergy assessment and will be 2 days after ID injection of IP (visit 
2) (Visit 3 is a phone visit). Subjects with negative allergy test result on ID injection will proceed to Part 
II-A and Part II-B baseline visit. On baseline visit for Part II-A and Part II-B, SC injection will be 
conducted. 
4. 
Written informed consent must be obtained from subjects prior to any study-related procedures. 
5. 
Sex, date of birth, skin conditions (presence of tattoo, acne, dermatitis, pigmentation, lesion and skin 
damage on the administration site), smoking (non-smoker, former smoker or current smoker), alcohol 
intake (non-drinker, former drinker or current drinker), weekly alcohol intake, and daily smoking amount 
will be investigated. 
6. 
Only for subjects considering participating in Part II-A, information about history of whole blood donation 
within 60 days, apheresis within 14 days, and transfusion within 14 days prior to Visit 4 will be 
investigated. 
7. 
Medical history (surgical history within 3 years prior to screening visit, past medical history within 1 year 
prior to screening visit and current medical conditions) and past drug allergy will be checked.  
8. 
At screening visit, medications administered within 3 months prior to screening will be investigated, and 
after screening visit, any change of medications investigated in the previous visit will be reviewed. 
9. 
After taking a rest for at least 5 minutes, sitting systolic and diastolic blood pressures, respiratory rate, 
Period 
Screening1 
Baseline1 
Follow up 
(Wash-out before Part II) 
Visit 
Visit 1 
Visit 22 
Visit 3 
(Phone Visit) 
Visit 43 
Day (D) 
–14D ~ –1D 
0D 
1D 
2D 
Written informed consent4 
X 
 
 
 
Assigning screening number 
X 
 
 
 
Demographic data5 
X6 
 
 
 
Medical history7 
X 
X 
 
 
Prior/Concomitant medications8 
X 
X 
 
X 
Vital signs9 
X 
X 
 
X 
ECG 
X 
 
 
 
Physical examination10 
X 
 
 
 
Laboratory test11 
X 
X 
 
X 
Immunogenicity test 
X 
 
 
 
Pregnancy test12 
X 
 
 
 
Check of inclusion/exclusion 
criteria 
X 
X 
 
 
Randomization  
 
X 
 
 
ID injection of the IP 
 
X 
 
 
Observation of administration site 
following ID injection of the IP13 
 
X 
9 
X 
Photographing of the ID injection 
site of the IP 14 
 
X 
() 
X 
Monitoring of AEs15 
 
X 
 
X 

 
 
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pulse rate and temperature will be measured, before performing other tests. 
10. Height, weight, and BMI measurements will be performed, and allergy, cardiovascular system, 
respiratory system, gastrointestinal system/hepatobiliary system, metabolism/endocrine system, 
renal/urinary system, reproductive system, musculoskeletal system, skin and connective tissues, 
nervous system, psychiatric system and other body organs will be examined. 
11. Tests results for baseline (Visit 2) may be replaced with the results obtained within 7 days prior to 
baseline visit (Visit 2), if available. 
12. Pregnancy test (serum β hCG or urine hCG) will be performed in women of child-bearing potential who 
have not received a sterilization procedure, or women who have the last menstrual period within 12 
months or are not diagnosed with menopause only. 
13. After the ID injection, injection site reaction and skin in the administration site will be observed for any 
abnormality. In a phone visit (Visit 3), subjects will report findings after observation on the ID injection 
site to the investigator via telephone, message or social media, etc. together with adverse events. 
14. Photograph of administration site following ID injection of the IP will be taken at Visit 2 and Visit 4. For 
details about photographing method, follow a separate manual. Through a phone visit (Visit 3), 
photograph of the administration site taken by the subject will be sent to the investigator, if necessary. 
15. AEs occurring following ID injection of the IP will be collected. Later, presence of AEs will be monitored 
for specified visits.  
 
 

 
 
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2.2.2 
Part II-A (PK assessment) 
 
Period 
Baseline 
Follow up 
USV 
Visit 
Visit 4 
Visit 
5 
Visit 6 
Day (D) 
0D 
1D 
28D 
Visit Window: day 
- 
- 
±2days 
PK Sampling Timepoint1 
Pre-dose2  Administ
ration 
30 min 
post-
dose 
3h post-
dose 
24h 
post-
dose 
- 
Concomitant medications3 
X 
 
 
 
X 
X 
(X) 
Vital signs4 
X 
 
 
X 
X 
X 
(X) 
ECG 
 
 
 
 
 
X 
(X) 
Physical examination5 
 
 
 
 
 
X 
(X) 
Laboratory test 
X 
 
 
X 
X 
X 
(X) 
Immunogenicity test 
 
 
 
 
 
X 
 
Pregnancy test6 
 
 
 
 
 
X 
(X) 
Blood collection for PK (total 
15 times) 
①~③ 
 
④~⑥ 
⑦~⑭ 
⑮ 
 
 
Assigning allocation number 
 
X 
 
 
 
 
 
SC injection of the IP 
 
X 
 
 
 
 
 
Observation of administration 
site following SC IP injection7 
 
X 
X 
X 
X 
X 
(X) 
Photographing of the SC 
injection site of the IP8 
 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Monitoring of AEs9 
 
X 
X 
X 
X 
X 
(X) 
USV: Unscheduled visit 
 
1. 
Blood collection time points for PK assessment (total 15 times) 
Pre-dose (3 times) 
Pre-dose 
①2 h (-2h±10mins), ②1 h (-1h±10mins), ③0 h (0h-
10mins, blood collection before SC injection) 
Post-dose (12 
times) 
 30min post-dose 
④10 min (0.17h±3mins), ⑤20 min (0.3h±3mins), ⑥30 
min (0.5h±3mins) 
3h post-dose 
⑦40 min (0.7h±3mins), ⑧50 min (0.8h±3mins), ⑨1 h 
(1h±5mins), ⑩1 h 15 min (1.25h±5mins), ⑪1 h 30 min 
(1.5h±5mins), ⑫2 h (2h±5mins), ⑬2 h 30 min 
(2.5h±10mins), ⑭3 h (3h±10mins) 
24h post-dose 
⑮24 h (24h±60mins) 
2. 
Vital signs and laboratory tests will be performed before the SC injection of the IP and may be replaced 
with the results obtained at Visit 4 of Part I. 
3. 
Any change of medications investigated in the previous visit will be reviewed. 
4. 
After taking a rest for at least 5 minutes, sitting systolic and diastolic blood pressures, respiratory rate, 
pulse rate and temperature will be measured, before performing other tests. 
5. 
Cardiovascular 
system, 
respiratory 
system, 
gastrointestinal 
system/hepatobiliary 
system, 
metabolism/endocrine system, renal/urinary system, reproductive system, musculoskeletal system, 
skin and connective tissues, nervous system, psychiatric system and other body organs will be 

 
 
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Protocol No.: ALT-BB4-01 
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examined. 
6. 
Pregnancy test (serum β hCG or urine hCG) will be performed in women of child-bearing potential 
who have not received a sterilization procedure, or women who have the last menstrual period within 
12 months or are not diagnosed with menopause only. 
7. 
After the SC injection, injection site reaction and skin in the administration site will be observed for 
any abnormality.  
8. 
Photographing will be performed when there is any visible abnormality in the administration site (if 
necessary). For details about photographing method, follow a separate manual. Through a phone visit, 
photograph of the administration site taken by the subject will be sent to the investigator. 
9. 
AEs occurring before IP injection will be collected. Later, presence of AEs will be monitored for 
specified visits. 
 
 
 

 
 
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2.2.3 
Part II-B (Safety assessment) 
Period 
Baseline 
Follow up 
USV 
Visit 
Visit 41 
Visit 5 
Visit 6 
(Phone Visit) 
Visit 7 
Day (D) 
0D 
2D 
14D 
28D 
Visit Window: day 
- 
±1day 
±2days 
±2days 
Concomitant medications2 
X 
X 
 
X 
(X) 
Vital signs3 
X 
X 
 
X 
(X) 
ECG 
 
 
 
X 
(X) 
Physical examination4 
 
 
 
X 
(X) 
Laboratory test 
X 
X 
 
X 
(X) 
Immunogenicity test 
 
 
 
X 
 
Pregnancy test5 
 
 
 
X 
(X) 
Randomization6 
X 
 
 
 
 
SC injection of the IP7 
X 
 
 
 
 
Observation of administration 
site following SC injection of the 
IP8 
X 
X 
7 
X 
(X) 
Photographing of the SC 
injection site of the IP9 
X 
X 
() 
(X) 
(X) 
Monitoring of AEs10 
X 
X 
 
X 
(X) 
Intensive monitoring of AEs10 
 
: 1D~5D 
 
 
 
USV: Unscheduled visit 
 
1. 
Vital signs and laboratory tests will be performed before the SC injection of the IP and may be replaced 
with the results obtained at Visit 4 of Part I. 
2. 
Any change of concomitant medications investigated in the previous visit will be reviewed. 
3. 
After taking a rest for at least 5 minutes, sitting systolic and diastolic blood pressures, respiratory rate, 
pulse rate and temperature will be measured, before performing other tests. 
4. 
Cardiovascular 
system, 
respiratory 
system, 
gastrointestinal 
system/hepatobiliary 
system, 
metabolism/endocrine system, renal/urinary system, reproductive system, musculoskeletal system, 
skin and connective tissues, nervous system, psychiatric system and other body organs will be 
examined. 
5. 
Pregnancy test (serum β hCG or urine hCG) will be performed in women of child-bearing potential who 
have not received a sterilization procedure, or women who have the last menstrual period within 12 
months or are not diagnosed with menopause only. 
6. 
Randomization into the study group or control group for SC injection in Part II-B. 
7. 
SC injection site and presence of injection reaction will be observed for 30 minutes after SC injection 
of the IP and subjects without any symptom will be discharged. More information about adverse events 
following SC injection will be given by the principal investigator. 
8. 
After the SC injection, injection site reaction and skin in the administration site will be observed for any 
abnormality. In a phone visit (Visit 6), subjects will report findings after observation on the SC injection 
site to the investigator via telephone, message or social media, etc. together with adverse events. 
9. 
Photograph of administration site following SC injection of the IP will be taken at Visit 4 and Visit 5. For 
details about photographing method, follow a separate manual. In other visits, photographing will be 
performed when there is any visible abnormality in the administration site (if necessary), and subjects 
will photograph the administration site and send it to the investigator. 
10. Adverse events before and after SC injection will be monitored. In particular, other adverse events 
following SC injection will be monitored, other than those found during observation of injection site 
reactions and skin in the SC injection site. Later, presence of AEs will be monitored for specified visits. 
However, investigator will contact subjects by telephone every day up to Day 5 after SC injection to 
check presence of AEs.  

 
 
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Protocol No.: ALT-BB4-01 
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2.3 Diagram 
 
Part I (Allergy assessment), Part II-A (PK assessment)/ Part II-B (Safety assessment) 
 
 
 

 
 
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3. Introduction 
3.1. Study Background 
Hyaluronic acid (HA) is composed of disaccharide repeats of N-acetylglucosamine and glucuronic 
acid joined alternately in a form of chain. HA is a natural high-molecular weight substance in the 
extracellular matrix (ECM) and exists in various parts, such as animal vitreous humor, cartilage, 
synovia, and placenta1). HA fills any empty space in the cells or fiber tissues, stimulating collagen 
regeneration and inhibiting germ invasion2). Regarding drug administration, HA binds to water, 
forming jelly-like form and thus inhibiting dispersion and absorption of a drug subcutaneously 
injected3), which limits a dose per subcutaneous (SC) injection4). SC injection may improve 
patient’s quality of life compared to intravenous (IV) injection, but it is associated with many 
problems due to limited dosage per injection and relatively lower bioavailability, increasing 
required amount of a drug, which increases viscosity of the drug during administration. Increased 
SC injection frequency with low dose causes a concern of immunogenicity. To overcome these 
problems, collagen, one composition of ECM, was degraded and used to enhance penetration of 
a drug subcutaneously injected. However, with known half-life of approx. 15 years, collagen is 
associated with concerns of permanent skin tissue changes. On the contrary, HA has relatively 
short half-life of 15-20 hours, compared to collagen. Thus, hyaluronidase, an enzyme that can 
degrade HA, has drawn attention. 
Hyaluronidase is an endoglycosidase that degrades the glycosidic bond of HA which is a 
disaccharide, between N-acetylglucosamine and glucuronic acid into monosaccharide5), which is 
found in various species, such as bacteria, fish and mammal6). Human hyaluronidase is classified 
into 6 subtypes, Hyaluronidase 1-4, PH20 and Hyaluronoglucosaminidase pseudogene 1 
(HYALP1), among which Hyaluronidase 1 is known to play the most important role5). A subtype 
of hyaluronidase, Sperm adhesion molecule 1 (SPAM 1, PH20), is located on the surface of sperm, 
degrading HA on the surface of egg at the time of conception, and thus increasing penetration 
into the egg3). Focusing on such functions, bovine or chicken-derived hyaluronidase has been 
used to enhance the absorption and dispersion of a drug since 1940s7). However, animal-derived 
hyaluronidase causes concerns of allergic reactions, such as anaphylaxis, and spread of 
zoonosis, such as variant Creutzfeldt-Jakob disease4). To solve the problems, Halozyme based 
in the US developed an allogenic human-derived hyaluronidase, called HYLENEX, which was 
approved in 2005 by the US Food and Drug Administration (FDA) and has been sold until now. 
Active ingredient of HYLENEX is the 5th subtype of human hyaluronidase, PH20 and the most 
frequently reported adverse events were mild local adverse events, such as erythema or pain, 
with incidence rate of allergic reaction less than 0.1%.8) HYLENEX has been approved and used 
as a spreading factor for anticancer agents or immunodeficiency treatment, for example HyQvia 
and Herceptin SC. 
Human-derived hyaluronidase allows change of drug administration route from IV to SC, which 
can enhance patient’s quality of life and improve problems, such as limited dosage or increased 
dosing frequency with the existing SC injection. However, no human-derived hyaluronidase has 
been developed until now since Halozyme’s HYLENEX. ALT-B4 is the second human-derived 
hyaluronidase developed after HYLENEX and provides additional benefits, increasing heat 
stability for increased protein stability, while maintaining the same mechanism of action and 
enzyme activity as the human-derived hyaluronidase of Halozyme. 
Adverse drug reaction (ADR) refers to any undesirable and untoward events occurring at any 
dose of an IP, where causality to the IP cannot be denied. Based on expectedness, ADRs are 
classified into Type A reactions, which are predictable, and Type B reactions, which are not 
predictable.9), 10) In Type B reaction, an ADR caused by immune mechanism is called drug 
allergy.11), 12) Drug allergy can be classified into immediate allergic reaction, mediated by 
immunoglobulin E (IgE) and appearing within 1 hour after drug administration, and non-immediate 
(delayed) allergic reaction, mediated by T cells and appearing within 1-24 hours or at the latest 
48 hours after drug administration13), 14). Medications known to have high prevalence of drug 
allergy include penicillins antibiotics, cephalosporins antibiotics, non-steroidal anti-inflammatory 
drugs, narcotic analgesics and anticonvulsants15), and frequent drug use, high single dose, and 
administration route of injection, instead of oral, and patients with asthma, sinusitis and chronic 
urticaria are known factors of allergic reaction15). Estimated prevalence of drug allergy is about 5-

 
 
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Protocol No.: ALT-BB4-01 
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10%, and drug allergy may increase mortality and economic burden10). 
 
3.2. Study Rationale 
The investigational product (IP), ALT-BB4, is a drug product containing ALT-B4 as an active 
ingredient. The active ingredient, ALT-B4, is a hyaluronidase that facilitates dispersion of other 
medications and reconstructs the dermal layer, which has the same enzyme activity as the 
commercial hyaluronidase available in the market. Enzyme activities of ALT-B4 are measured by 
turbidimetric assay and/or viscometric assay. Turbidimetric assay uses ultrasound-visible light 
spectrophotometer to measure turbidity due to HA-albumin complex, which uses a principle where 
degradation of HA into hyaluronidase reduces the amount of HA-albumin complex, decreasing 
absorbance at 600 nm wavelength (A600 nm). Enzyme activities of the sample are calculated by 
using A600 nm and external calibration curve of the standard hyaluronidase. When the 
turbidimetric assay established by Alteogen was applied to commercially available drug products 
containing recombinant human hyaluronidase (rHuPH20) (Herceptin SC, and Mabthera SC), it 
was found that they were very similar to activities specified in each product insert. The assay is 
in vitro assessment of the same mechanism as found in the human body, and thus can be applied 
to existing indications. Viscometric assay measures a change of viscosity caused by degradation 
of HA with hyaluronidase, which is applied to animal testis-derived hyaluronidase commercially 
available. Like turbidimetric assay, it was proven to provide very similar activity results to those 
specified in each product insert. 
Mechanism related to clearance of hyaluronidase administered is not well known. However, it is 
known that hyaluronidase is inactivated in blood of many mammals. Known half-life of bovine 
testis-derived hyaluronidase in human serum is 3.2 minutes. Reconstruction level of damaged 
skin barrier due to intradermal (ID) injection of hyaluronidase at 0.02-~20 Units/mL was checked 
for at 24 hours, which showed that the recovery of barrier was incomplete and was in inverse 
proportion to the amount of enzyme administered. Complete recovery was found at 48 hours. 
Hyaluronidase has been administered as a mixture with anticancer agents to enhance absorption 
of anticancer agents subcutaneously injected. 
 
3.3. Risk and Benefit Assessment 
Summary of results from non-clinical trials is provided. For details, refer to the Investigator’s 
Brochure (IB). 
 
3.3.1. In Vitro Pharmacology 
1. 
Immunogenicity Assessment 
By checking for its capability of inducing stimulus reaction in CD4 and CD8+ cells, potential 
immunogenicity of ALT-B4 was confirmed and ALT-BB4 and PH20 were compared at the 
concentration of 1.5 and 15 μg/mL (n=10 Per condition). As shown in Figure 3.3.1, majority of the 
ALT-BB4 and PH20 results had stimulation index (SI) no more than 2 (positive threshold). 
Especially for ALT-BB4, all results did not exceed the positive threshold, except one result in 
CD8+ cells at the concentration of 15 μg/mL. 

 
 
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Protocol No.: ALT-BB4-01 
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Figure 3.3.1 Stimulatory Potential of ALT-B4 
 
 
2. 
Enzyme Activity 
Alteogen, Inc. performed enzymatic kinetics assay (Morgan-elson assay) to determine enzyme 
kinetic parameters of ALT-B4, compared to PH20. Colorimetric morgan-elson assay is based 
upon a reducing N-acetyl-D-glucosamine terminus generated by cleavage reaction of enzyme 
(Figure 3.3.2). 
Figure 3.3.2 Morgan-Elson Reaction 
 
Morgan-Elson assay results are provided in Table 3.3.1 and Figure 3.3.3. The results showed 
that ALT-B4 has approximately 3-fold higher catalytic efficiency, compared to PH20. 
Table 3.3.1 Enzyme Kinetic Parameters at 37°C, pH 5.7 
Parameter 
PH20 
ALT-B4 
KM (mg/mL) 
1.59 
1.21  
kcat (/sec) 
13.65 
30.77 
kcat/KM (mL/mg sec) 
8.58 
25.40 
kcat=Catalytic rate constant; KM=Substrate concentration at which the reaction rate is half of Vmax; Vmax=Maximum rate 
achieved. 
 
 

 
 
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Protocol No.: ALT-BB4-01 
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Figure 3.3.3 Michaelis-Menten Plot of ALT-B4 and PH20 
 
 
3.3.2. In Vivo Pharmacology 
1. 
Study Y01934 
To evaluate efficacy of enzymes that help dispersion and absorption of other medications, dye 
dispersion study was performed, and sequential administration of ALT-B4 followed by Trypan 
blue was compared to concurrent administration of ALT-B4 and Trypan blue by mixing. The study 
results are provided in Table 3.3.2 and Figure 3.3.4. Dispersion of Trypan blue was proportional 
to ALT-B4 concentration, and shorter injection interval between ALT-B4 and Trypan blue was 
associated with larger area of dispersion. Mortality and adverse events were not observed 
following ID injection of ALT-BB4. 
Table 3.3.2 Diffusion Test 
Cohort 
No. 
Admin. 
ALT-
B4 
Conc. 
(U/mL) 
Time 
Between 
ALT-B4 
and 
Trypan 
blue 
0 min 
2.5 min 
5 min 
20 min 
AVG 
(mm2) SEM AVG 
(mm2) SEM AVG 
(mm2) SEM AVG 
(mm2) SEM 
1 
Co-mix 
0 
0 min 
30.7 
2.1 
74.5 
3.8 
94.9 
8.5 
110.4 
4.9 
2 
Seq. 
1 min 
49.3 
1.3 
53.7 
1.9 
59.0 
2.7 
70.1 
5.4 
3 
Co-mix 
100 
0 min 
29.6 
2.0 
63.2 
3.1 
81.3 
4.9 
102.7 
3.9 
4 
500 
35.0 
0.9 
16.3 
3.7 
91.4 
4.6 
125.5 
7.3 
5 
5,000 
31.9 
1.2 
87.7 
4.2 
107.8 
6.9 
145.6 
5.1 
6 
Seq. 
100 
1 min 
56.6 
3.3 
98.5 
3.5 
118.0 
2.5 
147.8 
5.3 
7 
100 
20 min 
56.6 
2.4 
98.8 
5.0 
107.7 
4.5 
123.1 
4.9 
8 
100 
4 hrs 
49.3 
2.7 
90.8 
4.8 
102.4 
6.1 
119.3 
5.8 
9 
100 
24 hrs 
41.3 
1.6 
76.0 
3.0 
90.9 
5.1 
106.1 
5.0 
10 
100 
48 hrs 
36.6 
1.9 
67.9 
4.2 
78.0 
5.1 
97.2 
7.0 
AVG=Average; Co-mix=Concomitant; SEM=Standard error of the mean; Seq=sequential. 
 
 
 

 
 
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Protocol No.: ALT-BB4-01 
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Figure 3.3.4 Diffusion Test 
 
 
3.3.3. Nonclinical Pharmacokinetics 
1. 
Study 2519133 
Single dose pharmacokinetic (PK) study of ALT-B4 was performed in Sprague dawley rats. Single 
dose of ALT-B4 at 1 mg/kg was injected intravenously in 4 male rats, and blood collection was 
performed pre-dose, and 1, 2, 3, 4, 8, 16, 30, and 60 minutes post-dose. Half-life in plasma was 

 
 
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Protocol No.: ALT-BB4-01 
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6.9 (±3.8) min. Mortality and adverse events were not observed. 
 
2. 
Study 2089-034 (ANC028) 
The study evaluated PK of ALT-B4 following single IV and SC injections of ALT-B4 in Cynomolgus 
monkeys. In Phases 1 and 2, single dose of ALT-B4 at 0.3, 3, 15 or 30 mg/kg was injected 
intravenously at 3 mL per kg to each animal, while in Phases 3 and 4, a single dose of ALT-B4 at 
1, 3, 10 or 30 mg/kg was injected subcutaneously at 3 mL per kg. Blood samples for PK analysis 
were collected at 0 (pre-dose), 1, 3, 5, 10, 15, 30, and 60 minutes post-dose for IV injection, or 
0.25, 0.5, 1, 4, 8, and 24 hours post-dose for SC injection. 
Until Phase 3, no clinical adverse event was found after injections. 
Table 3.3.3 Study design of monkey PK 
Group 
Test 
Article 
No. of 
Males 
Dose 
Route 
Vehicle 
Dose Level 
(mg/kg) 
Dose Volume 
(mL/kg) 
Collection 
Intervals 
Phase 1 
1 
ALT-B4 
3 
IV 
Histidine buffer 
0.3 
3 
Blooda 
2 
ALT-B4 
3 
IV 
Histidine buffer 
3 
3 
Blooda 
Phase 2 
1 
ALT-B4 
3 
IV 
Histidine buffer 
15 
3 
Blooda 
2 
ALT-B4 
3 
IV 
Histidine buffer 
30 
3 
Blooda 
Phase 3 
1 
ALT-B4 
3 
SC 
Histidine buffer 
1 
3 
Bloodb 
2 
ALT-B4 
3 
SC 
Histidine buffer 
3 
3 
Bloodb 
Phase 4 
1 
ALT-B4 
3 
SC 
Histidine buffer 
10 
3 
Bloodc 
2 
ALT-B4 
3 
SC 
Histidine buffer 
30 
3 
Bloodc 
aPhase 1 and 2 (IV): Blood samples were collected at 0(pre-dose), 1, 3, 5, 10, 15, 30, and 60 minutes postdose 
bPhase 3 (SC): Blood samples were collected at 0.25, 0.5, 1, 4, 8, and 24 hours postdose 
cPhase 4 (SC): Blood samples were collected at 0.25, 0.5, 1, 4, 8, 24, and 48 hours postdose 
 
Following IV injection at 3-30 mg/kg, blood ALT-B4 concentration was considered not affected by 
baseline value. Cmax and AUC values showed dose-dependent results and half-life of ALT-B4 was 
found 0.3-0.87 hours, confirming rapid elimination from the body (Figure 3.3.5, Table 3.3.4). 
However, all PK parameters did not show significant difference between groups in the dose range 
of 0.3-30 mg/kg (one-way ANOVA, p-value>0.05). 
 
 
 

 
 
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Figure 3.3.5 Plasma concentration vs. time profiles of ALT-B4 after IV injections (upper 
panel: rectilinear scale, lower panel: semi-logarithmic scale) 
 
 
 
 
 
 
0
200
400
600
800
1000
1200
0
0.25
0.5
0.75
1
ALT-B4 concentration (μg/mL)
Time (h)
0.3 mg/kg
3 mg/kg
15 mg/kg
30 mg/kg
1
10
100
1000
10000
0
0.25
0.5
0.75
1
ALT-B4 concentration (μg/mL)
Time (h)
0.3 mg/kg
3 mg/kg
15 mg/kg
30 mg/kg

 
 
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Protocol No.: ALT-BB4-01 
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Table 3.3.4 Average PK parameters of ALT-B4 after IV injections 
Parameter 
ALT-B4 dose 
0.3 mg/kg 
3 mg/kg 
15 mg/kg 
30 mg/kg 
t1/2 (h) 
1.13 ± 0.31 
0.3 ± 0.19 
0.75 ± 0.66 
0.87 ± 0.46 
C0 (μg/mL) 
4.27 ± 2.87 
7.52 ± 4.22 
6.02 ± 5.39 
13.9 ± 9.01 
AUClast (μg·h/mL) 
5.14 ± 3.07 
37.38 ± 10.71 
250.88 ± 97.82 
576.95 ± 223.42 
AUCinf (μg·h/mL) 
11.24 ± 7.87 
39.23 ± 10.3 
460.83 ± 323.39 
1155.31 ± 570.04 
Cl (mL/h/kg) 
42.87 ± 37.57 
80.22 ± 21.69 
52.99 ± 47.25 
32.98 ± 21.6 
Vss (mL/kg) 
53.29 ± 27.33 
27.57 ± 6.69 
31.16 ± 7.43 
33.87 ± 10.89 
t1/2: Elimination half-life, C0: Plasma concentration at time zero, AUClast and AUCinf: Area under the concentration, Cl: 
Clearance, Vss: Steady state volume 
 
For ALT-B4 subcutaneously injected at 1-30 mg/kg, PK parameters, such as Cmax and AUC were 
not dose dependent and in all animals, they couldn't be calculated due to effects of baseline 
values and absence of observation in the elimination phase. There was no meaningful difference 
in absorption kinetics among 1, 3, and 10 mg/kg groups. Considering increased ALT-B4 
concentration in blood at the highest dose (30 mg/kg), compared to lower doses, it is assumed 
that ALT-B4 has been absorbed into the body circulatory system. Although the finding that the 
highest concentration of ALT-B4 was reached within 0.25-1 hour after SC injection showed rapid 
absorption into body circulatory system (Figure 3.3.6), exposure to body circulatory system 
following SC injection of ALT-B4 was very low when comparing to IV injection group of the same 
dose. 
 
 
 

 
 
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Protocol No.: ALT-BB4-01 
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Figure 3.3.6 Plasma concentration vs. time profiles of ALT-B4 after SC injections (upper 
panel: rectilinear scale, lower panel: semi-logarithmic scale) 
 
 
 
 
3.3.4. Toxicology 
1. 
Study 2089-031 (ANC021) 
Non-GLP dose-finding and tolerability studies were performed following single dose. Vehicle, and 
0.04, 0.2, 2 mg/kg of ALT-B4 were administered at a dose of 0.5 mL/kg to all directions of the 
trunk of 4 male minipigs aged 5 months. 
There was no clinical or veterinary symptom related to local toxicity of ALT-B4, and skin evaluation 
did not show any ALT-B4-related symptoms. 
The study results showed good tolerability of single SC injection of ALT-B4 up to 2 mg/kg. 
 
2. 
Study 891-0003-TX (ANC026) 
A single dose rat study was performed for 7 days, where toxicity of ALT-B4 IV and SC injection 
was evaluated, and reversibility, persistence or delayed toxicity that can occur during 7-day 
0
5
10
15
20
25
30
35
0
6
12
18
24
30
36
42
48
ALT-B4 concentration (μg/mL)
Time (h)
1 mg/kg
3 mg/kg
10 mg/kg
30 mg/kg
1
10
100
0
6
12
18
24
30
36
42
48
ALT-B4 concentration (μg/mL)
Time (h)
1 mg/kg
3 mg/kg
10 mg/kg
30 mg/kg

 
 
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Protocol No.: ALT-BB4-01 
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recovery period were evaluated. 
Mortality was not observed, and there was no change related to ALT-B4 in aspects of clinical sign, 
weight, food intake, hematology, serum chemistry, coagulation, urinalysis, organ weight, gross 
pathology, and histopathology. 
Single IV and SC injection of ALT-B4 to rats at 0, 0.04, 0.2, and 2 mg/kg were found well tolerable 
and no observed adverse effect level (NOAEL) was determined 2 mg/kg. 
 
3. 
Study 891-0004-TX (ANC027) 
A GLP repeat dose toxicity rat study was performed for 29 days, where toxicity and toxicokinetics 
of repeat weekly SC injection of ALT-B4 were evaluated for 5 weeks (Days 1, 8, 15, 22, and 29). 
During the study, there was no mortality and no ALT-B4-related change was observed in aspects 
of clinical sign, weight, food intake, ophthalmic examinations, functional observational battery, 
clinical pathology (hematology, serum chemistry, coagulation, urinalysis), gross (necropsy) 
evaluation, organ weight, and histopathology. 
Exposure to body circulatory system following weekly SC injection of ALT-B4 to male and female 
rats at 0.04, 0.2, and 2 mg/kg for 5 weeks was clear in 2 mg/kg group only. In both male and 
female rats, Tmax was 0.5 hours after injection, and time for recovery to below the limit of 
quantification (BLQ) of mean blood ALT-B4 concentration was 8 hours after injection. 2 mg/kg 
group did not show noticeable difference in ALT-B4 exposure to body circulatory system by sex 
and there was no difference in exposure between Day 1 and Day 22. 
In conclusion, weekly repeat dose SC injection of ALT-B4 at 0, 0.04, 0.2, and 2 mg/kg/dose for 5 
weeks (Days 1, 8, 15, 22 and 29) to Sprague dawley rats showed good tolerability and there was 
no mortality and adverse events related to ALT-B4 in aspects of clinical sign, weight, food intake, 
ophthalmology, functional observational battery, clinical pathology, and pathology. NOAEL was 
determined 2 mg/kg/dose. 
 
4. 
Study 20219-234 (ANC020) 
A repeat dose toxicity study of ALT-B4 in Cynomolgus monkeys (ALT-B4-19001, weekly/total 5 
SC injections) was performed (Table 3.3.5). 
Table 3.3.5 Experimental design of monkey repeat dose toxicity study 
Group 
No. 
Test 
Material 
Dose Level 
(mg/kg/dose) 
Dose 
Volume 
(mL/kg)a 
Dose 
Concentration 
(mg/mL) 
No. of Animalsb 
Main Study 
Recovery Study 
Males Females Males Females 
1 
Control 
0 
1.0 
0 
3 
3 
2 
2 
2 
ALT-B4 
0.04 
1.0 
0.04 
3 
3 
- 
- 
3 
0.2 
1.0 
0.2 
3 
3 
- 
- 
4 
2 
1.0 
2 
3 
3 
2 
2 
- = Not applicable. 
aBased on the most recent body weight measurement. 
bMain Study animals were euthanized on Day 30. Recovery animals were euthanized on Day 43. 
 
SC injection in Cynomolgus monkeys showed good tolerability at all doses. Mortality was not 
observed and there were no clinical signs, ophthalmic effect, abnormal electrocardiogram (ECG) 
or any change in body weight and food intake related to ALT-B4. 
Despite difference in ALT-B4 injection concentrations by multiple folds among groups, there was 
no detectable additive effect in all blood samples collected, confirming absence of ALT-B4-related 
change in hyaluronidase activity in plasma. 
Final euthanasia found microscopic findings including mixed cell inflammation (minimal-mild), 
hemorrhage, regionally extensive myocyte degeneration in the SC injection site at all doses of 

 
 
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ALT-B4, which were not observed in recovery group. 
In the conclusion based on meaningful study findings, weekly SC injection of ALT-B4 at 0, 0.04, 
0.2, and 2 mg/kg/dose for total 5 weeks (total 5 injections) was well tolerable and NOAEL was 
determined 2 mg/kg/dose. 
 
3.4. Rationale for Dose Selection 
Allergy reactivity is assumed caused by animal-derived proteins mostly included in animal-derived 
hyaluronidase. Therefore, skin allergy reaction test should be performed by using a small amount 
of hyaluronidase before administration of hyaluronidase to identify patients who may develop 
hypersensitivity to hyaluronidase.  
Traditionally, skin allergy reaction test for hyaluronidase has been performed by using 4-8 units 
of hyaluronidase16). For skin allergy reaction test, a high dose of hyaluronidase is diluted and 
appropriate dosing volume is used. Recent study results showed that low doses of 10, 20, and 
30 units have a chance of false negative, and allergy reactivity was found dose-dependent17). 
Therefore, use of a higher dose of hyaluronidase is recently recommened19), for example directly 
using 1,500 IU/mL dose of hyaluronidase without dilution in the skin allergy reaction test18). 
A representative skin allergy reaction test for hyaluronidase is ID test20), 21), and following 
precautions are required. 
 
Bubbles must be removed carefully before injection to make sure that there will be no 
splash reaction. 
 
Caution should be taken not to burst capillaries under the epidermis while inserting the 
needle. 
 
A syringe should be inserted at 45 degrees. The sloping side of the needle should face 
the skin and the entire sloping side should be fully inserted into the skin before 
administering the reagent. 
 
0.01-0.05 mL will be injected to make a small blister with a diameter of 2-3 mm. 
 
Do not inject 0.05 mL or more. 
 
If injected subcutaneously, false negative develops. 
For an allergy test through ID test, it is mandatory to form appropriate size of blister by injecting 
appropriate volume of the study drug before observation of the reactions. Therefore, generally no 
more than 0.05 mL is injected and in other hyaluronidase clinical trials, this was reflected and 
0.02-0.05 mL of the study drug was injected for testing.  
Dosing volumes of study drugs in other hyaluronidase clinical trials are provided below:  
Study drug 
Dosing volume 
Amphdase 
0.02 mL 
Vitrase 
0.02 mL 
Hydase 
0.02 mL 
Hylenex 
0.10 mL 
 
As per the provided grounds, 
 
1,500 IU/mL of concentration is used to minimize a chance of false negative result, and 
 
0.02 mL is injected to form an appropriate blister for the ID test. 
This means that 30 IU/20 L of ALT-BB4 will be used as an ID injection dose for allergy test. 
 
For hyaluronidase products approved in Korea, dosage regimen of SC and intramuscular (IM) 
injections are provided below. Based on the clinical literatures reported in Korea,22),23) general 
clinical dose of hyaluronidase would be 1,500 IU or less. In addition, most animal-derived 
hyaluronidase products available in Korean market are lyophilized products with potency of 1,500 

 
 
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IU. These products are used after dissolving in 1 mL water for injection. Therefore, the safety of 
ALT-BB4 at 1,500 IU contained in a 1mL vial will be secured through a clinical study. 
<Dosage regimen of animal-derived hyaluronidase products approved in Korea> 
- 
SC infusion (mass SC injection) 
1,500 IU as hyaluronidase will be dissolved in 1 mL of water for injection or normal 
saline injection and administered into applicable site before starting SC infusion, or 
administered into a tube about 2 cm above the needle for infusion when starting the 
infusion. At administration of 500-1,000 mL of fluid, approximately 1,500 IU of this 
drug is appropriate. 
- 
SC injection or IM injection 
1,500 IU of this drug will be directly dissolved in the injection solution to be 
administered. 
- 
Extravasation 
In the event of extravasation, especially when there is diffusion, compared to 
regional case, 1,500 IU of this drug will be dissolved in 1 mL of water for injection 
or normal saline injection and infiltrated into the lesion area as soon as possible. 
- 
Hematoma 
1,500 IU of this drug will be dissolved in 1 mL of water for injection or normal saline 
injection and infiltrated into the applicable site. This drug and about 1 mL of water 
for injection will be dissolved in solution for injection immediately before use. 
Therefore, 1,500 IU was decided as a clinical dose. 
To check for toxicity of ALT-B4, it was injected IV in rats, and IV and SC in monkeys. In 
Cynomolgus monkeys, 0.3-30 mg/kg was injected after 4-5 times of washout period. Through PK 
studies, very low bioavailability and very rapid exposure to body circulatory system of ALT-B4 
were confirmed. In Minipigs, a dose-finding study was performed following single SC injection and 
no adverse event was found at the highest dose (2 mg/kg). In addition, single dose and repeat 
dose toxicity studies in rats were performed. In the single dose toxicity study, ALT-B4 was injected 
IV or SC at 0.04, 0.2, and 2 mg/kg, while it was injected SC in the repeat dose toxicity study. At 
the same dose, a repeat dose toxicity study was performed in Cynomolgus monkeys. In both 
animal species, no toxicity was found at all doses, and NOAEL was determined 2 mg/kg. 
As ALT-B4 was inactive at approx. 70,000 IU/mg, the highest dose in the non-clinical trials, 2 
mg/kg/dosage corresponds to approx. 140,000 IU/kg/dosage, which is 9,800,000 IU/dosage for a 
70-kg adult. Considering that the actual treatment dose is ≤1,500 IU (single fixed dose), the dose 
in the non-clinical trials is much higher than the expected clinical dose. 
 

 
 
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Protocol No.: ALT-BB4-01 
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4. Study Objectives 
4.1. Primary Objective 
To evaluate the allergic reactivity, safety and tolerability of ALT-BB4 (recombinant hyaluronidase) 
by checking for drug allergy following single ID injection of ALT-BB4 and monitoring systemic 
reactions and adverse events following its single SC injection  
 
4.2. Secondary Objectives 
To evaluate the PK profile of single dose of ALT-BB4 
 
4.3. Exploratory Objectives 
To evaluate the immunogenicity of ALT-BB4 (Visit 1 and End of Study visit) 
 

 
 
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Protocol No.: ALT-BB4-01 
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5. Study Population (Sample Size and Subject Selection) 
5.1. Sample Size 
1. 
Total number of subjects: At least 231 subjects (Maximum of 290 including subjects with 
allergic reactions, dropped out and excluded from analysis)  
2. 
Number of subjects for each Part 
 
Part I (Allergy assessment): At least 231 subjects 
 
Part II-A (PK assessment): Total 23 subjects (including 10% drop-out rate) 
 
Part II-B (Safety assessment): At least 208 subjects (Study group- at least 139 
subjects, Control group - at least 69 subjects; randomized in a 2:1 ratio) 
 
 

 
 
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5.2. Rationale for Sample Size Determination 
The minimum target number of subjects 231 was based on the empirical cases of previous clinical 
trials and statistical hypothesis. Based on the existing similar clinical trials of allergy 
hypersensitivity with clinically significant effect size of 10% and existing literature, the number of 
subjects was determined for the sample size. 
The number of subjects who participated in a previous similar clinical trial with an animal-
derived/human gene recombinant hyaluronidase was 100 for Hylenex, 162 for Amphadase and 
65 for Vitrase. In the study to check for allergenicity of hyaluronidase products approved by the 
FDA, incidence of positive skin reaction was 0% (0/100 subjects) with a recombinant 
hyaluronidase, called Hylenex, and 5% (8/162 subjects) with an animal-derived hyaluronidase, 
called Amphadase. In the recent Study NCT01689363, "Evaluation of the Allergenicity of 
AMPHADASE INJECTION (Hyaluronidase Injection USP) (H001-A2)", it was 0% (0/183 subjects) 
in PP group, 2.7% (7/253 subjects) in ITT group, and 0% (0/65 subjects) in Vitrase group. 
This study aims to evaluate the safety, tolerability and PK following administration of ALT-BB4 
(recombinant hyaluronidase) in healthy volunteers. The study consists of Part I for allergy 
assessment, Part II-A for PK assessment, and Part II-B for safety assessment. 
 
 
Part I 
Allergy incidence rate (%) is defined as proportion of subjects with positive allergy test result, 
which will be used to determine the target number of subjects. Statistical hypothesis is provided 
below: 
 
H0: pT ≥p0               vs.             HA: pT < p0  
 
pT: Allergy incidence rate in the study group (ALT-BB4) 
p0 : Allergy incidence rate recommended by the FDA 
 
Main purpose of Part I is to identify the allergy incidence rate in the study group (ALT-BB4) and 
to determine whether it is appropriate. Therefore, it is deemed inappropriate to determine the 
sample size under hypothesis where allergy incidence rates between the placebo-control group 
(saline) and the study group (ALT-BB4) would be statistically compared. Thus, a hypothesis to 
test the incidence rate of the study group by comparing with the target incidence rate was used 
to determine the target number of subjects. 
 
As suggested in the FDA medical review for Amphadase, the FDA-recommended allergy 
incidence rate is 10%. This was used as the test threshold (p0 =0.1) in the statistical hypothesis. 
In addition, allergy incidence rate in Study API-H001-CLN-A of Amphadase was 5% (7/142 
subjects). As the study group using a similar agent (ALT-BB4) is expected to show the same level 
of allergy incidence rate, pT was assumed 0.05. 
 
The target number of subjects using “Numeric Results for Testing One Proportion using the Exact 
Test” of PASS 15 with one-sided significance level of 2.5% and 80% power is 231 subjects. 
However, when the target number of subjects (231) required for analysis has not been met due 
to severe protocol deviation, additional subjects can be recruited to maintain statistical power, 
and up to 290 subjects may be enrolled. 
 
 
Part II-A 
Among the 231 subjects who have completed Part I, 23 subjects, which accounts for 10%, will be 
enrolled in Part II-A. Assuming 10% of drop-out rate, we can expect that at least 20 subjects will 

 
 
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Protocol No.: ALT-BB4-01 
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finally complete Part II-A. 
 
 
Part II-B 
Among 231 subjects who will complete Part I, 208 subjects will be enrolled in Part II-B, except 
23 subjects who will be enrolled in Part II-A. The sample size is considered sufficient to assess 
the safety in the Phase 1 study. 
 

 
 
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Protocol No.: ALT-BB4-01 
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5.3. Subject Selection 
5.3.1. Inclusion Criteria 
 
Common 
1. 
Healthy volunteers aged ≥19 years at the time of Screening (Visit 1) 
2. 
Female subjects or male subjects’ female partner must be menopausal or should have 
received a sterilization procedure or have agreed to use contraceptive methods during the 
study period, as defined below: 
 
Post-menopausal female subjects or male subjects’ female partners (non-drug induced 
amenorrhea for at least 12 months or confirmed diagnosis with menopause) 
 
Female subjects or male subjects’ female partners who have received a sterilization 
procedure (removal of ovary and/or uterus) 
 
Subjects who have agreed to practice total abstinence during the study period [For 
female subjects, periodic abstinence (e.g., ovulation, symptothermal, or post-ovulation 
methods) and withdrawal are not acceptable methods of contraception.] 
 
When female subjects or male subjects’ female partners are women of childbearing 
potential (WOCBP) who have not received a sterilization procedure, they must agree to 
use of following contraceptive methods: 
- 
Hormones (implantable, patch, and oral) 
- 
Intrauterine device (IUD) 
- 
Double barrier methods (concomitant use of two of following contraceptive methods: 
male condom, female condom, cervical cap, diaphragm, sponge, spermicide) 
(However, concomitant use of male condom and female condom is excluded from 
double barrier methods.) 
3. 
WOCBP or females who have the last menstrual period within 12 months must have a 
negative serum or urine pregnancy test at Screening (Visit 1). 
4. 
Subjects who have voluntarily decided to take part in the study and able to comply with the 
study protocol 
5. 
Subjects who have no tattoo, acne, dermatitis, pigmentation or lesion on the administration 
site and who have no damage in the skin, so that they can receive the IP and allergy test 
6. 
Subjects determined eligible for the study through Screening tests (vital signs, physical 
examination, medical history and surgery history, ECG, and laboratory tests) 
 
Part II-A 
7. 
Subjects with BMI no less than 18.5 kg/m2, no greater than 24.9 kg/m2 
 
5.3.2. Exclusion Criteria 
Below are the study exclusion criteria: 
 
Common 
1. 
Subjects who have received or treated with following medications within the specified 
timeframe prior to Baseline (Visit 2) or who are expected to receive them during the study 
period: 
 
Within 1 month: Hyaluronidase, Chemotherapeutic agent, penicillins antibiotics (e.g.: 
Amoxicillin, Ampicillin, etc.), cephalosporins antibiotics (e.g.: Cefaclor, Cefadroxil, 
Cefixime, etc.), sulfonamides antibiotics (e.g.: Sulfadiazine, Sulfamethoxazole, etc.), 
quinolones antibiotics (e.g.: Ciprofloxacin, Levofloxacin, etc.), Glucocorticosteroid, 
Immunosuppressive agent 
 
Within 14 days: Antihistamine (e.g.: Chlorpheniramine, Hydroxyzine, Ketotifen, etc.), 
non-steroidal anti-inflammatory drugs (NSAIDs; e.g.: Aspirin, Aceclofenac, etc.)24-6) 

 
 
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Protocol No.: ALT-BB4-01 
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2. 
Subjects who have current or prior history of clinically significant liver, kidney, gastrointestinal, 
cardiovascular, respiratory, endocrine, immune system, psychiatric/nerve system, and 
blood/oncological disorders 
3. 
Subjects with acute fever > 37.5℃ within 7 days from the expected IP administration or 
showing symptoms suspecting acute diseases within 14 days from the expected IP 
administration 
4. 
Subjects who are diagnosed with hypertension/hypotension or subjects who are with 
clinically significant blood pressure* 
*Hypertension=systolic blood pressure ≥ 140 mmHg and/or diastolic blood pressure ≥ 90 mmHg27) 
*Hypotension=systolic blood pressure ≤ 90 mmHg and/or diastolic blood pressure ≤ 60 mmHg28)-30) 
5. 
Subjects who persistently drink more than the weekly recommended alcohol units* 
14 g of alcohol content per unit is applied, which corresponds to 1 can (small bottle) of beer (5%), 350 
mL of draft beer (5%), 1 cup (300 mL) of makgeoli, 1 glass (150 mL) of wine (12%), 1/4 bottle (90 mL) 
of soju (20%), and 1 shot (45 mL) of liquor (40%). Moderate amount of alcohol by age and sex is 
provided below:31) 
Age 
Recommended unit per week 
Adults (≥ 19 and < 65 years) 
Male: 8 units/week 
Female: 4 units/week 
Elderly (≥ 65 years) 
Male: 4 units/week 
Female: 2 units/week 
 
6. 
Subjects of usual smoker (exceeding 10 cigarettes per day) 
7. 
Subjects who have a past history of autoimmune diseases (e.g.: rheumatoid arthritis, etc.) or 
active immune diseases that may affect the immune system [e.g.: flu, cancer, Human 
immunodeficiency virus (HIV), Hepatitis B virus (HBV), Hepatitis C virus (HCV) etc.], diabetes 
mellitus, heart diseases, asthma, sinusitis, chronic urticaria, dermographism, or any skin 
conditions that may affect post-IP administration assessments (e.g.: Dermatitis, 
dermatomycosis and other skin diseases or tattoo) 
8. 
Subjects with known allergic reactions or hypersensitivity to bee sting or other common 
allergens or known contraindications to hyaluronidase, thimerosal (disinfectants) and EDTA  
9. 
Subjects with hypersensitivity to the IP or its ingredients or a history of anaphylaxis 
10. Past history of drug abuse 
11. Subjects who have participated in other clinical trials within 6 months prior to the expected 
IP administration 
12. Others determined ineligible for the study participation in the opinion of investigator 
 
Part II-A 
13. Subjects who have been treated with a drug-metabolizing enzyme inducers and inhibitors*32), 
33) within 30 days prior to Visit 4 
* Example: Phenytoin, Carbamazepine, Barbiturates, Rifampicin, Griseofulvin, Cimetidine, Disulfiram, 
Erythromycin, Ketoconazole, Fluconazole, Itraconazole, Valproic acid, Isoniazid, Ciprofloxacin, 
Omeprazole, Clarithromycin, Quinidine, Sulfonamides, etc. 
14. Subjects who have significant bleeding or blood loss within 60 days prior to Visit 4 
15. Subjects who have donated whole blood within 60 days or donated blood by apheresis within 
14 days or received transfusion within 14 days prior to Visit 4 
16. Subjects who have consumed grapefruit juice34) within 7 days prior to Visit 4 
 
5.3.3. Withdrawal Criteria 
Subjects can withdraw their consent without providing any reason at any time during the study. 
In addition, subjects who are considered unable to continue the study due to their clinical 
conditions will be discontinued from IP administration by investigator. Subjects will discontinue 
the study or be withdrawn/dropped out from the study by investigator for following reasons: 

 
 
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Protocol No.: ALT-BB4-01 
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1. 
Voluntary consent withdrawal of subject (or subject’s legally acceptable representative) 
2. 
Subjects who cannot perform the study due to adverse events in the investigator’s opinion 
3. 
Subjects who cannot receive SC injection due to drug allergy following ID injection in the 
investigator’s opinion  
4. 
Major protocol deviation/violation (inclusion/exclusion criteria violation, use of forbidden 
concomitant medications, etc.) 
5. 
Subjects who have received a drug that may affect the IP assessments (allergy assessment, 
safety and tolerability assessment, and PK assessment) in the investigator’s opinion 
6. 
Subjects lost to follow-up (except those considered not affecting the PK and safety 
assessments) 
7. 
Females with confirmed pregnancy during the study period 
8. 
In the opinion of the investigator, a subject should discontinue the study participation 
 
5.3.4. Protocol Deviation/Violation 
Principal investigator and subinvestigator of the study must be fully aware of the protocol and 
strictly follow it to prevent any protocol deviation/violation. Principal investigator (or its designee) 
will take appropriate measures to ensure subjects can return the institution for their scheduled 
visits for IP administration and tests, for example providing written notice or phone call as a 
reminder of the next visit. In addition, investigator must inform the sponsor of any protocol 
deviation/violation occurring in a subject as soon as possible and determine whether the subject 
can continue the study participation. Drop-out due to protocol deviation/violation must be 
documented in the electronic case report form (eCRF). Following cases are considered as major 
protocol deviation/violations: 
 
1. 
Violations against the inclusion or exclusion criteria 
2. 
Use of forbidden concomitant medications that may affect the PK, safety and tolerability 
assessments of the IP 
3. 
Other cases considered as major protocol deviation/violations 
 
For minor deviation/violations that are considered not affecting interpretation of the study results, 
the level of deviation/violation or delay and reasons must be accurately documented in the eCRF 
and it will be comprehensively discussed in a blind meeting whether it has affected the study. 
 

 
 
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6. Study Design 
6.1. Study Period 
1. 
Entire Study Period 
Approximately 18 months from approval of the Institutional Review Board (IRB) (however, it 
may be changed depending on the subject enrollment rate.) 
 
2. 
Study Period for Individual Subjects 
 
Part I (Allergy assessment):  
- 
Screening period: Up to 2 weeks 
- 
Treatment with IP: 1 day (ID single dose) 
- 
Follow-up period: 2 days (48 hours) 
 
Part II-A (PK assessment): 
- 
Treatment with IP: 1 day (SC single dose) 
- 
PK Assessment: Total 15 times 
◼ 
Pre-dose: -2h, -1h, and 0h  
◼ 
Post-dose: 10 min (0.17h), 20 min (0.3h), 30 min (0.5h), 40 min (0.7h), 50 min 
(0.8h), 1h, 1h 15 min (1.25h), 1h 30 min (1.5h), 2h, 2 h 30 min (2.5h), 3h and 
24h 
- 
Follow-up period: 4 weeks 
 
 
Part II-B (Safety assessment) 
- 
Treatment with IP: 1 day (SC single dose) 
- 
Follow-up period: 4 weeks 
 
6.2. Study Group/Control Group 
1. 
Part I (Allergy assessment)  
Treatment group 
Study group 
Control group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
Comparator (0.9% NaCl) 
ID dose 
30 IU (0.02 mL) 
0.9% NaCl(0.02 mL) 
Administration site* 
Right or left forearm 
Forearm in the opposite side of the 
study drug administration 
* For example, when the study drug is intradermally injected into the left forearm, the comparator will be 
intradermally injected into the right forearm. 
 
2. 
Part II-A (PK assessment) 
Treatment group 
Study group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
SC Dose 
1,500 IU (1 mL) 
Administration site 
Right or left upper arm 
 
3. 
Part II-B (Safety assessment) 
Treatment group 
Study group 
Control group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
Comparator (0.9% NaCl) 
SC Dose 
1,500 IU (1 mL) 
1 mL 
Administration site 
Right or left upper arm 
Right or left upper arm 

 
 
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Protocol No.: ALT-BB4-01 
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6.3. Randomization 
1. 
Part I (Allergy assessment) and Part II-B (Safety assessment) 
Randomization will be used to ensure the scientific validity of the study and to avoid subjectivity 
of the investigator in the subject assignment to each treatment group. Biostatistician independent 
from the study will send the randomization list to an IP packaging staff and prepare and send a 
blinded envelop to the principal investigator of the institution. The list will be kept blinded. 
 
1. 
Allocation number will be generated by using SAS®  9.4 or higher Proc plan procedure. 
2. 
Subjects enrolled in Part I (allergy assessment) will be randomized to receive the study drug 
or comparator into the both arms in a ratio of 1:1 based on their allocation number. Allocation 
number will be assigned in a form of R00-000. For example, the allocation number of the first 
subject enrolled in Institution 01 will be R01-001. 
Part I allocation number: R@@-### (@@: Institution No., ###: Serial number, Example: 
R01-001) 
 
3. 
Subjects enrolled in Part II-B (Safety assessment) will be randomized into the study group 
and control group in a ratio of 2:1. Allocation number will be assigned in a form of B00-000. 
For example, the allocation number of the first subject assigned in Part II-B in Institution 01 
will be B01-001. 
 
Part II-B allocation number: B@@-### (@@: Institution No., ###: Serial number, Example: 
B01-001) 
 
The investigator will review the subject inclusion/exclusion criteria and assign a unique number 
to subjects determined eligible for the study; and will use this number to prescribe the IP. 
Until the end of the study, investigators, subjects, clinical pharmacists and people conducting the 
study should be kept blinded and when unblinding is required during the study, it will be performed 
based on the procedure as specified in Section 6.4.2 of the protocol. 
 
2. 
Part II-A (PK assessment) 
Part II-A of the study has a single arm and an open-labeled design, and randomization will not be 
separately performed. However, subject number (allocation number) will be sequentially assigned 
to subjects in Part II-A. For example, the subject number of the first subject assigned in Part II-A 
in Institution 01 will be A01-001. 
 
Part II-A allocation number: A@@-### (@@: Institution No., ###: Serial number, Example: 
A01-001) 
 
6.4. Blinding 
Part I and Part II-B will be performed based on a double-blinded design. In this study, double-
blinding will be performed in accordance with a separate blinding procedure to ensure that 
subjects in Part I and Part II-B cannot know whether they are receiving the study drug or 
comparator in the administration site. For Part II-A performed with a single arm, a separate 
blinding process will not be performed. 
 
6.4.1. Double-blinding 

 
 
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This is a Phase 1 study. To exclude subjective judgment of investigator and subjects, Part I and 
Part II-B will be performed based on a double-blinded design. This means that both investigator 
and subject will not know which IP is administered (a person who conducts administration will be 
unblinded). 
 
In Part I allergy assessment, subjects will be blinded to the IP administered and administration 
site, while unblinded staff administers the IP as randomized. 
In Part II-B safety assessment, subjects will be blinded to the type of IP administered, while 
unblinded staff administers the IP as randomized. 
 
To maintain double-blinding in Part I and Part II-B, unblinded administration staff will prepare the 
IP in an isolated place from subjects. At that time, subjects cannot see the IP preparation process 
in a syringe by administration staff, and will be asked to turn their head during administration not 
to see the administration process and administration site. Maintaining double-blinding for 
investigator and subjects will prevent bias in the safety and tolerability assessment by the 
investigator or subjects. To maintain double-blinding, only allocation number will be used to 
identify subjects and the list of allocation number assignment by group will not be disclosed until 
the end of the study. 
 
6.4.2. Unblinding 
When unblinding is deemed necessary due to reasons such as emergency situation threatening 
the subject safety, investigator must inform Alteogen, Inc. or its designated CRO’s monitor 
promptly. Monitor has responsibility for immediate contact to a representative of Alteogen, Inc. 
After receiving the information, the representative of Alteogen, Inc. will discuss with the 
investigator to determine whether to unblind and document the decision. After conferring with the 
sponsor, the investigator shall confirm the IP information of the relevant subject by opening the 
code breaking card box delivered at the kick-off meeting and sign it before keeping it in the 
investigator study file. When the sponsor, Alteogen, Inc. is not contactable immediately, 
investigator will conduct the unblinding process with the same procedure and must report the 
blind-breaking to Alteogen at the earliest time available and document the reason of conducting 
unblinding without discussion with the sponsor. When the investigator gets to know the subject’s 
randomized IP during the study, the investigator must take effort to exclude bias in the safety and 
tolerability assessments. 
After the end of the study, problems of all data will be solved out through query. Once integrity 
and accuracy of the database are confirmed, database is locked (DB lock) and the allocation 
number will be disclosed. Subsequent database change is only allowed with written consent of 
the sponsor and database manager.  
 
 
 
 
 

 
 
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7. Criteria for Study Termination and Early Termination 
7.1. Scheduled Termination 
Termination of entire study is defined as the study discontinuation of the last subject or 
discontinuation visit of the last subject. 
 
7.2. Early Termination 
Investigator may terminate the partial or entire study after discussion with the sponsor when 
continuation of the study is considered inappropriate based on the study findings.  
The sponsor can terminate the entire study or the study at a particular institution early. Reasons 
of study termination by the sponsor include: 
 
1. 
The institution fails in recruiting the target number of subjects 
2. 
Any information that may significantly affect the study continuation becomes available 
3. 
Deviation/violations of Good Clinical Practice (GCP), protocol or contract by the institution or 
investigator affecting continuation of the study 
4. 
Other administrative reasons that may significantly affect the study continuation 
 
In case of early termination or temporary termination of the study, investigator must inform the 
subjects immediately, make arrangement for appropriate actions and follow-up and report current 
study status and outcomes regarding subjects who had participated in the trial until the time of 
termination to the sponsor. In case of study termination, this must be reported to the IRB and the 
Ministry of Food and Drug Safety (MFDS). The entire study schedule may be suspended 
depending on the IRB or MFDS decision. 
 

 
 
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Protocol No.: ALT-BB4-01 
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8. Investigational Product Information and Management 
8.1. Investigational Product (IP) 
8.1.1. Study Drug 
1. 
Product name or Code name: ALT-BB4 (Recombinant Hyaluronidase) 
2. 
Active ingredient and Content: ALT-BB4 (recombinant hyaluronidase) 1,500 IU/mL 
3. 
Appearance and Formulation: Colorless, clear liquid injection 
4. 
Manufacturer: Alteogen, Inc. 
 
8.1.2. Comparator 
1. 
Product name or Generic name: Normal saline (0.9% NaCl)  
2. 
Active ingredient and Content: 0.9% NaCl 
3. 
Appearance and Formulation: Colorless, clear liquid injection 
4. 
Manufacturer: JW Pharmaceutical Corp.  
 
8.2. Production/Packaging and Labeling 
The sponsor will produce or purchase and then pack the IPs and provide them to the clinical 
pharmacist in an institution. IP labeling should be performed in accordance with Article 8.4 of 
[Attached Table 4-2] Good Manufacturing Practice for IPs of the Regulations on Safety of Drugs, 
Etc. and Article 7.7 of [Attached Table 11] IP Manufacturing of the Regulations on the Drug 
Manufacturing and Quality Control and should include the following:  
1. 
Statement that it can be used for clinical trial only (e.g. “For clinical trial use only”) 
2. 
Name or identification of the IP 
3. 
Batch number or code number that can identify the contents and packaging process 
4. 
Name, address and phone number of the sponsor (IND holder) 
5. 
Shelf-life (expiry date) 
6. 
Storage conditions 
7. 
Reference code that can identify the clinical trial (Protocol No.) 
8. 
Subject identification No., IP No., and Visit No. (can be omitted when documented) 
 
8.3. Investigational Product Management 
All IPs will be supplied to the institutions by the sponsor. IPs will be stored in a locked place with 
restricted access at refrigerated temperature (2-8℃). 
After receipt of IPs, the clinical pharmacist at an institution will check for the quantity and 
conditions of them and ensure that they cannot be used without prescription of the principal 
investigator or designated study staff, and store and manage them so that they can be used for 
the study purpose only. 
The clinical pharmacist must record accurate details, including quantity of IPs received from the 
sponsor and dispensed to subjects together with dispense date, in a form provided by the sponsor, 
while the monitor must examine the IP accountability by periodically monitoring the quantity kept 
by the investigator or clinical pharmacist and the storage condition. 
The principal investigator or clinical pharmacist at each institution must keep unused IPs until 
monitored by the sponsor’s monitor. At the end of the study or upon the sponsor’s request, unused 
IPs will be collected or discarded by the sponsor. (Used empty containers will be discarded at the 
institution in accordance with the institution’s SOP, if feasible.) 
The sponsor will check for the IP quantity and storage condition during the study period and make 
necessary measures to ensure smooth conduct of the study. 

 
 
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Protocol No.: ALT-BB4-01 
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9. Methods and Administration Plan 
9.1. Overall Methods 
This is a Phase 1 study to perform allergy assessment and PK profiling as well as to evaluate the 
safety and tolerability following administration of ALT-BB4 (recombinant hyaluronidase) in healthy 
volunteers. This study consists of Part I (Allergy assessment), Part II-A (PK assessment) and Part 
II-B (Safety assessment). In Part I, ALT-BB4 allergy assessment will be performed first. Then only 
in eligible subjects, PK of ALT-BB4 will be assessed in Part II-A, and the safety and tolerability 
assessment will be performed in Part II-B. 
 
Part I (Allergy assessment) 
For subjects who have agreed to participate by giving a written informed consent, Screening tests 
will be performed. By confirming the Screening test results and the inclusion/exclusion criteria, 
eligible subjects for participating in Part I of the study for the drug allergy assessment will be 
enrolled. Enrolled subjects will visit the institution and receive the IP intradermally. For up to 30 
minutes after the IP administration, subjects will be observed for cutaneous hypersensitivity of 
immediate allergic reaction and subjects without any adverse event will be discharged. Later at 
Visit 3, subjects will be contacted via phone to monitor concomitant medications and adverse 
events. At Visit 4, subjects will visit the institution for monitoring of cutaneous hypersensitivity of 
delayed allergic reaction occurring within 48 hours after the ID injection, blood collection, change 
of concomitant medications and adverse events, performing scheduled tests. ALT-BB4 allergy 
assessment will be conducted through scheduled tests and evaluations for each visit. 
 
Part II-A (PK assessment) 
Eligible subjects for Part II-A of the study will be enrolled only from subjects with drug allergy test 
negative in Part I. Enrolled subjects will receive the IP subcutaneously at the institution and 
perform scheduled tests and blood collection at designated time points. At 3 hours after the IP 
administration, scheduled tests will be completed and subjects without any adverse event will be 
discharged. In a case of adverse events, the subjects will be treated by investigator as required 
before discharge. Later at Visit 5, subjects will visit the institution for blood collection, review of 
any change in concomitant medications and adverse events, and scheduled tests, and at Visit 6 
(End of Study visit), subjects will visit the institution for review of any change in concomitant 
medications and adverse events and scheduled tests. ALT-BB4 PK assessment will be conducted 
through scheduled tests and blood collection for PK assessment for each visit. 
 
Part II-B (Safety assessment) 
Eligible subjects for Part II-B of the study will be enrolled only from subjects with drug allergy test 
negative in Part I. Subjects enrolled in Part II-B study will be randomized into the study group or 
control group at the institution and receive single dose of the IP subcutaneously into the right or 
left upper arm. For up to 30 minutes after the IP administration, subjects will be discharged if there 
is no adverse event when monitoring systemic and administration site adverse events. In a case 
of adverse events, the subjects will be treated by investigator as required before discharge. Later 
at Visit 5, subjects will visit the institution for blood collection, review of any change in concomitant 
medications and adverse events, and scheduled tests. Visit 6 is a phone visit, where concomitant 
medications and adverse events including systemic and administration site events will be 
monitored. At Visit 7, the End of Study visit, scheduled tests and safety assessment will be 
performed to evaluate the safety and tolerability of ALT-BB4. 
 
9.2. Investigational Product Dosage and Administration Method 
1. 
Part I (Allergy assessment) 
Subjects will be randomized into the study group or control group at Baseline visit (Visit 2), and 
receive single dose of the study drug or comparator intradermally into the right or left forearm.  

 
 
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Protocol No.: ALT-BB4-01 
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Treatment group 
Study group 
Control group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
Comparator (0.9% NaCl) 
ID dose 
30 IU (0.02 mL) 
0.9% NaCl (0.02 mL) 
Administration site* 
Right or left forearm 
Forearm in the opposite side of the 
study drug administration 
* For example, when the study drug is intradermally injected into the left forearm, the comparator will be 
intradermally injected into the right forearm. 
 
2. 
Part II-A (PK assessment) 
Part II-A PK assessment will be performed only in subjects with drug allergy test negative at Visit 
4, 2 days (48 hours) after the ID injection in Part I. All subjects participating in Part II-A will receive 
single dose of the study drug subcutaneously into the right or left upper arm.  
Treatment group 
Study group 
IP 
Study drug (ALT-BB4, 1,500 IU/mL) 
SC Dose 
1,500 IU (1 mL) 
Administration site 
Right or left upper arm 
 
3. 
Part II-B (Safety assessment) 
Part II-B safety assessment will be performed only in subjects with drug allergy test negative at 
Visit 4, 2 days (48 hours) after the ID injection in Part I, randomized into the study group and 
control group in a 2:1 ratio. Subjects assigned in each group of Part II-B will receive single dose 
of the study drug or comparator subcutaneously into the right or left upper arm. 
Treatment group 
Study group 
Control group 
IP 
Study drug (ALT-BB4, 1,500 
IU/mL) 
Comparator (0.9% NaCl) 
SC Dose 
1,500 IU (1 mL) 
1 mL 
Administration site 
Right or left upper arm 
Right or left upper arm 
 
9.3. Concomitant Medications and Therapies 
9.3.1. Permitted Concomitant Medications and Therapies 
In following cases, medications and therapies are allowed throughout the study period. 
1. 
Non-forbidden concomitant medications and therapies can be administered during the study 
period, but only the concomitant medications and therapies considered not affecting the 
study result interpretation are allowed at the discretion of investigator. 
2. 
Other medications and therapies for routine treatment of other diseases will be used 
concomitantly following discussion with the investigator. 
Details (product name, purpose of administration, dosage, and treatment duration, etc.) of all 
concomitant medications and therapies (including treatment for other diseases or adverse events) 
will be documented in the electronic case report form (eCRF). 
 
9.3.2. Forbidden Concomitant Medications and Therapies 
Following medications are forbidden during the study period: 
 
Hyaluronidase, Chemotherapeutic agent, Glucocorticosteroid, immunosuppressive agent, 
Antihistamine, NSAIDs (e.g.: Aspirin, Aceclofenac, etc.), penicillins antibiotics (e.g.: 
Amoxicillin, Ampicillin, etc.), cephalosporins antibiotics (e.g.: Cefaclor, Cefadroxil, Cefixime, 
etc.), sulfonamides antibiotics (e.g.: Sulfadiazine, Sulfamethoxazole, etc.), quinolones 
antibiotics (e.g.: Ciprofloxacin, Levofloxacin, etc.), ethical-the-counter drugs*, herbal 

 
 
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Protocol No.: ALT-BB4-01 
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medicines*, over-the-counter drug (OTC drug)*, health functional food*, Vitamins*, grapefruit 
juice**, drug-metabolizing enzyme-inducers and inhibitors**32),33) (e.g.: Phenytoin, 
Carbamazepine, Barbiturates, Rifampicin, Griseofulvin, Cimetidine, Disulfiram, Erythromycin, 
Ketoconazole, 
Fluconazole, 
Itraconazole, 
Valproic 
acid, 
Isoniazid, 
Ciprofloxacin, 
Omeprazole, 
Clarithromycin, 
Quinidine, 
Sulfonamides, 
etc.) 
* Medications started before Screening visit and continuously used are allowed, while new medications 
after Screening are not allowed. 
**For Part II-A only 
 
In addition, whole blood donation, apheresis, and transfusion are prohibited for subjects in Part II-A. 
 
9.4. Treatment Compliance 
Single dose of IP will be administered for Part I (Allergy assessment), Part II-A (PK assessment) 
and Part II-B (Safety assessment). IP administration will be performed under supervision of the 
principal investigator (or its designee) at the institution and the time of administration to a subject 
will be documented in the eCRF. 

 
 
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10. Study Procedures and Assessment 
10.1. Observations 
10.1.1. Written Informed Consent Form and Assignment of Screening 
Number 
Investigator must provide detailed explanation about the study purpose and what involved to 
subjects who want to take part in the study. Written informed consent must be obtained prior to 
any study procedures and a copy of signed informed consent form (ICF) will be provided to a 
subject by the investigator. 
After obtaining written informed consent from a subject, the investigator will assign a screening 
number in the order of obtaining ICF. Screening number will be assigned in a form of S00-000. 
For example, the screening number of the first subject who provides the written informed consent 
in Institution 01 will be S01-001.  
 
Screening number: S@@-### (@@: Institution No., ###: Serial number, Example: S01-001) 
 
For a subject who failed screening, the subject’s screening number will not be re-assigned to 
other subjects.  
 
10.1.2. Demographic Data 
Information about subject’s sex, date of birth, skin conditions (presence of tattoo, acne, dermatitis, 
pigmentation, lesion and skin damage on the administration site), smoking (non-smoker, former 
smoker or current smoker), alcohol intake (non-drinker, former drinker or current drinker), weekly 
alcohol intake, and daily smoking amount will be investigated. In addition, only for subjects 
considering participating in Part II-A, information about history of whole blood donation within 60 
days, apheresis within 14 days, and transfusion within 14 days prior to Visit 4 will be investigated. 
 
10.1.3. Investigation of Medication History and Medical History 
For medication history, information of medications will be recorded (product name or ingredient 
name, purpose of administration, dosage, and treatment duration, etc.). 
 
For medical history, presence of past medical history and current medical conditions, time of drug 
allergy onset (dd/mmm/yyyy), ongoing status and investigator’s opinion will be documented.  
 
10.1.4. Vital Sign 
Vital signs will be measured before other scheduled tests. After taking a rest for at least 5 minutes, 
sitting systolic and diastolic blood pressures, respiratory rate, pulse rate and temperature will be 
measured. 
 
10.1.5. Electrocardiogram (ECG) 
12-lead ECG will be performed after taking a rest for at least 3 minutes, and ventricular rate 
(beats/min), PR (msec), QRS (msec), QT (msec), and QTcF (msec) will be confirmed. 
 
10.1.6. Physical Examination 
Physical examination includes height (cm, whole number), weight (kg, one decimal place), and 
body mass index (BMI) (kg/m2, round to one decimal place) measurements and comprehensive 
evaluation 
of 
allergy, 
cardiovascular 
system, 
respiratory 
system, 
gastrointestinal 
system/hepatobiliary system, metabolism/endocrine system, renal/urinary system, reproductive 
system, musculoskeletal system, skin and connective tissues, nervous system, psychiatric 

 
 
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system and other body organs. 
Clinically meaningful symptoms will be recorded in the physical examination section in the eCRF. 
Any meaningful changes in physical examination findings that meet definitions of AEs after IP 
administration must be reported as AEs. 
 
10.1.7. Laboratory Test 
Tests results for baseline (Visit 2) may be replaced with the results obtained within 7 days prior 
to baseline visit (Visit 2), if available. Laboratory tests will be analyzed at the internal lab of the 
institution. Creatine phosphokinase in chemistry and serology will be performed at screening visit 
only. Subjects who do not meet inclusion/exclusion criteria based on the screening tests may 
undergo re-test once. Detailed tests are listed below: 
Hematology 
White blood cell (WBC), Red blood cell (RBC), Hemoglobin, Hematocrit, 
Platelet count, WBC diff count (Neutrophil, Lymphocyte, Monocyte, 
Eosinophil, Basophil) 
Chemistry† 
Na, K, Ca, Cl, Blood urea nitrogen (BUN), Creatinine, Uric acid, Total 
bilirubin, Albumin, Total protein, Alanine aminotransferase (ALT), 
Aspartate aminotransferase (AST), γ-GTP, Alkaline phosphatase (ALP), 
Glucose, Creatine phosphokinase (CPK) 
Serology‡ 
HBsAg, Venereal disease research laboratory (VDRL), Anti-HBs, Anti-
HIV, Anti-HCV 
Urinalysis 
pH, Specific gravity, Albumin, Bilirubin, Glucose, Urobilinogen, Ketone, 
Nitrite, Occult blood, Microscopy (Squamous cell, Urine RBC, urine 
WBC) 
†Creatine phosphokinase test will be performed at screening visit (Visit 1) only. 
‡For screening visit (Visit 1) only. 
 
10.1.8. Immunogenicity Test 
This test is for exploratory purpose and will be performed at screening visit (Visit 1 of Part 1) and 
the end of study visit (Visit 6 of Part II-A or Visit 7 of Part II-B)(total twice). Blood collection for 
immunogenicity test may be performed together with laboratory test. Details about blood 
collection, storage/disposition and analysis of the test will be described in a separate document. 
 
10.1.9. Pregnancy Test 
Pregnancy test will be performed in women of child-bearing potential who have not received a 
sterilization procedure and women who have the last menstrual period within 12 months or are 
not diagnosed with menopause only. Pregnancy test will be performed with blood or urine 
samples (serum β human Chorionic gonadotrophin (hCG) or Urine hCG). 
 
10.1.10. Randomization 
Only for subjects who are finally determined eligible, investigator will conduct randomization and 
assign allocation number for Part I (Allergy assessment) participation in the order of providing 
consent, as described in Section 6.3.  
 
For subjects enrolled in Part I (Allergy assessment), both of their forearms will be randomized into 
the study group or control group in a ratio of 1:1 for ID injection. 
Allocation number will be used as a subject forearm identification code during the study. 
After evaluation of cutaneous hypersensitivity of delayed allergic reaction occurring from 30 
minutes to 48 hours after IP administration at Visit 4 of Part I, only subjects with negative skin 
reaction of drug allergy can take part in Part II. Subjects will take part in Part II-A for PK 
assessment or Part II-B for safety assessment. Subjects enrolled in Part II-B (Safety assessment) 
will receive allocation numbers by randomization into the study group or control group in a ratio 
of 2:1 for SC injection.  
 

 
 
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10.1.11. PK Assessment 
Subjects participating in Part II-A will be evaluated. Blood collection for PK assessment will be 
performed pre-dose (-2h, -1h, and 0h) and 10 min (0.17h), 20 min (0.3h), 30 min (0.5h), 40 min 
(0.7h), 50 min (0.8h), 1h, 1h 15 min (1.25h), 1h 30 min (1.5h), 2h, 2h 30 min (2.5h), 3h and 24 h 
post-dose. PK parameters (Cmax, AUClast, t1/2, Tmax) of plasma ALT-BB4 will be calculated and 
evaluated. In addition, PK parameters adjusted with baseline (pre-dose) will be calculated and 
evaluated. 
Cmax 
Peak plasma drug concentration 
AUClast 
Area under plasma concentration-time curve calculated until the time of measurable 
last blood collection 
t1/2  
Drug elimination half-life 
Tmax 
Time to peak plasma drug concentration 
 
Below are acceptable windows. 
Scheduled time 
Window 
Visit 4 pre-dose (-2h, -1h, pre-dose) 
± 10 min 
Visit 4 pre-dose (0h pre-dose) 
- 10 min 
Visit 4 0.17h, 0.3h, 0.5h, 0.7h, 0.8h post-dose 
± 3 min 
Visit 4 1h, 1.25h, 1.5h, 2h post-dose 
± 5 min 
Visit 4 2.5h, 3h post-dose 
± 10 min 
Visit 4 24h post-dose 
± 60 min 
 
10.1.12. Investigational Product Administration 
At Baseline visit (Visit 2), cutaneous hypersensitivity due to drug allergy within 30 minutes after 
ID injection will be evaluated for Part I (drug allergy assessment). In the next visit, Visit 4, subjects 
with negative results evaluated from 30 minutes to 48 hours after IP administration can take part 
in Part II-A and Part II-B. Single dose of ALT-BB4 will be injected subcutaneously to the right or 
left upper arm of subjects participating in Part II-A and Part II-B, by principal investigator (or its 
designee). 
 
Presence of temperature exceeding 37.5℃ within 7 days prior to IP administration and presence 
of acute disease within 14 days prior to IP administration will be checked before IP administration. 
Subjects with temperature exceeding 37.5℃ or history of acute diseases within past 14 days 
cannot receive the IP. 
 
10.1.13. Monitoring of Adverse Event 
Subjects will be monitored for adverse events after IP administration. In case of adverse events, 
the symptoms, start date/stop date, severity, causality, action taken to IP, action taken to other 
items except for IP, outcome and seriousness must be documented in detail. 
For presence of drug allergy, immediate allergic reaction occurring within 30 (±5) minutes after 
ID injection of the IP at baseline visit (Visit 2) and delayed allergic reaction occurring from 30 
minutes to 48 (±2) hours after ID injection of the IP will be identified. Administration site will be 
observed every minute for up to 5 minutes after IP administration and then every 5 minutes from 
5 minutes to 30 minutes after IP administration. Subjects who show no immediate allergic reaction 
or whose symptoms disappear will be discharged, and then presence of delayed allergic reaction 
will be additionally monitored at 48 hours after IP administration. 
Presence of any adverse events, including drug allergy, will be monitored from the time of IP 
administration at each visit. Investigator will advise the subjects to visit the institution to check 
presence of delayed allergic reaction and provide appropriate observation and treatment, if 
necessary, if he/she notices any adverse events occurring within 48 hours after IP administration. 
 

 
 
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10.1.14. Photographing of Investigational Product administration Site 
Details about photographing method, collection/storage method of photographs taken, and quality 
assessment of collected photographs will be provided in a separate manual. 
 
 

 
 
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10.2. Efficacy Assessment 
10.2.1. Efficacy Endpoints 
The study aims to evaluate allergy, safety and tolerability and efficacy assessment will not be 
separately performed. 
 
10.3. Safety Assessment 
10.3.1. Safety Endpoints 
Part I study will evaluate the drug allergy and adverse events occurring following ID injection of 
the IP. Part II-A study will evaluate adverse events occurring following SC injection of the IP. Part 
II-B study will evaluate the safety and tolerability following SC injection of the IP.  
 
10.3.1.1. Primary Endpoints 
1) 
Incidence rate of drug allergy following ID injection of the IP* in Part I 
*Subjects developing either immediate or delayed allergic reaction are considered to have drug allergy. 
2) 
Safety and tolerability assessment following SC injection of the IP** in Part II 
** Incidence rate of adverse events, including systemic and administration site-related adverse events 
 
10.3.1.2. Secondary Endpoints 
1) 
Incidence rate of immediate allergic reactions† occurring within 30 minutes after the 
ID injection of the IP 
†defined as drug allergy associated with wheal ≥ 8 mm with or without pseudopod (elevation 
considered due to an injection is not considered wheal), erythema or localized itching 
occurring in the administration site within 30 minutes after IP administration 
 
2) 
Incidence rate of delayed allergic reactions‡ occurring within 48 hours after the ID 
injection of the IP 
‡drug allergy occurring from 30 minutes to 48 hours after the IP administration, including but 
not limited to redness, rash, urticaria, edema, and erythema in the administration site as well 
as acute generalized exanthematous pustulosis and erythema multiforme. Investigator will 
decide whether an adverse event is a delayed allergic reaction. 
 
3) 
Incidence rate of administration site events* at 30 min. and 48 hours after the IP 
administration 
* classified into wheal, erythema, localized itching and other adverse events, occurring after the IP 
administration, at each time point 
 
4) 
Size of wheal and erythema occurring at 30 min. and 48 hours after the IP 
administration 
 
10.3.1.3. Other Safety Endpoints 
1) 
Adverse Events 
2) 
Physical Examination, Vital Signs, and ECG 
3) 
Laboratory tests 
 
10.3.2. Safety Assessment Method 
Part I study will evaluate the drug allergy and adverse events occurring following ID injection of 
the IP. Part II-A study will evaluate adverse events occurring following SC injection of the IP. Part 
II-B study will evaluate the safety and tolerability following SC injection of the IP.  

 
 
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10.3.2.1. Primary Endpoints 
1) 
Incidence rate of drug allergy following ID injection of the IP* in Part I 
*Subjects developing either immediate or delayed allergic reaction are considered to have drug allergy. 
 
Incidence rate of drug allergy by treatment group following IP administration will be provided. 
 
2) 
Safety and tolerability assessment following SC injection of the IP** in Part II 
** Incidence rate of adverse events, including systemic and administration site-related adverse events 
 
Systemic and administration site-related AEs following IP administration will be provided.  
 
10.3.2.2. Secondary Endpoints 
1) 
Incidence rate of immediate allergic reactions† occurring within 30 minutes after the 
ID injection of the IP 
†defined as drug allergy associated with wheal ≥ 8 mm with or without pseudopod 
(elevation considered due to an injection is not considered wheal), erythema or localized 
itching occurring in the administration site within 30 minutes after IP administration 
 
2) 
Incidence rate of delayed allergic reactions‡ occurring within 48 hours after the ID 
injection of the IP 
‡drug allergy occurring from 30 minutes to 48 hours after the IP administration, including 
but not limited to redness, rash, urticaria, edema, and erythema in the administration site as 
well as acute generalized exanthematous pustulosis and erythema multiforme. Investigator 
will decide whether an adverse event is a delayed allergic reaction. 
 
Incidence rate of immediate allergic reaction and delayed allergic reaction at 30 minutes and 48 
hours after IP administration by treatment group will be provided. 
In case of immediate allergic reaction assessment, localized itching will be assessed together. 
Localized itching will be rated with a numeric rating scale (NRS) between 0-10, and subjects will 
be asked to indicate the degree of itching in a scale from 0 (no itching) to 10 (unimaginably severe 
itching)35). Severity based on NRS score will be assessed by referring to the table below. 
➢ 
Example of questions: Please tell us how serious itching have you experienced on a scale 
from 0 to 10. 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
No itching 
 
 
 
 
 
 
 
Unimaginable 
itching 
 
NRS Score 
Severity 
1-3 
Mild 
4-6 
Moderate 
7-10 
Severe 
 
3) Incidence rate of administration site events* at 30 min. and 48 hours after the IP 
administration 
* classified into wheal, erythema, localized itching and other adverse events, occurring after 
the IP administration, at each time point. Incidence rate of adverse events (urticaria, erythema, 
localized itching and other adverse events) at 30 minutes and 48 hours after IP administration 
by administration will be provided. 
 
4) 
Size of wheal and erythema occurring at 30 min. and 48 hours after the IP 

 
 
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administration 
Size of wheal and erythema at each time point after IP administration will be provided. To measure 
size of urticaria or erythema, the length of the long axis and the length of the short axis that 
orthogonally crosses the middle of the long axis will be measured. The mean value of the length 
of both axes will be provided as the size of urticaria or erythema. 
Figure 10.3.1 How to measure size of urticaria and erythema 
 
 
 
 
10.3.2.3. Other Safety Endpoints 
1) Adverse Events 
AEs will be collected from baseline visit (Visit 2) to the end of the study, and in case of AEs, AE 
symptoms, start date/stop date, duration, severity and causality must be documented in eCRF 
without omission. All AEs will be classified by system organ class (SOC) and preferred term (PT) 
using the most recent version of Medical dictionary for regulatory activities (MedDRA). 
 
2) Physical Examination, Vital Signs, and ECG 
• 
Physical examination 
For physical examination, cardiovascular system, respiratory system, gastrointestinal 
system/hepatobiliary system, metabolism/endocrine system, renal/urinary system, 
reproductive system, musculoskeletal system, skin and connective tissues, nervous 
system, psychiatric system and other body organs will be comprehensively evaluated. 
Significant symptoms and meaningful symptoms detected during screening period will 
be recorded in the physical examination section in the eCRF. Any significant changes 
in physical examination findings that meet definitions of AEs after IP administration 
must be reported as AEs. 
 
• 
Vital signs 
It will be measured before other scheduled tests. After taking a rest for at least 5 
minutes, sitting systolic and diastolic blood pressures, pulse rate and temperature will 
be measured. 
 
• 
ECG 
Any significant changes in ECG findings that meet definitions of AEs after IP 
administration must be reported as AEs. 
 
3) Laboratory tests 
Laboratory test results will be classified into normal, not clinically significant (NCS) and clinically 
significant (CS). Clinically meaningful laboratory results based on comparison between IP pre-
dose and post-dose will be reported as AEs.  
(Size of urticaria or 
redness= A+B/2) 

 
 
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10.4. Pharmacokinetic Assessment 
10.4.1. PK Endpoints 
1) 
Primary endpoints: Cmax, AUClast, t1/2, and Tmax of ALT-BB4 
2) 
Secondary endpoints: Baseline-adjusted Cmax, AUClast, t1/2, and Tmax of ALT-BB4 
 
10.4.2. PK Assessment Method 
1) 
Blood collection method, pretreatment and storage method of isolated plasma 
 
Blood collection will be performed in the forearm in the opposite side of IP administration 
and via a venipuncture or saline-locked angiocatheter inserted to the brachial vein. 
When saline-locked angiocatheter is used for blood collection, approximately 1 mL of 
blood will be drawn before collecting blood for PK assessment to fully remove any 
remaining normal saline in the blood collection set, and then 1 mL of saline will be 
injected to the catheter to prevent blood coagulation. 
 
For the treatment and storage of collected blood samples, follow a separate document. 
 
2) 
Sample Storage and Disposition 
Samples collected for drug concentration analysis will be kept in the institution's sample storage 
below -70℃ and after the end of the study, they will be transferred after discussion with the 
analysis facility. Disposition of samples after drug concentration analysis will be performed in 
accordance with Standard Operating Procedures (SOPs) of the analysis facility. After submission 
of the Clinical Study Report (CSR) to the IRB, back-up samples under storage will be disposed 
or transferred to the sponsor after discussion with the sponsor and the storage period can be 
extended upon agreement. If it is disposed at the institution, the disposition will be performed in 
accordance with the SOPs of the institution.  
 
3) 
Methods of analysis 
Analysis methods will be described in a separate document. 
 
10.5. Evaluation Criteria and Report of Adverse Events 
10.5.1. Definition of Safety-related Terms 
1) Adverse event (AE) 
Any untoward medical occurrence in a patient or clinical investigation subject administered a 
pharmaceutical product and which does not necessarily have a causal relationship with this 
treatment. An adverse event (AE) can therefore be any unfavourable and unintended sign 
(including an abnormal laboratory finding), symptom, or disease temporally associated with the 
use of a medicinal (investigational) product, whether or not related to the medicinal 
(investigational) product. 
 
2) Adverse drug reaction (ADR) 
All noxious and unintended responses to a medicinal product related to any dose should be 
considered adverse drug reactions. The phrase responses to a medicinal product means that a 
causal relationship between a medicinal product and an adverse event is at least a reasonable 
possibility, i.e., the relationship cannot be ruled out. 
 
3) Serious AE/ADR (SAE/SADR) 
Any untoward medical occurrence that at any dose:  
① results in death, or is life-threatening, 
② requires inpatient hospitalization or prolongation of existing hospitalization, 
③ results in persistent or significant disability/incapacity, or  

 
 
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④ is a congenital anomaly/birth 
⑤ results in any other medical event such as development of drug dependency or abuse 
or hematologic disease 
4) Unexpected adverse drug reaction 
It refers to any adverse drug reactions with different pattern or degree of harm, based on the 
related information available in the Investigator's Brochure or product insert. 
In addition to the matters listed above, any medically significant event that may substantially affect 
the subject’s well-being and health may be determined as a SAE based on the medical judgment 
of the study investigator (principal investigator or subinvestigator) and related experts and 
appropriate actions will be taken accordingly.  
 
10.5.2. Evaluation Criteria of Adverse Events 
10.5.2.1. Severity 
Severity of AEs will be determined based on following criteria. 
 
1) Mild: presence of conscious or objective symptoms, but not interfering with everyday activities 
2) Moderate: discomfort interfering with everyday activities 
3) Severe: preventing from performing normal everyday activities 
 
10.5.2.2. Causality to the IP 
Investigator will evaluate causality between the AE and the IP based on the following, and all AEs, 
except those determined as "Definitely not related, None" will be considered as adverse drug 
reactions. 
 
1) Definitely related 
① There is evidence of IP administration and plausible temporal relationship of AE onset 
② The AE is most plausibly explained by IP administration than any other reasons 
③ The AE disappears after the discontinuation of administration 
④ Rechallenge (if feasible) result is positive 
⑤ The AE shows consistent patterns with the information already known for the IP or one 
in the same class 
 
2) Probably related 
① There is evidence of IP administration and plausible temporal relationship of AE onset 
② The AE is most plausibly explained by IP administration than any other reasons 
③ The AE disappears after the discontinuation of administration 
 
3) Possibly related 
① There is evidence of IP administration and plausible temporal relationship of AE onset 
② The AE is judged to be attributable to IP administration to the same extent as other 
possible causes 
③ The AE disappears after the discontinuation of administration 
 
4) Unlikely, Probably not related 
① There is evidence of drug administration 
② There are more possible causes of the AE 
③ The result of the discontinuation of IP administration is negative or unclear 
④ Rechallenge result is negative or unclear 
 
5) Definitely not related, None 
① Subject did not receive the IP 

 
 
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② Temporal relationship of the IP administration and AE onset is not reasonable 
③ There is other clear cause of the AE 
 
6) Unknown, Unassessable 
① Lack of evidence to determine relationship 
② Poor quality of source data or inconsistent data 
 
10.5.2.3. Action taken and Outcome 
During the study, principal investigator and subinvestigator must ensure the subject safety. In 
case of SADRs, prompt and appropriate actions must be taken to minimize the adverse events 
and the principal investigator may discontinue the study after discussion with the sponsor. For all 
AEs occurring during the study, even if they are not related to the IP, the symptoms and signs, 
start date/stop date, duration, severity, action taken and outcome, causality to the IP and 
seriousness must be documented in detail in the eCRF. In addition, the AE must be observed as 
far as possible until it recovers to pre-dose condition or baseline or is considered normalized by 
principal investigator or subinvestigator or no further observation is required.  
 
Action taken to IPs for AEs, Action taken to other items except for IP and outcome will be 
completed as explained below. 
 
1) Action taken to IPs 
① Treatment stopped 
② Dose reduced 
③ Dose increased 
④ Dose not change 
⑤ Unknown 
⑥ Not applicable 
 
2) Action taken to other items except for IP 
① No action taken 
② Medication treatment 
③ Non-medication treatment 
④ Both medication and non-medication treatment 
 
3) Outcome 
① Recovered/resolved 
② Recovering/resolving 
③ Not recovered/not resolved 
④ Recovered with sequela/resolved with sequela 
⑤ Death 
⑥ Unknown 
 
10.5.3. Reporting Method for Adverse Events 
10.5.3.1. Reporting of AE 
Investigator will instruct the subjects (or their legally acceptable representative) to report all 
adverse events and symptoms that may occur after IP administration. 
For systemic symptoms or various symptoms associated with laboratory tests after IP use, 
investigator will record the type, start date/stop date, severity, actions and progression, and 
causality to the IP in the eCRF. In addition, the AEs will be followed up until termination (resolution 

 
 
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of AE or loss of follow-up). 
 
10.5.3.2. Expedited Reporting 
Principal investigator and subinvestigator must report all SAEs occurring during the IP 
administration period, regardless of causality to the IP, to the sponsor or its designee, within 24 
hours or by the next business day at the latest from the first recognition, via telephone, email or 
fax. Report to the IRB is also required in accordance with the institution regulations. 
As far as possible, all information in the SAE Form should be included at the initial report. The 
completed form should be sent to the sponsor and AE page on the eCRF must be filled out. 
Upon receipt of the initial report, the sponsor and the IRB must review the information and ask 
the subinvestigator for further information, if necessary. After review of the AE information, 
causality to the IP will be investigated. 
If necessary, follow-up report containing any new information about a SAE must be submitted to 
the sponsor in the SAE Form. 
The sponsor must report any suspected unexpected serious adverse reaction (SUSAR) to the 
investigator, Minister of Food and Drug Safety and if necessary, the IRB within the time frame 
based on the following classification: 
• 
Death or life-threatening SUSARs: Within 7 days from the date the sponsor first gets the 
report or aware of it. However, when any information in Form 77, ADR Report is missed, such 
as ADR name, final observation outcome, and ADR summary, the sponsor must provide 
additional report containing details about the ADR within 15 days from the date he/she first 
gets the report or aware of it. 
• 
Any other SUSARs must be reported within 15 days from the date he/she first gets the report 
or aware of it. 
 
Contact Information for SAE Report: 
 
• 
Contact: +82-70-4404-8793 
• 
Phone: +82-10-8934-9157 
• 
Email: shkim@alteogen.com 
• 
Fax:+82-42-384-8770 
 
10.5.3.3. Responsibilities of Staff in case of Serious Adverse Drug Reactions 
Principal investigator and subinvestigator must make sure of subject safety and minimize AEs by 
taking prompt and appropriate actions during the study. When a SADR occurs during the study, 
responsibilities of each role are as below. 
 
1) Principal Investigator 
Principal investigator must report SADRs occurring during the study to the sponsor and the IRB 
within 24 hours or by the next business day at the latest from the first recognition to determine 
whether the study can continue or should discontinue. 
 
2) Subinvestigator 
Subinvestigator must report SADRs occurring during the study to the principal investigator and 
the sponsor immediately. 
 
3) Institutional Review Board (IRB) 
In case of SADRs, the IRB must require the principal investigator to take necessary actions, such 
as suspension of partial or entire study. 
 
4) Sponsor 
When a SUSAR is reported by principal investigator or subinvestigator, the sponsor must attach 
the ADR summary, such as Council for international organizations of medical sciences (CIOMS)-
I Form to the ADR report and submit to the Minister of Food and Drug Safety and it must be 
immediately notified in accordance with the IRB regulations of the institution. 
 

 
 
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10.5.3.4. Pregnancy 
Women of child-bearing potential who have not received a sterilization procedure and women 
who have the last menstrual period within 12 months must have negative pregnancy test to 
participate in the study and must maintain medically acceptable contraception during the study 
period. Medically acceptable contraception include hormones (implantable, patch, and oral), IUD, 
and double barrier methods (concomitant use of two of following contraceptive methods: male 
condom, female condom, cervical cap, diaphragm, sponge, spermicide) (However, concomitant 
use of male condom and female condom is excluded from double barrier methods.). 
When a female subject or a male subject’s spouse or sexual partner gets pregnant during the 
study, it is not considered as a SAE, but should be reported with the same reporting method for 
the SAE. Investigator must report pregnancy of a female subject or a male subject’s spouse (or 
sexual partner) to the sponsor within 24 hours from the first recognition, and when the pregnancy 
is confirmed, follow-up will be performed for the female subject or male subject’s spouse (or 
sexual partner) who have provided written informed consent with ICF for collection of pregnancy 
information (Annex 4. Informed Consent Form for Use of Pregnancy Information) until birth or 
termination of pregnancy. 
Investigator will instruct the subjects to visit the investigator as soon as they become aware of the 
fact that they or their spouse (or sexual partner) got pregnant during the study for confirmation of 
the pregnancy. Female subjects with confirmed pregnancy will be dropped out from the study. 
 

 
 
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11. Data Analysis and Statistical Considerations 
11.1. Analysis Sets 
11.1.1. Safety Set 
This includes subjects who have received IP. 
Part I – Subjects receiving ID injection of the IP will be included. 
Part II – Subjects who showed allergy test negative result in Part I and have received SC injection 
of the IP will be included. 
 
11.1.2. Full Analysis Set 
The study aims to evaluate allergy, safety and tolerability and efficacy set will not be separately 
defined. 
 
11.1.3. PK Analysis Set 
PK set includes all subjects who have at least 1 measurements of PK primary endpoints for ALT-
BB4 after IP administration. 
 
11.2. Statistical Analysis Method 
11.2.1. General Principle of Result Analysis 
For all statistical analysis, SAS®  version 9.4 (or higher) will be used. Unless specified otherwise, 
two-sided test under significance level of 5% will be performed in principle. For continuous 
variables, descriptive statistics (sample size, mean, standard deviation, median, minimum, and 
maximum) will be presented, and for categorical variables, frequencies and percentages will be 
presented. For safety endpoints, missing data will not imputed and will be analyzed with OC 
(observed case) method. Analysis details are provided in the Statistical Analysis Plan.  
 
11.2.2. Demographic Data and Medical History 
For background and demographic data of subjects included in the safety set, descriptive statistics 
(sample size, mean, standard deviation, median, minimum, and maximum) for continuous 
variables (such as age) will be presented. Frequency and percentage (%) for categorical variables 
will be presented. 
 
11.2.3. Efficacy Endpoints 
The study aims to evaluate allergy, safety and tolerability and statistical test for efficacy analysis 
will not be separately performed. 
 
11.2.4. Safety Endpoints 
Part I study will evaluate the drug allergy and adverse events occurring following ID injection of 
the IP. Part II-A study will evaluate adverse events occurring following SC injection of the IP. Part 
II-B study will evaluate the safety and tolerability following SC injection of the IP.  
 
11.2.4.1. Primary Endpoints 
1) 
Incidence rate of drug allergy following ID IP administration* in Part I 
*Subjects developing either immediate or delayed allergic reaction are considered to have drug allergy. 
 
The number and percentage (%) of subjects showing any change of drug allergy from the IP 
administration to the end of study or early termination in the control group and study group will be 

 
 
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provided and as the primary analysis, the upper limit of binomial exact 95% two-sided confidence 
interval (97.5% one-sided confidence interval) < 10% is considered statistically significant. 
In addition, the analysis will be performed using the McNemar’s test to compare the incidence 
rate of drug allergy between the study group and the control group. 
 
2) 
Safety and tolerability assessment following SC injection of the IP** in Part II 
** Incidence rate of adverse events, including systemic and administration site-related adverse events 
 
To evaluate the safety and tolerability, systemic and administration site-related adverse events 
will be assessed following SC injection of the IP.  
In addition to the routine safety tests of physical examination, vital signs, ECG and laboratory 
tests, comprehensive data about clinical adverse events will be evaluated to determine 
relationship of the adverse drug reactions, including the nature, signs and type of adverse events, 
severity, frequency and duration, occurrence site and types of disease, and symptoms of AEs by 
system organ class (SOC). 
The number and percentage (%) of subjects developing adverse events will be provided and the 
analysis will be performed using the Pearson’s chi-square test or Fisher’s exact test to compare 
the incidence rate of AEs between the study group and the control group. 
 
11.2.4.2. Secondary Endpoints 
1) 
Incidence rate of immediate allergic reactions† occurring within 30 minutes after the 
ID injection of the IP 
†defined as drug allergy associated with wheal ≥ 8 mm with or without pseudopod 
(elevation considered due to an injection is not considered wheal), erythema or localized 
itching occurring in the administration site within 30 minutes after IP administration 
 
The number and percentage (%) of subjects showing any change of immediate allergic reaction 
occurring within 30 minutes after the IP administration in the control group and study group will 
be provided and analyzed using the McNermar’s test to compare the incidence of immediate 
allergic reaction between the study group and the control group. 
 
2) 
Incidence rate of delayed allergic reactions‡ occurring within 48 hours after the ID 
injection of the IP 
‡drug allergy occurring from 30 minutes to 48 hours after the IP administration, including 
but not limited to redness, rash, urticaria, edema, and erythema in the administration site as 
well as acute generalized exanthematous pustulosis and erythema multiforme. Investigator 
will decide whether an adverse event is a delayed allergic reaction. 
 
The number and percentage (%) of subjects showing any change of delayed allergic reaction 
occurring from 30 minutes to 48 hours after the IP administration in the control group and study 
group will be provided and analyzed using the McNermar’s test to compare the incidence of 
delayed allergic reaction between the study group and the control group. 
 
3) 
Incidence rate of administration site events* at 30 min. and 48 hours after the IP 
administration 
* classified into wheal, erythema, localized itching and other adverse events, occurring after 
the IP administration, at each time point 
 
For wheal, erythema, localized itching and other adverse events at 30 minutes and 48 hours after 
the IP administration in the control group and study group, the number and percentage (%) of 
subjects showing any change of each event will be provided and analyzed using the McNermar’s 
test in Part 1 and Pearson’s chi-square test or Fisher’s exact test in Part 2 to compare the 
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related AE.  
 
4) 
Size of wheal and erythema occurring at 30 min. and 48 hours after the IP 
administration 
For the size of wheal and erythema occurring at 30 minutes and 48 hours and change in the size 
of wheal and erythema occurring at 48 hours after the IP administration compared to 30 minutes 
after the IP administration in the control group and study group, descriptive statistics (mean, 
standard deviations, median, min and max) will be provided. The analysis will be performed using 
the Paired t-test or Wilcoxon signed-rank sum test to compare the change in the size of wheal 
and erythema occurring at 48 hours after the IP administration compared to 30 minutes, and for 
the comparison between the study group and control group, an analysis of covariance (ANCOVA) 
with the baseline as a covariate will be performed. If the assumption of normality is not satisfied, 
a rank ANCOVA will be performed. 
 
11.2.4.3. Other Safety Endpoints 
1) Adverse Events 
All AEs will be coded by using the most recent version of MedDRA. The number of subjects with 
adverse events and adverse drug reactions, incidence rate (%) and frequency by treatment group 
will be provided, along with its 95% confidence interval. 
In addition, the number of subjects with adverse events and adverse drug reactions, incidence 
rate and frequency by SOC, PT, and treatment group will be provided. 
 
2) Physical Examination, Vital Signs, ECG and Laboratory Tests 
For continuous data, descriptive statistics (mean, standard deviation, median, min and max) for 
absolute values and changes from baseline at each time point of visits and Paired t-test or 
Wilcoxon signed-rank test will be performed. For laboratory tests and ECG values, the frequency 
and proportion of subjects shifting from normal/clinically insignificant abnormal value before the 
IP administration (Screening) to clinically significant abnormal value after the IP administration 
will be provided and McNemar’s test will be performed. 
 
11.2.5. PK Endpoints 
PK endpoints are PK parameters of ALT-BB4, which will be provided with descriptive statistics. 
For PK parameters, statistical test is not separately performed. PK assessment results can be 
provided in a separate report. 
 
11.3. Planned Analysis Time 
In this study, interim analysis is not planned and analysis will be performed after completing 
observation in all subjects enrolled. 

 
 
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12. Data Management 
12.1. Record and Access 
Source documents are defined as outcomes of study data collection activities and observation. 
Source documents include medical records, electronic data and measurements by instrument as 
well as other records. All source documents available in the study will be recorded and kept by 
the investigator at the institution and only authorized person can access it. 
 
12.2. Data Collection 
All electronic data capture (EDC) systems used in the study will comply with 21 CFR part 11 
(Code of Federal Regulations). Only authorized person can access EDC system and all entries, 
corrections, saves and deletions in the eCRF will be tracked and recorded via EDC system. 
Investigator will ensure the eCRF data is accurate, complete and interpretable and timely through 
electronic signature. 
After the end of the study, a copy of eCRFs prepared through EDC system will be transferred to 
the institution via electronic storage media and will be kept under the same standards as for other 
essential documents. 
 
12.3. Record Protection and Storage 
In accordance with relevant regulations on study record storage, principal investigator must store 
various study-related data (including electronic documents), such as protocol, and IP 
manufacturing and management records for 3 years from the date of study completion (or early 
termination) or product approval under appropriate storage conditions (The storage period can 
be extended, when determined necessary by the sponsor). 
These documents will be subject to investigation in case of inspection of the sponsor or related 
regulatory authorities and investigator must not destroy any documents related to the study 
without written permission of the sponsor. Investigator must take appropriate measures to prevent 
any accidental damage or premature loss of these documents. 
 

 
 
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13. Ethical Considerations and Administrative Procedures 
13.1. Compliance with Regulations, including Korean Good Clinical 
Practice, and Ethical Standards 
This study will be conducted ethically and scientifically in accordance with the Korean Good 
Clinical Practice (KGCP) and all related regulations. In addition, based on the Helsinki Declaration, 
the study will respect human dignity and rights and will not cause any disadvantage to the subjects. 
 
13.2. Institutional Review Board (IRB) 
Investigator must obtain written approval for the protocol, ICFs, subject recruitment materials and 
procedures and subject information sheet (SIS) provided to the subjects from the IRB before the 
study starts. The IRB decision on the study implementation will be notified to the principal 
investigator and the sponsor in a written form before the study starts. 
Principal investigator will report the study progression, SAEs and life-threatening problems or 
death to the IRB and must inform the IRB of study termination. 
 
13.3. Informed Consent Process 
Investigator must obtain written informed consent from subjects (or their legally acceptable 
representative) after fully explaining study details, procedures, and effects and possible adverse 
events of the study drug to conduct the study and the written informed consent process must be 
performed prior to any other study procedures (Annex 2 Subject Information Sheet and Informed 
Consent Form). Investigator must provide a copy of signed ICF and SIS to subjects (or their legally 
acceptable representative), and the original copy must be kept in the investigator file. 
By signing the ICF, subjects will agree to study participation as well as collection and use of their 
personal information in relation to the study. The range of personal information to be collected in 
relation to the study includes personal information (date of birth, sex, etc.), demographic data, 
medical records (past medical history, treatment history, etc.) and study-related test results, and 
all data collected will be handled in accordance with Personal Information Protection Act, rules 
and regulations.  
 
13.4. Confidentiality 
All records that can identify subjects must be kept confidential. All study-related documents, 
including eCRFs, will be recorded and identified by using subject identification code, not name. 
When the study result is published, the subject identity will not be disclosed. 
The sponsor or monitor and auditor as well as the MFDS and the IRB may access the subject's 
medical records to verify the information collected and during such activities, confidentiality of the 
subject information must be kept. 
 
13.5. Subject Safeguards 
13.5.1. Actions Taken for Adverse Events 
When AEs occur due to the study, investigator must make arrangement for necessary tests and 
treatment without delay. In addition, AEs must be observed until the resolution of AE or loss of 
follow-up. 
 
13.5.2. Post-Clinical-Study Treatment and Therapy for Subjects 
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the end of the study, subjects will be treated with the optimal treatment option, as determined by 
the investigator. 
 
13.5.3. Patient Compensation Rule 
Annex 3. Patient Compensation Rule  
 
13.6. Quality Control and Reliability Assurance 
13.6.1. Monitoring of Institutions 
Monitoring will be performed to check for protection of subject rights and welfare and ensure that 
the study-related data reported is accurate, complete and verifiable, compared to the source 
documents and the study is conducted in accordance with the approved protocol, KGCP and 
related regulations. 
Study monitoring will be performed via periodic institution visits and contacts of the sponsor (or 
its designee) or monitor. Their visit schedule will be established through discussion between the 
investigator and monitor. 
Monitor must review the overall study process and basically check for source documents, IP 
management records and document storage status, etc. In addition, any issues found must be 
discussed with the investigator, and the investigator must cooperate with the process. 
 
13.6.2. Audit 
For reliability assurance during the study period, the sponsor may perform audit separately, in 
addition to routine monitoring. Audit will include confirmation that the study is conducted in 
accordance with the protocol, SOPs, KGCP and other related regulations and review of all source 
data, drug records and medical records, etc. The sponsor (or its designee) may ask access to the 
source documents and other essential documents for audit of the institution and the investigator 
must accept it and cooperate with the process. 
 
13.6.3. Inspection 
The MFDS may conduct inspection during or after the study and when an inspection is scheduled, 
the investigator must inform the sponsor immediately. The MFDS may ask access to the source 
documents and other essential documents for inspection of the institution and the investigator 
must accept it and cooperate with the process. 
 

 
 
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14. Sponsor’s Information, and Principal Investigator’s Name 
and Position 
14.1. Sponsor Information 
• 
Sponsor: Alteogen, Inc. 
• 
CEO: Soon Jae Park 
• 
Address: 62, Yuseong-daero 1628beon-gil, Yuseong-gu, Daejeon, Republic of Korea 
 
14.2. Institution, and Principal Investigator’s Name and Position 
 
Annex 1 List of Institutions and Principal Investigators 

 
 
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15. Other Matters required for Safe and Scientific Conduct of 
Study 
15.1. Protocol Amendment 
To make a change in the approved protocol, the protocol amendment must be approved by the 
IRB and if necessary, by the Minister of Food and Drug Safety. 
 
 
 

 
 
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16. References 
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of Chemically Sulfated-hyaluronic Acid from Streptococcus dysgalactiae, Journal of 
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Chip, Journal of biomedical engineering research 2017; 38(1): 1-8. 
3) K.W.Locke, DC Maneval, MJ LaBarre, ENHANZE®  drug delivery technology: a novel 
approach to subcutaneous administration using recombinant human hyaluronidase PH20, 
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subcutaneous drug and fluid administration, Expert opinion on drug delivery 2007; 4(4): 427-
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5) HW Jung, Hyaluronidase: An overview of its properties, applications, and side effects, Archive 
of plastic surgery 2020; 47(4): 297–300. 
6) N Labrou, Therapeutic Enzymes: Function and Clinical Implications, Springer 2019: 255.  
7) WS 
Atkinson, 
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11) AL Dunn, JE Heavner, G Racz, et al., Hyaluronidase: a review of approved formulations, 
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therapy 2010: 10(1):127-131. 
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single dose of recombinant human hyaluronidase injection: a double-blind, placebo-controlled 
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400-409. 
15) MS Yang, GY Ban, KH Lim, et al., KAAACI Standardization Committee Report on the 
procedures and applications of the diagnostic tests for drug allergy, Allergy Asthma Respir 
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16) Flynn, T. Hyaluronidase. Body Language, Issue 44. 
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cutaneous augmentation. Arch Facial Plast Surg. 2005;7(4):231-7. 
18) Leibovitch I, Tamblyn D, Casson R, Selva D. Allergic reaction to hyaluronidase: a rare cause 
of orbital inflammation after cataract surgery. Graefes Arch Clin Exp Ophthalmol. 
2006;244(8):944-9. 
19) King M, Convery C, Davies E. This month's guideline: The Use of Hyaluronidase in Aesthetic 
Practice (v2.4). J Clin Aesthet Dermatol. 2018;11(6):E61-E68. 
20) 손병관, 임대현. 알레르기 피부시험. Korean J Pediatr. 2007;50(5):409-15. 

 
 
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21) YJ Cho, Update on the diagnosis of drug allergy, The korean journal of medicine 2009; 76(3): 
282-290. 
22) 송정훈, 심혁, 2006, Hyaluronidase 를 이용하여 치료한 방사선 조영제 혈관 밖 유출 손상. 
23) 노낙경, 2013, 대한피부항노화연구회 Newsletter 2013 (2). 필러 시술 후 발생하는 
혈관합병증의 치료: 실질적인 가이드라인. 
24) Shinya I. Pharmacokinetics 101. Paediatrics & Child Health. 2011;16(9):535–6. 
25) Maureen Y. A Practical Approach to Discontinuing NSAID Therapy Prior to a Procedure. 2013. 
26) Michael EW. 임상약동학의 이해와 적용. 
27) The korean society of hypertension, 2018 년 고혈압 진료지침 
28) 통계로 보는 질병정보. 건강보험심사평가원. http://www.hira.or.kr/dummy.do?pgmid=HIRAA 
030502010000&ilnsCtgyNm=summer&sortSno=3. Accessed November 8, 2021. 
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20355465. Accessed November 8, 2021. 
31) SM Lee, JS Kim, JG Jung, et al., Korean alcohol guidelines for moderate drinking based on 
facial flushing, Korean journal of family medicine 2019; 40: 204-211. 
32) SS Kye, Clinically significant drug interactions, Korean journal of family medicine 2009; 30(11 
suppl): S168-S179. 
33) JS Lee, SH Ahn, Recent Trends in Drug Interaction Studies based on Drug Metabolism, 
Yakhak Hoeji 2020; 64(1): 8-20. 
34) 식품의약품안전평가원, 약물의 효능에 영향을 미치는 과일주스, 식품의약품안전평가원 2011. 
35) JI Gwak, SY Suh, The Assessment Tools in Palliative Medicine, Korean journal of hospice 
and palliative care 2009; 12(4): 177-193. 
 
 
 
 

 
 
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17. Appendix 
Appendix 1 Adverse Events of IP 
This study is planned to monitor expected adverse events. However, based on human 
hyaluronidase, the active ingredient of Hylenex, a similar drug to ALT-BB4, and non-clinical trial 
data, expected adverse events are as below.  
The most frequently reported adverse events with Hylenex were mild local adverse events, such 
as erythema or pain, and the reported incidence rate of allergic reaction was less than 0.1%.  
At the highest dose in Minipig (2mg/kg) used for dose-finding, no adverse event was reported. In 
single dose and repeat dose toxicity studies of rats and repeat dose toxicity studies of cynomolgus 
monkeys, expected adverse event was not separately reported. 
 

 
 
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18. Annex 
Annex 1 List of Institutions and Principal Investigators 
Annex 2 Subject Information Sheet and Informed Consent Form 
Annex 3 Patient Compensation Rule 
Annex 4 Informed Consent Form for Use of Pregnancy Information