Clinical trial NCT06170827
Conditions: Respiratory Disease
Sponsor: Syneos Health
Start date: 2023-11-21; Completion date: 2024-07-15
Enrollment: 16 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This goal of the open-label single dose study is to evaluate and compare the safety, tolerability, pharmacokinetic (PK), and immunogenicity of AIO-001 using two different formulations in 16 healthy volunteers.

Detailed description:
This is an open-label single dose, parallel group, 24-week, Phase 1 study in 16 healthy participants.

The study is designed to evaluate and compare the safety, tolerability, PK, and immunogenicity of AIO-001 using two different formulations (Formulation A and Formulation B) in 16 healthy volunteers (8 receiving each formulation).

The study will include a screening visit from Day -28 to Day -2. Eligible participants will be admitted to the clinical site on Day -1 and will be confined until completion of the assessments on Day 3. Participants will return to the clinical site for outpatient visits for study assessments and laboratory tests.

Arms: arm 1: 400 milligram (mg) of 100 milligrams per milliliter (mg/ml) AIO-001 Subcutaneous (SC) injection will be administered. arm 2: 400 mg of 182 mg/ml AIO-001 SC injection will be administered.
Interventions: intervention 1: AIO-001
Intervention descriptions: intervention 1: AIO-001 Solution for SC injection.

Protocol documents:
Prot_000.pdf: 
AIO-001-101 Solution for SC Injection Comparative Safety and PK 
Aiolos Bio, Inc. 
29-NOV-2023 
 
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Page 1 of 47 
PROTOCOL AIO-001-101 
 
OPEN-LABEL, SINGLE DOSE, PARALLEL GROUP, PHASE 1 STUDY IN HEALTHY 
VOLUNTEERS EVALUATING SAFETY, TOLERABILITY, PHARMACOKINETICS, 
AND IMMUNOGENICITY OF A 400 MG DOSE OF AIO-001 ADMINISTERED BY 
SUBCUTANEOUS INJECTIONS WITH TWO FORMULATIONS 
 
 
Sponsor: 
 
 
Aiolos Bio, Inc.  
c/o Latham & Watkins LLP 
200 Clarendon Street 
Boston, MA 02116 
USA 
 
Contract Research 
Organization: 
 
Protocol Version: 
Amendment 2 
Date: 
29-NOV-2023 
CONFIDENTIAL 
 
This document is strictly confidential. It was developed for Aiolos Bio, Inc. by Syneos Health. The information in 
this document may be disclosed only to persons involved in the study, ethics committees, regulatory agencies and 
study audit personnel. This document is copyrighted in favor of Syneos Health and cannot be reproduced, modified, 
or adapted, in part or in total, without prior written approval by the Sponsor and Syneos Health. 


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Protocol Historical File 
Version 
Brief description/summary of changes 
Date 
Final 
Version submitted to the Independent Ethics Committee 
(IEC). 
20-OCT-2023 
Amendment 1 
Secondary endpoints were slightly modified to be more 
concise and avoid repetition. To give some flexibility 
surrounding Public Holidays for outpatient visits, beginning at 
Day 8, an additional +1 day visit window may be granted with 
sponsor approval. The justifications of the dose and risks have 
been updated. The clinical laboratory assessments to be 
performed now include IgE. More detail about statistical PK 
population will be included in the PK analysis plan instead to 
be described in the protocol. 
27-OCT-2023 
Amendment 2 
See summary of changes below.  
29-NOV-2023 
 
Summary of Changes for Amendment 2  
 
In line with local regulations, tubal ligation is noted as an effective contraceptive method, 
therefore it is not considered a form of sterilization and has been updated. This implies changes 
in the following sections of the protocol: 
 
• 
Section 5.2 Inclusion Criteria #7 
• 
Section 7.1 Contraception 
 
Contraception restriction duration has been changed from 5 months to 5 half-lives +1 month. 
This implies changes in the following section of the protocol: 
 
• 
Section 7.1 Contraception 
 
Contact for safety reporting has been changed. This implies changes in the following section of 
the protocol: 
 
• 
Section 8.3.6.1 Serious Adverse Event Reporting to the Sponsor 
 
Total blood volume to be collected in the study has been corrected to be 550 mL. This implies a 
change in the following section of the protocol: 
 
• 
Section 8.1.1 Pharmacokinetic and Immunogenic Blood Sample Collection and 
Processing 
 
Unscheduled visit will be allowed as needed. This implies a change in the following section of 
the protocol: 
 


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29-NOV-2023 
 
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• 
Table 1 Schedule of Assessments. 
 
Tympanic temperature will be taken instead of oral temperature. Changes were made throughout 
the document. 
 
Finally, minor typographical edits and corrections were made throughout the document. 


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29-NOV-2023 
 
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Sponsor Signature Page 
Aiolos Bio, Inc.  
c/o Latham & Watkins LLP 
200 Clarendon Street 
Boston, MA 02116 
USA 
 
Sponsor’s representative: 
 
________________________________________  
______________________ 
Name: Tony Adamis 
Date 
Title: Chief Medical Officer  


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29-NOV-2023 
 
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Investigator Signature Page 
Study Title 
Open-Label, Single Dose, parallel group, Phase 1 Study in Healthy 
Volunteers Evaluating Safety, Tolerability, Pharmacokinetics, and 
Immunogenicity of a 400 mg Dose of AIO-001 Administered by 
Subcutaneous Injections with Two Formulations 
 
Protocol Number 
AIO-001-101 
Version 
Amendment 2; November 29, 2023 
 
I have carefully read this study protocol and agree that it contains all necessary information 
required to conduct this study. I agree to conduct the study according to this protocol (including 
any amendments) and in accordance with the clinical site’s Standard Operating Procedures 
(SOPs), ICH Good Clinical Practice (GCP), all other applicable regulations, and the 
recommendations laid down in the most recent version of the Declaration of Helsinki. 
 
________________________________________  
______________________ 
Investigator signature 
Date 
 
 
______________________________________ 
Investigator name  
 
 
_________________________________________ 
Name of clinical facility  
 
 
__________________________________________________ 
Location of facility 


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Table of Contents 
PROTOCOL HISTORICAL FILE ............................................................................................. 2 
INVESTIGATOR SIGNATURE PAGE..................................................................................... 5 
TABLE OF CONTENTS ............................................................................................................. 6 
LIST OF TABLES ........................................................................................................................ 8 
LIST OF ABBREVIATIONS ...................................................................................................... 9 
SYNOPSIS OF PROTOCOL..................................................................................................... 12 
1. 
INTRODUCTION ............................................................................................................. 19 
1.1 
BACKGROUND INFORMATION ........................................................................................................................... 19 
1.2 
SUMMARY OF NONCLINICAL STUDIES ............................................................................................................... 21 
1.3 
SUMMARY OF CLINICAL EXPERIENCE ............................................................................................................... 23 
1.4 
STUDY RATIONALE ........................................................................................................................................... 24 
1.5 
BENEFIT/RISK ASSESSMENT .............................................................................................................................. 26 
2. 
OBJECTIVES .................................................................................................................... 27 
3. 
ENDPOINTS ...................................................................................................................... 27 
4. 
STUDY DESIGN ............................................................................................................... 27 
5. 
STUDY POPULATION .................................................................................................... 28 
5.1 
NUMBER OF SUBJECTS ...................................................................................................................................... 28 
5.2 
INCLUSION CRITERIA ........................................................................................................................................ 28 
5.3 
EXCLUSION CRITERIA ....................................................................................................................................... 28 
5.4 
SUBJECT WITHDRAWAL AND REPLACEMENT .................................................................................................... 30 
6. 
STUDY TREATMENTS .................................................................................................. 30 
6.1 
DRUG SUPPLIES AND ACCOUNTABILITY ........................................................................................................... 30 
6.2 
IDENTIFICATION OF TREATMENTS ..................................................................................................................... 30 
6.3 
AIO-001 RANDOMIZATION AND BLINDING ....................................................................................................... 31 
6.4 
STUDY DRUG ADMINISTRATION ....................................................................................................................... 31 
7. 
STUDY RESTRICTIONS ................................................................................................ 31 
7.1 
CONTRACEPTION ............................................................................................................................................... 31 
7.2 
CONCOMITANT MEDICATIONS .......................................................................................................................... 32 
7.3 
DRUGS, NICOTINE, AND ALCOHOL .................................................................................................................... 33 
7.4 
DIET .................................................................................................................................................................. 33 
7.5 
POSTURE AND PHYSICAL ACTIVITY .................................................................................................................. 33 
8. 
STUDY PROCEDURES ................................................................................................... 34 
8.1 
PHARMACOKINETIC AND IMMUNOGENIC ASSESSMENTS ................................................................................... 34 
8.2 
SAFETY AND TOLERABILITY ASSESSMENTS ...................................................................................................... 35 
8.3 
ADVERSE EVENTS ............................................................................................................................................. 38 
8.4 
PREMATURE TERMINATION OF THE STUDY ....................................................................................................... 42 
9. 
STATISTICAL ANALYSES ............................................................................................ 42 
9.1 
ANALYSIS POPULATIONS .................................................................................................................................. 42 
9.2 
PHARMACOKINETIC ANALYSIS ......................................................................................................................... 43 
9.4 
PHARMACOKINETIC AND IMMUNOGENICITY STATISTICAL ANALYSIS ............................................................... 43 
9.5 
SAFETY AND TOLERABILITY ANALYSIS ............................................................................................................ 44 
10. 
DATA COLLECTION ...................................................................................................... 44 


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11. 
REGULATORY CONSIDERATIONS AND QUALITY ASSURANCE .................... 45 
11.1 
IEC APPROVAL OF PROTOCOL AND OTHER STUDY DOCUMENTS ...................................................................... 45 
11.2 
COMPLIANCE..................................................................................................................................................... 45 
11.3 
QUALITY ASSURANCE AND MONITORING ......................................................................................................... 45 
11.4 
CONFIDENTIALITY AND RETENTION OF STUDY RECORDS ................................................................................. 46 
12. 
REFERENCES .................................................................................................................. 47 
 


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List of Tables  
TABLE 1. 
SCHEDULE OF ASSESSMENTS ..................................................................................... 16 
TABLE 2. 
TIME WINDOWS FOR PK BLOOD SAMPLES ................................................................ 34 
TABLE 3. 
INJECTION SITE REACTION SCORES ............................................................................ 36 
TABLE 4. 
CLINICAL LABORATORY ASSESSMENTS ..................................................................... 37 
TABLE 5. 
SEVERITY OF ADVERSE EVENTS ................................................................................ 39 
TABLE 6. 
ADVERSE EVENT RELATIONSHIP CATEGORIES ........................................................... 40 


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List of Abbreviations 
ACT 
Asthma Control Test  
ADA 
Anti-drug antibody  
AE 
adverse event 
ALT 
alanine aminotransferase 
ANOVA 
analysis of variance 
aPTT 
activated partial thromboplastin time 
AST 
aspartate aminotransferase 
AUC0-inf 
area under the concentration-time curve from time zero to infinity 
(extrapolated) 
AUC0-t 
area under the concentration-time curve from time zero until the last 
observed concentration 
AUC0-τ 
area under the concentration-time curve for one dosing interval (τ) at 
steady-state 
BMI 
body mass index 
CDER 
Center for Drug Evaluation and Research 
CFR 
Code of Federal Regulations 
CHMP 
Committee for Medicinal Products for Human Use 
CI 
confidence interval 
CK 
creatine kinase 
Cl/F 
apparent clearance 
ClR 
renal clearance 
Cmax 
maximal observed concentration 
Cmin 
minimal observed concentration 
CRF 
case report form 
CRO 
contract research organization 
CV 
coefficient of variation 
DMP 
data management plan 
ECG 
electrocardiogram 
eGFR 
estimated glomerular filtration rate 
EMA 
European Medicines Agency 
EOS 
end of study 
ET 
early termination 
FDA 
Food and Drug Administration 
FeNO 
fractional exhaled nitric oxide  
FEV1 
forced expiratory volume  


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FSH 
follicle-stimulating hormone 
GCP 
Good Clinical Practice 
GGT 
gamma-glutamyl transferase 
GLM 
general linear model 
GLP 
Good Laboratory Practice 
GMP 
Good Manufacturing Practice 
HBsAg 
hepatitis B surface antigen 
HCV 
hepatitis C virus 
HEENT 
head, eyes, ears, nose, and throat 
HIV 
human immunodeficiency virus 
IB 
Investigator’s Brochure 
ICF 
informed consent form 
ICH 
International Council for Harmonisation of Technical Requirements for 
Pharmaceuticals for Human Use 
IEC 
Independent Ethics Committee 
IgE 
immunoglobulin E 
INR 
international normalized ratio 
IRB 
Institutional Review Board 
IV 
intravenous 
Kel 
terminal elimination rate constant 
LLN 
lower limit of normal 
MAD 
multiple ascending dose 
Max 
maximum 
MCH 
mean cell hemoglobin 
MCHC 
mean cell hemoglobin concentration 
MCV 
mean cell volume 
MDMA 
3,4-methylenedioxymethamphetamine 
MDRD 
modification of diet in renal disease 
MedDRA 
Medical Dictionary for Regulatory Activities 
Min 
minimum 
NOAEL 
no observed adverse effect level 
OTC 
over-the-counter 
PCP 
phencyclidine 
PK 
pharmacokinetic(s) 
PR 
PR interval 
PRN 
Pro re nata (as needed) 


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PT 
prothrombin time 
QA 
quality assurance 
QC 
quality control 
QD 
quaque die (once a day) 
QT 
QT interval 
QTcF 
Fridericia’s corrected QT interval 
RBC 
red blood cell 
SAE 
serious adverse event 
SAP 
statistical analysis plan 
SAS 
statistical analysis system 
SC 
subcutaneous  
SD 
standard deviation 
SOP 
standard operation procedure 
SUSAR 
suspected, unexpected, serious adverse reaction 
T½ el 
terminal elimination half-life 
T3 
triiodothyronine 
T4 
thyroxine 
TEAE 
treatment-emergent adverse event 
THC 
tetrahydrocannabinol 
Tmax 
time when the maximal concentration is observed 
TSH 
thyroid-stimulating hormone 
TSLP 
thymic stromal lymphopoietin 
ULN 
upper limit of normal 
Vz/F 
apparent volume of distribution 
WBC 
white blood cell 


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Synopsis of Protocol 
Project Number: 
Aiolos Bio, Inc. Project Number: AIO-001-101 
Title: 
Open-Label, Single Dose, Parallel Group, Phase 1 Study in Healthy Volunteers  
Evaluating Safety, Tolerability, Pharmacokinetics, and Immunogenicity of a 400 mg 
Dose of AIO-001 Administered by Subcutaneous Injections with Two Formulations 
Investigational Products: 
AIO-001 solution for subcutaneous (SC) injection at 100 mg/ml (Formulation A) 
AIO-001 solution for SC injection at 182 mg/ml (Formulation B) 
Study Phase and Type: 
Phase 1 – Comparative Safety and Pharmacokinetic (PK)  
Objectives: 
Primary objective: 
• 
To compare the safety and tolerability of 400 mg of AIO-001 as the new 
formulation (Formulation B) with a 400 mg dose of AIO-001 administered as 
the original formulation (Formulation A) in healthy volunteers. 
Secondary objectives: 
• 
To evaluate the PK of a single SC AIO-001 dose of Formulation A in healthy 
volunteers. 
• 
To evaluate the PK of a single SC AIO-001 dose of Formulation B in healthy 
volunteers. 
• 
To evaluate the immunogenicity of AIO-001 in healthy volunteers. 
Endpoints: 
Primary endpoints: 
• 
Adverse events (AEs), vital signs measurements (blood pressure, heart rate, 
respiratory rate, and tympanic temperature), 12-lead electrocardiogram 
(ECG) recordings, physical examinations, and clinical laboratory test 
results, including hematology, biochemistry, and urinalysis.  
Secondary endpoints: 
• 
PK endpoints may include but are not limited to AIO-001 concentrations, 
AUC0-last, AUC0-inf, Cmax, Tmax, T½  
• 
Incidence of anti-drug antibody (ADA) to AIO-001 
Study Design: 
This is an open-label single dose, parallel group, 24-week, Phase 1 study in 
16 healthy subjects. 
 
The study is designed to evaluate and compare the safety, tolerability, PK, and 
immunogenicity of SC AIO-001 400 mg using two different formulations (original 
Formulation A and new Formulation B) in 16 healthy volunteers (8 receiving each 
formulation). 
The study will include a screening visit from Day -28 to Day -2 (Visit 1). Eligible 
subjects will be admitted to the clinical site on Day -1 and will be confined until 
completion of the assessments on Day 3 (Visit 2). Subjects will return to the clinical 
site for outpatient visits for study assessments and laboratory tests as described in the 
Table 1. Schedule of Assessments.  
A staggered dosing schedule will be used for dosing of each cohort and will include 
2 sentinel subjects (1 for each treatment) dosed initially, and the remaining 14 
subjects dosed at least 24 hours later. 


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Study Population: 
It is planned to enroll up to 16 healthy subjects. 
Inclusion/Exclusion 
Criteria: 
Inclusion Criteria  
Subjects must meet all of the following criteria to be included in the study: 
1. 
Able to understand the study procedures and provide signed informed 
consent to participate in the study.  
2. 
Male or female. 
3. 
Non-smokers. Light smokers (no more than 5 cigarettes daily 
[approximately 50 to 60 mg of nicotine per day], or products with 
equivalent amount of nicotine within 3 months prior to screening) may 
be permitted. 
4. 
≥18 and ≤55 years of age. 
5. 
BMI >18.5 and <32.0 kg/m2 and body weight ≥45.0 kg. 
6. 
Healthy subjects as defined by: 
a. 
the absence of clinically significant illness and surgery within 
4 weeks prior to dosing. 
b. 
the absence of clinically significant history of neurological, 
endocrine, 
cardiovascular, 
respiratory, 
hematological, 
immunological, psychiatric, gastrointestinal, renal, hepatic, 
and metabolic disease. 
7. 
Female subjects of non-childbearing potential must be: 
a. 
post-menopausal (spontaneous amenorrhea for at least 12 
months prior to dosing) with confirmation by documented 
FSH levels ≥ 40 mIU/mL; or 
b. 
surgically 
sterile 
(bilateral 
oophorectoy, 
bilateral 
salpingectomy, or hysterectomy) at least 3 months prior to 
dosing. 
8. 
Sexually active female subjects of childbearing potential who are 
sexually active with the opposite sex and non-sterile male subjects must 
be willing to use an acceptable contraceptive method throughout the 
study as detailed in section 7.1 
Exclusion Criteria 
Subjects to whom any of the following applies will be excluded from the study: 
1. 
Any clinically significant abnormal finding at physical examination at 
screening. 
2. 
Clinically significant abnormal laboratory test results or positive 
serology test results for HBsAg, HCV antibody, or HIV antigen and 
antibody, or QuantiFERON®-TB test at screening. 
3. 
Positive pregnancy test or lactating female subject. 
4. 
Positive urine drug screen or alcohol breath test. 
5. 
History of anaphylaxis, or severe allergy. 


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6. 
Previous exposure to thymic stromal lymphopoietin antibody. 
7. 
Known allergic reactions to thymic stromal lymphopoietin antibody 
drugs and their excipients, or other biologic products or to any excipient 
in the formulation. 
8. 
Estimated Glomerular Filtration Rate (eGFR, using CKD-EPI 
Creatinine Equation) < 90 ml/min/1.73m2 during screening or baseline 
visit.   
9. 
Clinically significant ECG abnormalities (e.g., For male QTcF > 
450msec, for female QTcF > 470msec) or vital signs abnormalities 
(systolic blood pressure lower than 90 or over 140 mmHg, diastolic 
blood pressure lower than 40 or over 90 mmHg, or heart rate less than 
40 or over 100 bpm) at screening. 
10. 
History of drug abuse within 1 year prior to screening. 
11. 
History of alcohol abuse within 1 year prior to screening or regular use 
of alcohol within 6 months prior to screening that exceeds 10 units for 
women or 15 units for men of alcohol per week (1 unit = 375 mL of 
beer 3.5%, or 100 mL of wine 13.5%, or 30 mL of distilled alcohol 
40%). 
12. 
Use of medications within the timeframes specified in section 7.2. 
13. 
Previous participation in a clinical trial of AIO-001. 
14. 
Dosed in a clinical research study involving the administration of an 
investigational or marketed drug or device within 30 days prior to 
dosing, administration of a biological product in the context of a clinical 
research study within 90 days or 5 half-lives (whichever is longer) prior 
to dosing, concomitant participation in an investigational study 
involving no drug or device administration. 
15. 
Donation of plasma within 7 days prior to dosing or donation or loss of 
500 mL or more of whole blood within 4 weeks prior to dosing. 
16. 
Any reason which, in the opinion of the Investigator, would prevent the 
subject from participating in the study. 
17. 
Intolerance to venipuncture. 
18. 
Any scars, tattoos, etc., at the injection site. 
19. 
More than 5 cigarettes daily (or products with equivalent amount of 
nicotine) for 3 months prior to screening and unable to abstain from 
smoking during the trial. 
Study Treatments: 
Sixteen healthy volunteers (8 receiving each formulation) will receive one of the 
following treatments, according to the randomization scheme: 
Group A: 
400 mg of 100 mg/ml AIO-001 as Formulation A, given as 4 X 1.0 ml 
subcutaneous injections administered within approximately 5 
minutes.  
Group B: 
400 mg of 182 mg/ml AIO-001 as Formulation B, given as 2 X 1.1 ml 
subcutaneous injections administered within approximately 5 
minutes.  


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Study Procedures: 
Blood samples for PK and ADA analysis will be collected and safety procedures will 
be performed at pre-defined times throughout the study as specified in Table 1.
 
Schedule of Assessments. 
Subjects will be monitored throughout the study by the clinical staff for AEs and 
concomitant medication use. 
Statistical Analyses: 
Safety and tolerability analysis: 
Safety and tolerability of AIO-001 will be evaluated through the assessment of AEs 
(i.e., seriousness, severity, relationship to the study drug, outcome, duration, and 
management), vital signs, 12-lead ECGs, clinical laboratory tests, and physical 
examinations. Treatment-emergent adverse events (TEAEs) will be tabulated by 
treatment. Safety and tolerability data will be reported using descriptive statistics. 
PK analysis: 
The pharmacokinetic profile of AIO-001 will be established using non-
compartmental analysis of serum concentration data obtained at the timepoints 
outlined in the Schedule of Assessments. PK parameters including but not limited to 
AUC0-last, AUC0-inf, Cmax, Tmax, and T½  will be calculated if sufficient data are 
available for estimation, at the discretion of the pharmacokineticist. 
AIO-001 serum concentrations and PK parameters will be presented in summary 
tables for each dose cohort and in listings, as appropriate. Individual and mean AIO-
001 serum concentrations over time will be presented graphically. 
In addition, population pharmacokinetic modeling may be applied for PK analysis 
and will be reported separately. 
Immunogenicity:  
The incidence of ADA to AIO-001 will be summarized.  As feasible, the impact on 
PK will be assessed graphically and may be reported separately. 
Interim analyses: 
Interim analyses of preliminary data will be performed at Sponsor discretion. 


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Table 1. 
Schedule of Assessments 
Study Stage 
Screening Baseline 
Inpatient1 
Visit2 
EOS/
ET3 
Day 
-28 to -2 
-1 
1 
2 
3 
4 
5 
6 
8 
11 
15 
22 
29 
43 
57 
71 
85 113 141 169 
Hour 
 
 
0 
2 
4 
8 
12 
24 
48 
72 
96 120 168 
 
 
 
 
 
 
 
 
 
 
 
Visit Number 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
17 
Time Window ± 
 
 
Within 
60 min 
30 
min 
30 
min 
30 
min 
30 
min 
30 
min 
30 
min 2 h 2 h 2 h 4 h 8 h 8 h 1 d 1 d 2 d 2 d 2 d 2 d 2 d 2 d 2 d 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/exclusion 
criteria 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographic data 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical and 
medication history 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Admission to CRU 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Confinement 
 
 
X1 
X1 X1 
X1 
X1 
X1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Discharge 
 
 
 
 
 
 
 
 
X1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Outpatient visits 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Safety 
Physical 
examination4 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
X 
Body 
measurements 
(height, weight, 
BMI) 
X 
X5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X5 
Vital signs6 
X 
X 
Pre-
dose 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
12-lead ECG 
X 
X 
 
 
 
 
 
X 
 
 
 
 
X 
 
X 
 
 
X 
 
 
X 
X 
 
X 
QuantiFERON-TB 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serology tests7 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COVID 19 test8 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Hematology 
X 
X 
 
 
 
 
X 
X 
 
 
X 
 
 
 
X 
 
X 
X 
 
 
X 
X 
X 
X 


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Study Stage 
Screening Baseline 
Inpatient1 
Visit2 
EOS/
ET3 
Day 
-28 to -2 
-1 
1 
2 
3 
4 
5 
6 
8 
11 
15 
22 
29 
43 
57 
71 
85 113 141 169 
Hour 
 
 
0 
2 
4 
8 
12 
24 
48 
72 
96 120 168 
 
 
 
 
 
 
 
 
 
 
 
Visit Number 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
17 
Time Window ± 
 
 
Within 
60 min 
30 
min 
30 
min 
30 
min 
30 
min 
30 
min 
30 
min 2 h 2 h 2 h 4 h 8 h 8 h 1 d 1 d 2 d 2 d 2 d 2 d 2 d 2 d 2 d 
Clinical laboratory 
tests9 
X 
X 
 
 
 
 
 
X 
 
 
X 
 
X 
 
X 
 
X 
X 
 
 
X 
X 
 
X 
FSH (for post-
menopausal 
females)10 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy tests11 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
Drug and alcohol 
screens 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Monitoring and 
recording of AEs 
and 
prior/concomitant 
medication use 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Pharmacokinetics 
Blood samples for 
PK analysis12 
 
 
Pre-
dose 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Immunogenicity 
Blood samples for 
ADA 
 
 
Pre-
dose 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
X 
 
X 
 
X 
 
X 
X 
Study drug administration 
Treatment Group 
Assignment for 
Healthy Volunteers 
(Group A or Group 
B) 
 
 
Pre-
dose 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AIO-001 injection13 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Injection site 
evaluation 
 
 
Pre-
dose 
 
 
 
X 
X 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Abbreviations: ADA=, anti-drug antibody; AE = adverse event; BMI = body mass index; d = Day; ECG = electrocardiogram; EOS = end of study; ET = early 
termination; FSH = follicle-stimulating hormone; h = hour; HBsAg = hepatitis B surface antigen; HCV = hepatitis C virus; HIV = human immunodeficiency 
virus; PK = pharmacokinetic; TB= tuberculosis;  
1. Subjects will remain in the investigational study center from check-in on Day -1 until after the 48 hours PK samples and all other required assessments 
are completed on Day 3 (Visit 2). 
2. Beginning at Day 8 (Visit 6), for operational feasibility surrounding Public Holidays, an additional +1day visit window may be granted with sponsor 
approval. 
3. In case of ET, EOS procedures will be performed as soon as possible. 
4. A complete physical examination will be performed at screening. A brief physical examination will be performed on Day -1 (Visit 2), at Day 85 (Visit 
14), and at Day 169 (Visit 17) (EOS/ET). 
5. Weight and BMI only. 
6. Includes blood pressure, heart rate, respiration rate, and tympanic temperature. 
7. Serology tests include HBsAg, HCV antibody, and HIV antigen and antibody. 
8. COVID-19 screening will be performed according to clinical site’s procedure. 
9. Standard biochemistry, coagulation, lipid profile, thyroid function, and urinalysis tests will be performed.  
Note: IgE is not an eligibility determining assessment and therefore results are not needed prior to eligibility confirmation.  
10. FSH levels will be measured at screening to confirm the post-menopausal status. 
11. A serum pregnancy test will be performed at screening and at EOS/ET, and a urine pregnancy test will be performed on Day -1 (Visit 2). 
12. Refer to Table 2 for more details. 
13. Group A: 400 mg of 100 mg/ml AIO-001 as Formulation A, given as 4 X 1.0 ml subcutaneous injections administered within approximately 5 minutes. 
Group B: 400 mg of 182 mg/ml AIO-001 as Formulation B, given as 2 X 1.1 ml subcutaneous injections administered within approximately 5 minutes. 
Note:  
- When the timing of multiple procedures coincides, the following priority order should be adhered to, whenever possible: vital signs, ECGs, blood samples for 
PK/ADA analysis, clinical laboratory samples, physical examination. 
-Unscheduled visits are allowed, as needed.


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1. 
Introduction 
AIO-001 (formally SHR-1905) is a humanized anti-thymic stromal lymphopoietin (TSLP) 
monoclonal antibody (IgG1 subtype) developed by Hengrui Pharmaceuticals Co., Ltd, and 
acquired by Aiolos Bio, Inc. as of August 14, 2023. AIO-001 binds to TSLP and block the 
interaction between TSLP and receptor complex, thereby preventing TSLP-targeted immune cells 
from releasing proinflammatory cytokines to prevent asthma attack and improve asthma control. 
In non-clinical studies conducted to date, AIO-001 injection has strong in vitro activity and was 
well tolerated in toxicity studies up to 26-weeks duration in cynomolgus monkeys. Two clinical 
studies evaluating AIO-001 have been completed, one in China and one in Australia.  In addition, 
one is ongoing in China. The successful development of AIO-001 injection may provide new 
treatment options for a wide range of patients with severe uncontrolled asthma, including those 
with non-type 2 immune responses (Th2)-driven asthma.1 
1.1 
Background Information 
Asthma is a common chronic respiratory disease worldwide. The main symptoms include airway 
stenosis, airway hyperresponsiveness, inflammatory cell infiltration, increased mucus discharge, 
as well as symptoms caused by airway remodeling such as wheezing, tachypnea, chest tightness, 
and paroxysmal cough. There are approximately 339 million asthma patients in the world, of which 
5-10% have severe asthma. These patients receive high-dose standard care, but their conditions 
are still poorly controlled. The persistence of symptoms and frequent life-threatening deterioration 
seriously affect the quality of life. For 20-60% of patients with poorly controlled asthma, chronic 
oral corticosteroid treatment is required, but there is also a series of infectious, cardiovascular, 
metabolic, mental, and gastrointestinal side effects. 1 
 
Inflammation is the main component of asthma pathology. Thus, anti-inflammation has always 
been an important direction for research and development of asthma treatment. However, the 
progress is relatively slow. One-half of patients with severe asthma are Th2 inflammation-driven. 
It is characterized by increased Th2 inflammatory biomarkers, infiltration of inflammatory cells 
such as eosinophils, mast cells, and activated CD4+ T cells, as well as increased Th2 cytokines 
such as interleukin-4 (IL-4), IL-5, and IL-13. Antigen-specific Th2 cells interact with Th2 
cytokines, eventually leading to pathological conditions such as mucosal thickening of airway 
tissue, tracheal stenosis, and airway hyperresponsiveness, resulting in severe asthma and even 
glucocorticoid tolerance. Therefore, blocking the Th2 inflammatory pathway is a direction for the 
treatment of severe asthma. However, asthma is a complex disease that exhibits different 
phenotypes due to the interaction of multiple molecules. In addition to Th2 inflammation, innate 
immune inflammation also plays certain pathogenic role. Currently marketed biologics for the 
treatment of asthma, including omalizumab for asthma pathogenic immunoglobulin E (IgE), 
Mepolizumab, Reslizumab, and Benralizumab targeting IL-5/IL-5R, as well as dupilumab 
targeting IL-4Rα, only act on specific inflammatory. molecules that drive asthma inflammation 
and are only suitable for certain types of patients with severe asthma, i.e., subgroup patients, such 
as eosinophilic asthma and allergic asthma. 1 
 
The mechanism of TSLP inducing innate and Th2 immune inflammation has only recently been 
elucidated. TSLP is an IL-7-like cytokine first discovered in the conditioned medium of mouse 
thymic stromal cells. TSLP is mainly expressed in lung, skin, and intestinal epithelial cells. TSLP 


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receptor is a complex consisting of two parts: TSLPR and IL-7Rα. TSLP first binds to TSLPR 
with a relatively low affinity, then recruits IL-7Rα with high affinity, and finally induces the 
activation of signal pathways such as signal transducer and activator of transcription 5 (STAT5), 
leading to the maturation of dendritic cells (DCs) and the differentiation of T cells. 1 
 
Bone marrow-derived dendritic cells (mDCs) are the most important effector cells of TSLP. TSLP 
acts on immature mDCs such that mDCs secrete cytokines including IL-8, eotaxin-2, thymus and 
activation-regulated chemokine (TARC), and macrophage-derived chemokine (MDC), and highly 
express ligand OX40 (OX40L). In the absence of IL-12, OX40L binds to naïve CD4+ T cells, 
making them differentiate into Th2 cells, promoting Th2 cells to secrete Th2 cytokines such as IL-
5, IL-4, IL-13, and TNF, and inducing Th2 inflammatory response in the body. In addition, TSLP 
can induce DCs to produce IL-8 to recruit neutrophils, resulting in an innate inflammatory state of 
neutrophils. TSLP also induces DCs to produce eotaxin-2 to recruit eosinophils and acts together 
with IL-5 to quickly enter the inflammatory state of eosinophil infiltration. TSLP acts on mast cells 
and natural killer cells, and mediates innate inflammation by producing IL-4, IL-6, IgE, etc. In 
summary, TSLP can induce innate inflammation and Th2 inflammation simultaneously, thereby 
increasing tissue mucus, airway remodeling, and severe cell fibrosis, gradually evolving into 
asthma.1 
 
However, there is also a short form TSLP (sf TSLP) in the human body sf TSLP is expressed in 
oral, skin and intestinal epithelial cells and salivary glands, and the expression of sf TSLP is down-
regulated under inflammation. sf TSLP can inhibit the production of cytokines by DC cells and 
maintain immune homeostasis. The specific receptor of sf TSLP is still unclear, although this 
specific receptor does not bind to TSLPR. Therefore, it is a potentially effective strategy for the 
treatment of asthma by selectively blocking TSLP without blocking sf TSLP. 1 
 
Tezepelumab, an anti-TSLP antibody, has recently been approved for all moderate-to-severe 
patients with poorly controlled severe asthma. The phase 2b trial (PATHWAY) showed that 
compared with patients receiving placebo, the annual asthma attack rate of patients who received 
Tezepelumab was significantly reduced, and their lung function was significantly improved. 
Moreover, unlike IL-5, which only targets patients with eosinophilic asthma, Tezepelumab was 
effective in both patients with or without elevated eosinophil counts, suggesting that Tezepelumab 
is suitable for a wider range of patients. Because of this feature, Tezepelumab was granted the 
breakthrough therapy designation by the FDA in 2018 for the treatment of severe asthma patients 
with no eosinophilic phenotype. Tezepelumab’s phase 3 trial (NAVIGATOR) reached the primary 
endpoint in patients with severe uncontrolled asthma. Compared with the combination of placebo 
and standard of care (SoC), the combination of Tezepelumab and SoC significantly decreased 
patients' annual asthma exacerbation rate (AAER) at Week 52 (with statistical significance and 
clinical significance). In the subgroup of patients with baseline eosinophil count less than 300 
cells/μL, Tezepelumab also reached the primary endpoint. A similar decrease in AAER was also 
observed in the subgroup of patients with baseline eosinophil count less than 150 cells/μL, 
indicating that Tezepelumab can significantly reduce the acute exacerbation of severe asthma in a 
wide range of patients. Also, there was no clinically significant difference in the tolerability of 
Tezepelumab compared with the safety of the placebo group.1 
 


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In November 2020, AZ and Amgen announced that Tezepelumab reached the primary endpoint in 
the phase 3 study (NAVIGATOR) in patients with severe uncontrolled asthma. Compared with 
the combination of placebo and standard of care (SoC), the combination of Tezepelumab and SoC 
significantly decreased patients' annual asthma exacerbation rate (AAER) at Week 52 (with 
statistical significance and clinical significance). In the subgroup of patients with baseline 
eosinophil count less than 300 cells/μL, Tezepelumab also reached the primary endpoint. A similar 
decrease in AAER was also observed in the subgroup of patients with baseline eosinophil count 
less than 150 cells/μL, indicating that Tezepelumab can significantly reduce the acute exacerbation 
of severe asthma in a wide range of patients. Also, there was no clinically significant difference in 
the tolerability of Tezepelumab compared with the safety of the placebo group. 1  
1.2 
Summary of Nonclinical Studies 
1.2.1 
Nonclinical Pharmacology 
1.2.2 
Nonclinical Safety Pharmacology 
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1.2.3 
Nonclinical Pharmacokinetics and Product Metabolism in Animals 
1.2.4 
Nonclinical Toxicology 
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1.3 
Summary of Clinical Experience 
1.3.1 
Clinical Safety 
As of 09 May 2023 (data cut-off date), Hengrui Pharmaceuticals Co., Ltd has initiated 3 clinical 
studies (two Phase I and one Phase II) of AIO-001 (SHR-1905). Two clinical studies (SHR-1905-
101 and SHR-1905-201) were performed in China, and the other one (SHR-1905-I-101-AUS) was 
conducted in Australia. A total of 147 subjects received AIO-001, including 100 healthy subjects 
were exposed to AIO-001 (n=80) or placebo (n=20) at a single dose of 50 - 600 mg, 12 patients 
with mild asthma received a single subcutaneous dose of 200 mg AIO-001 (n=8) or placebo (n=4), 
24 patients with moderate or severe asthma received a single subcutaneous dose of 100/600 mg 
AIO-001 (n=16) or placebo (n=8), and 11 patients with severe asthma received multiple 
subcutaneous doses of 
 AIO-001 or placebo.1 
 
Among 100 healthy subjects exposed to AIO-001/placebo, 63 subjects (63.0%) experienced 
TEAEs, which occurred in ≥5% of the subjects included blood thyroid stimulating hormone 
increased, blood uric acid increased and blood bilirubin increased (7.0% each), bilirubin 
conjugated 
increased, 
aspartate 
aminotransferase 
increased, 
headache 
and 
alanine 
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aminotransferase increased (6.0% each), and haemoglobin decreased and blood creatine 
phosphokinase increased (5.0% each).1 
 
Among 47 patients with asthma who were exposed to AIO-001/placebo, the incidence of TEAEs 
was 31.9% (15/47), including alanine aminotransferase increased, blood creatine phosphokinase 
increased, blood uric acid increased and triiodothyronine free increased (2 subjects each), and the 
others were seen in single subjects.1 
 
All TEAEs above were mild or moderate in severity, which can be controlled with or without 
medical intervention. No subjects experienced serious adverse events (SAEs) or fatal TEAEs. No 
subjects withdrew from studies due to TEAEs.1 
1.3.2 
Pharmacokinetics and Immunogenicity in Humans 
Following a single SC injection of AIO-001 from 50 mg to 600 mg in Australian healthy subjects 
(SHR-1905-I-101-AUS) (N=40), the serum concentrations of AIO-001 increased along with the 
increased dose level. 
 
 
To explore the dose proportionality of AIO-001 in tested dose range (50 mg to 600 mg) in healthy 
subjects, Cmax, AUC0-last, and AUC0-inf were further examined against dose using power model. 
The slopes of 
 indicating 
that the exposure of AIO-001rs to increase in a slightly greater-than-dose-proportional manner 
with increased dose from 50 mg to 600 mg. The slopes of 
 
, indicating that the exposure of AIO-001 appears to increase in a 
slightly greater-than-dose-proportional manner with increased dose from 50 mg to 600 mg.1   
 
In all the Australian healthy subjects (N=50) in this study, treatment-induced ADA positive 
subjects were reported in both placebo group 
 and AIO-001 treatment group 
 
. Treatment-induced ADA positive subjects with AIO-001 were:
 
. There was  
 subject in the placebo group who had 
a pre-existing ADA sample at baseline. The observed earliest time point of treatment-induced 
ADA after AIO-001 Treatment started from 
 in the 200 mg and 600 mg group, and 
 in 
the 400 mg group. There was no obvious effect observed in corresponding PK concentrations in 
all the ADA positive subjects.1 
1.4 
Study Rationale 
Existing safety data from China and Australia Phase 1 single ascending dose studies of AIO-001 
are from subjects who received SC injections with a 100 mg/ml formulation requiring multiple 
1 ml injections to achieve doses up to 600 mg. The study in China also included subjects with 
asthma.  
 
In a proposed phase 2 trial, Aiolos Bio will be utilizing a new formulation and subjects will receive 
two SC injections of 1.1 ml at AIO-001 concentration of 182 mg/ml to achieve a 400 mg dose. 
The primary purpose of the present study is to compare the safety, tolerability, and PK parameters 
of 400 mg of AIO-001 as the new formulation (Formulation B) with a 400 mg dose of AIO-001 
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administered as the original formulation (Formulation A) in healthy volunteers. Pharmacokinetic 
parameters and other primary and secondary endpoints will be described and compared 
quantitatively. 
1.4.1 
Rationale for the Dose  
1.4.2 
Rationale for the Study Population 
A healthy volunteer population has been selected for the study because healthy subjects with no 
concomitant diseases and using no concomitant medications represent a homogenous population 
allowing for proper evaluation of the PK, safety, and tolerability of a drug without confounding 
factors.  
The effects of AIO-001 on fertility, pregnancy and lactation in human have not been investigated 
so far. Since there have been no studies with pregnant women, it is uncertain whether there is 
human fetal risk associated with the use of AIO-001. Therefore, only male and non-pregnant, non-
lactating female subjects will be included in the study. Non-sterile males and females of 
childbearing potential will be included if they use appropriate methods of contraception. 
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1.5 
Benefit/Risk Assessment 
The inclusion and exclusion criteria have been chosen to select subjects who are known to be free 
from any significant illness, history of autoimmune diseases, and from any condition that could 
impact their safety or interfere with meeting the study objectives. The proposed safety screening 
and monitoring assessments are deemed to be sufficient to monitor potential risks of AIO-001 
administration. There is no anticipated therapeutic benefit for the healthy subjects in this study. 
 
So far, as mentioned before, a total of 147 subjects received AIO-001/placebo, including 100 
healthy subjects were exposed to AIO-001 or placebo (n=20) at a single dose of 50 – 600 mg, 12 
patients with mild asthma received a single subcutaneous dose of 200 mg AIO-001 (n=8) AIO-
001 or placebo (n=4), 24 patients with moderate or severe asthma received a single subcutaneous 
dose of 100/600 mg AIO-001 (n=16) or placebo (n=8), and 11 patients with severe asthma received 
multiple subcutaneous doses of 
 mg AIO-001 or placebo. 
 
 
  
 
All the safety data obtained so far was with AIO-001 using Formulation A. Although this is the 
first time the new Formulation (Formulation B) will be tested in humans, this study will evaluate 
Formulation B, and a similar safety profile is expected. All excipients are generally recognised as 
safe. The information obtained in this study may help in the treatment of future patients with 
severe, uncontrolled asthma. 
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2. 
Objectives 
Primary objective: 
• To compare the safety and tolerability of 400 mg of AIO-001 as the new formulation 
(Formulation B) with a 400 mg dose of AIO-001 administered as the original formulation 
(Formulation A) in healthy volunteers. 
Secondary objectives: 
• To evaluate the PK of a single SC AIO-001 dose of Formulation A in healthy volunteers. 
• To evaluate the PK of a single SC AIO-001 dose of Formulation B in healthy volunteers. 
• To evaluate the immunogenicity of AIO-001 in healthy volunteers. 
3. 
Endpoints 
Primary endpoints: 
• Adverse events (AEs), vital signs measurements (blood pressure, heart rate, respiratory 
rate, and tympanic temperature), 12-lead electrocardiogram (ECG) recordings, physical 
examinations, and clinical laboratory test results, including hematology, biochemistry, 
and urinalysis.  
Secondary endpoints: 
• PK endpoints may include but are not limited to AIO-001 concentrations, AUC0-last, 
AUC0-inf, Cmax, Tmax, T½ . 
• Incidence of anti-drug antibody (ADA) to AIO-001. 
4. 
Study Design 
This is an open-label single dose, parallel group, 24-week phase 1 study in 16 healthy subjects. 
The study is designed to evaluate and compare the safety, tolerability, pharmacokinetics, and 
immunogenicity of subcutaneously (SC) administered AIO-001 at a total dose of 400 mg using 
two different formulations (original Formulation A and new Formulation B) in 16 healthy 
volunteers (8 receiving each formulation). 
The study will include a screening visit from Day -28 to Day -2 (Visit 1). Eligible subjects will be 
admitted to the clinical site on Day -1 and will be confined until completion of the assessments on 
Day 3 (Visit 2). Subjects will return to the clinical site for outpatient visits for study assessments 
and laboratory tests as described in the Table 1. Schedule of Assessments.  
A staggered dosing schedule will be used for dosing of each cohort and will include 2 sentinel 
subjects (1 for each formulation) dosed initially, and the remaining 14 subjects dosed at least 
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The total duration of study participation for each subject from screening through the EOS is 
anticipated to be approximately 28 weeks. 
5. 
Study Population 
5.1 
Number of Subjects 
It is planned to enroll up to 16 healthy subjects for participation in this study. The proposed number 
of subjects is in line with the sample sizes commonly used in clinical studies of this nature and is 
considered sufficient to achieve the study objectives. 
5.2 
Inclusion Criteria  
Subjects must meet all of the following criteria to be included in the study: 
1. Able to understand the study procedures and provide signed informed consent to participate 
in the study.  
2. Male or female. 
3. Non-smokers. Light smokers (no more than 5 cigarettes daily [approximately 50 to 60 mg of nicotine 
per day], or products with equivalent amount of nicotine within 3 months prior to screening) may be 
permitted. 
4.  ≥18 and ≤55 years of age. 
5. BMI >18.5 and <32.0 kg/m2 and body weight ≥45.0 kg. 
6. Healthy subjects as defined by: 
a. the absence of clinically significant illness and surgery within 4 weeks prior to dosing. 
b. the absence of clinically significant history of neurological, endocrine, cardiovascular, 
respiratory, hematological, immunological, psychiatric, gastrointestinal, renal, hepatic, and 
metabolic disease. 
7. Female subjects of non-childbearing potential must be: 
a. post-menopausal (spontaneous amenorrhea for at least 12 months prior to dosing) with 
confirmation by documented FSH levels ≥40 mIU/mL; or 
b. surgically sterile (bilateral oophorectoy, bilateral salpingectomy, or hysterectomy) at least 
3 months prior to dosing. 
8. Sexually active female subjects of childbearing potential who are sexually active with the 
opposite sex and non-sterile male subjects must be willing to use an acceptable contraceptive 
method throughout the study as detailed in section 7.1. 
5.3 
Exclusion Criteria 
Subjects to whom any of the following applies will be excluded from the study: 
1. Any clinically significant abnormal finding at physical examination at screening. 


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2. Clinically significant abnormal laboratory test results or positive serology test results for 
HBsAg, HCV antibody, or HIV antigen and antibody, or QuantiFERON®-TB test at screening. 
3. Positive pregnancy test or lactating female subject. 
4. Positive urine drug screen or alcohol breath test. 
5. History of anaphylaxis, or severe allergy. 
6. Previous exposure to thymic stromal lymphopoietin antibody. 
7. Known allergic reactions to thymic stromal lymphopoietin antibody drugs and their excipients, 
or other biologic products or to any excipient in the formulation. 
8. Estimated Glomerular Filtration Rate (eGFR, using CKD-EPI Creatinine Equation) < 
90 ml/min/1.73m2 during screening or baseline visit.   
9. Clinically significant ECG abnormalities (e.g., For male QTcF > 450msec, for female QTcF > 
470msec) or vital signs abnormalities (systolic blood pressure lower than 90 or over 
140 mmHg, diastolic blood pressure lower than 40 or over 90 mmHg, or heart rate less than 
40 or over 100 bpm) at screening. 
10. History of drug abuse within 1 year prior to screening. 
11. History of alcohol abuse within 1 year prior to screening or regular use of alcohol within 
6 months prior to screening that exceeds 10 units for women or 15 units for men of alcohol per 
week (1 unit = 375 mL of beer 3.5%, or 100 mL of wine 13.5%, or 30 mL of distilled alcohol 
40%). 
12. Use of medications within the timeframes specified in section 7.2. 
13. Previous participation in a clinical trial of AIO-001. 
14. Dosed in a clinical research study involving the administration of an investigational or 
marketed drug or device within 30 days prior to dosing, administration of a biological product 
in the context of a clinical research study within 90 days or 5 half-lives (whichever is longer) 
prior to dosing, concomitant participation in an investigational study involving no drug or 
device administration. 
15. Donation of plasma within 7 days prior to dosing or donation or loss of 500 mL or more of 
whole blood within 4 weeks prior to dosing. 
16. Any reason which, in the opinion of the Investigator, would prevent the subject from 
participating in the study. 
17. Intolerance to venipuncture. 
18. Any scars, tattoos, etc., at the injection site. 
19. More than 5 cigarettes daily (or products with equivalent amount of nicotine) for 3 months 
prior to screening and unable to abstain from smoking during the trial. 


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5.4 
Subject Withdrawal and Replacement 
Subjects will be advised that they are free to withdraw from the study at any time. Over the course 
of the study, the Sponsor and the Investigator or designee may withdraw any subject from the 
study for one of the reasons described below; subject withdrawal will be done in accordance with 
the local guidance at the clinical site: 
• Safety reason; 
• Non-compliance with protocol requirements; 
• Significant protocol deviation; 
• Positive pregnancy test, drug screen or alcohol breath test. 
Subjects who withdraw prior to dosing may be replaced automatically. In the event that the number 
of drop-outs exceeds initial expectations, subjects who withdraw or are withdrawn from the study 
after dosing, for reasons other than safety and tolerability, may be replaced after consultation 
between the Investigator and the Sponsor. In such case, the total number of subjects dosed will 
remain within a maximum of 10 subjects for Groups A and B. 
Subjects who withdraw or are withdrawn on Day-1 to Day 3 will be asked to remain at the clinic 
until the Investigator or designee agrees that the subject is fine and can be discharged. A PK blood 
sample may be collected at the time of withdrawal if deemed required by the Investigator. End of 
study EOS/ET procedures will be performed at the time of withdrawal from the study or as soon 
as possible thereafter. 
6. 
Study Treatments 
6.1 
Drug Supplies and Accountability 
It is the responsibility of the Sponsor to ensure that study drugs provided for this study are 
manufactured under Good Manufacturing Practice (GMP) and are suitable for human use. The 
Sponsor is responsible to ship a sufficient amount of dosage units to allow the clinical site to 
maintain an appropriate supply for the study. 
The study drugs will be stored at the clinical site as per GCP and local requirements in a locked, 
temperature-controlled room with restricted access. IP vials and any secondary packaging (if 
applicable) will be labeled according to applicable local regulations. 
For full detail on study drug dispensation, storage, handling, and disposal, please refer to the 
Pharmacy Manual. 
6.2 
Identification of Treatments 
Sixteen healthy volunteers (8 receiving each formulation) will receive one of the following 
treatments, according to the randomization scheme: 
Group A: 
400 mg of 100 mg/ml AIO-001 (as Formulation A), AIO-001 antibody as the active 
ingredient
 
 
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Group B: 
400 mg of 182 mg/ml AIO-001 (as Formulation B): AIO-001 antibody as the active 
ingredient
 
 
6.3 
AIO-001 Randomization and Blinding 
This study will be an open-label study due to the objective nature of the data. Subjects will be 
administered each treatment according to the block randomization scheme. Randomization will be 
performed on Day 1 by the pharmacy personnel as per site standard process and will be conducted 
manually via a printed randomization schedule/scheme provided to site. 
6.4 
Study Drug Administration 
The study drug (AIO-001 injection Formulation A or B) will be injected subcutaneously on Day 1. 
For safety reasons, subjects will be required to remain seated or semi-reclined during dosing. 
• Formulation A: Subjects from Group A will receive 400 mg of 100 mg/ml AIO-001, given 
as 4 X 1.0 ml subcutaneous injections administered within approximately 5 minutes. The 
four injection sites are two injection sites on each upper arm separated from each other by 
at least 5 cm. 
• Formulation B: Subjects from Group B will receive 400 mg of 182 mg/ml AIO-001 given 
as 2 X 1.1 ml subcutaneous injections administered within approximately 5 minutes. The 
2 injection sites are one on each upper arm. 
 
IP should not be administered on skin disfigurements or lesions such as scars, tattoos, wounds, 
bruises, redness and hardening. Please refer to Pharmacy Manual for more details.  
The start time of the first injection will be called “0” hour. 
7. 
Study Restrictions 
7.1 
Contraception 
Female subjects who are of non-childbearing potential will not be required to use contraception. 
Female subjects of childbearing potential who are sexually active with a non-sterile male partner 
(sterile male partners are defined as men vasectomized for at least 3 months prior to dosing) must 
be willing to use one of the following acceptable contraceptive methods throughout the study and 
for 5 half-lives +1 month after the last dose: 
• Simultaneous use of hormonal contraceptive (e.g., oral, depot injection, implant, vaginal 
ring, intrauterine device) or non-hormonal intrauterine device used for at least 4 weeks 
prior to dosing (must agree to use the same contraceptive throughout the study) and 
condom for the male partner. 
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• Bilateral tubal occlusion or ligation, and condom for the male partner, started at least 21 
days prior to dosing. 
• Abstinence 
Female subjects of childbearing potential must not donate ova during the study and for at least 90 
days after the dosing. 
Male subjects who are not surgically sterile for at least 3 months prior to dosing, and who are 
sexually active with a female partner of childbearing potential must be willing to use one of the 
following acceptable contraceptive methods from the first dose and for 5 half-lives +1 month after 
the last dose: 
• Simultaneous use of condom and hormonal contraceptive (e.g., oral, patch, depot 
injection, implant, vaginal ring, intrauterine device) or non-hormonal intrauterine device 
used for at least 4 weeks prior to sexual intercourse for the female partner. 
• Abstinence 
Male subjects (including men who have had a vasectomy) with a pregnant partner must agree to 
use a condom from the first dose and for 5 half-lives +1 month after the last dose. Male subjects 
must also be willing not to donate sperm and for 5 half-lives +1 month after the last dose. 
Sexual abstinence is considered a highly effective method only if defined as refraining from 
heterosexual intercourse during the entire period of risk associated with the study treatment. The 
reliability of sexual abstinence needs to be evaluated in relation to the duration of the study and 
the preferred and usual lifestyle of the subject.  
7.2 
Concomitant Medications  
Subjects will be required to avoid using any non-study necessitated medications for the 
timeframes specified below: 
• Depot injection or implant of any drug (other than hormonal contraceptives) for 3 months 
prior to dosing. 
• Prescription medications for 4 weeks prior to dosing. 
• Any vaccine, including COVID-19 vaccine and live attenuated vaccination, for 30 days 
prior to dosing. 
• Over-the-counter (OTC) medications and natural health products (including herbal 
remedies such as St. John’s wort, homeopathic and traditional medicines, probiotics, food 
supplements such as vitamins, minerals, amino acids, essential fatty acids, and protein 
supplements used in sports) for 14 days prior to dosing (with the exception of the 
occasional use of acetaminophen up to 2 g daily), throughout the study. 
No concomitant medications will be allowed during the study, with the exception of hormonal 
contraceptives, medications required for the medical management of an AE, and medications 


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exempted by the Investigator on a case-by-case basis that are judged unlikely to affect the PK 
profile of the study drug or subject safety (e.g., topical drug products without significant systemic 
absorption). 
If a vaccination is required for any reason, it must first be discussed with and exempted by the 
Investigator on a case-by-case basis to ensure that it does not compromise the PK profile of the 
study drug or the subject safety. 
Medications taken by subjects before dosing will be documented as prior medications and 
medications taken by subjects after dosing through the last study day will be documented as 
concomitant medications. Any prior or concomitant medication use, other than the allowed 
medications stated above, will be reviewed, and evaluated on a case-by-case basis by the 
Investigator to determine if they affect a subject’s eligibility or continued participation in the study, 
or for potential impact on the study results. 
7.3 
Drugs, Nicotine, and Alcohol 
Subjects will be required to abstain from: 
• Drugs of abuse and illicit use of recreational drugs from screening through study end; 
• Any nicotine product within 48 hours prior to admission until the end of inpatient stay 
(Day 3), and 2 hours prior to each outpatient visit. No more than equivalent of 5 cigarettes 
per day will be allowed.  
• Alcohol-based products from 24 hours prior to admission until discharge. 
7.4 
Diet 
Subjects will be required to abstain from: 
• Food containing poppy seeds from 24 hours prior to admission. 
While confined at the study site, subjects will receive a standardized diet at scheduled times that 
will not conflict with other study-related activities. 
7.5 
Posture and Physical Activity 
For safety reasons, subjects will be required to remain seated or semi-reclined and avoid lying 
down or sleeping for the first 4 hours after the start of study treatment administration. 
Failure of subjects to comply with these requirements does not constitute a deviation from the 
protocol if it is medically necessary, required for procedures, or to go to the bathroom. When 
appropriate, subjects will be accompanied by a staff member while walking.  
Subjects will be required to refrain from strenuous exercise at least 3 days before admission and 3 
days prior each follow-up visit. 


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8. 
Study Procedures 
Subjects must provide written informed consent prior to initiation of any study procedures. 
Unless otherwise specified, study procedures will be conducted in accordance with local guidance 
at the clinical site. From screening through the EOS, subjects will undergo study procedures at 
pre-defined times as specified in Table 1. 
Schedule of Assessments and as described in 
sections 8.1 and 8.2. 
Every effort will be made to schedule and perform the procedures within the specified time 
window, giving consideration to appropriate posture conditions, practical restrictions, and other 
procedures to be performed at the same time point. 
Unless otherwise specified or for subject safety, when the timing of multiple procedures coincides, 
the following priority order should be adhered to, whenever possible: vital signs, ECGs, blood 
samples for PK/ADA analysis, clinical laboratory samples, physical examination. 
8.1 
Pharmacokinetic and Immunogenic Assessments 
8.1.1 
Pharmacokinetic and Immunogenic Blood Sample Collection and Processing 
Blood samples for PK and ADA analysis will be collected via an intravenous catheter or by direct 
venipuncture at the time points indicated in Table 1. Schedule of Assessments. Applicable time 
windows for PK and ADA blood samples are defined in the table below.  
Table 2 
Time Windows for PK and ADA Blood Samples 
Day 
Hour 
Visit 
Number 
Timepoints for 
Serum PK 
Timepoints for 
Serum ADA 
Time 
window 
1 
Pre-dose 
2 
X 
X 
Within 60 
minutes 
12 hours post-dose 
 
X 
 
±30 minutes 
2 
24 hours post-dose 
 
X 
 
±30 minutes 
3 
48 hours post-dose 
 
X 
 
±30 minutes 
4 
72 hours post-dose 
3 
X 
 
±2 hour 
5 
96 hours post-dose 
4 
X 
 
±2 hour 
6 
120 hours post-dose 
5 
X 
 
±2 hour 
8* 
168 hours post-dose 
6 
X 
X 
± 4 hours 
11* 
240 hours post-dose 
7 
X 
 
±8 hours 
15* 
336 hours post-dose 
8 
X 
X 
±8 hours 
22* 
504 hours post-dose 
9 
X 
X 
±1 day 
29* 
672 hours post-dose 
10 
X 
X 
±1 day 


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43* 
1008 hours post-
dose 
11 
X 
 
±2 days 
57* 
1344 hours post-
dose 
12 
X 
X 
±2 days 
71* 
1680 hours post-
dose 
13 
X 
 
±2 days 
85* 
2016 hours post-
dose 
14 
X 
X 
±2 days 
113* 
2688 hours post-
dose 
15 
X 
 
±2 days 
141* 
3360 hours post-
dose 
16 
X 
X 
±2 days 
169* 
4032 hours post-
dose 
17 
X 
X 
±2 days 
* Beginning at Day 8, for operational feasibility surrounding Public Holidays, an additional +1 day visit window 
may be granted with sponsor approval. 
The planned volume of blood to be collected for this study, including that collected for eligibility 
and safety purposes, should not exceed 550 mL. Additional tests or blood draws could be 
performed, if deemed required by the Investigator or study staff. 
Procedures for collection, processing, and shipping of PK blood samples will be detailed in the 
Bioanalytical Laboratory Manual. 
Serum concentrations of the AIO-001, as well as anti-drug antibodies will be determined using 
validated analytical methods. Details of the analytical methods will be provided in a separate 
document. 
8.2 
Safety and Tolerability Assessments 
Subjects will be monitored throughout the study by the clinical staff for AEs. Adequate medical 
surveillance will be assured during the confinement period and the Investigator or designee will 
be available on call at all times. If necessary, the Investigator or designee at the clinical site or a 
healthcare professional in a nearby hospital will administer treatment for any AE(s). A crash cart 
or emergency bag containing the necessary rescue material and appropriate medications will be 
available in the clinic to allow rapid intervention in case of emergency. 
Safety parameters will be assessed by the Investigator or designee, using the clinical site 
acceptance ranges as suggested guidelines in making the medical assessment. 
For eligibility purposes, abnormal vital signs measurements or clinical laboratory test results may 
be repeated once if an abnormal result is observed at the initial reading. Moreover, abnormalities 
found in the ECG may need to be confirmed by repeated measurements. In the event that the 
participation of a subject in the study is delayed and some screening procedures had been 
performed outside of the prescribed screening window, outdated screening procedures can be 
repeated. 


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Safety assessments scheduled during the study will be repeated according to the clinical site local 
guidance or upon request from the Investigator or designee. Any abnormal repeated measurement 
will be evaluated and repeated, if judged necessary. Further action may be taken upon the 
Investigator or designee’s request. 
8.2.1 
Physical Examination 
Physical examinations will be performed at the times specified in Table 1. Schedule of 
Assessments A complete physical examination will include assessments of the following: head, 
eyes, ears, nose, throat (HEENT), neck, chest, lungs, abdomen, musculoskeletal, dermatological, 
cardiovascular/peripheral vascular, and general neurological examination. 
A brief physical examination will include assessments of the following: HEENT, chest, lungs, 
abdomen, dermatological, cardiovascular/peripheral vascular, and areas of note elicited from the 
subject. In the event of significant skin reactions appropriate informal photographs may be taken 
of the affected area. 
8.2.2 
Body Measurements 
Body measurements will be performed at the times specified in Table 1. Schedule of 
Assessments and will include body weight and height measurements, as well as BMI calculation. 
8.2.3 
Injection Site Evaluation 
Local reactions at the injection site will be evaluated by a trained observer before dosing on Visit 
2 (Day 1 and approximately 1, 12, 24 hours (Day 2), 48 hours (Day 3)), 72 hours (Day 4, Visit 3) 
and 96 hours (Day 5, Visit 4) post-dose. A global severity rating for injection site reactions will be 
included in the assessment of AEs. An injection site examination post-dose will consist of 
examining the injection site and making note of the presence of any skin reactions, disturbances, 
or discolorations which will be graded and reported as per the table below: 
Table 3. 
Injection Site Reaction Scores 
Local Reaction to 
Injectable Product 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Pain 
Does not interfere with 
activity 
Repeated use of non-
narcotic pain reliever > 
24 hours or interferes 
with activity 
Any use of narcotic pain 
reliever or prevents daily 
activity 
Tenderness 
Mild discomfort to 
touch 
Discomfort with 
movement 
Significant discomfort at rest 
Erythema/Redness* 
2.5-5 cm 
5.1-10 cm 
> 10 cm 
Induration/Swelling** 
2.5-5 cm and does not 
interfere with activity 
5.1- 10 cm or interferes 
with activity 
> 10 cm or prevents daily 
activity 


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* In addition to grading the measured local reaction at the greatest single diameter, the measurement should be 
recorded as a continuous variable. 
** Induration/Swelling should be evaluated and graded using the functional scale as well as the actual 
measurement. 
8.2.4 
Vital Signs 
Blood pressure, heart rate, respiratory rate, and tympanic temperature will be measured after the 
subjects have been resting for at least 5 minutes in a sitting position at the times specified in 
Table 1. Schedule of Assessments. On Day 1 (Visit 2), vital signs will be performed: within 60 min 
of dosing, at 2, 4, 8, and 12 hours (±30 min) post-dose. 
8.2.5 
12-lead ECG 
Standard 12-lead ECG will be recorded after the subjects have been resting for at least 5 minutes 
in a semi-recumbent or supine position at the times specified in Table 1. Schedule of Assessments. 
The examination includes: heart rate, PR interval, QT interval, corrected QT (QTcF using 
Fridericia’s formula) interval and QRS, and should be performed by qualified personnel. 
8.2.6 
Laboratory Assessments 
Blood and urine samples for clinical laboratory assessments will be collected according to the 
clinical site SOPs at the times specified in Table 1. Schedule of Assessments. The clinical 
laboratory assessments to be performed are listed in the table below. 
Table 4. 
Clinical Laboratory Assessments 
Biochemistry1 
Hematology 
Urinalysis 
Albumin 
Alkaline phosphatase 
ALT 
AST 
Calcium 
Chloride 
Creatine kinase 
Creatinine 
eGFR (calculated using the CKD-
EPI equation) 
GGT 
Glucose 
IgE 
Lactate dehydrogenase  
Phosphorus 
Potassium  
Sodium 
Hematocrit 
Hemoglobin 
MCH 
MCHC 
MCV 
Platelet count 
RBC count 
RBC distribution width 
WBC count and differential: 
• 
Basophils 
• 
Eosinophils 
• 
Lymphocytes 
• 
Monocytes 
• 
Neutrophils 
Bilirubin 
Blood (occult) 
Color and appearance 
Glucose 
Ketones 
Leukocyte esterase 
Nitrite 
pH 
Protein 
Specific gravity 
Urobilinogen 
Microscopic examination (in 
the event of abnormal 
findings) 
Coagulation 
Endocrinology 


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Total, direct and indirect bilirubin 
Total protein 
Troponin-I 
Urea 
Uric acid 
 
Lipid profile: 
Total cholesterol 
Triglycerides 
aPTT 
PT 
INR 
FSH 
TSH 
T3 
T4 (free) 
Serum pregnancy test 
Urine pregnancy test 
Serology 
Drug and alcohol screens 
Hormone panel - females 
only 
HBsAg 
HCV antibody 
HIV antigen/antibody 
QuantiFERON-TB 
COVID screening1 
 
Amphetamines/methamphetamines (MET) 
Barbiturates 
Benzodiazepines 
Cocaine 
MDMA 
Methadone 
Opiates 
PCP 
THC 
Alcohol breath test 
FSH (post-menopausal 
females only) 
Serum pregnancy test1 
Urine pregnancy test1 
1 Note: Biochemistry assessment will be done after 8 hours fasting period. Serum pregnancy test will be performed 
at screening and at EOS. A urine pregnancy test will be performed on Day -1 (Visit 2). COVID screening will be 
performed according to clinic site’s procedure. 
2 Note: IgE is not an eligibility determining assessment and therefore results are not needed prior to eligibility 
confirmation. 
Abbreviations: ALT = alanine aminotransferase; aPTT = activated partial thromboplastin time; AST = aspartate 
aminotransferase; eGFR = estimated glomerular filtration rate; FSH = follicle-stimulating hormone; GGT = 
gamma-glutamyl transferase; HBsAg = hepatitis B surface antigen; HCV = hepatitis C virus; HIV = human 
immunodeficiency virus; IgE = immunoglobulin E; INR = international normalized ratio; LDH=Lactate 
dehydrogenase; MCH = mean cell hemoglobin; MCHC = mean cell hemoglobin concentration; MCV = mean cell 
volume; MDMA = 3,4- methylenedioxymethamphetamine; MDRD = modification of diet in renal disease; PCP = 
phencyclidine; PT = prothrombin time; RBC = red blood cell; T3 = triiodothyronine; T4 = thyroxine; THC = 
tetrahydrocannabinol; TSH = thyroid-stimulating hormone; WBC = white blood cell. 
8.3 
Adverse Events 
8.3.1 
Definition of Adverse Event 
An AE is defined as any untoward medical occurrence in a patient or clinical trial subject 
administered a pharmaceutical product and which does not necessarily have a causal relationship 
with the treatment. An AE can, therefore, be any unfavorable and unintended sign (including an 
abnormal laboratory finding, for example), symptom, or disease temporally associated with the 
use of a medicinal (investigational) product, whether or not considered related to the medicinal 
(investigational) product. 


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8.3.2 
Recording of Adverse Events 
AEs will be recorded and evaluated for their seriousness, severity, and relationship to the study 
drug. AEs will be collected and documented from the time of signing the ICF and throughout the 
study. AEs will be followed-up until complete resolution, or until the Investigator or designee 
judges safe to discontinue follow-up. The severity of AEs and relationship to the study drug will 
be classified according to the sections below. 
8.3.3 
Assessment of Severity 
The severity of AEs will be described and documented using the following definitions: 
Table 5. 
Severity of Adverse Events 
Severity 
Description 
Mild (Grade 1) 
Awareness of signs and symptoms but are easily tolerated; are of minor irritant type; 
causing no limitations of usual activities. Signs or symptoms may require minor 
action. 
Moderate (Grade 2) 
Discomfort severe enough to cause some limitations of usual activities and may 
require action. 
Severe (Grade 3) 
Incapacitating with inability to carry out usual activities or significantly affects 
clinical status and requires specific action and/or medical attention. 
 
8.3.4 
Assessment of Relationship to Study Drug 
Each AE must be classified based on medical judgment and according to the following relationship 
categories: probable, possible, remote/unlikely, and unrelated. The definitions for the relationship 
categories are as follows: 


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Table 6. 
Adverse Event Relationship Categories 
Category 
Description 
[Related] 
Probable 
(must have first three 
points) 
This category applies to AEs that are considered, with a high degree of certainty, to 
be related to the investigational product. An AE may be considered probable, if: 
1. It follows a reasonable temporal sequence from the administration of the drug. 
2. It cannot be reasonably explained by the known characteristics of the subject’s 
clinical state, environmental or toxic factors or other modes of therapy 
administered to the subject. 
3. It disappears or decreases on cessation or reduction in dose (there are important 
exceptions when an AE does not disappear upon discontinuation of the drug, yet 
drug relatedness clearly exists; e.g. (1) bone marrow depression, (2) tardive 
dyskinesias). 
4. It follows a known pattern of response to the suspected drug.  
5. It reappears upon re-challenge. 
[Related] 
Possible 
(must have first two 
points) 
This category applies to AEs in which the connection with the investigational product 
administration appears unlikely but cannot be ruled out with certainty. An AE may be 
considered possible if, or when: 
1. It follows a reasonable temporal sequence from the administration of the drug. 
2. It may have been produced by the subject’s clinical state, environmental or toxic 
factors, or other modes of therapy administered to the subject. 
3. It follows a known pattern of response to the suspected drug. 
Remote/Unlikely 
(must have first two 
points) 
In general, this category is applicable to an AE that meets the following criteria: 
1. It does not follow a reasonable temporal sequence from the administration of the 
investigational product. 
2. It may readily have been produced by the subject’s clinical state, environmental 
or toxic factors, or other modes of therapy administered to the subject. 
3. It does not follow a known pattern of response to the suspected drug. 
4. It does not reappear or worsen when the investigational product is 
re-administered. 
Unrelated 
This category is applicable to AEs that are judged to be clearly and incontrovertibly 
due only to extraneous causes (disease, environment, etc.), and do not meet the 
criteria for medication relationship listed under remote/unlikely, possible, or 
probable. 
 
8.3.5 
Definition of Serious Adverse Event 
A SAE is defined as any untoward medical occurrence that, at any dose: 
• Results in death; 
• Is life-threatening; 
• Requires inpatient hospitalization or prolongation of existing hospitalization; 
• Results in persistent or significant disability/incapacity; 
• Is a congenital anomaly/birth defect, or; 
• Is otherwise considered to be an important medical event that may not be immediately life-
threatening or result in death or hospitalization but may jeopardize the patient or may require 


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intervention to prevent one of the other outcomes listed in the definition above. Medical and 
scientific judgement should be exercised in deciding whether an event should be considered as 
an Important Medical Event. Examples of such medical events include: 
o Allergic bronchospasm requiring intensive treatment in an emergency room or at home; 
o Blood dyscrasias or convulsions that do not result in hospitalization; 
o Development of drug dependency or drug abuse. 
8.3.6 
Reporting of Adverse Events 
8.3.6.1 
Serious Adverse Event Reporting to the Sponsor 
Any SAE will be reported to the Sponsor (or representative) within 24 hours of learning of the 
event. The notification must be directed to: 
Aiolos Bio Clinical Safety 
Email: clinical.safety@aiolosbio.com 
Information on SAEs will be recorded on the SAE Report Form. Blank copies are included in the 
study Investigator’s file. It is not acceptable for the Investigator to send photocopies of the patient’s 
medical records to the Sponsor company or its representative in lieu of completion of the 
appropriate AE eCRF page or SAE Report Form. However, there may be instances when copies 
of medical records for certain cases are requested by the Sponsor or its representative. In this 
instance, all subject identifiers will be blinded on the copies of the medical records before 
submission to the Sponsor or its representative.  
The completed SAE Form and SAE cover sheet should be sent via e-mail immediately upon 
completion to Aiolos Bio Clinical Safety. If the Investigator does not have all information 
regarding an SAE, he/she will not wait to receive additional information before completing and 
sending the form. 
Additional relevant information or clinical follow-up should be sent via e-mail to Aiolos Bio 
Clinical Safety as soon as it becomes available. The Investigator should follow the subject with 
the event until resolution or stabilization of the condition. Follow-up reports (as many as required) 
should be completed and faxed/e-mailed following the same procedure above. 
A final report is required once the condition is resolved or stabilized and no more information 
about the event is expected. The final report should be completed and e-mailed following the same 
procedure above. 
The Investigator must keep a copy of all documentation related to the event in the clinical site files. 
If the Investigator learns of any SAE, including death, at any other time after a subject completes 
the study, and he/she considers the event reasonably related to the study drug, the Investigator will 
promptly notify the Sponsor/Medical Monitor. 


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8.3.6.2 
Serious Adverse Event Reporting to the IEC and Regulatory Agency(ies) 
It is the responsibility of the clinical site to report suspected, unexpected, serious adverse reactions 
(SUSARs) to the IEC responsible for the study per their policies. Report of fatal or life-threatening 
SUSARs must be made as soon as possible, but no later than 7 calendar days after first knowledge 
of the event. Report of SUSARs that are neither fatal nor life-threatening must be made as soon as 
possible, but no later than 15 calendar days after first knowledge of the event. 
The Sponsor (or representative) is responsible for notifying the regulatory agency(ies) of SUSARs 
observed during the study conduct per their regulations, as soon as possible but no later than 
7 calendar days after becoming aware of the information when fatal or life-threatening, or 
15 calendar days when neither fatal nor life-threatening. The Sponsor (or representative) is 
responsible to comply with any other applicable regulatory requirement(s) related to the reporting 
of SAE to other regulatory authority(ies). 
8.3.7 
Pregnancy 
In the event a dosed female subject or the female partner of a dosed male subject received at least 
one dose of the investigational product) becomes pregnant during or shortly after participation in 
the study, this pregnancy will be reported to the Sponsor (or representative) within 24 hours of 
first knowledge of the event. Any subject who becomes pregnant during the study will be 
immediately withdrawn. Follow-up information regarding the course and outcome of the 
pregnancy will be documented (after obtaining the consent of the female partner) as per site’s SOP. 
If the outcome of the pregnancy meets the criteria of reportable event, reporting of the event to the 
IEC responsible for the study and/or to applicable regulatory agency(ies) will be performed as per 
site’s SOP. 
8.4 
Premature Termination of the Study 
The study may be prematurely terminated by the Investigator following consultation with the 
Sponsor, by the Sponsor or by the regulatory authorities. Following a decision to discontinue the 
trial, the Investigator will promptly inform the active study subjects and the IEC responsible for 
this trial, stating the reasons for discontinuation of the study. It is the responsibility of the Sponsor 
(or representative) to report the premature termination of the study to the regulatory authority(ies), 
when required by the applicable regulatory requirement(s). 
9. 
Statistical Analyses 
A complete description of the statistical analyses to be performed on PK and ADA as well as safety 
and tolerability data will be presented in a statistical analysis plan (SAP). 
9.1 
Analysis Populations 
9.1.1 
Safety Population 
The safety population is defined as all subjects who receive at least one dose of the study drug. 


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9.1.2 
Pharmacokinetic Population 
The PK population will include all subjects who have at least 1 measured PK concentration 
following dosing. If any participants have incomplete data, protocol deviations, or clinical events 
that affect PK, a decision will be made on a case-by-case basis as to their inclusion in the analysis. 
9.1.3 
Immunogenicity Population 
The immunogenicity population will include all subjects who have received any amount of AIO-
001 and have at least one post-dose ADA measurement.  
9.2 
Pharmacokinetic Analysis 
Serum samples collected at the PK timepoints in Table 1 and Table 2 will be analyzed to determine 
concentrations of AIO-001. The actual start and end times of AIO-001 administration and the exact 
timing of blood sampling must be recorded in the CRF. Samples collected for analysis of AIO-001 
PK may additionally be used to evaluate safety and efficacy to address concerns arising during or 
after the study period or for further development and characterization of PK and immunogenicity 
assays. Instructions for handling of blood samples and shipping of samples for storage or analyses 
will be provided in the Laboratory Manual (provided separately). 
The following PK parameters will be calculated based on the PK population using standard 
non-compartmental analysis methods and may include but are not limited to: 
• AUC0-last: Area under the concentration-time curve from time zero until the last observed 
concentration 
• AUC0-inf: Area under the concentration-time curve from time zero to infinity (extrapolated) 
• Cmax: Maximal observed concentration 
• Tmax: Time when the maximal concentration is observed 
• T½ : Terminal elimination half-life 
9.3 
Immunogenicity Analysis 
Anti-AIO-001 antibodies will be evaluated in serum samples collected from all participants at the 
timepoints listed in Table 1/Table 2. Serum samples will be screened for antibodies binding to 
AIO-001 and the titer of confirmed positive samples will be reported. Incidence of treatment-
emergent ADA will be reported. Other analyses may be performed to further characterize the 
immunogenicity of AIO-001. Immunogenicity analyses may be conducted on PK samples 
collected at other timepoints, if deemed necessary. 
9.4 
Pharmacokinetic and Immunogenicity Statistical Analysis 
The pharmacokinetic profile of AIO-001 will be established using non-compartmental analysis of 
serum concentration data obtained at the timepoints outlined in the Schedule of Assessments. PK 
parameters including but not limited to AUC0-last, AUC0-inf, Cmax, Tmax, and T½  will be calculated 
if sufficient data are available for estimation, at the discretion of the pharmacokineticist. 


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AIO-001 serum concentrations and PK parameters will be presented in summary tables for each 
dose cohort and in listings, as appropriate. Individual and mean AIO-001 serum concentrations 
over time will be presented graphically. 
In addition, population pharmacokinetic modeling may be applied for PK analysis and will be 
reported separately. 
Individual and mean serum concentration versus time curves will be presented for both linear and 
semi-log scales. Descriptive statistics (arithmetic and geometric means, standard deviation [SD], 
CV%, minimum [Min], maximum [Max], and median) of the serum concentrations versus time 
will be presented as well for the PK parameters. 
Additional PK statistical analysis may be performed. 
9.5 
Safety and Tolerability Analysis 
Demographic parameters will be summarized descriptively. 
Safety and tolerability analysis will be performed for all subjects in the safety population. No 
inferential statistical analysis of safety data is planned. 
Safety and tolerability of AIO-001 will be evaluated through the assessment of AEs (i.e., 
seriousness, severity, relationship to the study drug, outcome, duration, and management), vital 
signs, 12-lead ECGs, clinical laboratory tests, and physical examinations. AEs will be coded using 
the latest version of the Medical Dictionary for Regulatory Activities (MedDRA). 
TEAEs will be tabulated by treatment. Changes from baseline values in vital signs, 12-lead ECGs, 
and clinical laboratory tests will be evaluated. Safety and tolerability data will be reported using 
descriptive statistics. 
9.5.1 
Interim Analyses 
 Interim analyses of preliminary data will be performed at Sponsor discretion. 
10. Data Collection 
The electronic source data capture system is the primary data collection instrument for the study. 
Collection screen in the electronic source data capture system will be utilized for the collection of 
all data. Data will be entered using the English language and should be kept current to enable the 
monitor to review the subjects’ status throughout the course of the study. 
Source data should be attributable, legible, contemporaneous, original, accurate, and complete. 
Source documents will be maintained in order to maintain data integrity. The Investigator and/or 
the clinical staff have the responsibility of ensuring the accuracy, completeness, legibility, and 
timeliness of the source data. 
Details on the data management process will be described in a data management plan (DMP). 


AIO-001-101 Solution for SC Injection Comparative Safety and PK 
Aiolos Bio, Inc. 
29-NOV-2023 
 
Confidential 
Page 45 of 47 
11. Regulatory Considerations and Quality Assurance 
11.1 IEC Approval of Protocol and Other Study Documents 
The Investigator(s) agree to provide the IEC with all appropriate documents, including a copy of 
the protocol/amendments, ICFs, advertising text (if any), Investigator’s Brochure (if any), and any 
other written information provided to study subjects. The trial will not begin until the Investigators 
have obtained the IEC favourable written approvals for the above-mentioned study documents. A 
properly executed written ICF shall be read, signed, and dated by each subject prior to entering the 
trial or prior to performing any study procedure. The original signed and dated ICF will be kept at 
the clinical site and a copy will be given to the subject. 
In the event that the protocol is amended, the revised protocol must be approved by the IEC prior 
to its implementation, unless the changes involve only logistical or administrative aspects of the 
trial. If a revised ICF is introduced during the study, each subject’s further consent must be 
obtained. The new version of the ICF must be approved by the IEC, prior to subsequently obtaining 
each subject’s consent. 
The Investigator and the Sponsor’s representative must sign the protocol and its amendments (if 
any) before initiating the study. 
It is the Sponsor’s responsibility to submit the protocol and its amendments (if any), and the ICFs 
to regulatory authorities when necessary. 
11.2 Compliance 
This study will be conducted in compliance with the protocol, GCP, the requirements of Directive 
2001/83/EC Annex I, as amended by Directive 2003/63/EC and Directive 2001/20/EC, and all 
applicable regulations, including the Federal Food, Drug and Cosmetic Act, U.S. applicable Code 
of Federal Regulations (title 21), and any IEC requirements relative to clinical studies. The study 
will also be conducted in compliance with the recommendations laid down in the most recent 
version of the Declaration of Helsinki, with the exception that registration of such Phase 1 trials in 
a publicly accessible database is not mandatory.  
11.3 Quality Assurance and Monitoring 
The Sponsor or its designee will perform the quality assurance and quality control activities of this 
study; however, responsibility for the accuracy, completeness, security, and reliability of the study 
data presented to the Sponsor lies with the Investigator generating the data. When applicable, 
quality assurance procedures will be performed according to the site SOPs. 
The study will be monitored according to the site monitoring plan and SOP to ensure that it is 
conducted and documented properly according to the protocol, GCP, and all applicable regulatory 
requirements. The Sponsor is entitled to access information about the status of the study and to 
review the original documents of the study. 
The Sponsor may arrange audits as part of the implementation of quality assurance to ensure that 
the study is being conducted in compliance with the protocol, SOPs, GCP, and all applicable 


AIO-001-101 Solution for SC Injection Comparative Safety and PK 
Aiolos Bio, Inc. 
29-NOV-2023 
 
Confidential 
Page 46 of 47 
regulatory requirements. In such case, audits will be independent of and separate from the routine 
monitoring and quality control functions. 
11.4 Confidentiality and Retention of Study Records 
This document contains trade secrets and commercial information that is confidential and may not 
be disclosed to third parties. Persons to whom this study protocol is disclosed must be informed 
that all the information herein is confidential and may not be further divulged. These restrictions 
will apply as well to all future communications if deemed privileged or confidential. Publication 
of the study results may only be allowed with written permission from the Sponsor. 
All information on a subject obtained during the conduct of the study will be kept confidential. 
Subjects will be identified by an anonymized identifier on all samples and study records provided 
to the Sponsor or designee. In compliance with ICH GCP, the Sponsor’s authorized 
representatives, monitor(s), auditor(s), IEC, and regulatory authority(ies) will be granted direct 
access to the subject’s original trial-related records for verification of clinical trial procedures 
and/or data, without violating the confidentiality of the subject, to the extent permitted by the 
applicable laws and regulations. Consent from the subject for disclosure of such information will 
be obtained in writing in the ICF. In addition, should a subject require medical care or 
hospitalization during the course of the study, the clinical site may contact the treating physician 
with the subject’s consent, except that consent may not be requested if there is an emergency 
situation. If the results of the study are published, the subject’s identity will remain confidential. 
The clinical site will maintain adequate study records according to applicable regulatory 
requirements. The Sponsor will be notified prior to the destruction of study records. 


AIO-001-101 Solution for SC Injection Comparative Safety and PK 
Aiolos Bio, Inc. 
29-NOV-2023 
 
Confidential 
Page 47 of 47 
12. References 
1 
AIO-001. Investigator’s Brochure. Aiolos Bio. Inc . Version No. 3.1, 20 Oct. 2023. 
2 
van Dijk, B.C.P., Svedsater, H., Heddini, A. et al. Relationship between the Asthma 
Control Test (ACT) and other outcomes: a targeted literature review. BMC Pulm Med 20, 
79 (2020). https://doi.org/10.1186/s12890-020-1090-5 

SAP_001.pdf: 
 
 
Page 1 of 24 
Statistical Analysis Plan for Interventional Studies (Early Phase) 
 
 
Sponsor Name: Aiolos Bio, Inc. 
 
Protocol Number: AIO-001-101 
 
Protocol Title: Open-Label, Single Dose, Parallel Group, Phase 1 Study in Healthy Volunteers 
Evaluating Safety, Tolerability, Pharmacokinetics, and Immunogenicity of a 400 mg Dose of 
AIO-001 Administered by Subcutaneous Injection with Two Formulations  
 
Protocol Version and Date: Amendment II and 29 Nov 2023 
 
Syneos Health Project Code: 7053709 
 
Author: 
 Sr Biostatistician 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Notice of Confidential and Proprietary Information: 
The information contained in this document is confidential belonging to Aiolos Bio, Inc.. 
Acceptance of this document constitutes agreement by the recipient that no information contained 
herein will be published or disclosed without prior written authorization from an official of Aiolos 
Bio, Inc.. However, this document may be disclosed to appropriate Institutional Review Board and 
ethics committees or duly authorized representatives of a national regulatory authority under the 
condition that they are requested to keep it confidential. In the event of an actual or suspected 
breach of this obligation, Syneos Health should be notified promptly. 
 
PPD


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
SAP Version: 1.0. 22 Apr 2024 
Controlled Document ID: 2805A.00, Effective Date 29-Jul-2019 
Filing requirements: TMF 
Page 2 of 24 
Revision History 
Version 
Date 
(DD-Mmm-YYYY) 
Document Owner 
Revision Summary 
0.1 
06 Mar 2024 
Initial draft 
0.2 
20 Mar 2024 
2nd Draft version post sponsor 
review comments 
0.3 
04 Apr 2024 
3rd Draft version post sponsor 
review comments 
1.0 
22 Apr 2024 
Final  
 
 
PPD


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
SAP Version: 1.0. 22 Apr 2024 
Controlled Document ID: 2805A.00, Effective Date 29-Jul-2019 
Filing requirements: TMF 
Page 3 of 24 
Approvals  
I confirm that I have reviewed this document and agree with the content. 
Syneos Health Approval 
 Sr 
Biostatistician  
  
 
 
Name, Title 
Lead Biostatistician  
 Signature 
 
Date  
(DD-Mmm-YYYY) 
 Principal 
Biostatistician 
  
 
 
Name, Title 
Reviewing Biostatistician 
 Signature 
 
Date  
(DD-Mmm-YYYY) 
 Sr 
Pharmacokineticist 
  
 
 
Name, Title 
Lead Pharmacokineticist 
 Signature 
 
Date  
(DD-Mmm-YYYY) 
 Mgr, Statistical Prog 
  
 
 
Name, Title 
Lead Programmer  
 Signature 
 
Date  
(DD-Mmm-YYYY) 
 Medical Writer II 
  
 
 
Name, Title 
Lead Medical Writer 
 Signature 
 
Date  
(DD-Mmm-YYYY) 
Aiolos Bio, Inc.Approval 
 Director, Statistics 
  
 
 
 GSK on behalf of Aiolos Bio, Inc. 
Sponsor Contact 
 Signature 
 
Date  
(DD-Mmm-YYYY) 
 
 
PPD
PPD
PPD
PPD
PPD
PPD


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
SAP Version: 1.0. 22 Apr 2024 
Controlled Document ID: 2805A.00, Effective Date 29-Jul-2019 
Filing requirements: TMF 
Page 4 of 24 
Table of Contents 
Revision History ..............................................................................................................................2 
Approvals .........................................................................................................................................3 
Table of Contents .............................................................................................................................4 
Glossary of Abbreviations ...............................................................................................................6 
1. 
Purpose .....................................................................................................................................8 
1.1 Responsibilities ....................................................................................................................8 
1.2 Timings of Analyses ............................................................................................................8 
2. 
Study Objectives .......................................................................................................................9 
3. 
Study Description .....................................................................................................................9 
Subject Selection ..........................................................................................................................9 
3.1 Determination of Sample Size .............................................................................................9 
3.2 Treatment Assignment .........................................................................................................9 
3.3 Randomization ...................................................................................................................10 
3.4 Blinding .............................................................................................................................10 
3.5 Subject Withdrawal and Replacement ...............................................................................10 
4. 
Endpoints ................................................................................................................................10 
Analysis Populations ......................................................................................................................11 
4.1 Randomized Population .....................................................................................................11 
4.2 Safety Population ...............................................................................................................11 
4.3 Pharmacokinetic Population ..............................................................................................11 
4.3.1 Pharmacokinetic (PK) Concentration Population .......................................................11 
4.3.2 Pharmacokinetic (PK) Parameter Population .............................................................11 
4.4 Immunogenicity Population ..............................................................................................11 
5. 
General Aspects for Statistical Analysis .................................................................................11 
5.1 General Methods ................................................................................................................11 
5.2 Summary Statistics: ...........................................................................................................12 
5.3 Key Definitions ..................................................................................................................13 
5.4 Missing Data ......................................................................................................................13 
6. 
Study Population .....................................................................................................................14 
6.1 Subject Disposition ............................................................................................................14 
6.2 Protocol Deviations ...........................................................................................................14 
6.3 Inclusion and Exclusion Criteria .......................................................................................14 
6.4 Demographics and Other Baseline Characteristics ...........................................................14 
6.5 Medical History .................................................................................................................15 
6.6 Medications .......................................................................................................................15 
6.7 Drug, and Alcohol Screens ................................................................................................15 
6.8 Pregnancy Screening and FSH Testing .............................................................................15 
6.9 Additional Screening Tests ................................................................................................15 
7. 
Pharmacokinetic (PK) Analyses .............................................................................................15 
7.1 Data Presentation ...............................................................................................................16 
7.2 PK Sampling Schedule ......................................................................................................16 
7.3 PK Parameters ...................................................................................................................16 
7.4 PK Comparison Between Formulation A Versus Formulation B .....................................17 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
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Filing requirements: TMF 
Page 5 of 24 
8. 
Immunogenicity Analysis .......................................................................................................18 
9. 
Safety ......................................................................................................................................18 
9.1 Exposure ............................................................................................................................18 
9.2 Adverse Events (AEs) .......................................................................................................18 
9.1 Laboratory Evaluations ......................................................................................................19 
9.2 Vital Signs .........................................................................................................................19 
9.3 Electrocardiograms (ECGs) ...............................................................................................19 
9.4 Physical Examination ........................................................................................................19 
9.5 Injection Site Evaluation ...................................................................................................20 
10. 
Changes from Analysis Planned in the Protocol .................................................................20 
11. 
Programming Considerations ..............................................................................................20 
11.1 General Considerations ....................................................................................................20 
11.2 Table, Listing, and Figure Format ...................................................................................20 
11.2.1 General ......................................................................................................................20 
11.2.2 Headers and Footers ..................................................................................................21 
11.2.3 Display Titles ............................................................................................................21 
11.2.4 Column and Row Headings ......................................................................................21 
11.2.5 Body of the Data Display ..........................................................................................21 
11.2.6 Footnotes ...................................................................................................................23 
12. 
Quality Control ...................................................................................................................23 
13. 
Reference List .....................................................................................................................24 
 
 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
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Filing requirements: TMF 
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Glossary of Abbreviations 
 
Abbreviation 
Description 
AE 
adverse event 
ANOVA 
analysis of variance 
ATC 
anatomical therapeutic chemical 
AUC 
area under the concentration-time curve 
AUC0-last 
area under the concentration-time curve from time zero until the last 
observed concentration 
BLQ 
below the lower limit of quantification  
BMI 
body mass index 
CI 
confidence interval 
Cmax 
maximum observed concentration 
CSR 
clinical study report 
CV 
coefficient of variation 
ECG  
electrocardiogram 
eCRF 
electronic case report form 
FSH 
follicle stimulating hormone 
ICH 
International Council for Harmonisation of Technical Requirements for 
Pharmaceuticals for Human Use 
Kel  
terminal elimination rate constant 
Kel lower 
lower terminal elimination rate constant 
Kel upper 
upper terminal elimination rate constant 
Ln 
natural logarithm 
LS 
least squares 
max 
maximum 
MedDRA® 
Medical Dictionary for Regulatory Activities 
min 
minimum 
N 
number of subjects 
n 
number of observations 
N/A 
not applicable 
PK 
pharmacokinetic(s) 
PT 
preferred term 
p-value 
probability value 
R2 
R-squared 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
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Filing requirements: TMF 
Page 7 of 24 
Abbreviation 
Description 
SAP 
statistical analysis plan 
SC 
subcutaneous 
SD 
standard deviation 
SOC 
system organ class 
SOP 
standard operating procedure 
T1/2 el 
terminal elimination half-life 
TBD 
to be determined 
TEAE 
treatment-emergent adverse event 
TFLs 
tables, figures, and listings 
Tmax 
time of maximum observed concentration 
T½. 
terminal elimination half-life 
vs 
versus 
WHODrug  
World Health Organization Global Drug Dictionary 
 
 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
This document is confidential. 
SAP Version: 1.0. 22 Apr 2024 
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Filing requirements: TMF 
Page 8 of 24 
1. Purpose 
The purpose of this statistical analysis plan (SAP) is to ensure that the data listings, summary 
tables, and figures which will be produced, and the statistical methodologies that will be used, are 
complete and appropriate to allow valid conclusions regarding the study objectives. 
This SAP is based on the following documents: 
• Protocol AIO-001-101 2.0, dated 29 Nov 2023 
• Electronic case report form (eCRF) version 3.0, dated 04 Mar 2024  
The plan may change due to unforeseen circumstances; any changes made after the plan has been 
finalized will be documented in the associated clinical study report (CSR). No revision to the SAP 
is required for changes which do not affect the statistical analysis methods, definitions, or rules 
defined in this document. If additional analyses are required to supplement the planned analyses 
described in the SAP, the changes and justification for the changes will be outlined in the 
associated (CSR). No change will be made without prior approval of the Sponsor.  
When applicable, all methodologies and related processes will be conducted according to Syneos 
Health’s standard operating procedures (SOPs), as appropriate. Shells for all statistical tables, 
listings, and figures referred to in this SAP will be presented in a separate document. 
1.1 
Responsibilities 
Syneos Health will perform the statistical analyses and are responsible for the production and 
quality control of all tables, figures, and listings (TFLs). 
1.2 
Timings of Analyses 
Interim Analysis 
 An interim analysis will be performed at the end of 12 weeks, after all subjects complete Day 85, 
safety, tolerability, pharmacokinetic (PK), and immunogenicity data will be analyzed for primary 
and secondary endpoints for an interim report 
Final Analysis 
The final safety, tolerability, PK, and immunogenicity analysis will be completed after all subjects 
complete the final study visit or terminate early from the study. 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
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This document is confidential. 
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2. Study Objectives 
• Primary objective: 
o To compare the safety and tolerability of 400 mg of AIO-001 as the new 
formulation (Formulation B) with a 400 mg dose of AIO-001 administered as the 
original formulation (Formulation A) in healthy volunteers.  
• Secondary Objectives: 
o To evaluate the PK of a single subcutaneous (SC) AIO-001 dose of Formulation A 
in healthy volunteers.  
o To evaluate the PK of a single SC AIO-001 dose of Formulation B in healthy 
volunteers.  
o To evaluate the immunogenicity of AIO-001 in healthy volunteers.  
 
3. Study Description  
This is an open-label single dose, parallel group, 24-weeks, Phase 1 study in 16 heathy subjects. 
The study is designed to evaluate and compare the safety, tolerability, PK, and immunogenicity of 
SC AIO-001 400 mg using two different formulations (original Formulation A and new 
Formulation B) in 16 healthy volunteers (8 receiving each formulation). 
Subject Selection 
Up to approximately 16 healthy adult males or females aged 18 to 55 years, with body mass index 
(BMI) >18.5 and <32.0 kg/m2, non-smokers, will be enrolled in this study.  
3.1 
Determination of Sample Size 
The proposed number of subjects is in line with the sample sizes commonly used in clinical studies 
of this nature and is considered sufficient to achieve the study objectives. No formal sample size 
calculation was performed for this Phase 1 study.  
3.2 
Treatment Assignment 
Sixteen healthy volunteers (8 receiving each formulation) will receive one of the following 
treatments, according to the randomization scheme. The planned dose levels are listed in Table 
3.3-1, below. 
Table 3.3-1: Treatments 
Group  
Number of 
Subjects 
Treatments 
Planned Dose  
A 
8 
AIO-001 as Formulation A 
400 mg of 100 mg/ml, given 
as 4 X 1.0 ml 
B 
8 
AIO-001 as Formulation B 
400 mg of 182 mg/ml,  
given as 2 X 1.1 ml 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
Sponsor: Aiolos Bio, Inc.; Protocol: AIO-001-101 
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SAP Version: 1.0. 22 Apr 2024 
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Page 10 of 24 
3.3 
Randomization  
Randomization schemes will be generated using SAS® for Windows software, prior to study 
execution. 
Subjects will be randomized prior to dosing, after confirming continued eligibility. Subjects 
eligible for participation will be randomized to receive formulation A or formulation B in a 1:1 
ratio, for a total of 8 subjects receiving formulation A and 8 subjects receiving formulation B. 
The study will include 2 sentinel subjects (1 for each treatment) dosed initially, and the remaining 
14 subjects dosed no sooner than 24 hours. 
Each subject will be randomly assigned a 4-digit randomization number beginning with “1”. Each 
randomization number corresponds to a treatment assignment on the randomization scheme. 
Sentinel subjects will be assigned to the first 2 randomization numbers (i.e., 1001, 1002). 
3.4 
Blinding 
This is an open-label study; blinding is not applicable. 
3.5 
Subject Withdrawal and Replacement 
Subjects will be advised that they are free to withdraw from the study at any time. Over the course 
of the study, the Sponsor and the Investigator or designee may withdraw any subject from the 
study for one of the reasons described below; subject withdrawal will be done in accordance with 
the local guidance at the clinical site: 
 
• Safety reason;  
• 
Non-compliance with protocol requirements;  
• 
Significant protocol deviation;  
• 
Positive pregnancy test, drug screen or alcohol breath test.  
 
Subjects who withdraw prior to dosing may be replaced automatically. In the event that the number 
of dropouts exceeds initial expectations, subjects who withdraw or are withdrawn from the study 
after dosing, for reasons other than safety and tolerability, may be replaced after consultation 
between the Investigator and the Sponsor. In such case, the total number of subjects dosed will 
remain within a maximum of 10 subjects for groups A and B. 
4. Endpoints 
• Primary Endpoints: 
o Adverse events (AEs), vital sign measurements (blood pressure, heart rate, 
respiratory rate, and tympanic temperature), 12-lead electrocardiogram (ECG) 
recordings, physical examinations, and clinical laboratory test results, including 
hematology, biochemistry, and urinalysis.  
• Secondary Endpoints: 
o PK endpoints may include but are not limited to AIO-001 concentrations, AUC0-
last, AUC0-inf, Cmax, Tmax, T½.  


Statistical Analysis Plan for Interventional Studies (Early Phase) 
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Page 11 of 24 
o Incidence of anti-drug antibody (ADA) to AIO-001.  
 
Analysis Populations 
All subjects’ inclusion status into each analysis population will be determined after database lock. 
Subjects will be analyzed according to the treatment received. 
4.1 
Randomized Population 
The randomized population will include all randomized subjects. The randomized population will 
be used for subject listings. 
4.2 
Safety Population 
The safety population is defined as all subjects who receive at least one dose of the study drug. 
4.3 
Pharmacokinetic Population 
The PK population will include all subjects who have at least 1 measured PK concentration 
following dosing. If any participants have incomplete data, protocol deviations, or clinical events 
that affect PK, a decision will be made on a case-by-case basis as to their inclusion in the analysis. 
Before the final analysis, the Pharmacokineticist, in agreement with the Sponsor, will make the 
final decision of which subjects will be included in the PK populations, based on the datasets 
received. 
4.3.1 Pharmacokinetic (PK) Concentration Population 
The PK concentration population will include all subjects who receive AIO-001 and have at least 
one quantifiable post-dose PK concentration. The PK concentration population will be used for all 
PK concentration summaries and analyses. 
4.3.2 Pharmacokinetic (PK) Parameter Population 
The PK parameters population will include all subjects who receive AIO-001 and have sufficient 
post-dose concentration data to support accurate estimation of at least one PK parameter. The PK 
parameters population will be used for all PK parameter summaries and analyses. 
4.4 
Immunogenicity Population 
The immunogenicity population will include all subjects who have received any amount of AIO-
001 and have at least one post-dose ADA measurement. 
5. General Aspects for Statistical Analysis 
5.1 
General Methods 
SAS for Windows software will be used to perform all statistical analyses. All data in the database 
will be presented in the data listings. Unless otherwise stated, all listings will be sorted by 
treatment, subject number, and assessment date/time.  
The following labels for treatment will be used on all tabulations where the results are displayed 
by treatment, in the following order: 
• 400 mg of 100 mg/ml AIO-001 Formulation A 


Statistical Analysis Plan for Interventional Studies (Early Phase) 
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• 400 mg of 182 mg/ml AIO-001 Formulation B 
 
5.2 
Summary Statistics: 
Unless otherwise stated, continuous variables will be summarized using the number of 
observations (n), and the statistics mean, median, standard deviation (SD), minimum (min) and 
maximum (max). The min and max values will be presented to the same number of decimal places 
as recorded in the eCRF, mean and median will be presented to one more decimal place than the 
raw data, and the SD will be presented to two more decimal places than the raw data.  
Summaries of change from baseline variables will include only subjects who have both a baseline 
value and corresponding post baseline value at the timepoint of interest. Categorical and binary 
variables will be summarized with frequency counts and percentages. Percentages will be rounded 
to one decimal place, with the denominator being the number of subjects (N) in the relevant 
population, unless otherwise stated. 
For PK data, values will be rounded to two decimal places in the listings and tables, except for the 
following situations: 
• Terminal elimination rate constant (Kel) and R-squared (R2) adjusted data shall be rounded 
off to four decimal places. 
• PK parameters related to time, such as Tmax, lower terminal elimination rate constant (Kel 
Lower), and upper terminal elimination rate constant (Kel Upper), must be reported with the 
same precision as the actual sampling time, rounded to three decimal places. 
• Concentration versus time data, as well as Cmax shall be reported as they appear in the 
corresponding dataset. 
• Summary statistics, including geometric mean and coefficient of variation (CV%), will be 
presented to the number of decimal places is contingent on the number of places in the raw 
data plus one additional decimal place. The geometric mean and CV% will not be 
calculated for Tmax, T1/2, and Kel. 
Only data from protocol scheduled (“nominal”) visits will be included in the summary tables. Data 
from unscheduled visits will not be included in the summary tables (unless they were used as 
baseline) but will be included in the listings and figures. 
In the case of a repeat test, both assessments will be presented in the listings. If a repeat 
measurement was performed due to the first measurement being outside the normal range for the 
given assessment and the second assessment confirms the first measurement, then only the first 
measurement will be used for analysis and the second assessment will be considered an 
unscheduled timepoint. In the same case, if the second assessment falls within normal ranges, then 
the second assessment will be used for analysis of that timepoint. In all other cases, the first 
measurement will be taken as the assessment to be used for the analysis. 


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5.3 
Key Definitions 
Baseline: 
Unless stated otherwise, baseline will be defined for each subject and will be defined as the last 
non-analy measurement (including repeated and unscheduled assessments) obtained prior to study 
drug administration. Post baseline will be considered as all measurements collected after study 
drug administration. “Unknown”, “Not Done”, “Not Applicable” and other classifications of 
missing data will not be considered when calculating baseline observations unless the finding is a 
valid categorical observation. 
Study Day: 
Study day will be calculated using first study drug administration date as the reference date. If the 
date of interest occurs on or after the first study drug administration date, study day will be 
calculated as (date of interest – first study drug administration date) + 1. If the date of interest 
occurs prior to the first study drug administration date, study day will be calculated as (date of 
interest – first study drug administration date). There will be no study day 0. 
Prior and Concomitant Medication: 
All medications taken by subjects after screening and through to dosing will be documented as 
prior medications. All medications taken by subjects after dosing through to the last study day will 
be documented as concomitant medications. 
5.4 
Missing Data 
There will be no imputation for missing data, unless otherwise specified. Missing data shall be 
presented in subject listings as either “-” (unknown or not evaluated) or “N/A” (not applicable), 
with the corresponding definition in the footnotes. Missing descriptive statistics, or probability 
values (p-values), which cannot be estimated shall be presented as “-”. 
For inclusion in concomitant medication and AE tables, incomplete start and stop dates on the 
eCRF will be imputed as follows: 
• If the stop date is incomplete, the following rules will be applied: 
o Missing day: Assume the last day of the month 
o Missing day and month: Assume the last day of the year 
o Missing day, month, and year: Assume that the event/medication is continuing 
o In the case of the death of a subject, and if the imputed end date is after the date of 
death, the end date will be imputed as the date of death 
• If the stop date is incomplete, imputed end date will be used instead of reported end date. 


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• If the start date is incomplete, the following rules will be applied: 
o Missing day: Assume the first day of the month; however, if the partial date and the 
date of first study drug administration lie within the same month and year and the 
date of first study drug administration is not after the stop date of the 
event/medication, set to the date of study drug administration. Otherwise, set to the 
stop date of the event/medication. 
o Missing day and month: Assume January 1st; however, if the partial date and the 
date of first study drug administration lie within the same year and the date of first 
study drug administration is not after the stop date of the event/medication, set to 
the date of first study drug administration. Otherwise, set to the stop date of the 
event/medication. 
o Missing day, month, and year: Assume date of first study drug administration if it 
is not after the stop date for the event/medication. Otherwise, set to the stop date 
for the event/medication. 
In the case of withdrawal of consent, all data from subjects who withdraw from the study will be 
included in all summaries up to the time of withdrawal. For all other withdrawals, all data captured 
will be included in the safety summaries. 
For PK analysis, only observed concentration data will be used in the data analysis except for 
concentration values below the lower limit of quantification (BLQ). No attempt will be made to 
extrapolate or interpolate estimates for missing data. 
 
6. Study Population 
6.1 
Subject Disposition 
The number of subjects who were enrolled, who were dosed, who completed the study, and who 
were discontinued (post dose) from the study, along with reasons for discontinuation, will be 
summarized. The data will be presented by treatment and overall (frequency and the percentage of 
subjects) and presented by subject in a data listing. 
6.2 
Protocol Deviations 
Subject data will be examined for evidence of protocol deviations. All protocol deviations will be 
categorized and presented by subject in a data listing.  
6.3 
Inclusion and Exclusion Criteria 
All recorded inclusion and exclusion criteria status will be presented by subject in a data listing. 
Subjects who were screen-failed will not include in this listing. Each subject’s inclusion or 
exclusion from each analysis population will also be presented in a data listing. 
6.4 
Demographics and Other Baseline Characteristics 
All demographics and baseline body measurements will be summarized by treatment and 
presented by subject in a data listing. 


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Descriptive statistics (n, mean, SD, min, median, and max) will be calculated for continuous 
variables using the last results obtained prior to study drug administration. Frequency counts and 
percentages will be tabulated for categorical and binary variables.  
6.5 
Medical History 
Medical history will be presented by subject in a data listing. The latest version of the Medical 
Dictionary for Regulatory Activities (MedDRA) will be used to classify medical history terms by 
system organ class (SOC) and preferred term (PT). Output data will include the MedDRA version 
26.1. 
6.6 
Medications 
Prior and concomitant medications will be presented by subject in a data listing. The latest version 
of the World Health Organization Global Drug Dictionary (WHODrug) will be used to classify 
medications by anatomical therapeutic chemical (ATC) classification code (2nd level) and 
preferred name. When 2nd level classification code is not available, 1st level classification will be 
used instead. Output data will include the WHODrug version used in the study. 
Concomitant medications will be summarized by treatment and overall (frequency and the 
percentage of subjects). This summary will include the percentage of subjects with at least one 
concomitant medication. 
6.7 
Drug, and Alcohol Screens 
The results of drug, and alcohol screens will be presented by subject in data listings. 
6.8 
Pregnancy Screening and FSH Testing 
Pregnancy tests will be administered to females. The follicle stimulating hormone (FSH) level will 
be tested in postmenopausal females. All results will be presented by subject in data listings. 
6.9 
Additional Screening Tests 
The results of serology tests will be presented by subject in data listings. 
 
7. Pharmacokinetic (PK) Analyses 
Phoenix® WinNonlin® software version 8.3.4 (Certara USA, Inc., Princeton, NJ) will be used for 
all PK analyses. Statistical analyses will be performed using SAS® for Windows, Release 9.4 
(SAS® Institute Inc., Cary, NC, USA). Bioanalysis of all samples should be completed prior to the 
initiation of the PK and statistical analyses. 
All PK concentration and PK parameter analyses will be conducted on the PK concentration 
population and PK parameter population, respectively.  
PK concentrations will be listed and summarized by treatment, analyte, and nominal sampling 
timepoint using descriptive statistics (n, mean, SD, CV%, min, max, and median). Individual and 
mean (±SD) concentration versus time curves will be presented for both linear and semi-log scale 
by treatment. 


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7.1 
Data Presentation 
For all PK analyses, concentration values BLQ that occur before the first measurable concentration 
of the study drug will be set to “0.00”; BLQ values that occur after first measurable concentration 
will be set to “missing”. No imputations will be made on BLQ concentrations. 
Invalid concentration values (due to bioanalytical or clinical issue) that occur prior to dosing will 
be replaced by “0.00”. Invalid concentration values that occur after dosing will be set to “missing” 
for tabulation, graphical representation, and calculation purposes. 
The actual clock time for dosing and the actual clock time for each PK sample collection will be 
recorded. For all sampling times, the actual sampling duration will be calculated as the difference 
between the sample collection actual clock time and the actual clock time of dosing. The actual 
post-dose sampling times, expressed in hours and rounded off to three decimal places, will be used 
to calculate the PK parameters. Pre-dose sampling times will always be reported as zero (0.000), 
regardless of the time difference. Nominal sampling times will be used in concentration tables and 
mean graphs, while actual sampling times for post-dose samples will be used in the individual 
graphs. Actual sampling times for post-dose samples also will be used for PK parameter derivation, 
unless the actual sampling time is missing, in which case, the nominal time will be used. 
7.2 
PK Sampling Schedule  
A total of 19 blood samples will be drawn from each subject for PK analyses of AIO-001. 
Specifically, will be collected before (pre-dose) and after dosing on day 1 (12 hours) administration 
(within 30 minutes) and at, 24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 168 hours, 240 
hours, 336 hours, 504 hours, 672 hours, 1008 hours, 1344 hours, 1680 hours, 2016 hours, 2688 
hours, 3360 hours and 4032 hours post-dose.  
7.3 
PK Parameters 
All PK parameters will be presented in data listings and summarized by treatment and analyte 
using descriptive statistics (n, arithmetic and geometric means, SD, CV%, min, max, and median). 
The PK parameters shown in Table 8.3-1, below, will be calculated by standard 
non-compartmental methods.  
Table 8.3-1: PK Parameters 
Parameter  Definition  
AUC0-last 
area under the concentration-time curve from time zero until the last observed 
concentration 
AUC0-inf 
area under the concentration-time curve from time zero to infinity (extrapolated) 
Residual 
area  
Percentage of AUC0-inf due to extrapolation from the time of the last observed 
concentration to infinity, calculated as [1 - (AUC0-t / AUC0-inf)] x 100  
Cmax  
maximum observed concentration 
Tmax 
time when the Cmax is observed 
T1/2 el 
terminal elimination half-life, calculated as ln(2)/Kel  
Kel  
Terminal elimination rate constant  


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Area under the concentration-time curve (AUC) parameters will be calculated using the linear up 
log down trapezoidal method, where the linear trapezoidal rule is used any time the concentration 
data are increasing, and the logarithmic trapezoidal rule is used any time that the concentration 
data are decreasing. 
Kel will be the negative of the estimated slope of the linear regression of the ln-transformed plasma 
concentration versus time profile in the terminal elimination Phase. The best fit method will be 
used to calculate the Kel from at least three concentration data points, excluding Cmax. R2 adjusted, 
the goodness of fit statistic for the terminal elimination Phase, adjusted for the number of points 
used in the estimation of Kel must be ≥ 0.8. If the R2<0.8, the PK parameters derived from Kel will 
be presented in listing(s) but excluded from descriptive statistics in tables. The timepoint where 
ln-linear Kel calculation begins (Kel Lower), the actual sampling time of the last measurable 
concentration used to estimate the Kel (Kel Upper), and the R2 adjusted for the ln-linear regression for 
the calculation of the elimination rate constant will be reported.  
If the Residual area is more than 20%, all the derived parameters depending on AUC0-inf (i.e., AUC0-
inf) will be flagged accordingly. 
7.4 
PK Comparison Between Formulation A Versus Formulation B 
For the PK comparison of the relative bioavailability between formulation A and formulation B, 
an ANOVA model will be used to compare the log transformed PK parameters AUC0-last, AUC0-inf, 
and Cmax . The estimated ratio (B/A) of model -adjusted geometric means will be used to perform 
this comparison. To estimate the geometric mean ratio, a linear mixed effects model will be used 
to the data. The response variable will be the natural log transformed PK parameter, and the model 
will include treatment as fixed effect and subject as random effect. The model will be fit using 
restricted maximum likelihood, and the Kenward--Roger degrees of freedom approximation. From 
this model, the adjusted means will be estimated for each treatment (A and B). The difference in 
geometric means will be estimated, accompanied by the corresponding two one-sided 90% 
confidence interval (CI). The estimates and CIs will be exponentiated using base e in order to 
estimate the geometric mean ratio and associated two one-sided 90% CI..  
 
The SAS code for the analysis model for ANOVA will follow the format given below (using the 
Mixed Procedure to fit the linear regression). The input variables, datasets, and labels are depicted 
in italicized red text and have been given generic names. 
proc mixed data= dataset; 
class subject treatment; 
model ln_PK_parameter = treatment / ddfm= kr solution; 
random subject; 
lsmeans treatment; 
estimate ‘formualtion A vs Formulation B’ treatment 1 -1/cl alpha=0.1; 
run; 
 


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8. Immunogenicity Analysis 
Anti-AIO-001 antibodies will be evaluated in serum samples collected before and after dosing at 168 
hours, 336 hours, 504 hours, 672 hours, 1344 hours, 2016 hours, 3360 hours, and 4032 hours post-
dose. The results will be presented in a data listing by subject. 
A subject with any ADA is defined as subject with at least one baseline or post-baseline ADA-positive 
sample. A pre-existing ADA subject is defined as subject who has a baseline ADA-positive sample. 
ADA-negative subject is defined as subject without treatment-induced or treatment-boosted ADA-
positive sample during the treatment or follow-up observation period. ADA-positive subject is defined 
as subject with at least one treatment-induced or treatment-boosted ADA-positive sample at any time 
during the treatment or follow-up observation period. Treatment-induced ADA-positive is defined as 
subject who has baseline ADA-negative sample and at least one post-baseline ADA-positive sample, 
while treatment-boosted ADA positive is defined as subject who has both baseline and post-baseline 
ADA-positive samples, and the titer of the post-baseline sample is equal to or more than 4-fold of the 
baseline titer. 
The frequency and percentage of the following information will be summarized; subjects with any 
ADA, pre-existing ADA subjects, ADA-positive subjects and ADA-negative subject. 
All immunogenicity analyses will be conducted on the immunogenicity population. 
The frequency and percentage of ADA results will be summarized by treatment and overall.  
In addition, the adverse event overview by subject with ADA status will be presented 
9. Safety 
Safety and tolerability analysis will be performed for all subjects in the safety population. No 
inferential statistical analysis of safety data is planned. 
9.1 
Exposure 
Study drug administration will be listed by subject.  
9.2 
Adverse Events (AEs) 
AEs will be coded using the latest version of the MedDRA. Output data will include the MedDRA 
version 26.1 AEs will be grouped by SOC and PT and summarized by actual treatment and by 
overall. The summary tables will present the number and percentage of total subjects and number 
of events by SOC and by PT. 
All AE summaries will be restricted to treatment-emergent AEs (TEAEs), defined as AEs that 
commence on or after the time of study drug administration. AEs without an onset date or time, or 
AEs with an onset date of the date of study drug administration but without an onset time, will be 
defined as treatment-emergent, unless an incomplete date (e.g., month and year) clearly indicates 
that the event started prior to administration of study drug, or the AE stop date indicates that the 
event started and stopped prior to administration of study drug. 
The number and percentage of subjects experiencing TEAEs and the number of TEAEs will be 
tabulated. Subjects who experience the same TEAE (in terms of MedDRA PT) more than once 
will only be counted once per treatment; however, the total number of events will be counted per 
category. This also applies to sub-categories displayed in the summaries. 


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The following summaries will be presented: 
• Overall summary of TEAEs 
• TEAEs by SOC and PT 
• TEAEs by SOC, PT, and severity 
• TEAEs by SOC, PT, and relationship to study drug 
• Serious TEAEs by SOC and PT 
• Serious TEAEs by SOC, PT and relationship to study drug 
All AEs will be listed. The following listings will be included: Non-TEAEs, TEAEs, and serious 
AEs. 
9.1 
Laboratory Evaluations 
Laboratory data, including hematology, biochemistry, coagulation, and urinalysis (quantitative 
and categorical), will be listed by subject, and summarized by treatment and visit, categorical 
values less than x or greater than x will be summarized by frequency. Observed values and changes 
from baseline will be presented. 
In addition, a shift table representing the categorical change (normal, abnormal not clinically 
significant, or abnormal clinically significant) from baseline to each post baseline visit will be 
presented by treatment. 
Abnormal results will be flagged in the listings. 
9.2 
Vital Signs  
Vital sign measurements will be listed by subject and summarized by treatment, visit and 
timepoint. Observed values and changes from baseline will also be presented.  
In addition, a shift table representing the categorical change (normal, abnormal not clinically 
significant, or abnormal clinically significant) from baseline to each post baseline visit and 
timepoint will be presented by treatment. 
Abnormal results will be flagged in the listings. 
9.3 
Electrocardiograms (ECGs) 
Electrocardiogram (ECG) values will be listed by subject and summarized by treatment, visit and 
timepoint. Observed values and changes from baseline will be presented. 
In addition, a shift table representing the categorical change in overall interpretation (normal, 
abnormal not clinically significant, or abnormal clinically significant) from baseline to each post 
baseline visit and timepoint will be presented by treatment. 
Abnormal results will be flagged in the listings. 
9.4 
Physical Examination  
The results of physical examinations will be listed by subject. Abnormal results will be flagged in 
the listings. 


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9.5 
  Injection Site Evaluation 
The results of injection site evaluation will be listed by subject.  
 
10. Changes from Analysis Planned in the Protocol 
Inclusion of the randomized population  
 
11. Programming Considerations 
All TFLs and statistical analyses will be generated using SAS for Windows, release 9.4 
(SAS Institute Inc., Cary, NC, USA) software. Phoenix® WinNonlin®, version 8.3.4 (Certara USA, 
Inc., Princeton, NJ) will be used for all PK analysis. This software was validated by Syneos in 
compliance with US 21 CFR Part 11 regulation. 
11.1 General Considerations 
• One SAS program can create several outputs. 
• Each output will be stored in a separate file. 
• Output files will be delivered in rich text format that can be manipulated in MS Word. 
• Numbering of TFLs will follow International Council for Harmonisation of Technical 
Requirements for Pharmaceuticals for Human Use (ICH) guideline E3a .  
11.2 Table, Listing, and Figure Format 
11.2.1 General 
• TFLs will be produced in landscape format. The orientation may be changed to portrait, as 
necessary to allow additional rows to be presented. 
• TFLs will be produced using the Times New Roman font, size 10. The font size may be 
reduced as necessary to allow additional columns to be presented, but not at the expense of 
clarity.  
• The data displays for all TFLs will have a minimum blank 1-inch margin on all four sides. 
• Unless otherwise specified, TFLs will be in black and white (no color). 
• Specialized text styles, such as bolding, italics, borders, shading, and superscripted and 
subscripted text, will not be used in the TFLs, unless otherwise specified. On some 
occasions, superscripts 1, 2, or 3 may be used; see below. 
• Standard keyboard characters will be used in the TFLs. Special characters, such as 
non-printable control characters, printer-specific, or font-specific characters, will not be 
used. Hexadecimal-derived characters will be used, where possible, if they are appropriate 
to help display math symbols (e.g., μ). Certain subscripts and superscripts (e.g., cm2, Cmax) 
will be employed on a case-by-case basis. 
• TFLs will be produced using sentence case, unless otherwise specified. 


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11.2.2 Headers and Footers 
• Times New Roman font, size 10 will be used for TFL headers and footers. 
• All outputs will have the following at the top of each page: Aiolos Bio, Inc.Aiolos Bio, 
Inc.  Protocol  AIO-001-101. 
• All outputs will have page x of y at the top or bottom right corner of each page. TFLs are 
individually paginated in relation to total length (i.e., the page number appears sequentially 
as page x of y, where y is the total number of pages in the output). 
• The date and time the output was generated will appear, along with the program name, at 
the bottom of each page. 
11.2.3 Display Titles 
Each display title includes the appropriate designation (“Table”, “Figure”, or “Listing”) and a 
numeral, along with a descriptive name (e.g., Table 14.1-1 Subject Enrollment and Disposition). 
ICH E3 numbering is strongly recommended, but Sponsor preferences are obtained for final 
determination. Display titles are left aligned, single spaced, and presented in title case. A solid line 
spanning the margins will separate display titles from column headings.  
11.2.4 Column and Row Headings 
• Column and row headings are presented in title case, with the exception of complete 
sentences, which will be presented in sentence case.  
• Column and row headings will include “Unit” for numeric variables, as appropriate. 
• Column and row headings will include the number of subjects in the analysis population 
for each group, presented as (N=xx). This is different from the ‘n’ used in descriptive 
statistics, which represents the number of observations. 
• The order of treatments in the tables and listings will be Treatment A followed by 
Treatment B, with “overall” (if applicable) last. 
11.2.5 Body of the Data Display 
11.2.5.1 General Conventions 
Data in columns of a table or listing are formatted as follows: 
• Alphanumeric values are left aligned. 
• Whole numbers (e.g., counts) are right aligned. 
11.2.5.2 Table Conventions 
• Units will be included, where available. 


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• If the categories of a parameter are ordered, all categories between the maximum and 
minimum category are presented in the table, even if n=0 for all groups in a category 
between the minimum and maximum level for that parameter. See the example for the 
frequency distribution for symptom severity below. If percentages are presented in these 
tables, 0% will not be presented, therefore, counts of zero will be presented as “0”, not “0 
(0%)”. 
Severity Rating N 
Severe 
0 
Moderate 
8 
Mild 
3 
• Where the categories are not ordered (e.g., Reason for Discontinuation), only those 
categories for which there is at least one subject represented will be included. 
• An “Unknown” or “Missing” category will be added to each parameter for which 
information is unavailable for one or more subjects. 
• Probability values (p-values) are presented in the format: 0.xxxx, where xxxx is the value. 
If the p-value is less than 0.0001, it will be presented as “<0.0001.” If the p-value is >0.999, 
it will be presented as “>0.999.” 
• Percentage values are presented in parentheses with no spaces, one space after the count 
[e.g., 7 (12.8%), 13 (5.4%)]. Unless otherwise noted, for all percentages, the denominator 
will be the number of subjects in the analysis population for the group that has an 
observation. Percentages after zero counts are not displayed, and percentages equating to 
100% are presented as “100%” (without decimal places).  
• Unless otherwise noted, tabular displays of data for medical history, prior/concomitant 
medications, and AEs data are presented in alphabetical order.  
• The percentage of subjects is typically calculated as a proportion of the number of subjects 
assessed in the relevant group (or overall) for the analysis population presented; however, 
careful consideration is required in many instances, due to the complicated nature of 
selecting the denominator. Details of this will be presented in footnotes or programming 
notes. 
• In categorical summaries where a subject can be included in more than one category, a 
footnote or programming note will specify whether the subject is included in the summary 
statistics for all relevant categories or just one category and the criteria for selecting the 
category. 
• Where a category with a subheading (such as system organ class [SOC]) must be split over 
more than one page, present the subheading followed by “(cont.)” at the top of each 
subsequent page. The overall summary statistics for the subheading will only be presented 
on the first relevant page. 
11.2.5.3 Listing Conventions 
• Unless otherwise noted, listings will be sorted for presentation in order of subject number, 
visit/collection day, and visit/collection time. 


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• Dates are printed in SAS DATE9.format (e.g., “ddMMMyyyy”: 01JUL2000). Missing 
portions of dates are represented on subject listings as dashes (e.g.,--JUL2000). Dates that 
are missing because they are not applicable for the subject are presented as “N/A”, unless 
otherwise specified. 
• All observed time values are presented using a 24-hour clock HH:MM:SS or HH:MM 
format (e.g., 11:26:45, or 11:26). Time will only be reported if it was measured as part of 
the study. 
• Units will be included, where available. 
11.2.5.4 Figure Conventions 
• For safety figures, study visits will be displayed on the X-axis and endpoint (e.g., treatment 
mean change from baseline) values will be displayed on the Y-axis, unless otherwise 
specified. 
• Legends will be used for all figures with more than one variable, group, or item displayed. 
• Units will be included, where available. 
11.2.6 Footnotes 
• A solid line spanning the margins will separate the body of the data display from the 
footnotes.  
• All footnotes will be left aligned, with single spacing, immediately below the solid line 
beneath the data display. 
• Informational footnotes begin with “Note:”. Reference footnotes begin with a reference 
number or letter (e.g., 1, 2, 3 or a, b, c).  
• Each new footnote starts on a new line, where possible. 
• Subject-specific footnotes are avoided, where possible. 
• Footnotes will be used sparingly and add value to the table, figure, or data listing. If more 
than six lines of footnotes are planned, a cover page may be used to display footnotes, and 
only those essential to comprehension of the data will be repeated on each page.  
12. Quality Control 
SAS programs are developed to produce outputs such as analysis data sets, summary tables, data 
listings, figures, and statistical analyses. These are developed and undergo quality control in 
accordance with the latest versions of SOP 2800b  and SOP 2801c. 
 


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13. Reference List  
 
aInternational Council for Harmonisation of Technical Requirements for Pharmaceuticals for 
Human Use (ICH). (1996). Guideline for Industry, Structure and Content of Clinical Study 
Reports (ICH E3). 
bSyneos Health. Standard Operating Procedure, Developing Statistical Programming 
Specifications for Early Phase Studies (SOP 2800). 
cSyneos Health. Standard Operating Procedure, Developing Statistical Programs for Early Phase 
Studies (SOP 2801).  
 
End of document