Conditions: Healthy
Allocation: NON_RANDOMIZED | Intervention model: SEQUENTIAL
Enrollment: 8
Healthy volunteers: yes

Brief summary:
A clinical study to measure the Safety, Tolerability, and Pharmacokinetics of OP-101 After Subcutaneous Administration in Healthy Volunteers

Detailed description:

Arms:
arm 1: Subjects in Cohort 1 will be administered 4 mg/kg OP-101 as a subcutaneous (SC) injection. arm 2: Subjects in Cohort 2 will be administered 8 mg/kg OP-101 as a SC injection.

Interventions: intervention 1: OP-101
Intervention descriptions: intervention 1: Subcutaneous injection of OP-101 in healthy volunteers

Protocol document:
Prot_000.pdf: 
CLINICAL STUDY PROTOCOL
A Phase 1 Open-Label Single-Ascending Dose Study to Evaluate the 
Safety, Tolerability, and Pharmacokinetics of OP-101 (dendrimer 
n-acetyl-cysteine) After Subcutaneous Administration in Healthy
Volunteers
Investigational Product: OP-101 (dendrimer n-acetyl-cysteine)
Protocol Number: OP-101-003
Sponsor:
Orpheris, Inc.
1235 Radio Road, Suite 200
Redwood City, CA 94065
Telephone: 650-505-5048
Version Number: 2.0
Date: 17 March 2020
Confidentiality Statement
The information in this document is confidential and is not to be disclosed without the written 
consent of Orpheris, Inc., hereinafter Orpheris, except to the extent that disclosure would be 
required by law and for the purpose of evaluating and/or conducting a clinical study for Orpheris.
You are allowed to disclose the contents of this document only to your Institutional Review 
Board and study personnel directly involved with conducting this protocol. Persons to whom the 
information is disclosed must be informed that the information is confidential and proprietary to 
Orpheris and that it may not be further disclosed to third parties.
NCT04321980


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
2
SIGNATURE PAGE
STUDY TITLE: A Phase 1 Open-Label Single-Ascending Dose Study to Evaluate the 
Safety, Tolerability, and Pharmacokinetics of OP-101 (dendrimer n-acetyl-cysteine) After 
Subcutaneous Administration in Healthy Volunteers
We, the undersigned, have read this protocol and agree that it contains all necessary information 
required to conduct the study.
Signature
Date
Orpheris, Inc. 
Medical Monitor


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
3
INVESTIGATOR AGREEMENT
By signing below I agree that:
I have read this protocol. I approve this document and I agree that it contains all necessary details 
for carrying out the study as described. I will conduct this study in accordance with the design and 
specific provision of this protocol and will make a reasonable effort to complete the study within 
the time designated. I will provide copies of this protocol and access to all information furnished 
by Orpheris to study personnel under my supervision. I will discuss this material with them to 
ensure they are fully informed about the study product and study procedures. I will let them know 
that this information is confidential and proprietary to Orpheris and that it may not be further 
disclosed to third parties. I understand that the study may be terminated or enrollment suspended 
at any time by Orpheris, with or without cause, or by me if it becomes necessary to protect the best 
interests of the study subjects.
All documentation for this study that is supplied to me and that has not been previously 
published will be kept in the strictest confidence.  This documentation includes this study 
protocol, Investigator's Brochure(s) (IB), Case Report Forms (CRFs), and other scientific data.
The study will not be commenced without the prior written approval of a properly constituted 
Human Research Ethics Committee (HREC). No changes will be made to the study protocol 
without the prior written approval of the Sponsor and the Ethics Committee, except where 
necessary to avert an immediate hazard to the participants.
I have read the protocol and agree that the study will be conducted in compliance with the 
protocol and in accordance with the principles of the current version of the Declaration of 
Helsinki (Ethical Principles for Medical Research Involving Human Subjects), and with the 
National Health and Medical Research Council (NHMRC) National Statement on Ethical 
Conduct in Human Research (2007, updated 2018). The conduct of the study will be in 
accordance with the Integrated Addendum to ICH E6 (R1), Guideline for GCP ICH E6 (R2), 
annotated with comments by the Australian Therapeutic Goods Administration (TGA) (2018).
I acknowledge that I am responsible for the overall study conduct.  I agree to personally conduct 
or supervise the described clinical study. I agree to ensure that all associates, colleagues and 
employees assisting in the conduct of the study at my site are informed about their obligations. 
Mechanisms are in place to ensure that site staff receive the appropriate information throughout
the conduct of the study.
______________________________________________
____________________
Investigator’s Signature
Date
_____________________________________________
Investigator’s Printed Name 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
4
SYNOPSIS
TITLE:
A Phase 1 Open-Label Single-Ascending Dose Study to Evaluate the Safety, 
Tolerability, and Pharmacokinetics of OP-101 (dendrimer n-acetyl-cysteine) After Subcutaneous
Administration in Healthy Volunteers
PROTOCOL NUMBER: OP-101-003
INVESTIGATIONAL PRODUCT: OP-101 (dendrimer n-acetyl-cysteine)
PHASE: 1
INDICATION(S): Treatment of children with childhood cerebral adrenoleukodystrophy who 
are not eligible for hematopoietic stem cell transplantation.
OBJECTIVES:
The primary objective is to evaluate the safety and tolerability of OP-101 after single subcutaneous
(SC) doses in healthy subjects.
The secondary objective is to determine the pharmacokinetic (PK) profile of OP-101 after single 
SC doses in healthy subjects.
POPULATION:
The population for this study includes men and women aged 18 to 65 years, inclusive, who are in 
good health as determined by screening medical history, a physical examination, vital sign
measurements, and clinical laboratory tests (chemistry, hematology, and urinalysis). Subjects are 
to have a body mass index between 18 and 32 kg/m2, inclusive. 
STUDY DESIGN AND DURATION:
This is a Phase 1, open-label, single-ascending dose study to evaluate the safety, tolerability, and 
PK of OP-101 after SC administration in up to 8 healthy subjects at 1 site in Australia.
This study will consist of a 21-day Screening Period, a 2-day Treatment Period, and Follow-up 
Visits on Days 8 and 15. Subjects will sign an informed consent form before any study procedures 
are performed and must meet all of the inclusion and none of the exclusion criteria to participate 
in the study. Eligible subjects will be admitted to the investigational site on Day -1, dosed on 
Day 1, and discharged on Day 3.
There will be 2 separate and sequential SC cohorts (4 mg/kg and 8 mg/kg) with 4 subjects in each 
cohort who complete Day 15. Cohort 1 will enroll 4 subjects administered a single SC dose of 
4 mg/kg.  Dose escalation from Cohort 1 (4 mg/kg) to Cohort 2 (8 mg/kg) will be determined by 
the safety review committee (SRC), consisting of the Medical Monitor, the Principal Investigator
(or delegate), and Sponsor representative after review of safety, tolerability, and any available PK 
data through Day 8 of Cohort 1. Cohort 2 will enroll 4 subjects administered a single SC dose of 
8 mg/kg.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
5
During the Treatment Period, plasma and urine samples will be collected for PK analysis. Plasma 
PK samples will be collected predose, at 30 minutes and at 1, 2, 4, 6, 8, 10, 12, 16, 24, and 
30-36 hours after dosing, as well as on Day 3 (~48 hr). Blood samples for PK analysis can be 
obtained within 5 minutes of the scheduled sampling time for sample times 1 hour, within 
10 minutes of the scheduled sampling time for sample times 12 hours, and within 2 hours of 
the scheduled sampling times for sample times >12 hours. 
Urine PK samples will be collected predose (spot sample) and between 0 to 4, 4 to 8, 8 to 12, 
12 to 18, 18 to 24, and 24 to 48 hours.
Safety assessments will include adverse event monitoring, clinical laboratory tests (hematology, 
chemistry, and urinalysis), physical examinations, and vital signs (pulse rate, blood pressure [BP], 
respiration rate, and temperature). Coagulation testing will be performed at screening. Pregnancy 
testing will be performed at screening and Day -1.
DOSAGE FORMS AND ROUTE OF ADMINISTRATION:
Subjects in Cohort 1 will be administered 4 mg/kg OP-101 as a SC injection.
Subjects in Cohort 2 will be administered 8 mg/kg OP-101 as a SC injection.
PHARMACOKINETIC VARIABLES: 
The PK variables will include the following:
Cmax – maximum plasma concentration,
tmax – time to maximum plasma concentration,
kel – apparent terminal rate constant, 
t½ – apparent elimination half-life, 
AUC0-t – area under the concentration-time curve based on the last measurable concentration,
AUC0-inf – area under the concentration-time curve from time zero to infinity, and
CL – clearance of study drug.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
6
SAFETY VARIABLES:
Safety assessments will include adverse event monitoring, clinical laboratory tests (hematology, 
chemistry, and urinalysis), physical examinations, and vital signs (pulse rate, BP, respiration rate, 
and temperature).
STATISTICAL ANALYSES:
The PK Concentration Population is defined as all subjects who received at least 1 dose of study 
drug and have at least 1 measurable concentration. The PK Parameter Population is defined as all 
subjects who received a dose of study drug and have at least 1 PK parameter. Plasma PK 
concentrations of OP-101 will be summarized with descriptive statistics (n, mean, standard 
deviation [SD], percent coefficient of variation [%CV], minimum, median, maximum, geometric 
mean, and %CV for the geometric mean) by nominal PK sampling time and treatment using the 
PK Concentration Population. Standard noncompartmental PK analysis will be used to calculate 
the PK parameters. Pharmacokinetic parameters will be summarized with descriptive statistics (n, 
mean, SD, %CV, minimum, median, maximum, geometric mean, and %CV for the geometric 
mean) by treatment using the PK Parameter Population.
The Safety Population is defined as all subjects who received at least 1 dose of study drug. Adverse 
events will be summarized overall and by system organ class and preferred term, and the 
relationship to the study drug received. A list of subjects who have SAEs and who discontinue 
from the study due to an adverse event will be provided. Deaths due to adverse events will also be 
provided. Safety laboratory tests will include chemistry, hematology, and urinalysis. Summary 
statistics for laboratory values will be provided at baseline, postbaseline, and for changes from 
baseline to postbaseline by treatment. Occurrence of significant laboratory abnormalities will be 
summarized by regimen for each treatment. Vital signs will be summarized in the same manner 
as clinical laboratory evaluations, and physical examination data will be listed.
SAMPLE SIZE DETERMINATION:
Up to 8 subjects will be enrolled and dosed in 2 cohorts with 4 subjects in each cohort who 
complete Day 15. The sample size in the study is based on clinical and practical considerations 
rather than statistical power.
SITES: 1 site in Australia.
SPONSOR:
Orpheris, Inc.
1235 Radio Road, Suite 200
Redwood City, CA 94065
Telephone:


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
7
TABLE OF CONTENTS
Signature Page ................................................................................................................................ 2
Investigator Agreement................................................................................................................... 3
Synopsis.......................................................................................................................................... 4
Table of Contents............................................................................................................................ 7
List of Figures............................................................................................................................... 11
List of Abbreviations and Definition of Terms............................................................................. 12
1
Introduction and Background Information ............................................................................. 14
1.1 
Rationale........................................................................................................................ 15
1.2 
Risk/Benefit................................................................................................................... 15
2
Study Objectives..................................................................................................................... 17
2.1 
Primary Objective ......................................................................................................... 17
2.2 
Secondary Objective ..................................................................................................... 17
3
Study Description.................................................................................................................... 18
3.1 
Summary of Study Design ............................................................................................ 18
3.2 
Study Stopping Rules.................................................................................................... 19
3.3 
Study Indication ............................................................................................................ 19
4
Selection and Withdrawal of Subjects .................................................................................... 20
4.1 
Inclusion Criteria........................................................................................................... 20
4.2 
Exclusion Criteria.......................................................................................................... 21
4.3 
Withdrawal Criteria....................................................................................................... 22
5
Study Treatments .................................................................................................................... 23
5.1 
Treatment Groups.......................................................................................................... 23
5.2 
Rationale for Dosing ..................................................................................................... 23
5.3 
Randomization and Blinding......................................................................................... 24
5.4 
Breaking the Blind ........................................................................................................ 24
5.5 
Drug Supplies................................................................................................................ 24
5.5.1
Formulation and Packaging............................................................................... 24
5.5.2
Study Drug Preparation and Dispensing ........................................................... 24
5.5.3
Study Drug Administration ............................................................................... 24
5.5.4
Treatment Compliance ...................................................................................... 25


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
8
5.5.5
Storage and Accountability ............................................................................... 25
5.6 
Prior and Concomitant Medications and/or Procedures................................................ 25
5.6.1
Excluded Medications and/or Procedures......................................................... 25
5.6.2
Restricted Medications and/or Procedures........................................................ 25
5.6.3
Restricted Cigarette Use.................................................................................... 25
5.6.4
Permitted Medications....................................................................................... 25
5.6.5
Documentation of Prior and Concomitant Medication Use.............................. 25
6
Study Procedures .................................................................................................................... 26
6.1 
Screening Visit (Days -21 to -1) ................................................................................... 26
6.2 
Check-in (Day -1).......................................................................................................... 26
6.3 
Treatment Period (Days 1 to 3)..................................................................................... 27
6.3.1
Day 1
27 
6.3.2
Day 2
27 
6.3.3
Day 3
27 
6.4 
Follow-up Visit (Day 8) ................................................................................................ 28
6.5 
Follow-up Visit (Day 15) .............................................................................................. 28
6.6 
Early Termination Visit and Withdrawal Procedures ................................................... 28
7
Pharmacokinetic Assessments ................................................................................................ 30
7.1 
Pharmacokinetic Parameters ......................................................................................... 30
8
Safety Assessments................................................................................................................. 31
8.1 
Adverse Events.............................................................................................................. 31
8.1.1
Adverse (Drug) Reaction .................................................................................. 31
8.1.2
Unexpected Adverse Drug Reaction................................................................. 31
8.1.3
Assessment of Adverse Events by the Investigator .......................................... 32
8.2 
Serious Adverse Events................................................................................................. 33
8.3 
Serious Adverse Event Reporting 
Procedures for Investigators................................ 34
8.4 
Pregnancy Reporting..................................................................................................... 34
8.5 
Expedited Reporting...................................................................................................... 34
8.6 
Clinical Laboratory Evaluations.................................................................................... 35
8.7 
Vital Signs..................................................................................................................... 35
8.8 
Physical Examinations .................................................................................................. 35
9
Statistics.................................................................................................................................. 36


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
9
9.1 
Analysis Populations..................................................................................................... 36
9.2 
Statistical Methods ........................................................................................................ 36
9.2.1
Analysis of Efficacy.......................................................................................... 36
9.2.2
Analysis of Pharmacokinetics ........................................................................... 36
9.2.3
Analysis of Safety ............................................................................................. 36
9.2.3.1
Adverse events.................................................................................... 36
9.2.3.2
Clinical laboratory evaluations ........................................................... 37
9.2.3.3
Vital signs ........................................................................................... 37
9.2.3.4
Physical examinations......................................................................... 37
9.2.4
Protocol Deviations........................................................................................... 37
9.2.5
Medical History................................................................................................. 37
9.2.6
Subject Disposition ........................................................................................... 37
9.2.7
Demographic and Baseline Characteristics....................................................... 37
9.2.8
Prior and Concomitant Medications.................................................................. 37
9.2.9
Interim Analysis ................................................................................................ 37
9.3 
Sample Size Determination........................................................................................... 37
10 Data Management and Record Keeping ................................................................................. 38
10.1 Data Management ......................................................................................................... 38
10.1.1 Data Handling ................................................................................................... 38
10.1.2 Computer Systems............................................................................................. 38
10.1.3 Data Entry ......................................................................................................... 38
10.1.4 Medical Information Coding............................................................................. 38
10.1.5 Data Validation ................................................................................................. 38
10.2 Record Keeping............................................................................................................. 38
11 Investigator Requirements and Quality Control ..................................................................... 39
11.1 Ethical Conduct of the Study ........................................................................................ 39
11.2 Institutional Ethics Committee (IEC) or Human Research Ethics Committee (HREC)39
11.3 Informed Consent.......................................................................................................... 39
11.4 Study Monitoring Requirements ................................................................................... 39
11.5 Disclosure of Data......................................................................................................... 40
11.6 Retention of Records..................................................................................................... 40
11.7 Publication Policy ......................................................................................................... 40


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
10
11.8 Financial Disclosure...................................................................................................... 41
12 Study Administrative Information.......................................................................................... 42
12.1 Protocol Amendments................................................................................................... 42
12.2 Address List................................................................................................................... 42
12.2.1 Sponsor.............................................................................................................. 42
12.2.2 Contract Research Organization........................................................................ 42
12.2.3 Serious Adverse Event Reporting ..................................................................... 42
12.2.4 Biological Specimens........................................................................................ 42
13 References............................................................................................................................... 43
Appendix A:  Schedule of Procedures.......................................................................................... 45
Appendix B:  Clinical Laboratory Analytes ................................................................................. 48


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
11
LIST OF FIGURES
Figure 1. OP-101 Single-Ascending Subcutaneous Dose Schedule – Day 1............................. 18


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
12
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
Abbreviation
Definition
%CV
Percent coefficient of variation
ALD
Adrenoleukodystrophy
ANOVA
Analysis of variance
AUC0-inf
Area under the concentration-time curve from time zero to infinity
AUC0-t
Area under the concentration-time curve based on the last 
measurable concentration
BP
Blood pressure
ccALD
Childhood cerebral adrenoleukodystrophy
CFR
Code of Federal Regulations
CL
Clearance of study drug
Cmax
Maximum plasma concentration
CNS
Central nervous system
CRA
Clinical Research Associate
ECG
Electrocardiogram
eCRF
Electronic case report form
EDC 
Electronic data capture
eGFR
Estimated glomerular filtration rate 
FSH 
Follicle-stimulating hormone
GI 
Gastrointestinal 
GLP
Good Laboratory Practice
GSH
Glutathione
HBsAg
Hepatitis B surface antigen
HCG
Human chorionic gonadotropin
HCVab
Hepatitis C virus antibody
HIV
Human immunodeficiency virus
HREC
Human Research Ethics Committee 
HSCT
Hematopoietic stem cell transplantation
ICF 
Informed consent form
ICH 
International Council for Harmonisation 
IEC
Institutional Ethics Committee 
IL
Interleukin
IV
Intravenous 
kel
Apparent terminal rate constant
KIM-1
Kidney injury molecule-1
NAC
N-acetyl-cysteine
NGAL
Neutrophil gelatinase-associated lipocalin
PAMAM
Polyamidoamine


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
13
Abbreviation
Definition
PD 
Pharmacodynamic(s)
PK
Pharmacokinetic(s)
ROS
Reactive oxygen species
SAE 
Serious adverse event 
SD 
Standard deviation
SRC 
Safety review committee 
t½
Apparent elimination half-life
TGA
Therapeutic Goods Administration
tmax
Time to maximum plasma concentration


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
14
1
INTRODUCTION AND BACKGROUND INFORMATION 
OP-101 is a new chemical entity consisting of a metabolically stable inactive dendrimer covalently 
coupled to n-acetyl-cysteine (NAC). Upon intravenous (IV) administration, OP-101 is selectively 
taken up by reactive microglia and reactive astrocytes in the brain through fluid phase endocytosis. 
The active component of OP-101, NAC, is released upon uptake into reactive microglia and 
astrocytes. Upon intracellular release from OP-101, NAC acts to reduce oxidative stress and 
neuroinflammation in the brain. Following release of NAC, the dendrimer component is excreted 
from the cells and cleared without further metabolism.
Specifically, OP-101 consists of a generation-4 hydroxyl-terminated polyamidoamine (PAMAM) 
dendrimer linked to NAC using a disulfide bond. PAMAM is a class of dendrimer made of 
repetitive reactions of methyl acrylate and ethylene diamine. The PAMAM dendrimer contains an 
ethylene diamine core, amidoamine repeat units ([CH2CH2CONHCH2CH2N]), and 64 hydroxyl 
end groups (chemical formula: C622H1184N186O188). Approximately 20 of the 64 hydroxyls are 
subsequently converted to amine groups and then to a thiol group for eventual conjugation with 
NAC using a disulfide bond. The active metabolite of OP-101, NAC, comprises approximately 
16% of the total mass of OP-101. 
In vitro and in vivo nonclinical studies have demonstrated that the PAMAM dendrimer is 
metabolically stable and rapidly excreted via the kidney. Upon IV administration, OP-101 is 
endocytosed by reactive microglia and astrocytes in animal models of neuroinflammation. After 
endocytosis, the reducing environment inside these cells results in release of NAC and a 
normalization of glutathione (GSH) levels. As the active metabolite of OP-101, NAC reduces 
oxidative stress and related proinflammatory cy
decreasing neuroinflammation and central nervous system (CNS) toxicity. Pharmacology studies 
support the specific targeting of OP-101 to microglia and the intracellular release of NAC 
(Kurtoglu 2009; Navath 2008), as well as the ability of OP-101 to significantly reduce a 
proinflammatory response as compared with unconjugated free NAC (Dai 2010; 
Kambhampati 2015; Kannan 2012; Lesniak 2013; Mishra 2014; Nance 2015; Nemeth 2017).  
NAC, the active metabolite of OP-101, is an approved product and marketed under Acetadote ,
and the putative mechanism of action of OP-101 is related to the chemical and biological properties 
of NAC. NAC has been approved for use as a mucolytic agent and for the treatment of
acetaminophen intoxication in addition to other conditions and disorders, including 
doxorubicin-induced 
cardiotoxicity, 
radio-contrast-induced 
nephropathy, 
and 
chemotherapy-induced toxicity (Samuni 2013). NAC has been shown to interact with several 
metabolic pathways including regulation of cell cycle and apoptosis, but the main clinical effect 
of NAC is attributed mainly to its ability to scavenge reactive oxygen species (ROS) and elevate 
cellular GSH levels. NAC is recognized to suppress microglial activation (Tsai 2009) and inhibit 
-mediated proinflammatory cytokines by macrophages and glial cells (Pahan 1998). 
Examples of CNS effects of NAC include reduced levels of proinflammatory cytokines tumor 
necrosis factor-alpha, interleukin (IL)-1beta in rodent models of traumatic brain injury and cerebral 
ischemia (Chen 2008; Khan 2004), and IL-6 and IL-10 in lipopolysaccharide-treated rat fetal brain 
(Beloosesky 2012). The human safety profile for NAC, based on the collective experience with 
Acetadote, supports human testing of OP-101.  
OP-101 is being developed as a possible treatment for patients with advanced childhood cerebral 
adrenoleukodystrophy (ccALD) who are not considered eligible for hematopoietic stem cell 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
15
transplantation (HSCT). Adrenoleukodystrophy (ALD) is an X-linked disease caused by mutations 
in the ABCD1 that result in oxidative stress and neuroinflammation in several critical CNS cell 
types (Deon 2016). The disease affects cerebral white matter, spinal cord, and peripheral nerves, 
with some pediatric phenotypes suffering rapid progression leading to death in early childhood 
(Moser 2000; Moser 2007). About 35% of all hemizygous males between 3 and 10 years of age 
develop a form of ALD that manifests with progressive neuroinflammation resulting in widespread 
demyelination and rapid neurological deterioration. This phenotype is referred to as ccALD and 
typically leads to death within 2 to 5 years after onset of symptoms. While HSCT may stabilize 
patients in the very early stages of ALD, there is no effective therapy for patients who present with 
advanced stages of ccALD where HSCT is contraindicated. Therefore, OP-101 is being 
investigated as a potential treatment for patients with advanced ccALD who present with signs of 
progressive neuroinflammation potentially reversible by OP-101 and who are not candidates for 
HSCT.
1.1
Rationale
A
first-in-human study has been completed to evaluate
the safety, tolerability, and 
pharmacokinetics (PK) of IV administered OP-101 in normal healthy humans. This study indicated 
that treatment with a single IV dose of OP-101 at 20 and 40 mg/kg was generally well tolerated in 
healthy subjects based on an assessment of clinical and laboratory adverse events. The half-life of 
OP-101 in this study (5.5 hr) demonstrated the need for development of a more convenient dosing 
route (e.g. subcutaneous).  This study is being conducted to enable selection of the starting dose 
and regimen in a planned Phase 1/2/3 study in patients with ccALD. Currently, ccALD patients 
with progressive disease not treatable with HSCT have no therapeutic options, and reactive 
microglia are a recognized cause of the neuroinflammation associated with progression in ccALD. 
OP-101 has been demonstrated to selectively target reactive microglia and astrocytes in animal 
models of neuroinflammation, and has also been shown ex vivo to be effective in normalizing 
activated macrophages cells derived from ccALD patients. By targeting the reactive microglia,
OP-101 has the potential to stop or slow neurodegeneration in ccALD patients through antioxidant 
and anti-inflammatory mechanisms and may provide clinical benefit to ccALD patients not eligible 
for HSCT. Reversing the neuroinflammation driving the white matter injury in ccALD patients 
could stabilize the disease course and preserve neurological, developmental, and cognitive 
function. 
The safety and PK data collected in this subcutaneous dosing study will permit identification of 
the starting dose and regimen for the Phase 1/2/3 study in ccALD and support further clinical 
development of OP-101.
1.2
Risk/Benefit
This study is a follow up to the completed Phase 1 single ascending IV dosing study in healthy 
volunteers. In the completed Phase 1 study, treatment with a single IV dose of OP-101 at 20 and 
40 mg/kg was generally well tolerated in healthy subjects based on an assessment of clinical and 
laboratory adverse events. There were no clinically meaningful changes from baseline in 
laboratory parameters except changes in the urinalysis parameters of proteinuria and the presence 
of granular urinary casts. Transient elevations in KIM-1 and NGAL were observed, and subjects 
who experienced an increase in KIM-1 or NGAL at Day 15 underwent additional follow-up, at the 
recommendation of the SRC. The additional measurements in either KIM-1 or NGAL were 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
16
conducted as a precautionary measure and the values subsequently returned to baseline. There 
were no clinically meaningful changes in vital signs or ECGs. OP-101 has also been tested in 
nonclinical studies with no adverse clinical changes observed across multiple species and models. 
The anticipated risks associated with OP-101 may be categorized as potentially related to the 
PAMAM dendrimer and/or NAC. In vitro studies have demonstrated that the dendrimer is not 
systemically metabolized in human hepatocytes or human plasma and is rapidly cleared by the 
kidney. Good Laboratory Practice toxicity studies with repeat dosing of OP-101 in juvenile and 
adult rats have shown partial dose-dependent changes of proximal tubular cells characterized by 
mild intracellular basophilia and mild single-cell necrosis, but no associated clinical adverse 
events, mortality, or change in electrolytes. Toxicology studies in rats with single-doses of OP-101
have demonstrated resolution of the single-cell necrosis by 2 weeks postdosing and no adverse 
clinical observations or changes in urinalysis parameters. In summary, the renal clearance of 
OP-101 and the effects on proximal tubular cells in nonclinical studies indicate that renal function 
should be carefully assessed in human studies.
The active metabolite of OP-101, NAC, is an approved product and marketed under Acetadote. 
NAC is only released from OP-101 under intracellular GSH levels, and lower levels of GSH in 
plasma are insufficient to release NAC systemically. Regardless, the total level of NAC, if 
completely released from the highest possible human OP-101 dose, is well below the total systemic 
NAC typically administered for acetaminophen toxicity. Thus, the risks associated with Acetadote 
are relevant but unlikely to manifest given the projected low systemic exposure to NAC, even if 
fully released upon administration. For completeness, the major adverse events associated with IV 
NAC administration have been categorized as anaphylactoid reactions, with most either mild or 
moderate in degree. The most common adverse effects associated with NAC infusion were minor 
anaphylactoid reactions, including rash, urticarial, hypotension, and wheezing or shortness of 
breath. Literature reports indicate that adverse reactions consistent with hypersensitivity have been 
easily treated with antihistamines, epinephrine, and/or slowing of the drug infusion. Other reported 
adverse events related to IV NAC administration include tachycardia, nausea, and emesis. The 
frequency of adverse events has been reported to be between 0.2% and 20.8% and most commonly 
occur during the initial IV infusion of Acetadote (Acetadote Prescribing Information, Cumberland 
Pharmaceuticals Inc.).
In summary, clinical risks associated with the proposed subcuntaneous (SC) doses of OP-101 are 
considered low based on the lack of clinical adverse events observed at 3-5 fold higher doses 
administered IV in healthy volunteers. In addition, the human safety profile established for NAC, 
which is the active moiety in OP-101, further predicts a favorable safety profile and low risk with 
human investigation. 
There is no expected benefit of IV OP-101 administration for normal healthy subjects. Although 
safety and efficacy of OP-101 have not yet been evaluated in clinical studies, the collective efficacy 
observed in nonclinical studies indicate the potential to reverse neuroinflammation by targeting 
the reactive microglia in patients with ccALD. Because of the central role of neuroinflammation 
in driving disease progression in ccALD, the potential benefit in the intended patient population 
could include stabilization of white matter injury, and therefore the preservation of motor, 
cognitive, and developmental functions similar to the treatment effect after successful HSCT.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
17
2
STUDY OBJECTIVES
2.1
Primary Objective
The primary objective is to evaluate the safety and tolerability of OP-101 after single SC doses in 
healthy subjects.
2.2
Secondary Objective
The secondary objective is to determine the pharmacokinetic (PK) profile of OP-101 after single 
SC doses in healthy subjects.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
18
3
STUDY DESCRIPTION
3.1
Summary of Study Design
This is a Phase 1, open-label, single-ascending dose study to evaluate the safety, tolerability, and 
PK of OP-101 after SC administration in up to 8 healthy subjects at 1 site in Australia.
This study will consist of a 21-day Screening Period, a 2-day Treatment Period, and Follow-up 
Visits on Days 8 and 15. Subjects will sign an informed consent form (ICF) before any study 
procedures are performed and must meet all of the inclusion and none of the exclusion criteria to 
participate in the study. Eligible subjects will be admitted to the investigational site on Day -1, 
dosed on Day 1, and discharged on Day 3.
Figure 1 presents the cohort dosing schematic. There will be 2 separate and sequential SC cohorts 
(4 mg/kg and 8 mg/kg) with 4 subjects in each cohort who complete Day 15. Cohort 1 will enroll 
4 subjects administered a single SC dose of 4 mg/kg.  Dose escalation from Cohort 1 (4 mg/kg) to 
Cohort 2 (8 mg/kg) will be determined by the safety review committee (SRC), consisting of the 
Medical Monitor, the Principal Investigator (or delegate), and Sponsor representative after review 
of safety, tolerability, and any available PK data through Day 8 of Cohort 1. Cohort 2 will enroll 
4 subjects administered a single SC dose of 8 mg/kg.
During the Treatment Period, plasma and urine samples will be collected for PK analysis. Plasma 
PK samples will be collected predose, at 30 minutes and at 1, 2, 4, 6, 8, 10, 12, 16, 24, and 30-36 
hours after dosing, as well as on Day 3 (~48 hr). Blood samples for PK analysis can be obtained 
within 5 minutes of the scheduled sampling time for sample times 1 hour, within 10 minutes 
of the scheduled sampling time for sample times 12 hours, and within 2 hours of the scheduled 
sampling times for sample times >12 hours. 
Urine PK samples will be collected predose (spot sample) and between 0 to 4, 4 to 8, 8 to 12, 
12 to 18, 18 to 24, and 24 to 48 hours.
Safety assessments will include adverse event monitoring, clinical laboratory tests (hematology,
chemistry, and urinalysis), physical examinations, and vital signs (pulse rate, blood pressure [BP], 
respiration rate, and temperature). Coagulation testing will be performed at screening. Pregnancy 
testing will be performed at screening and Day -1.
Figure 1. OP-101 Single-Ascending Subcutaneous Dose Schedule – Day 1
1.
Cohort enrollment will be separate and sequential with 4 subjects in each cohort who complete Day 15.
2.
Cohort 1 will enroll 4 subjects administered a single SC dose of 4 mg/kg OP-101. Dose escalation to Cohort 2 (8 mg/kg) 
will be determined by the SRC after review of safety, tolerability and any available PK data through Day 8 of Cohort 1.  
Cohort 2 will enroll 4 subjects administered a single SC dose of 8 mg/kg.
PK = pharmacokinetic(s); SRC = safety review committee.
Cohort 2 [1,2]
8 mg/kg OP-101
Cohort 1 [1,2]
4 mg/kg OP-101


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
19
3.2
Study Stopping Rules 
This study will stop and no further dosing and/or dose escalation will occur until safety information 
can be reviewed in the event that:
1 subject within a dose cohort has a drug-related Grade 3 adverse event or serious adverse 
event (SAE), or
2 subjects within the same dose cohort have a drug-related Grade 2 adverse event. 
3.3
Study Indication
OP-101 is being developed for the treatment of children with ccALD who are not eligible for 
HSCT.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
20
4
SELECTION AND WITHDRAWAL OF SUBJECTS 
4.1
Inclusion Criteria
Subjects who meet all of the following criteria will be eligible to participate in the study:
1. Is a healthy man or woman age 18 to 65 years, inclusive, at the Screening Visit;
2. Has the ability to understand and sign the written ICF and local medical privacy authorization 
forms, which must be obtained prior to any study related procedures being completed;
3. Body mass index (BMI) between 18 and 32 kg/m2, inclusive;
4. Is in general good health, based upon the results of a medical history assessment, physical 
examination, vital signs, and laboratory profile, as judged by the Investigator;
5. Female subjects of non-childbearing potential must be either surgically sterile (hysterectomy, 
bilateral tubal ligation, salpingectomy, and/or bilateral oophorectomy at least 26 weeks before 
the Screening Visit) or postmenopausal, defined as spontaneous amenorrhea for at least 
2 years, with follicle-stimulating hormone (FSH) in the postmenopausal range at screening,
based on the central laboratory’s ranges;
6. Female subjects of childbearing potential (ie, ovulating, premenopausal, and not surgically 
sterile) and all male subjects must use a medically accepted contraceptive regimen (including 
hormonal contraceptives) during their participation in the study and for 30 days after the last 
administration of study drug. Medically accepted contraceptive methods are defined as those 
with 90% or greater efficacy;
7. Acceptable methods of contraception for male subjects enrolled in the study include the 
following:
Condoms or surgical sterilization of subject at least 26 weeks before the Screening Visit 
(vasectomy);
Acceptable methods of contraception for female subjects enrolled in the study include the 
following:
Surgical sterilization of subject at least 26 weeks before the Screening Visit (includes 
hysterectomy or bilateral tubal ligation, oophorectomy, or salpingectomy);
Intrauterine device for at least 12 weeks before the Screening Visit;
Hormonal contraception (oral, implant, injection, ring, or patch) for at least 12 weeks 
before the Screening Visit; or
Diaphragm;
8. If male, subjects must agree to abstain from sperm donation through 90 days after 
administration of the last dose of study drug; 
9. Female subjects may not be pregnant, lactating, or breastfeeding;
10. Female subjects of childbearing potential must have negative result for pregnancy test at 
screening and Check-in;


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
21
11. Subjects must have a negative test result for hepatitis B surface antigen (HBsAg), hepatitis C 
virus antibody (HCVab), and human immunodeficiency virus (HIV) antibody at screening;
12. Subjects must have an estimated glomerular filtration rate (eGFR) of 90 mL/min/1.73m2 at 
screening;
13. Subjects must have a negative urine test for drugs of abuse (opiates, benzodiazepines, 
amphetamines, cannabinoids, cocaine, barbiturates, and phencyclidine), cotinine, and breath 
alcohol test at screening and Check-in; and
14. Subjects must be willing and able to abide by all study requirements and restrictions.
4.2
Exclusion Criteria
Subjects who meet any of the following criteria will be excluded from participation in the study:
1. Evidence of clinically significant hematologic, renal, endocrine, pulmonary, cardiac, 
gastrointestinal (GI), hepatic, psychiatric, neurologic, immunologic, allergic disease (including 
multiple or clinically significant drug allergies), or any other condition that, in the opinion of 
the Investigator, might significantly interfere with the absorption, distribution, metabolism, or 
excretion of study drug, or place the subject at an unacceptable risk as a participant in this 
study;
2. History of malignancy (other than successfully treated basal cell or squamous cell skin cancer);
3. History or presence of an abnormal ECG that, in the opinion of the Investigator, is clinically 
significant;
4. Laboratory results (serum chemistry, hematology, coagulation, and urinalysis) outside the 
normal range at screening and Check-in and considered clinically significant in the opinion of 
the Investigator. Any elevation of aspartate transaminase (AST) and alanine transaminase 
(ALT) above the upper limit of normal at screening and/or Check-in is exclusionary. One retest 
of an exclusionary laboratory result is allowed at the discretion of the Investigator;
5. Has had an acute illness considered clinically significant by the Investigator within 30 days 
prior to screening;
6. History of alcoholism or drug abuse within 2 years prior to screening;
7. Has used any product containing nicotine within 90 days prior to screening or intends to use 
any product containing nicotine during the course of the study;
8. Has had any immunizations (live vaccines) in the 4 weeks prior to screening;
9. Has used medications that affect GI motility or gastric emptying; such as metoclopramide, 
proton pump inhibitors, and H2 blockers; within 30 days prior to Day 1;
10. Has used any prescription or over-the-counter medication (with exception of acetaminophen),
vitamins/herbal supplements (with the exception of hormonal contraceptives) within 14 days 
prior to Day 1;
11. Has used any other study drug within 30 days or 5 half-lives of the drug (whichever is longer) 
prior to Day 1;
12. Has lost or donated >450 mL of whole blood or blood products within 30 days prior to 
screening;


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
22
13. Investigator has reason to believe that the subject may be unable to fulfill the protocol visit 
schedule or requirements;
14. Has any finding that, in the view of the Investigator or Medical Monitor, would compromise 
the subject’s safety requirements; or
15. Is employed by the Sponsor, the Contract Research Organization (CRO), or the study site 
(permanent, temporary contract worker, or designee responsible for the conduct of the study), 
or is a family member (spouse, parent, sibling, or child) of the Sponsor, CRO, or study site 
employee.
4.3
Withdrawal Criteria
Participation of a subject in this clinical study may be discontinued for any of the following 
reasons:
The subject withdraws consent or requests discontinuation from the study for any reason;
Occurrence of any medical condition or circumstance that exposes the subject to substantial 
risk and/or does not allow the subject to adhere to the requirements of the protocol;
Any SAE, clinically significant adverse event, severe laboratory abnormality, intercurrent 
illness, or other medical condition which indicates to the Investigator that continued 
participation is not in the best interest of the subject;
Pregnancy;
Requirement of prohibited concomitant medication;
Subject failure to comply with protocol requirements or study-related procedures;
Termination of the study by the Sponsor or the regulatory authority; or 
Subject noncompliance. 
If a subject withdraws prematurely from the study due to the above criteria or any other reason, 
study staff should make every effort to complete the full panel of assessments scheduled for the 
Early Termination Visit. The reason for subject withdrawal must be documented in the electronic 
case report form (eCRF).
If a subject is withdrawn from the study due to an adverse event, the subject will be asked to return 
to the site for, at a minimum, the evaluations scheduled for the Early Termination Visit. If the 
adverse event has still not resolved, additional follow-up will be performed, as appropriate, based 
on the Investigator’s judgment, and documented in the subject’s medical records. As a minimum 
requirement, SAEs should be followed for 30 days after the subject’s last dose of study drug.
Withdrawn subjects will be replaced if the subject is withdrawn prior to dosing. Withdrawn 
subjects may be replaced after dosing if adequate PK data is not available for that subject at the 
discretion of the Investigator and Medical Monitor.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
23
5
STUDY TREATMENTS
5.1
Treatment Groups
There will 2 separate and sequential SC cohorts with 4 subjects in each cohort. The population will 
include healthy men and women aged 18 to 65 years, inclusive. 
Dose escalation from Cohort 1 to Cohort 2 will be determined by the SRC, consisting of the 
Medical Monitor, the Principal Investigator (or delegate), and Sponsor representative after review 
of safety, tolerability, and any available PK data through Day 8 of Cohort 1.
Dosing with OP-101 is planned for the cohorts as follows: 
Cohort 1: Single SC dose of 4 mg/kg, and 
Cohort 2: Single SC dose of 8 mg/kg.
5.2
Rationale for Dosing
The safety of OP-101 is supported by the completed Phase 1 IV dosing study in healthy volunteers 
at 20 and 40 mg/kg (3-5 fold higher than the planned SC doses in this study).  In addition, a series 
of nonclinical studies previously conducted with the active agent in OP-101, NAC, and numerous 
clinical studies that have been performed with NAC provide further support for the proposed 
dosing. The overall toxicological profile of OP-101 is expected to be comparable to NAC, which 
has been evaluated in nonclinical toxicity studies in support of the approval of Acetadote.
Good Laboratory Practice toxicology studies conducted with OP-101 in juvenile and adult rats, 
incorporating repeat IV doses up to 2000 mg/kg or single IV doses up to 1000 mg/kg, have shown 
no adverse clinical signs or mortality. The predominant finding in the toxicology studies was 
microscopic changes in the kidney that were generally localized to the proximal tubule. The 
findings included tubular degeneration/regeneration, mineralization, basophilic granules/tubules, 
and/or single cell necrosis, which increased in severity with increasing dose. While a defined 
NOAEL was not identified in the repeat-dose studies, the single-dose study in adult male rats, 
demonstrated a NOAEL of 200 mg/kg based upon the absence of single cell necrosis by 2 weeks 
after dosing and minimal-to-mild severity of this finding noted at earlier time points. Of note, no 
effects were observed for urinalysis or urine chemistry parameters, and no changes in other urinary 
biomarkers (kidney injury molecule-1, neutrophil gelatinase-associated lipocalin, albumin, 
c
-microglobulin) were detected.
From the completed IV dosing study in healthy volunteers it was determined that the clearance of 
OP-101 was rapid with no detectable drug in the plasma after 24 hours.  There were also no clinical 
adverse events in subjects administered IV doses of 20 or 40 mg/kg OP-101.  To enable chronic 
dosing, SC dosing is being evaluated in this study.  The SC dosage formulation has been optimized 
to enable a single SC dose of 400-500 mg.  Higher doses may require two SC injections to achieve 
the desired dose if necessary. The selection of 4 and 8 mg/kg SC doses enable the majority of the 
subjects to receive only one injection and is 3-5 fold lower dose than the previous IV study in 
healthy volunteers.  Exposure to NAC at the proposed starting dose and subsequent dose cohorts 
in this study is orders of magnitude less than that demonstrated to be safe in humans following 
oral or IV administration.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
24
5.3
Randomization and Blinding
This open-label study consists of 2 separate and sequential SC cohorts. Randomization and 
blinding methods will not be used in this study. 
Cohort 1 will enroll 4 subjects administered a single SC dose of 4 mg/kg.  Dose escalation from 
Cohort 1 (4 mg/kg) to Cohort 2 (8 mg/kg) will be determined by the safety review committee 
(SRC), consisting of the Medical Monitor, the Principal Investigator (or delegate), and Sponsor 
representative after review of safety, tolerability, and any available PK data through Day 8 of 
Cohort 1. Cohort 2 will enroll 4 subjects administered a single SC dose of 8 mg/kg.
If an eligible subject is not used in the first cohort and is outside the screening window for a future 
cohort, the subject may be rescreened.
5.4
Breaking the Blind
This is an open-label study. Study drug assignment will be known by subjects, investigational site 
staff, and the Sponsor.
5.5
Drug Supplies
5.5.1
Formulation and Packaging
OP-101 study drug is provided as lyophilized powder in Schott Glass, 5 cc/13 mm Type 1 clear 
glass vials with West, 13 mm, single vent lyophilizer grey butyl chloride stoppers, and West 
13 mm flip-top matte white caps. Each vial is filled to deliver 500 ± 50 mg OP-101.  Each vial is 
reconstituted 0.7 mL sterile water for injection to yield 400 mg/mL OP-101 in 20 mM histidine, 
pH 5.5 and 80 mg/mL trehalose. Vials of study drug will be supplied in boxes labeled for this 
study. Vials will be reconstituted with sterile water for injection prior to use.
OP-101 will be stored at 2 to 8°C and must not be frozen. Reconstituted OP-101 is stable at room
temperature for up to 7 days if the vial seal is not compromised.
5.5.2
Study Drug Preparation and Dispensing
Trained and qualified site personnel will administer OP-101 to subjects within the clinical facility. 
Site personnel should ensure that the study drug is stored in accordance with the environmental 
conditions (temperature, light, and humidity) as determined by Orpheris and outlined in the 
Pharmacy Manual. Instructions for OP-101 preparation are documented in the pharmacy manual.
5.5.3
Study Drug Administration
Study drug will be administered on Day 1 as a SC injection under the supervision of the site 
personnel.
Dose Escalation 
Cohort 1 will enroll 4 subjects administered a single SC dose of 4 mg/kg.  Dose escalation from 
Cohort 1 (4 mg/kg) to Cohort 2 (8 mg/kg) will be determined by the safety review committee 
(SRC), consisting of the Medical Monitor, the Principal Investigator (or delegate), and Sponsor 
representative after review of safety, tolerability, and any available PK data through Day 8 of 
Cohort 1. Cohort 2 will enroll 4 subjects administered a single SC dose of 8 mg/kg.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
25
5.5.4
Treatment Compliance
Study drug will be administered at the investigational site by site staff.
5.5.5
Storage and Accountability
Study drug will be stored in a secure, temperature-controlled location. Study drug will only be 
prepared and dispensed by authorized site personnel at the clinical site.
Documentation will be performed for the receipt, storage, dispensing, and retention of study drug 
during the conduct of study to ensure complete accountability. Used study drug vials will be 
retained for monitoring purposes.
5.6
Prior and Concomitant Medications and/or Procedures
5.6.1
Excluded Medications and/or Procedures
Subjects who have lost or donated >450 mL of whole blood or blood products within 30 days prior 
to screening will be excluded. 
See Section 4.2 for exclusion criteria. 
5.6.2
Restricted Medications and/or Procedures
Subjects are not permitted to take prescription or non-prescription drugs (including vitamins and
dietary or herbal supplements) within 14 days (14 days if the drug is a potential enzyme inducer) 
or 5 half-lives (whichever is longer) prior to the first dose of study medication until completion of 
the Day 15 Follow-up Visit, unless, in the opinion of the Investigator and Sponsor, the medication 
will not interfere with the study or is required out of medical necessity.
See Section 4.2 for exclusion criteria.
5.6.3
Restricted Cigarette Use
Subjects are not permitted to use any product containing nicotine during the course of this study 
and may not have used any product containing nicotine within 90 days prior to screening.
5.6.4
Permitted Medications
Nonsteroidal anti-inflammatory drugs (eg, ibuprofen or naproxen) may be considered for use 
during the study on a case-by-case basis by the Investigator in consultation with the Orpheris
Medical Monitor if required. 
Hormonal contraception (oral, implant, injection, ring, or patch) for at least 12 weeks before the 
Screening Visit is permitted. 
5.6.5
Documentation of Prior and Concomitant Medication Use
All medications administered during the study must be documented on the concomitant medication 
eCRF.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
26
6
STUDY PROCEDURES
A 30-minute window for vital signs and all postdose procedures is allowed.
When multiple procedures are scheduled at the same time point, the order of procedures should be 
as follows: obtain vital signs, collect blood sample (drawn at nominal time), and obtain urine 
sample.
Blood samples for PK analysis can be obtained within 5 minutes of the scheduled sampling time 
for sample times 1 hour, within 10 minutes of the scheduled sampling time for sample times 
12 hours, and within 2 hours of the scheduled sampling times for sample times >12 hours.
6.1
Screening Visit (Days -21 to -1)
The following procedures will be performed at screening: 
Obtain informed consent;
Evaluate inclusion/exclusion criteria;
Obtain demographics and medical history;
Record prior and concomitant medications;
Perform physical examination including height and weight;
Measure height and weight and calculate BMI;
Record vital signs;
Perform breath alcohol test;
Collect urine sample for urine drug screen, cotinine, and urinalysis; 
Obtain fasting blood sample for safety laboratory tests, coagulation (activated partial 
thromboplastin time [aPTT], partial thromboplastin time [PTT], and international normalized 
ratio [INR]) serum (beta human chorionic gonadotropin [HCG]) pregnancy test (women of 
childbearing potential only), FSH (only for women of non-childbearing potential who are not 
surgically sterile), HIV, HBsAg, and HCVab screen; and 
Assess adverse events.
6.2
Check-in (Day -1)
The following procedures will be performed at Day -1:
Admit subject to investigational site;
Record prior and concomitant medications;
Perform physical examination including weight for dose determination;
Record vital signs;
Perform breath alcohol test;


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
27
Collect urine sample for urine drug screen, cotinine, urinary albumin, urinary creatinine,
kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL) and 
urinalysis; 
Collect urine sample for pregnancy test women of childbearing potential only;
Obtain fasting blood sample for safety laboratory tests; and
Assess adverse events. 
6.3
Treatment Period (Days 1 to 3)
6.3.1
Day 1
The following procedures will be performed on Day 1:
Record prior and concomitant medications;
Record vital signs (predose, 30 and 60 minutes and at 2, 4, and 12 hours after SC dosing);
Collect urine sample for urinalysis predose;
Obtain fasting blood sample for safety laboratory tests within 30 minutes prior to dosing;
Administer study drug;
Collect plasma PK samples (collected within 30 minutes prior to dosing, and at 30 minutes and
1, 2, 4, 6, 8, 10, 12, and 16 hours);
Collect urine PK sample (within 60 to 30 minutes prior to dosing [spot sample] and between 0 
to 4, 4 to 8, 8 to 12, 12 to 18, and 18 to 24 hours after dosing);
Assess adverse events.
6.3.2
Day 2
The following procedures will be performed on Day 2:
Record prior and concomitant medications;
Perform physical examination;
Record vital signs at 24 hours;
Collect urine sample for urinary albumin, urinary creatinine, KIM-1, NGAL and urinalysis at 
24 hours;
Obtain fasting blood sample for safety laboratory tests at 24 hours;
Collect plasma PK sample (24 and 30-36 hours after dosing);
Collect urine PK sample (between 24 to 48 hours after dosing); and
Assess adverse events.
6.3.3
Day 3
The following procedures will be performed on Day 3:


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
28
Record prior and concomitant medications;
Perform physical examination;
Record vital signs at 48 hours;
Collect urine sample for urinary albumin, urinary creatinine, KIM-1, NGAL and urinalysis at 
48 hours;
Collect plasma PK sample (48 hours after dosing);
Collect urine PK sample up to 48 hours;
Obtain fasting blood sample for safety laboratory tests at 48 hours;
Assess adverse events; and
Discharge subject from the investigational site.
6.4
Follow-up Visit (Day 8)
The following procedures will be performed at the Follow-up Visit on Day 8:
Record prior and concomitant medications;
Perform physical examination;
Record vital signs; 
Collect urine sample for urinary albumin, urinary creatinine, KIM-1, NGAL and urinalysis;
Obtain fasting blood sample for safety laboratory tests; and 
Assess adverse events.
6.5
Follow-up Visit (Day 15)
The following procedures will be performed at the Follow-up Visit on Day 15:
Record prior and concomitant medications;
Perform physical examination;
Record vital signs; 
Collect urine sample for urinary albumin, urinary creatinine, KIM-1, NGAL and urinalysis;
Obtain fasting blood sample for safety laboratory tests; and
Assess adverse events.
6.6
Early Termination Visit and Withdrawal Procedures
The end of treatment for subjects completing the study is Day 15. For subjects who are withdrawn 
from the study prior to completion, the following procedures will be performed at the Early 
Termination Visit:
Record prior and concomitant medications;


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
29
Perform physical examination;
Record vital signs;
Collect urine sample for urinary albumin, urinary creatinine, KIM-1, NGAL and urinalysis; 
Obtain fasting blood sample for safety laboratory tests; and 
Assess adverse events.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
30
7
PHARMACOKINETIC ASSESSMENTS
The actual PK blood sampling times will be captured on the eCRF. The actual dosing time will 
also be captured on the eCRF.
During the Treatment Period, plasma and urine samples will be collected for PK analysis. Plasma 
PK samples will be collected predose, at 30 min and 1, 2, 4, 6, 8, 10, 12, 16, 24, and 30-36 hours 
after SC injection, as well as on Day 3 (~48 hr). Blood samples for PK analysis can be obtained 
within 5 minutes of the scheduled sampling time for sample times 1 hour, within 10 minutes 
of the scheduled sampling time for sample times 12 hours, and within 2 hours of the scheduled 
sampling times for sample times >12 hours. 
Urine PK samples will be collected predose (spot sample) and between 0 to 4, 4 to 8, 8 to 12, 
12 to 18, 18 to 24, and 24 to 48 hours after dosing.
7.1
Pharmacokinetic Parameters
Pharmacokinetic parameters will include the following:
Cmax – maximum plasma concentration,
tmax – time to maximum plasma concentration,
kel – apparent terminal rate constant, 
t½ – apparent elimination half-life, 
AUC0-t – area under the concentration-time curve based on the last measureable concentration,
AUC0-inf – area under the concentration-time curve from time zero to infinity, and
CL – clearance of study drug.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
31
8
SAFETY ASSESSMENTS
8.1
Adverse Events
An adverse event is defined as any untoward medical occurrence in a clinical investigation subject 
administered a pharmaceutical product, which does not necessarily have a causal relationship with 
this treatment. An adverse event can therefore be any unfavorable and/or unintended sign 
(including an abnormal laboratory finding), symptom, or disease temporally associated with the 
use of an investigational medicinal product, whether or not related to the investigational medicinal 
product. All adverse events, including observed or volunteered problems, complaints, or 
symptoms, are to be recorded on the appropriate eCRF.
Adverse events, which include clinical laboratory test variables, will be monitored and 
documented from the time of informed consent until study participation is complete. Subjects 
should be instructed to report any adverse event that they experience to the Investigator. Beginning 
with the screening visit, the Investigator should make an assessment for adverse events at each 
visit and record the event on the appropriate adverse event eCRF.
Wherever possible, a specific disease or syndrome rather than individual associated signs and 
symptoms should be identified by the Investigator and recorded on the eCRF. However, if an 
observed or reported sign or symptom is not considered a component of a specific disease or 
syndrome by the Investigator, it should be recorded as a separate adverse event on the eCRF.
Additionally, the condition that led to a medical or surgical procedure (eg, surgery, endoscopy, 
tooth extraction, or transfusion) should be recorded as an adverse event, not the procedure. 
Any medical condition already present at screening should not be reported as an adverse event 
unless the medical condition or signs or symptoms present at baseline (pre-dose) changes in 
severity or seriousness at any time during the study. In this case, it should be reported as an adverse 
event.
Clinically significant abnormal laboratory or other examination findings that are detected during 
the study or are present at screening and significantly worsen during the study should be reported 
as adverse events. The Investigator will exercise his or her medical and scientific judgment in 
deciding whether an abnormal laboratory finding or other abnormal assessment is clinically 
significant. Clinically significant abnormal laboratory values occurring during the clinical study 
will be followed until repeat tests return to normal, stabilize, or are no longer clinically significant. 
Any abnormal test that is determined to be an error does not require reporting as an adverse event.
8.1.1
Adverse (Drug) Reaction
All noxious and unintended responses to a medicinal product related to any dose should be 
considered an adverse drug reaction. “Responses” to a medicinal product means that a causal 
relationship between a medicinal product and an adverse event is at least a reasonable possibility, 
(ie, the relationship cannot be ruled out).
8.1.2
Unexpected Adverse Drug Reaction
An unexpected adverse drug reaction is defined as an adverse reaction, the nature or severity of 
which is not consistent with the applicable product information in the Investigator’s Brochure.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
32
8.1.3
Assessment of Adverse Events by the Investigator
The Investigator will assess the severity (intensity) of each adverse event as mild, moderate, or 
severe, and will also categorize each adverse event as to its potential relationship to study drug 
using the categories of yes or no.
The severity of all adverse events should be graded according to the Common Terminology 
Criteria for Adverse Events (CTCAE) Version 5.0. These criteria can be found at 
http://ctep.cancer.gov/reporting/ctc.html. For those adverse events not listed in the CTCAE, the 
following grading system should be used:
Mild (CTCAE Grade 1): Transient symptoms, awareness of sign/symptom, but easily tolerated 
and no interference with subject’s daily activities.
Moderate (CTCAE Grade 2): Marked signs/symptoms that interfere with subject’s usual 
activities, but still acceptable.
Severe (CTCAE Grade 3): Incapacitating signs/symptoms which cause considerable 
interference with the subject’s daily activities, unacceptable.
Life-threatening (CTCAE Grade 4): Life threatening or disabling adverse event.
Death (CTCAE Grade 5): Death-related adverse event.
Causality Assessment
The relationship of an adverse event to the administration of the study drug is to be assessed 
according to the following definitions:
No (unrelated, not related, no relation) – The time course between the administration of study drug 
and the occurrence or worsening of the adverse event rules out a causal relationship and another 
cause (concomitant drugs, therapies, complications, etc.) is suspected.
Yes (related) – The time course between the administration of study drug and the occurrence or 
worsening of the adverse event is consistent with a causal relationship and no other cause 
(concomitant drugs, therapies, complications, etc.) can be identified.
The definition implies a reasonable possibility of a causal relationship between the event and the 
study drug. This means that there are facts (evidence) or arguments to suggest a causal relationship.
The following factors should also be considered:
The temporal sequence from study drug administration-
o
The event should occur after the study drug is given. The length of time from study drug 
exposure to event should be evaluated in the clinical context of the event.
Underlying, concomitant, intercurrent diseases-
o
Each report should be evaluated in the context of the natural history and course of the 
disease being treated and any other disease the subject may have.
Concomitant drug-
o
The other drugs the subject is taking or the treatment the subject receives should be 
examined to determine whether any of them might be recognized to cause the event in 
question.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
33
Known response pattern for this class of study drug-
o
Clinical and/or preclinical data may indicate whether a particular response is likely to be a 
class effect.
Exposure to physical and/or mental stresses-
o
The exposure to stress might induce adverse changes in the recipient and provide a logical 
and better explanation for the event.
The pharmacology and pharmacokinetics of the study drug-
o
The known pharmacologic properties (absorption, distribution, metabolism, and excretion) 
of the study drug should be considered.
8.2
Serious Adverse Events
An adverse event or adverse reaction is considered serious if, in the view of either the Investigator
or Sponsor, it results in any of the following outcomes:
Death,
A life-threatening adverse event,
o
NOTE: An adverse event or adverse reaction is considered “life-threatening” if, in view of 
either the Investigator or Sponsor, its occurrence places the subject at immediate risk of 
death. It does not include an event that, had it occurred in a more severe form, might have 
caused death.
Requires hospitalization or prolongation of existing hospitalizations,
o
NOTE: Any hospital admission with at least 1 overnight stay will be considered an 
inpatient hospitalization. An emergency room visit without hospital admission will not be 
recorded as a SAE under this criterion, nor will hospitalization for a procedure scheduled 
or planned before signing of informed consent. However, unexpected complications and/or 
prolongation of hospitalization that occur during elective surgery should be recorded as 
adverse events and assessed for seriousness. Admission to the hospital for social or
situational reasons (ie, no place to stay, live too far away to come for hospital visits) will 
not be considered inpatient hospitalizations. 
A persistent or significant disability/incapacity or substantial disruption of the ability to 
conduct normal life functions,
A congenital anomaly/birth defect, or
An important medical event.
o
NOTE: Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered an SAE when, based upon appropriate medical 
judgment, they may jeopardize the subject and may require medical or surgical intervention 
to prevent one of the outcomes listed above. Examples of such medical events include 
allergic bronchospasm requiring intensive treatment in an emergency room or at home, 
blood dyscrasias or convulsions that do not result in inpatient hospitalizations, or the 
development of drug dependency.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
34
8.3
Serious Adverse Event Reporting 
Procedures for Investigators
Initial Reports
All SAEs occurring from the time of informed consent until 30 days following the last
administration of study drug must be reported to safety@clinical.net.au within 24 hours of the 
knowledge of the occurrence (this refers to any adverse event that meets any of the aforementioned 
serious criteria). All SAEs that the Investigator considers related to study drug occurring after the 
30-day follow-up period must be reported to the Sponsor.
If the event meets serious criteria, send an email of the paper SAE form to safety@clinical.net.au
or call the CNS Clinical Research Associate (CRA) or Project Manager (PM) within 24 hours of 
awareness.  The SAE information must be entered within the EDC within 24 hours of the system 
becoming available.
Safety Contact Information: safety@clinical.net.au
Follow-Up Reports
The Investigator must continue to follow the subject until the SAE has subsided or until the 
condition becomes chronic in nature, stabilizes (in the case of persistent impairment), or the subject 
dies.
Within 24 hours of receipt of follow-up information, the Investigator must update the SAE form 
electronically in the EDC system for the study and submit the updated SAE Paper form and any 
supporting documentation (eg, subject discharge summary or autopsy reports) to CNS via e-mail
at safety@clinical.net.au.
8.4
Pregnancy Reporting
If the subject or partner of a subject participating in the study becomes pregnant during the study 
or within 30 days of discontinuing study drug, the Investigator should report the pregnancy to CNS
within 24 hours of being notified via safety@clnical.net.au and cc the CRA and PM. CNS will 
then forward the Pregnancy Notification Form to the Investigator for completion.
A subject becoming pregnant while on study drug will immediately be withdrawn from the study 
and early termination study procedures will be performed.
The subject or partner should be followed by the Investigator until completion of the pregnancy. 
If the pregnancy ends for any reason before the anticipated date, the Investigator should notify 
CNS Safety via safety@clinical.net.au and cc CRA and PM.. At the completion of the pregnancy, 
the Investigator will document the outcome of the pregnancy. If the outcome of the pregnancy 
meets the criteria for immediate classification as an SAE (ie, postpartum complication, 
spontaneous abortion, stillbirth, neonatal death, or congenital anomaly), the Investigator should 
follow the procedures for reporting an SAE.
8.5
Expedited Reporting
The Sponsor will report all relevant information about suspected unexpected serious adverse 
reactions that are fatal or life-threatening as soon as possible to the TGA, and in any case no later 
than 7 days after knowledge by the Sponsor of such a case, and that relevant follow-up information 
will subsequently be communicated within an additional 8 days. 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
35
All other suspected unexpected serious adverse reactions will be reported to the TGA as soon as 
possible but within a maximum of 15 days of first knowledge by the Sponsor.
The Sponsor will also inform the Investigator as required. 
8.6
Clinical Laboratory Evaluations
Standard clinical laboratory profiles for chemistry, hematology, and urinalysis will be evaluated 
at screening, Day -1, and on Days 1 (pre-dose), 2 & 3. Samples will also be collected on Day 8,
Day 15, and at the Early Termination Visit. Note, the eGFR will be determined for each time point 
above to assess any potentially acute effects on kidney clearance. 
See Appendix B for a list of clinical laboratory analytes.
Tests for HIV, HBsAg, and HCVab will be completed at screening to determine eligibility.
Coagulation (aPTT, PTT, and INR) will be assessed at screening. 
A serum (beta HCG) pregnancy test will be performed at screening for female subjects of 
childbearing potential only. A urine pregnancy test will be performed at Day -1 for female subjects 
of childbearing potential only. A test for FSH level will be performed at screening only for women 
of non-childbearing potential who are not surgically sterile.
A urine drug, cotinine, and breath alcohol test will be administered at screening and Check-in. 
8.7
Vital Signs
Vital signs (pulse rate, respiration rate, body temperature, and BP) will be recorded at all study 
visits and the Early Termination Visit after the subject has been seated or supine for 5 minutes. 
During the Treatment Period, vital signs will be measured predose, 30 and 60 minutes and at 2, 4,
12, and 24, and 48 hours after dosing. Vital signs will also be measured at the Follow-up Visits on 
Days 8 and 15.
8.8
Physical Examinations
Complete physical examinations will be performed at screening, on Day -1, Day 2, Day 3, Day 8,
and Day 15, or at the Early Termination Visit. Body weight and height will be measured at 
screening and used to calculate BMI.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
36
9
STATISTICS
9.1
Analysis Populations
The Safety Population is defined as all subjects who received at least 1 dose of study drug.
The PK Concentration Population is defined as all subjects who received at least 1 dose of study 
drug and have at least 1 measurable concentration. 
The PK Parameter Population is defined as all subjects who received a dose of study drug and have 
at least 1 PK parameter.
9.2
Statistical Methods
A detailed Statistical Analysis Plan (SAP) describing the methodology to be used in the final 
analysis will be prepared and finalized prior to database lock. Statistical methods described here 
may be changed based on advances in research. If changes in the statistical methods are made 
which change the principal features of the protocol, a protocol amendment will be required. Any 
deviation from the planned statistical analyses in the protocol will be fully described in the SAP.
9.2.1
Analysis of Efficacy
Not applicable.
9.2.2
Analysis of Pharmacokinetics
Plasma PK concentrations of OP-101 will be summarized with descriptive statistics (n, mean, 
standard deviation [SD], percent coefficient of variation [%CV], minimum, median, maximum, 
geometric mean, and %CV for the geometric mean) by nominal PK sampling time and treatment 
using the PK Concentration Population.
Standard noncompartmental PK analysis will be used to calculate the PK parameters.
Pharmacokinetic parameters will be summarized with descriptive statistics (n, mean, SD, %CV, 
minimum, median, maximum, geometric mean, and %CV for the geometric mean) by treatment 
using the PK Parameter Population.
9.2.3
Analysis of Safety
Safety will be assessed using the Safety Population. 
9.2.3.1
Adverse events 
Adverse events will be summarized overall and by system organ class and preferred term, and the 
relationship to the study drug received. A list of subjects who have SAEs and who discontinue 
from the study due to an adverse event will be provided. Deaths due to adverse events will also be 
provided. 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
37
9.2.3.2
Clinical laboratory evaluations
Safety laboratory tests will include chemistry, hematology, and urinalysis. Summary statistics for 
laboratory values will be provided at baseline, postbaseline, and for changes from baseline to 
postbaseline by treatment. Occurrence of significant laboratory abnormalities will be summarized 
by regimen for each treatment.
9.2.3.3
Vital signs 
Vital signs will be summarized in the same manner as clinical laboratory evaluations.
9.2.3.4
Physical examinations
Physical examination data will be listed.
9.2.4
Protocol Deviations 
Subjects with protocol deviations will be listed for the Safety Population.
9.2.5
Medical History
Medical history will be listed for each treatment for the Safety Population.
9.2.6
Subject Disposition
Subject disposition information will be tabulated for each treatment based on the enrolled subjects. 
Counts of subjects enrolled, as well as counts and percentages of subjects who completed each 
period and the study, and who withdrew early from the study will be presented. The primary 
reasons for early withdrawal will also be tabulated.
9.2.7
Demographic and Baseline Characteristics
Demographic and baseline characteristics data will be summarized for each treatment.
9.2.8
Prior and Concomitant Medications 
Concomitant medication will be summarized for each treatment. A listing of all medications 
including the reported term, dictionary term, start and stop dates, and other relevant data will be 
provided.
9.2.9
Interim Analysis
Not applicable. 
9.3
Sample Size Determination
8 subjects will be enrolled and dosed in 2 cohorts with 4 subjects in each cohort to complete 
Day 15. The sample size in the study is based on clinical and practical considerations rather than 
statistical power.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
38
10
DATA MANAGEMENT AND RECORD KEEPING
10.1
Data Management
10.1.1
Data Handling
Data will be recorded at the site on eCRFs and reviewed by the Clinical Research Associate (CRA)
during monitoring visits. The CRAs will verify data recorded in the EDC system with source 
documents. All corrections or changes made to any study data must be appropriately tracked in an 
audit trail in the EDC system. An eCRF will be considered complete when all missing, incorrect, 
and/or inconsistent data has been accounted for.
10.1.2
Computer Systems
Data will be processed using a validated computer system conforming to regulatory requirements.
10.1.3
Data Entry
Data must be recorded using the EDC system as the study is in progress. All site personnel must 
log into the system using their secure user name and password in order to enter, review, or correct 
study data. These procedures must comply with Title 21 of the Code of Federal Regulations 
(21 CFR Part 11) and other appropriate international regulations. All passwords will be strictly 
confidential.
10.1.4
Medical Information Coding
For medical information, the latest versions of the following thesauri will be used:
Medical Dictionary for Regulatory Activities for medical history and adverse events, and
World Health Organization Drug Dictionary for prior and concomitant medications.
10.1.5
Data Validation
Validation checks programmed within the EDC system, as well as supplemental validation 
performed via review of the downloaded data, will be applied to the data in order to ensure 
accurate, consistent, and reliable data. Data identified as erroneous, or data that are missing, will 
be referred to the investigative site for resolution through data queries.
The eCRFs must be reviewed and electronically signed by the Investigator.
10.2
Record Keeping
Records of subjects, source documents, monitoring visit logs, eCRFs, inventory of study product, 
regulatory documents, and other Sponsor correspondence pertaining to the study must be kept in 
the appropriate study files at the site. Source data is defined as all information in original records 
and certified copies of original records of clinical findings, observations, or other activities in a 
clinical study necessary for the evaluation and reconstruction of the clinical study. Source data are 
contained in source documents (original records or certified copies). These records will be retained 
in a secure file for the period as set forth in the Clinical Study Agreement. Prior to transfer or 
destruction of these records, the Sponsor must be notified in writing and be given the opportunity 
to further store such records. 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
39
11
INVESTIGATOR REQUIREMENTS AND QUALITY CONTROL 
11.1
Ethical Conduct of the Study
Good Clinical Practice (GCP) is an international ethical and scientific quality standard for 
designing, conducting, recording, and reporting studies that involve human subjects. Compliance 
with this standard provides public assurance that the rights, safety, and wellbeing of study subjects 
are protected, consistent with the principles that have their origin in the Declaration of Helsinki, 
and that the clinical study data are credible.
11.2
Institutional Ethics Committee (IEC) or Human Research Ethics Committee 
(HREC)
The Institutional Ethics Committee (IEC) or Human Research Ethics Committee (HREC) will 
review all appropriate study documentation in order to safeguard the rights, safety, and wellbeing 
of subjects. The study will only be conducted at sites where IRB approval has been obtained. The 
protocol, Investigator’s Brochure, ICF, advertisements (if applicable), written information given 
to the subjects, safety updates, annual progress reports, and any revisions to these documents will 
be provided to the IRB by the Investigator.
Federal regulations and International Council for Harmonisation (ICH) Guidelines require that 
approval be obtained from an IRB prior to participation of subjects in research studies. Prior to 
study onset, the protocol, any protocol amendments, ICFs, advertisements to be used for subject
recruitment, and any other written information regarding this study to be provided to a subject or 
subject’s legal guardian must be approved by the IRB.
No drug will be released to the site for dosing until written IRB authorization has been received 
by the Sponsor.
11.3
Informed Consent
The ICF and any changes to the ICF made during the course of the study must be agreed to by the 
Sponsor or designee and the IRB prior to its use and must be in compliance with all ICH GCP, 
local regulatory requirements, and legal requirements.
The Investigator must ensure that each study subject is fully informed about the nature and 
objectives of the study and possible risks associated with participation and must ensure that the 
subject has been informed of his/her rights to privacy. The Investigator will obtain written 
informed consent from each subject before any study-specific activity is performed and should 
document in the source documentation that consent was obtained prior to enrollment in the study. 
The original signed copy of the ICF must be maintained by the Investigator and is subject to 
inspection by a representative of the Sponsor, their representatives, auditors, the IRB and/or 
regulatory agencies. A copy of the signed ICF will be given to the subject.
11.4
Study Monitoring Requirements
It is the responsibility of the Investigator to ensure that the study is conducted in accordance with 
the protocol, Declaration of Helsinki, ICH GCP, and applicable regulatory requirements, and that 
valid data are entered into the eCRFs.
To achieve this objective, the Medical Monitor’s duties are to aid the Investigator and, at the same 
time, the Sponsor in the maintenance of complete, legible, well organized and easily retrievable 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
40
data. Before the enrollment of any subject in this study, the Sponsor or their designee will review 
with the Investigator and site personnel the following documents: protocol, Investigator’s 
Brochure, eCRFs and procedures for their completion, informed consent process, and the 
procedure for reporting SAEs.
The Investigator will permit the Sponsor or their designee to monitor the study as frequently as 
deemed necessary to determine that data recording and protocol adherence are satisfactory. During 
the monitoring visits, information recorded on the eCRFs will be verified against source 
documents and requests for clarification or correction may be made. After the eCRF data is entered 
by the site, the CRA will review the data for safety information, completeness, accuracy, and 
logical consistency. Computer programs that identify data inconsistencies may be used to help 
monitor the clinical study. If necessary, requests for clarification or correction will be sent to the
Investigator. The Investigator and his/her staff will be expected to cooperate with the monitor and 
provide any missing information, whenever possible.
All monitoring activities will be reported and archived. In addition, monitoring visits will be 
documented at the investigational site by signature and date on the study-specific monitoring log.
11.5
Disclosure of Data
Data generated by this study must be available for inspection by Regulatory authorities, the 
Sponsor or their designee, applicable foreign health authorities, and the IEC/HEC as appropriate. 
Subjects or their legal representatives may request their medical information be given to their 
personal physician or other appropriate medical personnel responsible for their welfare.
Subject medical information obtained during the study is confidential and disclosure to third 
parties other than those noted above is prohibited.
11.6
Retention of Records
To enable evaluations and/or audits from regulatory authorities or the Sponsor, the Investigator 
will keep records, including the identity of all participating subjects (sufficient information to link 
records, eg, eCRFs and hospital records), all original signed ICFs, copies of all eCRFs, SAE forms, 
source documents, and detailed records of treatment disposition. The records should be retained 
by the Investigator according to specifications in the ICH guidelines, local regulations, or as 
specified in the Clinical Study Agreement, whichever is longer. The Investigator must obtain 
written permission from the Sponsor before disposing of any records, even if retention 
requirements have been met.
If the Investigator relocates, retires, or for any reason withdraws from the study, the Sponsor should 
be prospectively notified. The study records must be transferred to an acceptable designee, such 
as another Investigator, another institution, or to the Sponsor.
11.7
Publication Policy
Following completion of the study, the data may be considered for publication in a scientific 
journal or for reporting at a scientific meeting. Each Investigator is obligated to keep data 
pertaining to the study confidential. The Investigator must consult with the Sponsor before any 
study data are submitted for publication. The Sponsor reserves the right to deny publication rights 
until mutual agreement on the content, format, interpretation of data in the manuscript, and journal 
selected for publication are achieved.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
41
11.8
Financial Disclosure
The Investigator is required to provide financial disclosure information to the Sponsor to permit 
the Sponsor to fulfill its obligations under 21 CFR Part 54. In addition, the Investigator must 
commit to promptly updating this information if any relevant changes occur during the study and 
for a period of 1 year after the completion of the study.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
42
12
STUDY ADMINISTRATIVE INFORMATION
12.1
Protocol Amendments
Any amendments to the study protocol will be communicated to the Investigator by Medpace or 
the Sponsor. All protocol amendments will undergo the same review and approval process as the 
original protocol. A protocol amendment may be implemented after it has been approved by the 
IEC/HREC, unless immediate implementation of the change is necessary for subject safety. In this 
case, the situation must be documented and reported to the IEC/HREC within 5 working days.
12.2
Address List
12.2.1
Sponsor
Orpheris, Inc.
1235 Radio Road, Suite 200
Redwood City, CA 94065
Telephone: 
12.2.2
Contract Research Organization
Clinical Network Services Pty Ltd
Level 2 / 381 MacArthur Ave
Hamilton QLD 4007
Australia
12.2.3
Serious Adverse Event Reporting
safety@clinical.net.au
12.2.4
Biological Specimens
TBC


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
43
13
REFERENCES
Acetadote (acetylcysteine) Injection Prescribing Information. 15 February 2006. Manufactured 
for Cumberland Pharmaceuticals Inc. 
Beloosesky R, Weiner Z, Ginsberg Y, et al. Maternal N-acetyl-cysteine (NAC) protects the rat 
fetal brain from inflammatory cytokine responses to lipopolysaccharide (LPS). J. Matern Fetal 
Neonatal Med. 2012 Aug;25(8):1324-1328.
Chen G, Shi J, Hu Z, et al. Inhibitory effect on cerebral inflammatory response following 
traumatic brain injury in rats: a potential neuroprotective mechanism of N-acetylcysteine. 
Mediators Inflamm. 2008;2008:716458. doi: 10.1155/2008/716458.
Dai H, Navath RS, Balakrishnan B, et al. Intrinsic targeting of inflammatory cells in the brain by 
polyamidoamine dendrimers upon subarachnoid administration. Nanomedicine (Lond). 2010 
Nov;5(9):1317-29. doi: 10.2217/nnm.10.89.
Deon M, Marchetti DP, Donida B, et al. Oxidative stress in patients with X-linked 
adrenoleukodystrophy. Cell Mol Neurobiol. 2016 May;36(4):497-512. doi: 10.1007/s10571-015-
0234-2. Epub 2015 Jul 14.
Kambhampati SP, Clunies-Ross AJ, Bhutto I, et al. Systemic and intravitreal delivery of 
dendrimers to reactive microglia/macrophage in ischemia/reperfusion mouse retina. Invest 
Ophthalmol Vis Sci. 2015 Jul;56(8):4413-24. doi: 10.1167/iovs.14-16250.
Kannan S, Dai H, Navath RS, et al. Dendrimer-based postnatal therapy for neuroinflammation 
and cerebral palsy in a rabbit model. Sci Transl Med. 2012 Apr 18;4(130):130ra46. doi: 
10.1126/scitranslmed.3003162.
Khan M, Sekhon B, Jatana M, et al. Administration of N-acetylcysteine after focal cerebral 
ischemia protects brain and reduces inflammation in a rat model of experimental stroke. 
J Neurosci Res. 2004 May 15;76(4):519-27.
Kurtoglu YE, Navath RS, Wang B, et al. Poly(amidoamine) dendrimer-drug conjugates with 
disulfide linkages for intracellular drug delivery. Biomaterials. 2009 Apr;30(11):2112-21. doi: 
10.1016/j.biomaterials.2008.12.054. Epub 2009 Jan 25.
Lesniak WG, Mishra MK, Jyoti A, et al. Biodistribution of fluorescently labeled PAMAM 
dendrimers in neonatal rabbits: effect of neuroinflammation. Mol Pharm. 2013 Dec 
2;10(12):4560-71. doi: 10.1021/mp400371r. Epub 2013 Oct 30.
Mishra MK, Beaty CA, Lesniak WG, et al. Dendrimer brain uptake and targeted therapy for 
brain injury in a large animal model of hypothermic circulatory arrest. ACS Nano. 2014 Mar 
25;8(3):2134-47. doi: 10.1021/nn404872e. Epub 2014 Feb 19.
Moser HW, Loes DJ, Melhem ER, et al. X-Linked adrenoleukodystrophy: overview and 
prognosis as a function of age and brain magnetic resonance imaging abnormality. A study 
involving 372 patients. Neuropediatrics. 2000 Oct;31(5):227-39.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
44
Moser HW, Mahmood A, Raymond GV. X-linked adrenoleukodystrophy. Nat Clin Pract Neurol.
2007 Mar;3(3):140-51.
Nance E, Porambo M, Zhang F, et al. Systemic dendrimer-drug treatment of ischemia-induced 
neonatal white matter injury. J Control Release. 2015 Sep 28;214:112-20. doi: 10.1016/j.jconrel. 
2015.07.009. Epub 2015 Jul 13.
Navath RS, Kurtoglu YE, Wang B, et al. Dendrimer-drug conjugates for tailored intracellular 
drug release based on glutathione levels. Bioconjug Chem. 2008 Dec;19(12):2446-55. doi: 
10.1021/bc800342d.
Nemeth CL, Drummond GT, Mishra MK, et al. Uptake of dendrimer-drug by different cell types 
in the hippocampus after hypoxic-ischemic insult in neonatal mice: Effects of injury, microglial 
activation and hypothermia. Nanomedicine. 2017 Oct;13(7):2259-2369. doi: 10.1016/ 
j.nano.2017.06.014. Epub 2017 Jun 30.
Pahan K, Sheikh FG, Namboodiri AM, et al. N-acetyl cysteine inhibits induction of no
production by endotoxin or cytokine stimulated rat peritoneal macrophages, C6 glial cells and 
astrocytes. Free Radic Biol Med. 1998 Jan 1;23(1):39-48.
Samuni Y, Goldstein S, Dean OM, et al. The chemistry and biological activities of 
N-acetylcysteine. Biochim Biophys Acta. 2013 Aug;1830(8):4117-29. doi: 10.1016/ 
j.bbagen.2013.04.016. Epub 2013 Apr 22.
Tsai GY, Cui JZ, Syed H, et al. Effect of N-acetylcysteine on the early expression of 
inflammatory markers in the retina and plasma of diabetic rats. Clin Exp Ophthalmol. 2009 
Mar;37(2):223-31. doi: 10.1111/j.1442-9071.2009.02000.x. Epub 2009 Feb 3.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
45
APPENDIX A:  SCHEDULE OF PROCEDURES
Table 1.
Schedule of Procedures – Cohorts 1 and 2
Study 
Procedure
Screening
Period
Check-In
Treatment Period
Follow-Up Visits [1]
Early 
Termination 
Visit
Days -21
to -2
Day -1
Day 1
Day 2
Day 3
Day 8
Day 15
Informed 
consent 
X
Admit to 
investigational 
site
X
Inclusion/excl
usion criteria
X
X
Medical 
history 
X
Demographics
X
Prior/concomit
ant 
medications
X
X
X
X
X [3]
X
X
X
BMI 
calculation [2]
X
Physical 
examination 
(complete)
X
X
X
X [3]
X
X
X
Vital signs [4]
X
X
X
X 
X [3]
X
X
X
Breath alcohol 
screen
X
X
Pregnancy test 
[5]
X
X
FSH [5]
X
Urine drug and 
cotinine screen
X
X
Safety 
X
X
X
X 
X [3]
X
X
X


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
46
laboratory 
tests 
(chemistry, 
hematology, 
and urinalysis) 
[6]
aPTT, PTT, 
INR
X
HIV, HBsAg, 
HCVab screen
X
Plasma PK 
sampling [7]
X
X
X[3]
Urine PK 
sampling [8]
X
X 
X[3]
Administer 
study drug [9]
X
Assess adverse 
events
X
X
X
X
X[3]
X
X
X
Discharge 
from 
investigational 
site
X
Footnotes appear on the following page. 


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
47
A 30-minute window for vital signs and all postdose procedures is allowed.
When multiple procedures are scheduled at the same time point, the order of procedures should be as follows:  obtain vital signs, collect blood sample (drawn at nominal time), 
and obtain urine sample.
1.
There will be a 1-day visit window for Days 8 and 15.
2.
Body weight and height will be measured at screening and used to calculate BMI. 
3.
Assessments will be performed prior to discharge. 
4.
Vital signs (pulse rate, respiration rate, body temperature, and BP) will be recorded at all study visits after the subject has been seated or supine for 5 minutes. During the 
Treatment Period, vital signs will be measured predose, 30 and 60 minutes and at 2, 4, 12, 24 and 48 hours after SC dosing.
5.
A serum (beta HCG) pregnancy test will be performed at screening for female subjects of childbearing potential only. A urine pregnancy test will be performed at Day -1
for female subjects of childbearing potential only. A test for FSH level will be performed only for women of non-childbearing potential who are not surgically sterile.
6.
Safety laboratory tests will be performed under fasting conditions (at least 8 hours). Day 1 sample will be taken within 30 minutes prior to dosing. See Appendix B for the 
list of analytes assessed; eGFR will be assessed with each sample and will be based on the subject’s creatinine level, age, sex, and race. KIM-1 and NGAL will be 
evaluated from urine collected on Day -1, Day 2, Day 3, Day 8, Day 15 and on Early Termination. Additional chemistry parameters (i.e., glycosylated haemoglobin and 
homocysteine) will only be assessed at screening.
7.
Plasma PK samples will be collected within 30 minutes prior to dosing, and at 30 min and 1, 2, 4, 6, 8, 10, 12, 16, 24, and 30-36 hours after SC injection, as well as on Day 
3 (~48 hr). Blood samples for PK analysis can be obtained within 5 minutes of the scheduled sampling time for sample times 1 hour, within 10 minutes of the 
scheduled sampling time for sample times 12 hours, and within 2 hours of the scheduled sampling times for sample times >12 hours. 
8.
Urine PK samples will be collected 60 to 30 minutes prior to dosing (spot sample) and between 0 to 4, 4 to 8, 8 to 12, 12 to 18, 18 to 24, and 24 to 48 hours after dosing.
9.
Cohort dosing will be separate and sequential. Dose escalation from Cohort 1 (4 mg/kg) to Cohort 2 (8 mg/kg) will be determined by the SRC after review of safety, 
tolerability up to Day 8, and any available PK data from Cohort 1.
aPTT = activated partial thromboplastin time; BMI = body mass index; BP = blood pressure; eGFR = estimated glomerular filtration rate; FSH = follicle-stimulating hormone; 
HBV = hepatitis B virus; HCG = human chorionic gonadotropin; HCV = hepatitis C virus; HIV = human immunodeficiency virus; INR = international normalized ratio; 
PTT = partial thromboplastin time; SRC = safety review committee.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
48
APPENDIX B:  CLINICAL LABORATORY ANALYTES
Standard Safety Chemistry Panel
Alanine aminotransferase
Albumin
Alkaline phosphatase
Amylase
Aspartate aminotransferase
Bicarbonate
Blood urea nitrogen
Calcium
Chloride
Creatine kinase
Creatinine [1]
Estimated glomerular filtration rate (eGFR)
Gamma-glutamyl transferase
Glucose
Inorganic phosphorus
Lactate dehydrogenase
Lipase
Potassium
Sodium
Total bilirubin
Total protein
Uric acid
1.
eGFR will be assessed with each safety chemistry panel according to a mathematically derived formula based on the 
subject’s serum creatinine level, age, sex, and race. 
Additional Chemistry Parameters
Glycosylated hemoglobin 
Homocysteine
Endocrinology
Follicle-stimulating hormone [1]
1.
Follicle-stimulating hormone (only for female subjects of non-childbearing potential who are not surgically sterile).
Hematology
Hematocrit
Hemoglobin
Platelets
Red blood cell count
White blood cell count and differential 
Coagulation
Activated partial thromboplastin time
International normalized ratio
Partial thromboplastin time
Urinalysis
Bilirubin
Blood
Glucose
Ketones
Kidney injury molecule-1 (KIM-1)
Leukocyte esterase 
Microscopy [1]
Nitrite
Neutrophil gelatinase-associated lipocalin (NGAL)
pH
Protein (calculation of eGFR)
Specific gravity
Urobilinogen
1.
Microscopy is performed only as needed based on positive dipstick test results.


Orpheris, Inc.
Clinical Study Protocol OP-101-003
Confidential & Proprietary
Version 2.0, 17 March 2020
49
Pregnancy Test 
A serum (beta human chorionic gonadotropin) pregnancy test will be performed for female subjects of childbearing 
potential at screening only. A urine pregnancy test will be performed at Day -1 for female subjects of childbearing 
potential only. 
Urine drug, cotinine, and Breath Alcohol Tests
Urine drug, cotinine, and breath alcohol tests will be performed at screening and Check-in.
Amphetamine
Barbiturates
Benzodiazepines
Cocaine
Cannabinoids
Opiates
Phencyclidine
Serology 
Hepatitis B, hepatitis C, and human immunodeficiency virus at screening only.



SAP_001.pdf: 
NCT04321980