Clinical trial NCT03988023
Conditions: Severe Osteoarthritis of the Knee
Keywords: Osteoarthritis osteoarthritis of the knee
Sponsor: Ampio Pharmaceuticals. Inc.
Start date: 2019-06-24; Completion date: 2021-07-13
Enrollment: 1043 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The purpose of this study is to confirm the safety and efficacy of Ampion for the treatment of pain and function in patients with severe osteoarthritis of the knee.

Detailed description:
This study is a randomized, double-blind, single dose design.

The co-primary trial objectives are to evaluate the greater efficacy for pain improvement and function improvement of 4 mL Ampion versus saline intra-articular (IA) injection when applied to patients suffering from severe osteoarthritis of the knee (OAK). Efficacy will primarily be assessed with WOMAC A pain and WOMAC C function scores.

Mean change in the WOMAC A weekly pain scores from Baseline to Week 12 will be compared between Ampion and saline control. Mean change in WOMAC C function score will be compared between Ampion and saline control. This will test whether Ampion is superior to saline in improving pain and function.

Arms: arm 1: 4 mL injection of Ampion arm 2: 4 mL injection of placebo
Interventions: intervention 1: Ampion intervention 2: Saline
Intervention descriptions: intervention 1: 4 mL injection of Ampion intervention 2: Saline solution, 4 mL, single intra-articular injection

Protocol documents:
Prot_000.pdf: 
 
 
 
 
A RANDOMIZED, CONTROLLED, DOUBLE‐
BLIND STUDY TO EVALUATE THE EFFICACY 
AND SAFETY OF AN INTRA‐ARTICULAR 
INJECTION OF AMPION™ IN ADULTS WITH 
PAIN DUE TO SEVERE OSTEOARTHRITIS OF 
THE KNEE 
 
STUDY PROTOCOL 
 
STUDY NUMBER: AP‐013 
 
NCT 03988023 
 
7 JUNE 2019 


Ampion™ 
 
Ampio Pharmaceuticals, Inc. 
Clinical Study Protocol: AP-013 
 
Version 1.0 7 June 2019 
 
CONFIDENTIAL 
Page 1 
 
CLINICAL STUDY PROTOCOL 
 
A RANDOMIZED, CONTROLLED, DOUBLE-BLIND STUDY TO 
EVALUATE THE EFFICACY AND SAFETY OF AN INTRA-
ARTICULAR INJECTION OF AMPION™ IN ADULTS WITH PAIN 
DUE TO SEVERE OSTEOARTHRITIS OF THE KNEE 
 
STUDY NUMBER: AP-013 
Drug Development Phase: 
Phase 3 
Investigational Product: 
Ampion™ 
Indication: 
Osteoarthritis of the knee 
Sponsor: 
Ampio Pharmaceuticals, Inc. 
373 Inverness Parkway, Suite 200 
Englewood, CO 80112 
Lead Investigators: 
John Schwappach, MD 
Date: 
Version 1.0 7 June 2019 
Conduct: In accordance with the ethical principles that originate from the Declaration of 
Helsinki and that are consistent with International Conference on Harmonisation (ICH) 
guidelines on Good Clinical Practice (GCP) and regulatory requirements as applicable. 
 
 
 
 
Confidential Information 
This document is the sole property of Ampio Pharmaceuticals, Inc. This document and any 
and all information contained herein has to be considered and treated as strictly confidential.  
This document shall be used only for the purpose of the disclosure herein provided. No 
disclosure or publication shall be made without the prior written consent of 
Ampio Pharmaceuticals, Inc. 
 


Ampion™ 
 
Ampio Pharmaceuticals, Inc. 
Clinical Study Protocol: AP-013 
 
Version 1.0 7 June 2019 
 
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PROTOCOL SIGNATURE PAGE 
I have read and understand the contents of this clinical protocol for Study AP-013 dated 7 
June 2019 and agree to meet all obligations of Ampio Pharmaceuticals, Inc. as detailed in all 
applicable regulations and guidelines.  
 
Signed By: 
 
<Study personnel signature> 
 
 
 
<Enter date> 
 
 
 
 
 


Ampion™ 
 
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Clinical Study Protocol: AP-013 
 
Version 1.0 7 June 2019 
 
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PROTOCOL SYNOPSIS 
Sponsor: 
Ampio Pharmaceuticals, Inc. 
Investigational Product: 
Ampion™ 
Developmental Phase: 
Phase 3 
Title of Study:   
A Randomized, Controlled, Double-Blind Study to Evaluate the Efficacy and  
Safety of an Intra-Articular Injection of Ampion in Adults with Pain Due to Severe 
Osteoarthritis of the Knee  
Protocol Number:   
AP-013 
Study Center(s):   
Approximately 20 sites  
Lead Investigators:   
John Schwappach, MD 
Indication:   
Pain of severe osteoarthritis of the knee (OAK) 
Number of patients:  
This trial is designed for 1,034 patients, 517 per study arm, randomized 1:1 for Ampion and 
saline intra-articular (IA) injection. There will be an interim analysis at 724 patients, 362 per 
study arm, randomized 1:1, Ampion and saline. A promising zone design is used with the 
possibility to increase the sample size by 1.5 times the original sample size up to 1,552 
patients at the interim analysis. 
Objectives:  
The co-primary trial objectives are to evaluate the greater efficacy for pain improvement and 
function improvement of 4 mL Ampion versus saline intra-articular (IA) injection, when 
applied to patients suffering from severe osteoarthritis of the knee (OAK). Efficacy will 
primarily be assessed with WOMAC A pain and WOMAC C function scores. 
The secondary trial objectives include the following evaluations of an intra-articular injection 
of Ampion, as compared to saline, when administered to patients suffering from severe 
OAK: assessment of safety; improvement in the Patient Global Assessment; and the amount 
of rescue analgesia. 
 


Ampion™ 
 
Ampio Pharmaceuticals, Inc. 
Clinical Study Protocol: AP-013 
 
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Sponsor: 
Ampio Pharmaceuticals, Inc. 
Investigational Product: 
Ampion™ 
Developmental Phase: 
Phase 3 
Exploratory objectives are to analyze the effect, if any, of the efficacy of Ampion, as 
compared to saline, on stiffness; Outcome Measures in Rheumatology Clinical Trials and 
Osteoarthritis Research Society International (OMERACT-OARSI) responder status; 
proportion of treatment failures; change in pain and function as a cumulative distribution 
function (CDF) plot; and a Bayesian meta-analysis for WOMAC A pain sub score including 
KL 4 patients from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013. 
  
Methods:   
Study AP-013 is a randomized, controlled, double-blind study with a 28-day screening 
period for each patient followed by a 24-week participation period. This trial will use an 
adaptive design with promising zone method for sample size re-estimation.  
This trial is designed for 1,034 patients, 517 per study arm, randomized 1:1 for Ampion and 
saline IA injection. There will be an interim analysis to allow for sample size re-estimation 
up to 1.5 times the original trial size for a maximum of 1,552 patients, 776 per study arm, 
randomized 1:1 for Ampion and saline IA injection.  
The clinical effects of treatment on severe OAK will be evaluated at 2, 4, 6, 8, 10, 12, and 24 
weeks, using the Western Ontario and McMaster Universities Arthritis Index (WOMAC®) 
osteoarthritis Index 3.1, and the Patient’s Global Assessment of disease severity (PGA). 
Average daily WOMAC A pain scoring will be collected for seven days using a daily diary 
beginning one week prior to clinic visits at Week 0 (baseline) and Week 12, in order to 
determine a WOMAC A weekly pain score for Baseline and Week 12. 
Safety will be assessed by recording adverse events (through 24 hours post-dose and at all 
follow up contacts) and physical examination and vitals (Baseline, Weeks 6, 12, and 24). 
Diagnosis and Main Criteria for Inclusion:  
1. Able to provide written informed consent to participate in the study 
2. Willing and able to comply with all study requirements and instructions of the site study 
staff 
3. Male or female, 40 years to 85 years old (inclusive) 
4. Must be ambulatory 
5. Index knee must be symptomatic for greater than 6 months with a clinical diagnosis of 
OAK and supported by radiological evidence (Kellgren-Lawrence Grade 4) that is not 
older than 6 months prior to the date of screening 
6. Moderate to moderately-severe OA pain in the index knee (rating of at least 1.5 on the 
WOMAC Index 3.1 as measured by 5-point Likert Pain Subscale) assessed at screening 
7. Ability to temporarily discontinue nonsteroidal anti-inflammatory drug (NSAID) for 48 
hours prior to scheduled clinical efficacy evaluations 
8. No analgesia (including acetaminophen [paracetamol]) taken 12 hours prior to an 
efficacy measure 


Ampion™ 
 
Ampio Pharmaceuticals, Inc. 
Clinical Study Protocol: AP-013 
 
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Sponsor: 
Ampio Pharmaceuticals, Inc. 
Investigational Product: 
Ampion™ 
Developmental Phase: 
Phase 3 
9. No known clinically significant liver abnormality (e.g. cirrhosis, transplant, etc.).  
Main Criteria for Exclusion: 
1. As a result of medical review and screening investigation, the Principal Investigator 
considers the patient unfit for the study 
2. A history of allergic reactions to human albumin (reaction to non-human albumin such as 
egg albumin is not an exclusion criterion) 
3. A history of allergic reactions to excipients in 5% human albumin (N-acetyltryptophan, 
sodium caprylate) 
4. Presence of tense effusions 
5. Inflammatory or crystal arthropathies, acute fractures, history of aseptic necrosis or joint 
replacement in the affected knee, as assessed locally by the Principal Investigator 
6. Isolated patella femoral syndrome, also known as chondromalacia 
7. Any other disease or condition interfering with the free use and evaluation of the index 
knee for the duration of the trial (e.g. cancer, congenital defects, spine osteoarthritis) 
8. Major injury to the index knee within the 12 months prior to screening  
9. Severe hip osteoarthritis ipsilateral to the index knee 
10. Any pain that could interfere with the assessment of index knee pain (e.g. pain in any 
other part of the lower extremities, pain radiating to the knee) 
11. Any pharmacological or non-pharmacological treatment targeting OA started or changed 
during the 4 weeks prior to randomization or likely to be changed during the duration of 
the study 
12. Use of the following medications: 
a. No IA injected pain medications in the study knee during the study 
b. No analgesics containing opioids. NSAIDs may be continued at levels preceding the 
study, however may not be used 48 hours prior to efficacy evaluations, and 
acetaminophen is available as a rescue medication during the study from the provided 
supply  
c. No topical treatment on osteoarthritis index knee during the study  
d. No significant anticoagulant therapy (e.g. Heparin or Lovenox) during the study 
(treatment such as Aspirin and Plavix are allowed) 
e. No systemic treatments that may interfere with safety or efficacy assessments during 
the study 
f. No immunosuppressants 
g. No use of corticosteroids > 10 mg prednisolone equivalent per day 
h. If corticosteroid use is ≤ 10 mg prednisolone equivalent per day, and if clinically 
indicated, subjects should be allowed to decrease their corticosteroid 
use. Additionally, some subjects may need to increase their steroid dose to treat 
worsened symptoms in the treated knee, and subjects who increase their 
corticosteroid dose above their starting dose of corticosteroid during the study will be 
treated as “treatment failures” for efficacy analysis 
13. No human albumin treatment in the 3 months before randomization or throughout the 
duration of the study 


Ampion™ 
 
Ampio Pharmaceuticals, Inc. 
Clinical Study Protocol: AP-013 
 
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Sponsor: 
Ampio Pharmaceuticals, Inc. 
Investigational Product: 
Ampion™ 
Developmental Phase: 
Phase 3 
 
Test Product, Dose and Mode of Administration: 
Ampion, 4 mL, single intra-articular injection in the knee  
Reference Therapy, Dose and Mode of Administration: 
Saline, 4 mL, single intra-articular injection in the knee 
Study Duration:   
24 weeks 
Criteria for Evaluation: 
Safety:  
Adverse events, physical examination, and vitals 
Efficacy: 
WOMAC osteoarthritis Index 3.1 and PGA 
Statistical Methods:   
The co-primary efficacy endpoints are the change from Baseline to Week 12 in the WOMAC 
A pain score and the WOMAC C function score. The primary efficacy analysis will use the 
intent-to-treat (ITT) analysis population with the per-protocol (PP) analysis population as 
supportive. WOMAC A pain and WOMAC C function scores will be analyzed using mixed-
effects models with repeated measures (MMRM) with treatment assignment, week, and 
baseline WOMAC A pain scores and WOMAC C function scores, respectively, as 
covariates, with comparisons between Ampion and saline.  
 
All treatment emergent adverse events (TEAEs) will be reported by randomization 
assignment. There will be no statistical tests for incidence and severity of TEAEs. 
 


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Clinical Study Protocol: AP-013 
 
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TABLE OF CONTENTS 
PROTOCOL SYNOPSIS ..............................................................................................................3 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS .............................................11 
1 
INTRODUCTION ..............................................................................................................13 
1.1 Study Drug .......................................................................................................................14 
1.2 Background to the Disease ..............................................................................................14 
1.3 Previous Human Experience ...........................................................................................15 
1.4 Preclinical Data ...............................................................................................................15 
1.4.1 Pharmacology Studies .........................................................................................15 
1.4.2 Toxicity and Safety Studies .................................................................................16 
1.5 Clinical Experience .........................................................................................................16 
2 
RATIONALE FOR THE STUDY ....................................................................................19 
2.1 Rationale for the Doses and the Dosing Regimen ...........................................................19 
3 
STUDY DESIGN ................................................................................................................20 
3.1 Study Design Overview ...................................................................................................20 
3.2 Study Objectives ..............................................................................................................21 
3.2.1 Primary Objective ................................................................................................21 
3.2.2 Secondary Objectives ..........................................................................................21 
3.2.3 Exploratory Objectives ........................................................................................21 
3.3 Safety Monitoring Committee .........................................................................................21 
3.4 Stopping Rules .................................................................................................................22 
3.5 Study Endpoints ...............................................................................................................22 
3.5.1 Co-Primary Endpoint ...........................................................................................22 
3.5.2 Secondary Endpoints ...........................................................................................22 
3.5.3 Exploratory Endpoints .........................................................................................22 
3.6 Blinding and Randomization ...........................................................................................23 
3.7 Code-breaking .................................................................................................................23 
4 
SELECTION OF PATIENTS ...........................................................................................24 
4.1 Number of Patients ..........................................................................................................24 
4.2 Recruitment Methods ......................................................................................................24 
4.3 Inclusion Criteria .............................................................................................................25 
4.4 Exclusion Criteria ............................................................................................................25 
4.5 Inclusion of Patients Incapable of Giving Informed Consent .........................................26 
5 
STUDY PLAN AND PROCEDURES ..............................................................................27 
5.1 Description of Study Visits .............................................................................................27 
5.1.1 Visit -1 (Day -28 through Day 0, in-clinic): Screening .......................................27 
5.1.2 Visit 0 (Day -6, telephone contact): Pre-Baseline Daily Assessments ................28 
5.1.3 Visit 1 (Day 0, in-clinic): Baseline/Randomization/Treatment ...........................28 
5.1.4 Visit 2 (Day 1, telephone contact) .......................................................................28 
5.1.5 Visit 3 (Week 2 ± 7 days, telephone contact) ......................................................28 
5.1.6 Visit 4 (Week 4 ± 7 days, telephone contact) ......................................................29 


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Clinical Study Protocol: AP-013 
 
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5.1.7 Visit 5 (Week 6 ± 7 days, in-clinic) ....................................................................29 
5.1.8 Visit 6 (Week 8 ± 7 days, telephone contact) ......................................................29 
5.1.9 Visit 7 (Week 10 ± 7 days, telephone contact) ....................................................29 
5.1.10 Visit 8 (Week 11 ± 7 days, telephone contact): Pre-Week 12 Daily 
Assessments ...............................................................................................29 
5.1.11 Visit 9 (Week 12 ± 7 days, in-clinic)...................................................................30 
5.1.12 Visit 10 (Week 24 ± 7 days, in-clinic).................................................................30 
5.1.13 Early Termination Visit .......................................................................................30 
5.1.14 Unscheduled Visits ..............................................................................................30 
5.1.15 Missed Visits .......................................................................................................31 
6 
METHODS OF ASSESSMENT .......................................................................................32 
6.1 Efficacy Assessments ......................................................................................................32 
6.1.1 WOMAC® Osteoarthritis Index (Bellamy 1988) ................................................32 
6.1.2 Patient’s Global Assessment of Disease Severity (PGA) ....................................33 
6.2 Safety Parameters ............................................................................................................33 
6.2.1 Vital Signs ...........................................................................................................33 
7 
DISCONTINUATION CRITERIA ..................................................................................35 
7.1 Early Discontinuation of the Study .................................................................................35 
7.2 Early Discontinuation of Individual Patients ..................................................................35 
8 
TREATMENT ....................................................................................................................36 
8.1 Dosing and administration of study medication ..............................................................36 
8.2 Drug Storage and Accountability ....................................................................................36 
8.3 Concomitant Treatments and Rescue Medication ...........................................................37 
8.4 Treatment Compliance ....................................................................................................37 
9 
ADVERSE EVENTS ..........................................................................................................38 
9.1 Definition of an adverse event .........................................................................................38 
9.2 Definition of a Serious Adverse Event ............................................................................38 
9.3 Recording of adverse events and serious adverse events ................................................39 
9.3.1 AE Follow-up ......................................................................................................39 
9.3.2 Overdose ..............................................................................................................39 
9.4 Serious Adverse Event Reporting ...................................................................................40 
9.4.1 Reporting Requirements ......................................................................................40 
9.4.2 SAE Contact Information ....................................................................................40 
10 
STATISTICAL METHODS ..............................................................................................41 
10.1 
General Considerations ...............................................................................................41 
10.2 
Study Objectives .........................................................................................................41 
10.3 
Analysis Populations ..................................................................................................41 
10.3.1 Safety Analysis Population: ................................................................................41 
10.3.2 Intent-to-treat Population: ...................................................................................41 
10.3.3 Per Protocol Population: ......................................................................................41 
10.3.4 Statistical Analysis of Co-Primary Effectiveness Endpoints ..............................42 
10.3.5 Primary Hypotheses .............................................................................................42 


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10.3.6 Definition of Study Visits ....................................................................................43 
10.3.7 Number of patients to receive study drug ............................................................43 
10.3.8 Disposition of patients .........................................................................................43 
10.3.9 Interim analysis ...................................................................................................43 
10.3.10 
Blinding and randomization .......................................................................44 
10.3.11 
Data presentation .......................................................................................44 
10.3.11.1 
Demographic and Baseline Characteristics .................................44 
10.3.11.2 
Medical History and Physical Examination ................................44 
10.3.11.3 
Concomitant Medications or Treatments ....................................44 
10.3.11.4 
Safety data ...................................................................................45 
10.3.11.5 
Efficacy data: ...............................................................................45 
10.4 
STUDY ENDPOINTS ................................................................................................46 
10.4.1 Co-Primary Endpoints .........................................................................................46 
10.4.2 Secondary Endpoints ...........................................................................................46 
10.4.3 Exploratory Endpoints .........................................................................................47 
10.4.4 Subset Analyses ...................................................................................................48 
10.4.5 Sensitivity Analysis .............................................................................................48 
10.5 
Missing and Spurious Data .........................................................................................48 
11 
REGULATORY, ETHICAL AND LEGAL OBLIGATIONS .......................................49 
11.1 
Declaration of Helsinki ...............................................................................................49 
11.2 
Good Clinical Practice ................................................................................................49 
11.3 
Institutional Review Boards/Ethics Committees ........................................................49 
11.4 
Regulatory Authority Approval ..................................................................................49 
11.5 
Informed Consent .......................................................................................................49 
11.6 
Patient Confidentiality and Disclosure .......................................................................49 
11.7 
Collection, Monitoring and Auditing Study Documentation, and Data 
Storage ......................................................................................................................50 
11.7.1 Collection of Data and Monitoring Procedures ...................................................50 
11.7.2 Auditing Procedure ..............................................................................................51 
11.7.3 Retention of Documents ......................................................................................51 
11.8 
Disclosure of Information ...........................................................................................51 
11.9 
Discontinuation of the Study ......................................................................................51 
11.10 
Study Report, Publication Policy and Archiving of Study Documentation .............51 
11.10.1 
Study Report and Publication Policy .........................................................51 
11.10.2 
Study Documents .......................................................................................52 
11.10.3 
Archiving of Documents ............................................................................52 
12 
REFERENCES ...................................................................................................................53 
13 
CONTACT LIST ................................................................................................................56 
 
LIST OF IN-TEXT TABLES 
Table 1.1  Summary of Ampion nonclinical studies ...................................................................13 


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Clinical Study Protocol: AP-013 
 
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Table 1.2  Summary of the Studies Conducted with Ampion Administered as an Intra 
Articular Injection to the Knee ...................................................................................16 
Table 1.3  Summary of Adverse Events ......................................................................................18 
Table 6.1  Schedule of Assessments and Procedures ..................................................................34 
Table 9.1 
Definitions of AE Severity .........................................................................................38 
Table 9.2  Guidelines for Determining the Relationship (if any)  Between Adverse 
Event and the Study Drug ...........................................................................................38 
 


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Clinical Study Protocol: AP-013 
 
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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
Abbreviation 
Definition 
C 
degrees Celsius 
F 
degrees Fahrenheit 
g 
microgram 
AE 
adverse event 
BP 
Blood pressure 
CFR 
Code of Federal Regulations 
CI 
confidence interval 
CRF 
case report form 
CRO 
Clinical Research Organization 
Da 
dalton 
DA-DKP 
aspartyl-alanyl diketopiperazine 
DBP 
diastolic blood pressure 
DMP 
Data Management Plan 
eCRF 
electronic case report form 
eDC 
electronic data capture 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
HSA 
human serum albumin 
IA 
intra-articular 
ICF 
informed consent form 
ICH 
International Conference on Harmonisation 
IEC 
Independent Ethics Committee 
IND 
Investigational New Drug 
IRB 
Institutional Review Board 
ITT 
intent-to-treat 
kDa 
kilodalton 
mg 
milligram 
mL 
milliliter 
MMRM 
Mixed-effects model with repeated measures 
NA 
not applicable 
NSAID 
non-steroidal anti-inflammatory drug 
OA 
osteoarthritis 
OAK 
osteoarthritis of the knee 
OMERACT-OARSI 
outcome measures in rheumatology clinical trials and osteoarthritis 
research society international 
PGA 
patient’s global assessment of disease severity 
PP 
per protocol population 
REB 
Research Ethics Board 
SAE 
serious adverse event 
SAP 
Statistical Analysis Plan 
SBP 
systolic blood pressure 
SD 
standard deviation 
SEM 
standard error of the mean 
SMC 
Safety Monitoring Committee 
SOP 
standard operation procedure 


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Clinical Study Protocol: AP-013 
 
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Abbreviation 
Definition 
TEAE 
treatment-emergent adverse event 
WBC 
white blood cell 
WOMAC 
Western Ontario and McMaster Universities Arthritis Index 
 


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Clinical Study Protocol: AP-013 
 
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1 
INTRODUCTION 
The sole starting material of Ampion, human serum albumin (HSA), has a long history of 
clinical use as a colloid replacement therapy, dating back over 60 years. Currently, HSA is 
approved for intravenous administration for the indications of hypovolemia, hypoalbuminemia, 
prevention of central volume depletion after paracentesis due to cirrhotic ascites, ovarian 
hyperstimulation, adult respiratory distress syndrome (ARDS), acute nephrosis, hemolytic 
disease of the newborn and burns. In addition to its effects on oncotic pressure, HSA has 
pharmacological effects including decreased inflammation (Quinlan 2005), decreased vascular 
permeability (Evans 2002) and has no adverse effect on cardiac safety (Vincent JL 2004). The 
low molecular weight (< 5 kilodalton (kDa)) ultrafiltrate of 5% HSA, Ampion, is being 
developed as an intra-articular (IA) injection to provide relief for the pain due to severe 
osteoarthritis of the knee (OAK).  
There are no currently FDA-approved drugs for the indication of pain from severe (Kellgren 
Lawrence [KL] Grade 4) OAK, for which Ampion is indicated. The active components in 
Ampion, aspartyl-alanyl diketopiperazine (DA-DKP), N-acetyl tryptophan (NAT), and caprylic 
acid (Caprylate), play a role in Ampion’s biological activity and have been shown to reduce pro-
inflammatory cytokine tumor necrosis factor-alpha (TNFα). DA-DKP is naturally present in 
human serum and is a cyclized dipeptide derived from the two N-terminal amino acids of 
albumin, aspartic acid and alanine. These amino acids are cleaved by a dipeptidase, and the 
resulting dipeptide is cyclized to form a diketopiperazine (aspartyl-alanyl diketopiperazine or 
DA-DKP). The formation of DA-DKP occurs during the HSA pasteurization process (Bar-Or 
2013). The two other active components, NAT and Caprylate, are added during the manufacture 
of pharmaceutical HSA as stabilizers to protect albumin from heat denaturation and oxidation.  
In vitro nonclinical studies, which use cellular models designed for the study of osteoarthritis, 
evaluate the role of Ampion in regulating inflammatory responses with cytokine tumor necrosis 
factor-alpha (TNFα), and other inflammatory markers, in immune cells (peripheral blood 
mononuclear cells, monocytes, macrophages), and support the clinical application of Ampion. A 
high-level summary of these studies is shown in Table 1.1. 
Table 1.1  
Summary of Ampion nonclinical studies 
Study Objective 
Summary of Findings 
Anti-inflammatory effects 
of HSA and DA-DKP 
(TNFα, IFNγ) 
HSA inhibited the production of pro-inflammatory cytokines (IFNγ, TNFα) in 
activated immune cells; however, when the low-molecular weight ultrafiltrate was 
removed from HSA, no anti-inflammatory effects were observed. DA-DKP was 
detected in HSA from multiple vendors, and different concentrations of synthetic 
DA-DKP (25-500 µM) were shown to inhibit TNFα release in a dose-dependent 
manner. This study identified early anti-inflammatory properties of Ampion (< 5 
kDa ultrafiltrate of HSA) and of the active component DA-DKP. These results were 
published in the journal of Critical Care Medicine (Bar-Or 2006). 


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Study Objective 
Summary of Findings 
DA-DKP activity on 
cytokine production 
(TNFα, IFNγ, IL-8) 
DA-DKP decreased production of pro-inflammatory cytokines (IFNγ, TNFα) in T-
lymphocytes. DA-DKP also increased phosphorylation of the GTPase Rap-1, which 
in turn, decreased phosphorylated transcription factors involved in the regulation of 
TNFα and IFNγ gene transcription. Release of pro-inflammatory cytokine (IL-8) 
was not impacted by DA-DKP, suggesting that DA-DKP induction of active Rap-1 
impacted the phosphorylation of only selected transcription factor pathways and was 
not a global effect. These results were published in the Journal of Trauma, Injury, 
Infection, and Critical Care (Shimonkevitz 2008) 
Ampion anti-inflammatory 
activity (TNFα inhibition) 
Ampion inhibited pro-inflammatory cytokine TNFα in activated immune cells by an 
average of 39%. Individual components contribute to activity: DA-DKP (100 μM) 
17% TNFα inhibition, NAT (3.0 mM) 17% TNFα inhibition, and Caprylate (0.6 
mM) 19% TNFα inhibition and is published in the Journal of Immunoassay and 
Immunochemistry (Thomas 2016). 
Dose responses of Ampion 
and individual components 
(TNFα inhibition). 
Ampion and its components have been shown to inhibit TNFα in a dose-dependent 
manner in activated immune cells (IND 15345).  
This in vitro data translates to in vivo use in humans, as it has been reported that synovial levels 
of TNFα correlate directly with Western Ontario and McMaster Universities Arthritis Index 
(WOMAC) measures of pain, stiffness, and functional disability, while other studies have 
reported that TNFα levels are associated with pain while standing (Stannus 2013).  
1.1 
Study Drug 
Ampion is the < 5 kDa ultrafiltrate of 5% HSA. 
1.2 
Background to the Disease 
Osteoarthritis (OA) remains a leading contributor to global disability, with both hip and knee 
OA accounting for over 17 million years lived with disability (Cross 2014). OA is the most 
common form of arthritis and affects up to 38 million adults in the US alone. OA is caused by 
inflammation of the soft tissue and bony structures of the joint which worsens over time and 
leads to progressive thinning of articular cartilage, narrowing of the joint space, synovial 
membrane thickening, osteophyte formation and increased density of subchondral bone. These 
changes eventually result in chronic pain and disability and deterioration of the joint and may 
require eventual surgery for total joint replacement. OA of the knee is defined in stages as 
noted by Kellgren-Lawrence (KL) grading: Grade 0 (normal knee; no osteophytes or joint 
space narrowing), Grade 1 (possible osteophytic lipping and doubtful narrowing of joint space), 
Grade 2 (definite osteophytes and possible narrowing of joint space), Grade 3 (moderate 
multiple osteophytes, definite narrowing of joint space and some sclerosis, and possible 
deformity of the bone ends) and Grade 4 (large osteophytes, marked narrowing of joint space, 
severe sclerosis, and definite deformity of bone ends). 
 
The pain associated with OA is now thought to be multifactorial. The pathogenesis of pain from 
OA is comprised of both nociceptive and neuropathic pain pathways resulting from severely 
damaged cartilage, exposed subchondral bone, joint innervation, and less defined pathways 
including the synovium. The balance between nociceptive and neuropathic pain shifts towards 


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neuropathic pain as the severity of the condition increases (Akinci 2016). The multifactorial 
nature of the pain of OA may make pain control more difficult, particularly in patients with 
severe OAK (KL 4). Patients with severe OAK have few treatment options for pain management. 
Difficult-to-treat pain conditions, such as severe OAK, are a challenge for both medical doctors 
and patients. FDA has acknowledged an unmet medical need for patients with severe OAK. 
The presence of synovitis in KL 4 patients indicates active ‘inflammatory’ disease process. 
These findings were supported with a linear observation of an increasing trend correlated with 
the severity of synovitis related to advancing KL grades. Additionally, KL 4 patients were 
shown to have lesions that fluctuate with pain (i.e., fluctuating bone marrow lesions and 
synovitis) related to a high grade of cartilage loss in KL 4 patients (Guermazi 2015). These 
findings were further supported using a unique in vivo imaging technique that detected activated 
and not resting macrophages (Kraus 2016). In a patient cohort that included patients with severe 
(KL 4) OAK, the quantity of knee-related activated macrophages was statistically associated 
with more severe knee pain and radiographic severity of OAK. 
These data demonstrate that greater macrophage mediated inflammation is a cause of more 
severe pain in KL 4 OAK and provides a rationale whereby Ampion, which has demonstrated 
in vitro anti-inflammatory and anti-macrophage activity, may mediate symptomatic relief in 
this patient group. These data further demonstrate the need to examine patients by disease 
severity and supply treatment options for severely diseased patients (Guermazi 2015). Ampion 
has shown anti-inflammatory activity in vitro against macrophages, monocytes, and immune 
cells (Bar-Or 2015, Frederick 2016, Thomas 2016, Thomas 2016b). 
1.3 
Previous Human Experience 
In addition to the extensive HSA clinical experience, the sole starting material of Ampion, the 
Company has completed numerous clinical studies to evaluate the effect of Ampion in adults 
with OAK. These studies are summarized in Section 1.5. 
1.4 
Preclinical Data 
1.4.1 
Pharmacology Studies 
In vitro pharmacology studies demonstrated that the low molecular weight ultrafiltrate  
of pharmaceutical preparations of HSA, and the cyclized dipeptide, DA-DKP, contained in that 
filtrate have a range of anti-inflammatory properties, which may be expected to ameliorate the 
symptoms of inflammation, including pain, in humans (Bar-Or 2006). Further, in vitro studies 
support anti-inflammatory properties for the < 5 kDa ultrafiltrate of HSA (Ampion) and its 
components (DA-DKP, NAT, Caprylate) with the reduction of pro-inflammatory cytokines 
(Shimonkevitz 2008, Thomas 2016, IND 15345). No pharmacokinetics studies were performed 
as the plasma levels of the constituents of the < 5 kDa preparation, including the active 
components (DA-DKP, NAT, Caprylate), after intra-articular (IA) injection of a therapeutic dose 
in humans or in laboratory animals would be below the limits of detection. For more 
information, consult the Investigator Brochure. 


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1.4.2 
Toxicity and Safety Studies 
HSA and other components of serum albumin preparations are species specific and may be 
expected to be immunogenic if repeatedly injected into non-human species, even at very low 
concentrations. For this reason, it was not possible to perform formal toxicological studies in 
animals. 
1.5 
Clinical Experience 
Ampio has conducted multiple clinical studies to evaluate the treatment effect and safety of 
Ampion in subjects with OAK. Ampion has been administered across six clinical studies as a 
single IA injection or as three IA injections administered every two weeks. Four single-dose 
pivotal Phase 3 studies, two multi-dose studies, and one multiphase single-dose Phase 1b study 
have been completed. An overview of the efficacy studies is provided in the Table 1.2. 
 
For the clinical development program, all patients were required to have x-ray findings 
demonstrating KL 2, 3, or 4 disease severity. All studies enrolled patients with KL 4 disease. In 
addition, eligible subjects had at least moderate pain at Baseline (defined as a score of at least 1.5 
on the WOMAC Index 3.1, as measured by the 5-point Likert pain subscale at screening) in the 
study knee, which was to have been symptomatic for greater than six months with a clinical 
diagnosis of OA confirmed by radiological evidence. The clinical effects of treatment on OA 
pain were evaluated during clinic visits, with phone call follow-ups according to the study visit 
schedule. 
Table 1.2  
Summary of the Studies Conducted with Ampion Administered as an Intra 
Articular Injection to the Knee 
Study 
Phase 
N 
N: 
KL 2, 3, 4 
Route 
Primary 
outcome 
Primary 
endpoint 
Additional 
outcomes 
Test product 
Single-Injection Phase 3 Studies in Support of Efficacy & Safety 
AP-003-A 
3 
329 
115 / 139 / 75 
Single IA 
injection 
WOMAC A 
pain, 5- point 
Likert scale 
Week 12 
WOMAC B 
WOMAC C 
PGA 
OMERACT 
-OARSI 
Rescue 
analgesia 
Ampion, 
4 mL and 
10mL 
AP-004 
3 
538 
133 / 195/ 
210 
Single IA 
injection 
WOMAC A 
pain, 5- point 
Likert scale 
Week 12 
WOMAC C 
PGA 
WOMAC A 
pain over 12 
weeks 
Ampion, 
4 mL 
 


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Study 
Phase 
N 
N: 
KL 2, 3, 4 
Route 
Primary 
outcome 
Primary 
endpoint 
Additional 
outcomes 
Test product 
AP-003-C 
3 
168 
0 / 0 / 168 
Single IA 
injection 
OMERACT-
OARSI 
responder 
Week 12 
WOMAC A, 
B, C and 
PGA 
Ampion,  
4 mL 
AP-003- B 
3 
480 
101 / 247 / 
132 
Single IA 
injection 
WOMAC A 
pain, 5- point 
Likert scale 
Week 12 
WOMAC B 
WOMAC C 
PGA 
Rescue 
analgesia 
WOMAC A 
pain with 
movement, 
pain at rest 
Ampion,  
4 mL 
Additional Clinical Studies in Support of Safety 
AP-007 
1 / 2 
47 
2 / 23/ 15 
Multiple IA 
injection; 
3 injections, 
every 2 
weeks 
WOMAC A 
pain, 5- point 
Likert scale 
Week 20 
WOMAC B 
WOMAC C 
PGA 
Physical 
activity 
Radiologic 
changes 
Ampion,  
4 mL 
AP-008 
3 
342 
0 / 114 / 228 
Multiple IA 
injection; 
3 injections, 
every 2 
weeks 
WOMAC A 
pain, 5- point 
Likert scale 
Week 20 
WOMAC B 
WOMAC C 
PGA 
Physical 
activity 
WOMAC A 
pain with 
movement 
Ampion,  
4 mL 
AIK 
1 / 2 
103 
33 / 69 / 1 
Single IA 
injection 
Pain, 10- point 
NRS 
Day 3, 8, 
30 and 
84 
WOMAC 
total, parts 
A, B, C 
Range in 
motion 
Rescue 
analgesia 
Ampion  
10 mL, alone or 
in solution with 
betamethasone 
and/or 
lidocaine 
and/or 
saline 
 
In all four well-controlled, single-injection studies, Ampion demonstrated a larger improvement 
in pain in KL 4 patients compared to saline, indicating a reproducible and consistent response. 
Analysis of the combined data demonstrate a statistically significant result in this underserved 
patient population.  
In addition to the efficacy analysis described in this section, Ampion has also 
demonstrated a robust safety profile across the four well-controlled, single-injection 
studies, as well as the multiple-injection and early phase trials (Table 1.3). No drug-
related Serious Adverse Events (SAEs) have been reported in 2,007 patients treated 
during the clinical development program. The Adverse Event (AE) profile is similar for 


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Ampion and saline, with a majority of the AEs of minor or moderate severity and 
unrelated to treatment. 
Table 1.3  
Summary of Adverse Events 
Adverse event (AE) 
Overall population 
KL 4 population 
 
Ampion 
N=1,076 
Saline 
N=931 
Ampion  
N=475 
Saline  
N=361 
One or more AE 
381 (35.4%) 
386 (41.5%) 
150 (30.1%) 
140 (38.8%) 
One or more related AE 
11 (1.0%) 
19 (2.0%) 
3 (0.6%) 
6 (1.7%) 
AE by severity* 
 
  Mild 
264 (24.5%) 
253 (27.2%) 
100 (21.2%) 
86 (23.8%) 
  Moderate 
162 (15.1%) 
190 (20.4%) 
58 (12.3%) 
74 (20.5%) 
  Severe 
35 (3.3%) 
32 (3.4%) 
17 (3.6%) 
14 (3.9%) 
Serious AE (SAE) 
15 (1.4%) 
20 (2.1%) 
5 (1.1%) 
7 (1.9%) 
AE leading to withdrawal 
0 (0%) 
0 (0%) 
0 (0%) 
0 (0%) 
AE leading to death 
0 (0%) 
0 (0%) 
0 (0%) 
0 (0%) 
*Subjects could have AEs in more than one severity category 
CONCLUSIONS 
 
Clinical efficacy observed in previous single-injection studies suggest that Ampion safely 
relieves pain due to severe (KL 4) osteoarthritis of the knee, the most severe form of OAK, for 
which there is no FDA approved treatment. Currently, when KL 4 patients are treated for OAK, 
they are receiving treatment using IA therapy (such as viscosupplementation) that were not 
studied, nor required in their device approvals to demonstrate efficacy in the KL 4 population 
(Rutjes 2012). Historical treatments for OAK pain also involve opioid-based treatments or 
surgical intervention. Ampion treatment offers clinically meaningful pain relief for an unmet 
medical need in severely diseased KL 4 patients with a single IA injection. 
 


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2 
RATIONALE FOR THE STUDY 
This is a Phase 3 randomized study designed to confirm the efficacy of an IA injection of 
Ampion and is designed to support the completed single injection Ampion studies. 
2.1 
Rationale for the Doses and the Dosing Regimen 
This trial will use the 4 mL volume of Ampion used in the single-injection studies (AP-003-A, 
AP-004, AP-003-B and AP-003-C).  


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3 
STUDY DESIGN 
3.1 
Study Design Overview 
This is a randomized, saline-controlled, double-blind study with a 28-day screening period for 
each patient followed by a 24-week participation period. This trial will use an adaptive design 
with promising zone method for sample size re-estimation at the interim analysis. Daily 
assessments of WOMAC A pain using a subject diary will be assessed for seven days beginning 
one week prior to the clinic visits at Week 0 (baseline) and Week 12 to allow for calculation of a 
WOMAC A weekly pain score, which will be used for conducting the primary efficacy analysis.  
This trial is designed for 1,034 patients, 517 per study arm, randomized 1:1 for Ampion and 
saline IA injection. There will be an interim analysis at 724 patients, 362 per study arm, 
randomized 1:1, Ampion and saline. After the interim analysis, enrollment will either a) continue 
up to a total of 1,034 patients, 517 patients per study arm, randomized 1:1 for Ampion and saline 
IA injection; b) allow for sample size re-estimation using the promising zone method (Pocock, 
2011) up to 1.5 times the original trial size for a total of 1,552 patients, 776 per study arm, 
randomized 1:1 for Ampion and saline IA injection.  
The clinical effects of treatment on severe OAK will be evaluated at baseline 2, 4, 6, 8, 10, 12, 
and 24 weeks, using the Western Ontario and McMaster Universities Arthritis Index 
(WOMAC®) osteoarthritis Index 3.1, and the Patient’s Global Assessment of disease severity 
(PGA).  
The WOMAC® and PGA are validated scoring systems and sets the standard for the patient 
response. In order not to bias the collection of data, only questions from the validated WOMAC 
Index 3.1 and the PGA will be asked of patients.  
 
The trial has co-primary hypotheses and requires success on both for a successful trial.  First, 
pain reduction must be demonstrated for Ampion vs. saline control. Next, a function 
improvement must be shown on Ampion vs. saline control. 
 
The value for clinical benefit is derived from the minimal clinically important improvement 
(MCII), which is defined as the smallest change that signifies an important improvement to the 
patient (Tubach 2005). The Initiative on Methods, Measurement, and Pain Assessment in 
Clinical Trials (IMMPACT) advocates a provisional benchmark of 10-20% for minimally 
important improvement (Dworkin 2008). In addition, two studies have reported results of a 
survey of key opinion leaders and clinicians to determine value was best for MCII, suggesting a 
20% MCII may be useful in patients with OAK (Tubach 2005), including severe OAK (Salottolo 
2018). Clinical benefit will be determined as observed 20% improvements from baseline in pain 
and function at Week 12 to determine improvements are clinically important and not just 
statistically significant. 
 
Clinical meaningfulness will be determined by the end results of this trial, specifically by the 
apparent clinical benefit versus any adverse events or any increased apparent risk. Safety will be 
assessed by recording adverse events (through 24 hours post-dose and at all follow up contacts) 
and physical examination and vitals (Baseline, Weeks 6, 12, and 24). 


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3.2 
Study Objectives 
3.2.1 
Primary Objective 
The co-primary trial objectives are to evaluate the greater efficacy for pain improvement and 
function improvement of 4 mL Ampion versus saline IA injection when applied to patients 
suffering from severe OAK. Efficacy will primarily be assessed with WOMAC A pain and 
WOMAC C function scores. 
Mean change in the WOMAC A weekly pain scores from Baseline to Week 12 will be compared 
between Ampion and saline control.  Mean change in WOMAC C function score will be 
compared between Ampion and saline control. This will ensure that Ampion is superior to saline 
in improving pain and function. 
3.2.2 
Secondary Objectives 
The secondary trial objectives include the following evaluations of an intra-articular injection of 
Ampion, as compared to saline, when administered to patients suffering from severe OAK: 
improvement in pain utilizing a 100 mm Visual Analogue Scale (VAS) scale; assessment of 
safety; improvement in the Patient Global Assessment; and the amount of rescue analgesia.  
3.2.3 
Exploratory Objectives 
Exploratory objectives include analyzing the effect, if any, of the efficacy of Ampion, as 
compared to saline, on stiffness; Outcome Measures in Rheumatology Clinical Trials and 
Osteoarthritis Research Society International (OMERACT-OARSI) responder status; proportion 
of treatment failures; change in pain and function as a cumulative distribution function (CDF) 
plot. 
 
Analysis with biomarkers, such as those for analgesic use and chemokines and cytokines 
associated with inflammation and cartilage regeneration, may be analyzed to compare 
differences between treatment groups using urine samples collected at Week 12.  
 
Additionally, a Bayesian meta-analysis will be performed for WOMAC A Likert (LK) pain sub 
score using all KL 4 patients in studies AP-003A, AP-003-B, AP-003-C, AP-004, and AP-013. 
3.3 
Safety Monitoring Committee 
A Safety Monitoring Committee (SMC) will be established to review the safety of Ampion as 
the study progresses. The SMC will consist of independent clinicians and statisticians not 
involved in the conduct of the clinical trial. The Medical Monitor will attend open sessions of the 
SMC meetings. The SMC will be primarily responsible for reviewing any serious Adverse Event 
(SAE) and other clinically important safety findings (e.g., discontinuations due to AEs) that may 
occur during the study. Prior to unblinding any treatment assignments, a charter will be 
developed to detail the SMC review responsibilities, the frequency of meetings, and data 
evaluation plans. 


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A promising zone analysis will be run after outcomes are available for 724 patients (70% of final 
sample size).  The promising zone analysis will be run for WOMAC C since function will be the 
predominant determinant of sample size. 
The SMC, with the exclusion of the Medical Monitor, will also be tasked with evaluating 
whether sample size increase criteria has been met for the interim efficacy analysis. The SMC 
committee will review the interim analyses and report the next steps to the Sponsor, which will 
include one of the following prespecified actions: (1) continue to the a total of 1,034 patients, 
517 patients per study arm, randomized 1:1 for Ampion and saline IA injection; (2)  sample size 
re-estimation up to 1.5 times the original trial size for a total of 1,552 patients, 776 per study 
arm, randomized 1:1 for Ampion and saline IA injection, has occurred using the promising zone 
and the new sample size will be reported.  
3.4 
Stopping Rules 
There are no stopping rules for early success stopping.  The trial will enroll 1,034 patients, or if 
the promising zone triggers an expansion, up to 1,552 patients. 
The Safety Monitoring Committee may recommend trial termination if there is a safety concern. 
3.5 
Study Endpoints 
3.5.1 
Co-Primary Endpoint 
The trial will test whether Ampion can produce a superior improvement in pain and function vs. 
saline control. This goal is achieved using two co-primary endpoints: 
 
Change in the WOMAC A pain sub score between Baseline and Week 12  
Change in the WOMAC C physical function sub score between Baseline and Week 12 
 
Mean 12-week change in the WOMAC A weekly pain scores determined at Baseline and Week 
12 will be compared between Ampion and saline control. Mean 12-week change in WOMAC C 
physical function sub score will be compared between Ampion and saline control. 
3.5.2 
Secondary Endpoints 
• Change in PGA between Baseline and Week 12  
• Change in the 100 mm VAS Pain scale between Baseline and Week 12 
• Use of rescue analgesia (amount of acetaminophen used between Baseline and Week 12) 
• Incidence and severity of treatment-emergent adverse events (TEAEs) 
 
3.5.3 
Exploratory Endpoints 
• Change in WOMAC B stiffness sub score between Baseline and Weeks 2, 4, 6, 8, 10,  
12, and 24 


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• Response status based on the OMERACT-OARSI criteria at Weeks 2, 4, 6, 8, 10, 12 and 
24  
• Proportion of treatment failures at Week 12 
• Change in pain and function analysis as a cumulative distribution function (CDF) plot 
• A Bayesian meta-analysis for LK WOMAC A pain sub score including KL 4 patients 
from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013 
• Change in WOMAC A pain sub score between Baseline and Weeks 2, 4, 6, 8, 10, and 24  
• Change in WOMAC C function sub score between Baseline and Weeks 2, 4, 6, 8, 10, and 
24  
• Change in WOMAC C function sub score between Baseline and Weeks 2 through 12 and 
Weeks 2 through 24  
• Change in PGA between Baseline and Weeks 2, 4, 6, 8, 10, and 24 
• Change in WOMAC C physical function sub score between Baseline and Weeks 2, 4, 6, 
8, and 10 
• If collected, difference in urine analysis between treatment groups at Week 12 
3.6 
Blinding and Randomization 
Patients will be assigned to treatment by a randomization schedule developed and maintained by 
an independent statistician. Patients, the investigator, Sponsor, and all study staff having a role in 
the day-to-day conduct of the study will remain blinded to the treatment assignment. 
Ampion and saline will be provided as blinded study vials labeled with the appropriate 
information. To maintain study blinding, vial labels will be designed to adequately cover vial 
contents and prevent discrimination between active and placebo. Syringes, needles, and rescue 
medication (acetaminophen) will be provided to the sites.  
A comprehensive presentation of the Data Management Plan (DMP) and Statistical Analysis 
Plan (SAP) will be approved by Ampio Pharmaceuticals, Inc. prior to unblinding of study data. 
The SMC will be unblinded to perform the interim analysis.  
3.7 
Code-breaking 
Where required, safety personnel and/or investigator may be unblinded to a particular patient’s 
treatment assignment to meet reporting requirements to Regulators. Unblinding may occur by 
contacting Ampio Pharmaceuticals, Inc.  


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4 
SELECTION OF PATIENTS 
4.1 
Number of Patients 
This trial will use an adaptive design with promising zone method for sample size re-estimation. 
This trial is designed for 1,034 patients, 517 per study arm, randomized 1:1 for Ampion and 
saline IA injection. There will be an interim analysis at 724 patients, 362 per study arm, 
randomized 1:1, Ampion and saline.  
Following the interim analysis, enrollment will either a) continue up to a total of 1,034 patients, 
517 patients per study arm, randomized 1:1 for Ampion and saline IA injection; b) allow for 
sample size re-estimation up to 1.5 times the original trial size for a total of 1,552 patients, 776 
per study arm, randomized 1:1 for Ampion and saline IA injection.  
The Ampion effect was estimated as the average change in pain from Baseline to Week 12 for 
KL 4 subjects receiving Ampion from all prior single injection studies randomized 1:1 with 
saline control. Saline effect was estimated as the average change in pain from Baseline to Week 
12 for KL 4 subjects receiving saline from all prior single injection studies randomized 1:1 for 
Ampion and saline.  The sample size for assessing the WOMAC A Pain score alone would be 
690 total patients for 90% power.   
The Ampion effect was estimated as the average change in function from Baseline to Week 12 
for KL 4 subjects receiving Ampion from all prior single injection studies randomized 1:1 with 
saline control. Saline effect was estimated as the average change in function from Baseline to 
Week 12 for KL 4 subjects receiving saline from all prior single injection studies randomized 1:1 
for Ampion and saline.  The sample size for assessing the WOMAC C Function score alone 
would be 984 patients for 90% power.   
The total sample size is chosen as the larger of the two values. To be conservative sample sizes 
were estimated with a 5% dropout so 1,034 patients will be enrolled to achieve 984 total patients.  
All ITT patients will be included in the statistical analysis. At the assumed effect size there is a 
90% probability of success at 1,034 patients. 
The study design of 1,034 total patients achieves 97% power assuming a WOMAC A pain score 
with mean pain decreases of -0.83 for Ampion and -0.62 for saline and a standard deviation of 
0.85. 
 As described in Section 10, a Promising Zone method will be employed for a sample size re-
estimation and the trial may increase to 1,552 patients. 
4.2 
Recruitment Methods 
Patients will be recruited from the population being seen by Investigators at the clinical sites 
participating in the study. In addition, notifications about the opportunity for patients to 
participate in a clinical trial will be sent to referring physicians. A description of the clinical trial 
will also be posted at ClinicalTrials.gov, and advertisements and/or other notices may be 
produced to advise potential study patients on how they may obtain information about study 


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participation. All such materials will be reviewed and approved by the Institutional Review 
Board (IRB) prior to their publication or dissemination. 
4.3 
Inclusion Criteria 
It is recommended that patients should have a WOMAC A pain sub score of < 1.5 as measured 
by the 5-point Likert Pain Subscale in the contralateral knee, which is assessed at screening. 
Patients should fulfill the following inclusion criteria:  
1. Able to provide written informed consent to participate in the study 
2. Willing and able to comply with all study requirements and instructions of the site study staff 
3. Male or female, 40 years to 85 years old (inclusive) 
4. Must be ambulatory  
5. Index knee must be symptomatic for greater than six months with a clinical diagnosis of 
OAK and supported by radiological evidence (Kellgren-Lawrence Grade 4) that is not older 
than six months prior to the date of screening 
6. Moderate to moderately-severe OA pain in the index knee (rating of at least 1.5 on the 
WOMAC Index 3.1 as measured by the 5-point Likert Pain Subscale,) assessed at screening 
7. Ability to temporarily discontinue NSAID for 48 hours prior to scheduled clinical efficacy 
evaluations  
8. No analgesia (including acetaminophen [paracetamol]) taken 12 hours prior to an efficacy 
measure 
9. No known clinically significant liver abnormality (e.g. cirrhosis, transplant, etc.) 
4.4 
Exclusion Criteria 
Patients fulfilling one or more of the following criteria may not be enrolled in the study: 
1. As a result of medical review and screening investigation, the Principal Investigator 
considers the patient unfit for the study 
2. A history of allergic reactions to human albumin (reaction to non-human albumin such as egg 
albumin is not an exclusion criterion) 
3. A history of allergic reactions to excipients in 5% human albumin (N-acetyltryptophan, 
sodium caprylate) 
4. Presence of tense effusions 
5. Inflammatory or crystal arthropathies, acute fractures, history of aseptic necrosis or joint 
replacement in the affected knee, as assessed locally by the Principal Investigator 
6. Isolated patella femoral syndrome, also known as chondromalacia 
7. Any other disease or condition interfering with the free use and evaluation of the index knee 
for the duration of the trial (e.g. cancer, congenital defects, spine OA) 
8. Major injury to the index knee within the 12 months prior to screening  
9. Severe hip OA ipsilateral to the index knee 
10. Any pain that could interfere with the assessment of index knee pain (e.g. pain in any other 
part of the lower extremities, pain radiating to the knee) 
11. Any pharmacological or non-pharmacological treatment targeting OA started or changed 
during the four weeks prior to randomization or likely to be changed during the duration of 
the study  


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12. Use of the following medications: 
a. No IA injected pain medications in the study knee during the study 
b. No analgesics containing opioids. NSAIDs may be continued at levels preceding the 
study, however may not be used 48 hours prior to efficacy evaluations, and 
acetaminophen is available as a rescue medication during the study from the provided 
supply  
c. No topical treatment on osteoarthritis index knee during the study  
d. No significant anticoagulant therapy (e.g. Heparin or Lovenox) during the study 
(treatment such as Aspirin and Plavix are allowed) 
e. No systemic treatments that may interfere with safety or efficacy assessments during the 
study 
f. No immunosuppressants 
g. No use of corticosteroids > 10 mg prednisolone equivalent per day  
h. If corticosteroid use is ≤ 10 mg prednisolone equivalent per day, and if clinically 
indicated, subjects should be allowed to decrease their corticosteroid use. Additionally, 
some subjects may need to increase their steroid dose to treat worsened symptoms in the 
treated knee, and subjects who increase their corticosteroid dose above their starting dose 
of corticosteroid during the study will be treated as “treatment failures” for efficacy 
analysis 
13. Any human albumin treatment in the three months before randomization or throughout the 
duration of the study. 
4.5 
Inclusion of Patients Incapable of Giving Informed Consent 
No patient incapable of giving informed consent may be enrolled in the study. 


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5 
STUDY PLAN AND PROCEDURES 
Patients will be screened for inclusion into the clinical study, which will include the ability to 
record pain on a daily basis for seven days prior to randomization. Patients will be randomized 
into a study arm and receive an IA injection of Ampion or saline. Telephone contact will be 
made with the patient 24 hours after the IA injection. The patient will be followed for 24 weeks 
and the clinical effects of treatment on OAK pain will be evaluated during clinic visits at 6, 12, 
and 24 weeks, and during telephone contacts at 2, 4, 8 and 10 weeks, using the WOMAC 
osteoarthritis Index 3.1 (pain sub score, stiffness sub score and function sub score) and an overall 
global severity assessment (PGA).  
Safety will be assessed by recording adverse events (through 24 hours post-dose and at all 
follow-up in-clinic visits and telephone contacts), vital signs (Baseline, and Weeks 6 and 12), 
recording prior and concomitant medications including start/stop dates, indication, dose and 
frequency (through 24 hours post-dose and at all follow-up in-clinic visits and telephone 
contacts) and physical examination (Baseline and Weeks 6, 12, and 24). The assessments and 
procedures performed at each patient visit or contact are described in Section 5 and in Table 6.1. 
5.1 
Description of Study Visits 
5.1.1 
Visit -1 (Day -28 through Day 0, in-clinic): Screening 
The following procedures will be performed: 
• Obtain written informed consent before the start of any study specific procedure. 
• Review medical history including all previous treatments for OA 
• Record prior and concomitant medications including start/stop dates, indication, dose and 
frequency 
• Record demographic data including year of birth, age, gender and race 
• Measure and record height and weight 
• Perform and record physical examination 
• Record KL Grade from radiological assessment that is not older than 6 months prior to 
screening. Patient may have an x-ray at screening in order to satisfy this requirement 
• Record pain in the index knee with the WOMAC Index 3.1 using both the 5-point Likert 
scale and the 100mm VAS scale 
• Record pain in the contralateral knee with the WOMAC Index 3.1 pain subsection using 
both the 5-point Likert scale and the 100mm VAS scale 
• Record vital signs. (Measure body temperature, systolic blood pressure [SBP], diastolic 
blood pressure [DBP] and pulse rate) 
• Evaluate all inclusion and exclusion criteria to ensure that patients meet all inclusion 
criteria and none of the exclusion criteria 
• Review collection instructions of daily diary pain scores with subject 
 
 
 


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5.1.2 
Visit 0 (Day -6, telephone contact): Pre-Baseline Daily Assessments 
The following procedures will be performed:  
• Review requirements for daily collection of WOMAC A pain evaluation using both the 
5-point Likert scale and the 100mm VAS scale 
• Confirm Baseline visit date 
• It is possible, depending on the Baseline visit date and successful completion of 
Screening criteria defined above, that the first Pre-Baseline Daily Assessment may occur 
during the Screening visit. 
5.1.3 
Visit 1 (Day 0, in-clinic): Baseline/Randomization/Treatment 
The following procedures will be performed: 
• Confirm eligibility (review inclusion/exclusion criteria) 
• Record concomitant medications 
• Conduct pregnancy test, as applicable 
• Perform and record physical examination 
• Record vital signs (Measure body temperature, SBP, DBP, and pulse rate) pre- and post-
injection 
• Perform and/or review the final Pre-Baseline WOMAC A pain evaluation using both the 
5-point Likert scale and the 100mm VAS scale 
• Perform WOMAC B and C prior to treatment  
• Perform PGA evaluations prior to treatment 
• Randomize patient to study arm 
• Perform intra-articular injection of blinded study treatment  
• Record post-injection AEs if observed 
• Issue rescue medication (acetaminophen [paracetamol]) 
5.1.4 
Visit 2 (Day 1, telephone contact) 
The following procedures will be performed: 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.5 
Visit 3 (Week 2 ± 7 days, telephone contact) 
The following procedures will be performed: 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 


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5.1.6 
Visit 4 (Week 4 ± 7 days, telephone contact) 
The following procedures will be performed: 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.7 
Visit 5 (Week 6 ± 7 days, in-clinic) 
The following procedures will be performed: 
• Perform and record physical examination 
• Record vital signs. (Measure body temperature, SBP, DBP and pulse rate) 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
 
5.1.8 
Visit 6 (Week 8 ± 7 days, telephone contact) 
The following procedures will be performed: 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.9 
Visit 7 (Week 10 ± 7 days, telephone contact) 
The following procedures will be performed: 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.10 
Visit 8 (Week 11 ± 7 days, telephone contact): Pre-Week 12 Daily Assessments 
The following procedures will be performed:  
• Review requirements for daily collection of WOMAC A pain scores using both the 5-
point Likert scale and the 100mm VAS scale 
• Confirm Week 12 visit date 
 


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5.1.11 
Visit 9 (Week 12 ± 7 days, in-clinic) 
The following procedures will be performed: 
• Collect a urine sample for testing 
• Perform and record physical examination 
• Record vital signs. (Measure body temperature, SBP, DBP and pulse rate) 
• Perform and/or review the final WOMAC A pain evaluation using both the 5-point Likert 
scale and the 100mm VAS scale 
• Perform WOMAC B and C evaluations 
• Perform PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.12 
Visit 10 (Week 24 ± 7 days, in-clinic) 
The following procedures will be performed: 
• Perform and record physical examination 
• Record vital signs (Measure body temperature, SBP, DBP and pulse rate) 
• Perform WOMAC and PGA evaluations of the study knee 
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
 
5.1.13 
Early Termination Visit 
The following procedures will be performed at Early Termination Visit: 
• Perform and record physical examination 
• Record vital signs. (Measure body temperature, SBP, DBP and pulse rate) 
• Perform WOMAC and PGA evaluations of the study knee using both the 5-point Likert 
scale and the 100mm VAS scale  
• Record AEs 
• Record concomitant medications 
• Review rescue medication use 
5.1.14 
Unscheduled Visits 
Additional visits may be scheduled at the discretion of the Investigator, for example as part of 
follow-up of AEs. Unscheduled Visits will follow the visit structure described in  
Section 5.1.7 for the Week 6 visit.  


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5.1.15 
Missed Visits 
Patients unable to complete a study visit as scheduled should be re-scheduled for a replacement 
visit as soon as possible. If a subject misses any scheduled follow-up visit and cannot be seen 
within the +/- 7-day visit window, as described in Table 6.1, the visit is considered missed.  


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6 
METHODS OF ASSESSMENT 
Demographic Data 
At Visit 1(Screening), patient demographic data will be collected. These include year of birth, 
age, gender and race. 
Medical History 
At Visit 1(Screening) a complete medical history, including prior interventions to the study knee, 
will be obtained from each patient. 
Concomitant Medications 
Detailed history of medications will be documented for each patient at Screening and Baseline. 
Concomitant medications (especially any changes in medication) will be documented for each 
patient at each scheduled visit. 
Physical Examination and Vital signs 
Height in feet and inches will be measured at Screening. 
Body weight in pounds (lb) will be measured at Screening. 
Body temperature (deg F) will be measured at each visit. 
SBP and DBP and pulse rate will be measured with the patient in a seated position.  
The full physical examination will consist of examining the following body systems: 
cardiovascular, respiratory, abdominal, skin and musculoskeletal other than the knee. The 
physical examination of the target knee will consist of evaluating the knee joint for effusion and 
tenderness on palpation. Table 6.1 below lists the various study activities and timing. 
Biomarkers 
At Week 12 urine samples will be collected and may be analyzed to compare differences 
between treatment groups for biomarkers, such as those for analgesic use and chemokines and 
cytokines associated with inflammation and cartilage regeneration. 
6.1 
Efficacy Assessments 
Note: Efficacy questionnaire questions will be asked “with reference to study knee” i.e. to obtain 
scores specific for the treated knee. 
6.1.1 
WOMAC® Osteoarthritis Index (Bellamy 1988) 
The clinical effects of treatment on OA pain and function will be evaluated during clinic visits at 
6, 12, and 24 weeks, and telephone contacts at 2, 4, 8 and 10 weeks, using the WOMAC Index 


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3.1, and the PGA. All patients are required to take at least five minutes to complete the 
questionnaire. 
Patients are asked about their pain, stiffness, and function in the knee (study joint) due to arthritis 
during the last 24 hours. 
In addition to WOMAC scoring during clinic visits, daily WOMAC A pain sub scores will be 
collected for seven days up to and including Baseline and Week 12 to allow for calculation of 
WOMAC A weekly pain scores, which will be used for conducting the primary efficacy analysis. 
Patients respond to each subscale by using a 5-point adjectival Likert score (0 = None, 1 = Mild, 
2 = Moderate, 3 = Severe, 4 = Extreme) and 100mm VAS. 
The WOMAC B and WOMAC C sub scores will be collected at Weeks 0 (Baseline), 2, 4, 6, 8, 
10, 12, and 24 using the 5-point Likert scale. Patients respond to each subscale by using a 5-point 
adjectival Likert score (0 = None, 1 = Mild, 2 = Moderate, 3 = Severe, 4 = Extreme). 
6.1.2 
Patient’s Global Assessment of Disease Severity (PGA) 
The PGA should be completed by patients at Screening, pre-dose Day 1 (prior to injection), and 
at Weeks 2, 4, 6, 8, 10,12 and 24. 
Patients are asked the following question: “Considering all the ways in which your arthritis 
affects you, please indicate how you are doing.” 
Patients respond by using a 5-point adjectival Likert score (0 = Very Well, 1 = Well, 2 = Fair, 3 
= Poor, 4 = Very Poor). 
6.2 
Safety Parameters 
Based on results of a Phase 1b and a Phase 2 randomized, controlled, repeat-blind study of a 
single IA injection of Ampion < 5 kDa ultrafiltrate of 5% HSA in adults with OAK (AIK study), 
in which no clinically significant differences between active and placebo were found in either 
electrocardiogram or blood parameters, safety will be assessed by recording adverse events, vital 
signs, results of physical examination, and by recording prior and concomitant medications.  
6.2.1 
Vital Signs 
Vital signs (radial pulse rate, SBP, DBP, and body temperature) should be recorded at Screening, 
pre- and post–treatment Day 1, and then on visits at Weeks 6,12 and 24. 
Vital signs should be taken after the patient has rested in a seated position for at least 
five minutes.


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Table 6.1  
Schedule of Assessments and Procedures 
 
Screening 
Pre-
Baseline 
(phone) 
Baseline 
24-hours 
(phone) 
Week 2 
(phone) 
Week 4 
(phone) 
 
Week 6 
 
Week 8 
(phone) 
Week 10 
(phone) 
Pre-Week 
12 
(phone) 
Week 12 
 
Week 24 
 
Early 
Term. 
Assessments 
& Procedures 
Visit -1 
Day-28 to 0 
Visit 0 
Day -6 to 0 
Visit 1 
Day 0 
Visit 2 
Day 1 
Visit 3 
Day 14 ± 7 
Visit 4 
Day 28 ± 7 
Visit 5 
Day 42 ± 7 
Visit 6 
Day 56 ± 7 
Visit 7 
Day 70 ±7 
Visit 8 
Day 77 ±7 
Visit 9 
Day 84 ± 7 
Visit 10 
Day 174 ± 7 
 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/ 
exclusion criteria 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
Medical history/prior 
medications 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
Concomitant 
medications 
X 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Urine sample 
 
 
X** 
 
 
 
 
 
 
 
X 
 
 
Physical examination 
X 
 
X 
 
 
 
X 
 
 
 
X 
X 
X 
Vital Signs 
X 
 
X 
 
 
 
X 
 
 
 
X 
X 
X 
Randomization 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Review Diary 
Completion Instructions 
X 
 
 
 
 
 
 
 
X 
 
 
 
 
Phone Contact Daily 
Collection Reminder  
 
X 
 
 
 
 
 
 
 
X 
 
 
 
WOMAC A (Likert) 
X 
X* 
X*** 
 
X 
X 
X 
X 
X 
X* 
X*** 
X 
X 
WOMAC A (VAS) 
X 
X* 
X*** 
 
 
 
 
 
 
X* 
X*** 
 
X 
WOMAC B and C 
(Likert) 
X 
 
X 
 
X 
X 
X 
X 
X 
 
X 
X 
X 
PGA 
X 
 
X 
 
X 
X 
X 
X 
X 
 
X 
X 
X 
X-ray1 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Study treatment 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Dispense Rescue 
medication 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Review Rescue 
medication 
 
 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Adverse Events 
 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Visits are in clinic except for Day 1 and Weeks 2, 4, 8 and 10 when patients will be contacted by telephone 
1X-ray may be acquired at Screening to satisfy inclusion criteria, “Index knee must be symptomatic for greater than six months with a clinical diagnosis of OA and supported by 
radiological evidence (KL 4) that is not older than six months prior to the date of screening”.  
*Patients will complete 7 daily pain score assessments up to and including the Baseline and Week 12 visits to allow calculation of WOMAC A weekly pain scores. The Screening 
and first Pre-Baseline daily pain score collection may occur on the same day, in office or at home, as appropriate. 
**Urine will only be collected at Baseline if necessary to conduct a pregnancy test; collection to occur as applicable 
***The WOMAC may occur in office or at home, and may serve as the 7th WOMAC Daily Pain collection  


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7 
DISCONTINUATION CRITERIA 
7.1 
Early Discontinuation of the Study 
It is agreed that for reasonable cause, either the investigator or the Sponsor may terminate this 
study, provided a written notice is submitted at a reasonable time in advance of intended 
termination; if by the investigator notice is to be submitted to Ampio Pharmaceuticals, Inc., and 
if by the Sponsor, notice will be provided to each investigator. 
If a severe local reaction or drug-related SAE occurs at any time during the study, the Safety 
Monitoring Committee will review the case immediately. 
The study will be immediately suspended and no additional Ampion treatments administered 
pending review and discussion of all appropriate study data by the SMC if one or more patients 
develop any of the following adverse events deemed to be possibly, probably, or definitely 
related to Ampion by the Investigator and/or Medical Monitor, based upon close temporal 
relationship or other factors: 
• Death 
• Anaphylaxis (angioedema, hypotension, shock, bronchospasm, hypoxia, or 
respiratory distress) Induction of autoimmune arthritis  
• Hepatic failure 
• Aplastic anemia 
The study will not be restarted until all parties have agreed to the course of action to be taken and 
the IRB/EC has been notified. 
7.2 
Early Discontinuation of Individual Patients 
Patients are to be withdrawn from the study for any of the following reasons: 
• Withdrawal of informed consent 
• Patient is lost to follow-up 
Patients will also be withdrawn at any time if the investigator concludes that it would be in the 
patient’s best interest for any reason. Protocol violations do not lead to patient withdrawal unless 
they constitute a significant risk to the patient’s safety. 
Patients can voluntarily withdraw from the trial for any reason at any time. They are to be 
considered withdrawn if they state an intention to withdraw, fail to return for visits, or become 
lost to follow up for any reason. Patients withdrawing from the study because of an AE should be 
followed for at least 30 days, resolution of the AE or until no further improvement is expected, 
whichever comes first. 


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8 
TREATMENT 
Eligible patients will receive a single IA injection into one knee of either Ampion or a saline 
injection, dependent on study arm. 
Patients will be allocated to a treatment in accordance with the randomization schedule following 
confirmation of eligibility before treatment. 
8.1 
Dosing and administration of study medication 
Appropriately trained site personnel should administer the study treatment. 
If both knees are osteoarthritic, then the Investigator at Screening should select the knee that best 
satisfies the requirements for the study (see inclusion criteria) as the study knee. The study knee 
should be treated in accordance with the randomization schedule.  
It is recommended that the study treatment be administered into the knee joint space under sterile 
prep conditions (i.e. the knee should be cleaned with an antiseptic). The treatment area may be 
anesthetized with local anesthesia including a topical anesthesia, injection of lidocaine, or 
nothing, as determined by the Principal Investigator. The recommended needle of choice for this 
procedure is a 25-gauge needle that is 1.5 inches long. The needle may be passed obliquely 
towards the trochlear groove. If an effusion is present, aspiration should not be performed, and 
the effusion should be noted in the study source notes/eCRF. Injection should proceed easily. 
Failure to easily inject should be documented. The same sized needle should be used for both 
Ampion and saline administration and recorded in the study source notes/eCRF. The date of 
administration should be recorded in the study source notes/eCRF. 
8.2 
Drug Storage and Accountability 
Study drug should be stored at controlled room temperature (59º – 77ºF or 15º – 25ºC). Each site 
shipment of study drug will include temperature monitors and instructions to download the 
temperature data to Ampio Pharmaceuticals, Inc. If the study drug is stored at the clinical site, 
temperature should be monitored at least daily until use to ensure storage at the defined 
temperature. 
The Investigator, the clinical site pharmacist, or other personnel authorized to store and dispense 
investigational product is responsible for ensuring that the investigational product used in the 
clinical study is securely maintained as specified by the Sponsor and in accordance with the 
applicable regulatory requirements. 
All investigational product is to be dispensed in accordance with the Investigator’s prescription 
and it is the Investigator’s responsibility to ensure that an accurate record is maintained of 
investigational product issued and returned. 
If any quality issue is noticed upon the receipt or use of an investigational product (i.e. 
deficiencies in condition, packaging, appearance, associated documentation, labeling, etc.), 
Ampio Pharmaceuticals, Inc. must be promptly notified. 


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Under no circumstances may the Investigator supply investigational product to a third party, 
allow the investigational product to be used other than as directed by this clinical study protocol, 
or dispose of investigational product in any other manner. 
Ampio, or designated representative, will instruct sites on study kit returns and/or destruction of 
study drug. If returned, study kits must be returned to Ampio Pharmaceuticals, Inc., or 
designated representative after study completion.  
8.3 
Concomitant Treatments and Rescue Medication 
The following medications / therapies are NOT allowed during this clinical study:  
1. No IA injected pain medications in the study knee during the study. 
2. No analgesics containing opioids.  
3. No NSAID use during the 48 hours prior to efficacy evaluations. 
4. No clinically significant increase in NSAID use above baseline/pre-study dosing levels to 
treat worsened symptoms in the treated knee. 
a. NSAID use during the study may be continued at baseline/pre-study dosing levels 
and may be increased or decreased if clinically indicated; acetaminophen is also 
available as a rescue medication during the study from the provided supply.  
5. No non-pharmacological treatment targeting OA started or changed during the study. 
6. No topical treatment on osteoarthritis index knee during the study. 
7. No significant anticoagulant therapy (e.g. Heparin or Lovenox) during the study 
(treatment such as Aspirin or Plavix is allowed). 
8. No systemic treatments that may interfere with safety or efficacy assessments during the 
study. 
9. No immunosuppressants.   
10. No use of corticosteroids > 10 mg prednisolone equivalent per day. 
a. If corticosteroid use is ≤ 10 mg prednisolone equivalent per day, and if clinically 
indicated, subjects should be allowed to decrease their corticosteroid use. 
Additionally, some subjects may need to increase their steroid dose to treat 
worsened symptoms in the treated knee, and subjects who increase their 
corticosteroid dose above their starting dose of corticosteroid during the study 
will be treated as “treatment failures” for efficacy analysis. 
11. No human albumin treatment during the study. 
Any medication used during the study should be recorded. All concomitant medication start and 
stop dates, total daily dose, route and indication should to be recorded. 
The only allowed rescue medication is 500 mg of acetaminophen (paracetamol), one tablet every 
four hours as required by the patient. 
8.4 
Treatment Compliance 
The injection of study drug will be performed by the unblinded injector(s). A 5 mL syringe and 
25-gauge needle will be provided. Compliance with treatment is thus assured. 


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9 
ADVERSE EVENTS 
9.1 
Definition of an adverse event 
An adverse event (AE) is defined as any undesired medical occurrence in a patient or clinical 
investigation patient receiving a pharmaceutical product and which does not necessarily have a 
causal relationship with this treatment. An AE can therefore be any unfavorable sign or 
unintended sign (including an abnormal laboratory finding), symptom, or disease temporarily 
associated with the use of a study drug, whether or not it is related to the study drug. 
Assessment of severity of an AE will be rated according to categories listed in Table 9.1 . 
Table 9.1 
Definitions of AE Severity 
Grade 1 (MILD): 
The symptom is barely noticeable to the study patient and does not influence performance or functioning.  
Concomitant medication is not ordinarily indicated for relief of mild AEs. 
Grade 2 (MODERATE): 
The symptom is of sufficient severity to make the study patient uncomfortable and to influence performance of 
daily activities. Concomitant medication may be indicated for relief of moderate AEs. 
Grade 3 (SEVERE): 
The symptom causes severe discomfort, sometimes of such severity that the study patient cannot continue in the 
study. Daily activities are significantly impaired or prevented by the symptom. Concomitant medication may be 
indicated for relief of severe AEs. 
 
Determination of the relationship between the AE and the study drug will be made using the 
guidelines presented in Table 9.2. 
Table 9.2  
Guidelines for Determining the Relationship (if any)  
Between Adverse Event and the Study Drug 
Unrelated 
The adverse event is unlikely to have been caused by study drug. 
Possibly related 
It is unclear whether the adverse event may have been caused by study drug.  
Related 
The adverse event is likely to have been caused by study drug.  
 
9.2 
Definition of a Serious Adverse Event 
A Serious Adverse Event (SAE) is any untoward medical occurrence that occurs at any dose 
that: 
• Results in death 
• Is life-threatening (patient is at immediate risk of death from the event as it occurred) 
• Requires in-patient hospitalization (formal admission to a hospital for medical reasons) or 
prolongation of existing hospitalization 
• Results in persistent or significant disability/incapacity 
• Results in a congenital anomaly/birth defect 
Important medical events that may not result in death, are not life-threatening, or do not require 
hospitalization may be considered SAEs when, based on appropriate medical judgment, they 


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may jeopardize the patient and may require medical or surgical intervention to prevent one of the 
outcomes listed in this definition. Examples of such events include allergic bronchospasm 
requiring intensive treatment in an emergency room or at home, blood dyscrasias or convulsions 
that do not result in in-patient hospitalization, or the development of drug dependency or drug 
abuse. 
Hospitalizations for elective surgery or other medical procedures that are not related to a 
treatment-emergent AE are not considered SAEs. 
9.3 
Recording of adverse events and serious adverse events 
Recording and reporting of adverse events should be in accordance with the FDA final 
“Guidance for Industry and Investigators Safety Reporting Requirements for INDs and BA/BE 
Studies” of December 2012. 
Any AE is to be recorded in the eCRF. In order to avoid vague, ambiguous, or colloquial 
expressions, the AE should be recorded in standard medical terminology rather than the patient’s 
own words. Whenever possible, the investigator should combine signs and symptoms that 
constitute a single diagnosis. 
The existence of an AE may be concluded from a spontaneous report of the patient, from the 
physical examination, or from special tests (e.g. laboratory assessments, where applicable, or 
other study-specified tests [source of AE]). 
The reporting period begins from the time that the patient receives IA injection at the Baseline 
visit through including patient’s Final Visit at 24 weeks. Any events continuing at study exit will 
be followed for 30 days or to resolution, or until no improvement is expected, whichever comes 
first. Any SAE occurring after the reporting period must be promptly reported if a causal 
relationship to the investigational drug is suspected. If the patient begins a new therapy, the 
safety reporting period ends at the time the new treatment is started, however, death must always 
be reported when it occurs during the 24-week study period irrespective of intervening treatment. 
Each AE is to be evaluated for duration, severity, seriousness, and causal relationship to the 
investigational drug. Any action(s) taken, and the outcome must also be recorded. 
9.3.1 
AE Follow-up 
All AEs occurring during the study are to be followed up in accordance with good medical 
practice until they are resolved, stabilized or judged no longer clinically significant or, if a 
chronic condition, until fully characterized. Any AEs that are considered drug-related (possibly 
related, definitely related) must be followed for 30 days, or to resolution, or until no 
improvement is expected, whichever occurs first. 
9.3.2 
Overdose 
No information on treatment of overdose of Ampion is currently available. In the case of 
overdose, the patient should be followed as an AE and appropriate supportive medical treatment 
instigated. 


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9.4 
Serious Adverse Event Reporting 
9.4.1 
Reporting Requirements 
Unexpected serious suspected adverse reactions are subject to expedited reporting to FDA.  All 
SAEs must be entered into the eCRF within 24 hours of first knowledge of the event by study 
personnel. It is important that the investigator provide his/her assessment of relationship to study 
drug at the time of the initial report. Entry of an SAE into the eCRF triggers an automatic alert to 
the CRO safety team. The following information must be reported: 
• Protocol number 
• Site and/or Investigator number 
• Patient number 
• Demographic data 
• Brief description of the event 
• Onset date and time 
• Resolution date and time, if the event resolved 
• Current status, if event not yet resolved 
• Any concomitant treatment and medication 
• Investigator’s assessment of whether the SAE was related to Investigative product or not 
 
The CRO Safety Associate will contact the site for clarification of data entered onto the eCRF, or 
to obtain missing information. In the event of questions regarding SAE reporting, the site may 
contact the appropriate individual as in Section 9.4.2. 
9.4.2 
SAE Contact Information 
Ampio Main Line: 1 (720) 437-6500  
Ampio Pharmaceuticals, Inc., or their designee CRO, is responsible for submitting reports of 
AEs associated with the use of the drug that are both serious and unexpected to FDA according 
to 21 CFR 312.32 and the final guidance (2012). All investigators participating in ongoing 
clinical studies with the study medication will receive copies of these reports for prompt 
submission to their Institutional Review Board (IRB) or Ethics committee (EC). 


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10 STATISTICAL METHODS 
10.1 General Considerations 
This section describes the rules and conventions to be used in the presentation and analysis of the 
data. A comprehensive presentation of the data management and statistical analysis plan will be 
approved by Ampio Pharmaceuticals, Inc. prior to unblinding of study data.  
10.2 Study Objectives 
The co-primary trial objectives are to evaluate the greater efficacy for pain improvement and 
function improvement of 4 mL Ampion versus 4 mL saline IA injection when applied to patients 
suffering from severe OAK. Efficacy will primarily be assessed with WOMAC A pain and 
WOMAC C function scores. 
The secondary trial objectives include the following evaluations of an intra-articular injection of 
Ampion, as compared to saline, when administered to patients suffering from severe OAK: 
improvement in the 100 mm VAS Pain scale; assessment of safety; improvement in the Patient 
Global Assessment; the amount of rescue analgesia.  
 
Exploratory objectives are to analyze the effect, if any, of the efficacy of Ampion, as compared 
to saline, on stiffness; OMERACT-OARSI responder status; proportion of treatment failures; 
change in pain and function will be examined as a CDF plot for Ampion and saline treated 
patients; and a Bayesian meta-analysis for LK WOMAC A pain sub score including KL 4 
patients from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013. Analysis with 
biomarkers, such as those for analgesic use and chemokines and cytokines associated with 
inflammation and cartilage regeneration, may be analyzed to compare differences between 
treatment groups using urine samples collected at Week 12. 
10.3 Analysis Populations 
10.3.1 
Safety Analysis Population: 
The safety analysis population is defined as all patients who are randomized and receive study 
medication (Ampion or saline). Patients will be analyzed as treated. 
10.3.2 
Intent-to-treat Population: 
The intent-to-treat (ITT) analysis population is defined as all patients who are randomized. All 
efficacy analyses will be performed in the ITT population. Patients will be analyzed as 
randomized. 
10.3.3 
Per Protocol Population: 
The per protocol analysis population is defined as all patients included in the ITT analysis who 
met all entry criteria and had no major protocol violations. All efficacy analyses will be repeated 


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in the per-protocol population. These analyses will be supportive of the ITT analysis. Patients 
will be analyzed as treated. 
10.3.4 
Statistical Analysis of Co-Primary Effectiveness Endpoints 
A mixed-effects model with repeated measures (MMRM) will be used to test the main effect of 
change in WOMAC A weekly pain score (determined as the average of Daily Pain 
measurements taken at Baseline and Week 12) from Baseline of Ampion vs. saline at 12 weeks. 
The model will include change from Baseline measures at 2, 4, 6, 8, 10, and 12 weeks. 
Treatment assignment, week, and Baseline WOMAC A measure will be included as fixed 
effects.  Patient ID will be included to capture repeated measures using a compound symmetric 
covariance matrix for residuals.  The primary hypothesis test for pain is the change in pain at 12-
weeks at the one-sided  = 0.025 level. 
 
The same model will be used to test the effect of WOMAC C function change from Baseline of 
Ampion vs. saline at 12 weeks.  WOMAC C will be tested at one sided  = 0.025.  Because both 
endpoints have to achieve statistical significance, this pairing ensures the one-sided Type I error 
control at the 0.025 level. 
10.3.5 
Primary Hypotheses 
The trial has co-primary hypotheses – requiring success on both for a successful trial.  First, pain 
reduction must be demonstrated for Ampion vs. saline control.  Next a function improvement 
must be shown on Ampion vs. saline control. 
 
Reduction in Pain, as measured by a change in the WOMAC A pain sub score as measured by 
the 5-point Likert scale between Baseline and Week 12, will be greater in patients treated with 
Ampion than with saline: 
 
• Null Hypothesis: 
 
μA = μS 
• Alternate Hypothesis: 
μA < μS 
 
Where μA = mean pain reduction in Ampion arm; μS = mean pain reduction in saline arm. 
Lower mean change scores indicate greater pain reduction. 
 
Improvement in function, as measured by a reduction in the WOMAC C function sub score as 
measured by the 5-point Likert scale between Baseline and Week 12, will be greater in patients 
treated with Ampion than with saline: 
 
• Null Hypothesis: 
 
μA = μS  
• Alternate Hypothesis: 
μA < μS  
 
Where μA = mean function reduction in Ampion arm; μS = mean function reduction in saline 
arm.  


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10.3.6 
Definition of Study Visits 
This clinical trial has a total of 11 study visits, including telephone contacts, during the  
24-week study (see Table 6.1). The time-on-study for each patient observation will be defined 
relative to Day 0, the day of the initial dose. For analysis, the Baseline measure is the latest 
measure prior to initiation of treatment. For analysis of Baseline WOMAC A pain, the Baseline 
pain score is the average of all scores measured during days -6 to 0, prior to injection of 
treatment. For analysis of Week 12 WOMAC A pain, the Week 12 pain score is the average of 
all scores measured during the seven days prior to and including the Week 12 office visit. 
10.3.7 
Number of patients to receive study drug 
This study is designed to enroll 1,034 patients, 517 per study arm, randomized 1:1 for Ampion 
and saline. There will be an interim analysis at 724 patients, 362 per study arm, randomized 1:1, 
Ampion and saline.  
Following the interim analysis, the trial will either a) continue up to a total of 1,034 patients, 517 
patients per study arm, randomized 1:1 for Ampion and saline IA injection; b) allow for sample 
size re-estimation up to 1.5 times the original trial size for a total of 1,552 patients, 776 per study 
arm, randomized 1:1 for Ampion and saline IA injection. 
10.3.8 
Disposition of patients 
Disposition of patients, including study completion status and response to therapy as measured 
by WOMAC pain sub scores, will be summarized by treatment group (Ampion and saline), age 
group, race and gender for each of the analysis populations. 
10.3.9 
Interim analysis 
There will be one interim analysis once 724 subjects have reached the primary endpoint 
assessment at Week 12. 
At the interim analysis, the conditional probability based upon the observed treatment WOMAC 
C function score will be calculated.  The minimal conditional probability that allows a sample 
size increase without inflating Type I error under the promising zone for a sample size increase 
up to 150% is 0.391.  Any conditional power < 0.391 results in going to 1,034 patients, 
randomized 1:1 for Ampion and saline IA injection.  Sample size re-estimation according to the 
promising zone is done for conditional powers between 0.391 and 0.90 with a sample size 
increase capped to a maximum of 1,552 total patients, randomized 1:1 for Ampion and saline IA 
injection. A conditional power > 90% results in enrolling to 1,034 patients. 
The final analysis, defined by a sample size established using the promising zone, will use the 
critical value with one-sided  = 0.025 for WOMAC A (Ampion vs. saline control) and 
WOMAC C (Ampion vs. saline control).  


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10.3.10 Blinding and randomization 
Patients will be assigned to treatment by a randomization schedule generated by an independent 
statistician. Ampion and saline will be provided in study vials labeled with the appropriate 
information and packed into patient kits.  
Where required, safety personnel may be unblinded to a particular patient’s treatment assignment 
to meet reporting requirements to Regulators. Unblinding may occur by contacting Ampio 
Pharmaceuticals, Inc. 
A final Statistical Analysis Plan will be issued prior to unblinding at the interim analysis of data 
to adjust for any changes to the protocol or unexpected issues in study conduct and data that 
affect the planned analyses. 
10.3.11 Data presentation 
10.3.11.1 
Demographic and Baseline Characteristics 
Demographic (e.g., age, sex, race, ethnicity) and Baseline characteristics (e.g., weight, height) 
summarized using descriptive statistics, overall and by treatment group for the ITT analysis 
population. 
10.3.11.2 
Medical History and Physical Examination 
The number and percent of patients with past and current medical disorders at the time of 
randomization will be presented overall and by treatment group for the ITT analysis population. 
Results of any abnormalities documented from the abbreviated physical examination at Baseline 
and Week 12 and Week 24, will be summarized overall and by treatment group for the safety and 
ITT analysis populations. 
10.3.11.3 
Concomitant Medications or Treatments 
The number and percent of patients receiving concomitant medications or treatments prior to, 
during the study and at the final visit will be tabulated and presented overall and by treatment 
group for the ITT analysis population. Concomitant medications/treatments will be summarized 
using descriptive statistics and will be presented by type of drug (WHO drug classification) 
overall and by treatment group for the safety and ITT analysis populations. 
Treatment failures are defined as patients who receive additional medications or surgical 
procedures in order to treat worsened symptoms in the treated knee during the study; 
specifically, treatment failures are any patients who:  
• Increase their corticosteroid dose of ≤ 10 mg above their starting dose of corticosteroid 
during the study to treat worsened symptoms in the treated knee;  
• Increase use of NSAID pain medications by a clinically significant amount during the 
study to treat worsened symptoms in the treated knee; 
• Begin opioid treatment during the study to treat worsened symptoms in the treated knee; 
• Receive intra-articular injection of pain medicine in the treated knee during the study to 
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• Undergo total knee replacement or other surgical procedure in the treated knee prior to 
assessment of the primary efficacy endpoint. 
 
10.3.11.4 
Safety data 
Safety data will be evaluated by changes in vital sign measurements and the frequency and 
severity of AEs. Concomitant medication will be recorded for safety.  
Adverse events: 
The Investigator is responsible for monitoring the safety of patients who have enrolled in the 
study. All AEs considered to be possibly related to Ampion will be followed until the event 
resolves or stabilized without further change. Patients will be followed for the occurrence of AEs 
until 12 weeks after the first dose of study medication. 
Investigators are required to document all AEs occurring during the clinical trial, commencing 
with the first day of treatment and including the protocol-defined post-treatment follow-up 
period on the appropriate CRF pages. 
The severity of AEs (mild, moderate, severe), relatedness (related, possibly related, unrelated) 
along with the duration, action taken, and outcome (e.g., study withdrawal) will also be recorded. 
In addition, events meeting the criteria of a SAE) must be reported to the Sponsor within 24 
hours on the SAE reporting forms. 
10.3.11.5 
Efficacy data:  
All efficacy variables will be assessed at Baseline (Day 0), Week 2 (Day 14 ± 7), Week 4 
(Day 30 ± 7), Week 6 (Day 42 ± 7), Week 8 (Day 56 ± 7), Week 10 (Day 70 ± 7) and Week 12 
(Day 84 ± 7). For analysis of baseline WOMAC A pain, the baseline pain score is the average of 
all scores measured during days -7 to 0, prior to injection of treatment. For analysis of week 12 
WOMAC A pain, the week 12 pain score is the average of all scores measured during the seven 
days prior to and including the Week 12 office visit. 
 
All statistical tests will be two-sided, except the co-primary endpoints which require superiority 
in pain vs. saline control in pain and superiority to baseline in function. 
 
Unless otherwise specified, continuous variables will be summarized with the number of non-
missing observations, mean, standard deviation, median, minimum, and maximum displayed. 
Categorical data will be summarized as counts and percentages. All efficacy assessments will be 
summarized as the measured value and as the change from Baseline by treatment at each 
timepoint. Summary statistics will include number of observations, mean, standard deviation, 
median, minimum and maximum. Change from Baseline will also include a 95% confidence 
interval. 
Except where otherwise specified, missing data will not be estimated or carried forward in any of 
the descriptive analyses. No multiple comparison adjustment will be made for the secondary 


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efficacy analyses. Data transformation or use of rank-based tests may be used if endpoints depart 
substantially from a normal distribution. 
Baseline is defined as the last pre-treatment assessment.  
10.4 STUDY ENDPOINTS 
10.4.1 
Co-Primary Endpoints 
1) Change in the WOMAC Osteoarthritis Index 3.1 pain sub score as measured by the 5-
point Likert scale between Baseline and Week 12. 
 
The primary variable is the average score of the five WOMAC A (pain) subscale questions: 
In the last 12 hours, how much pain have you had in the study knee: 
1. When walking on a flat surface? 
2. When going up or down stairs? 
3. At night while in bed? (that is - pain that disturbs your sleep) 
4. While sitting or lying down? 
5. While standing? 
 
The primary analysis will include the repeated measures of WOMAC A pain sub score from 
Baseline to Weeks 2, 4, 6, 8, 10, and 12 and will use a mixed-effects model with repeated 
measures MMRM model, with Treatment assignment, Week, and Baseline WOMAC A pain sub 
score as fixed effects, and including patient ID as a repeated measure with a compound 
symmetric covariance matrix for residuals.  
2) Change in the WOMAC Osteoarthritis Index 3.1 function sub score as measured by the 
5-point Likert scale between Baseline and Week 12. 
 
The primary variable is the average score of the seventeen WOMAC C (function) subscale 
questions. 
The primary analysis will include the repeated measures of WOMAC C function sub score from 
Baseline to Weeks 2, 4, 6, 8, 10, and 12 and will use a mixed-effects model with repeated 
measures, with Week as a fixed effect, and including patient ID as a repeated measure with a 
compound symmetric covariance matrix for residuals.  
10.4.2 
Secondary Endpoints 
• Change in PGA between Baseline and Week 12  
• Change in the 100 mm VAS Pain Scale between Baseline and Week 12 
• Use of rescue analgesia (amount of acetaminophen used between Baseline and Week 12) 
• Incidence and severity of treatment-emergent adverse events (TEAEs) 
 


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A serial gatekeeping approach will be used to test the secondary endpoints in a sequential 
manner (i.e. PGA, 100 mm VAS, rescue analgesia).  
The difference between treatment groups in the mean change in PGA from Baseline to Week 12 
will be analyzed using a similar MMRM model, with Treatment assignment, Week, and Baseline 
value as fixed effects and including patient as a repeated measure with a compound symmetric 
covariance matrix for residuals. 
The difference between treatment groups in the mean change in 100 mm VAS from Baseline to 
Week 12 will be analyzed using a similar MMRM model, with Treatment assignment, and 
Baseline value as fixed effects and including patient as a repeated measure with a compound 
symmetric covariance matrix for residuals. 
The difference between treatment groups in the amount of acetaminophen used between Baseline 
and Week 12 will be analyzed with a Wilcoxon rank-sum test.  
All secondary efficacy analyses will be reported with one sided p-values and successively 
compared to a critical level of 0.025 if each preceding test was significant. 
10.4.3 
Exploratory Endpoints 
• Change in WOMAC B stiffness sub score between Baseline and Weeks 2, 4, 6, 8, 10, 12 
and 24 
• Response status based on the OMERACT-OARSI criteria at Weeks 2, 4, 6, 8, 10, 12 and 
24  
• Proportion of treatment failures at Week 12 
• Change in pain and function CDF plot for Ampion and saline patients 
• A Bayesian meta-analysis for LK WOMAC A pain sub score including KL 4 patients 
from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013 
• Change in WOMAC A pain sub score between Baseline and Weeks 2, 4, 6, 8, 10, and 24  
• Change in PGA between Baseline and Weeks 2, 4, 6, 8, 10, and 24 
• Change in WOMAC C physical function sub score between Baseline and Weeks 2, 4, 6, 
8, 10, and 24 
• If collected, difference in urine analysis between treatment groups at Week 12  
The Bayesian meta-analysis methodology is described in the Statistical Analysis Plan. 
 
Percent responders using the OMERACT-OARSI criteria will be analyzed using separate logistic 
regression models for each visit. 
 
Proportion of treatment failures will be analyzed using Fisher’s exact tests. 
 
Analysis with exploratory biomarkers, such as those for analgesic use and chemokines and 
cytokines associated with inflammation and cartilage regeneration may be analyzed to compare 
differences between treatment groups using urine samples collected at Week 12. 
 


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All other exploratory endpoints will be analyzed as the difference between treatment groups at 
each time point extracted from the MMRM model, adjusted for Baseline value.  
10.4.4 
Subset Analyses 
The two co-primary endpoints and three efficacy secondary endpoints will be re-analyzed 
separately for (a) ages <60, [60, 70), and ≥70, (b) men and women, (c) white and other, (d) Non-
Hispanic and Hispanic. 
10.4.5 
Sensitivity Analysis 
The two co-primary endpoints and three efficacy secondary endpoints will be re-analyzed 
separately in the per-protocol population to exclude treatment failures.  
10.5 Missing and Spurious Data 
All data collected under this study protocol will be included in the assessment of patient safety. 
Missing or incomplete AE data will assume greatest relationship to study drug and/or severity. 
For the co-primary effectiveness analysis of WOMAC pain change and WOMAC C function 
change, missing change scores will be imputed when the ITT analysis population is used. 
Missing data will be imputed by the fully conditional specification predictive mean matching 
multiple imputation approach of Berglund and Heeringa (2014). A sensitivity analysis will be 
conducted in which the missing 12-week endpoint is replaced by the Baseline WOMAC pain 
score. 
Per the WOMAC User Guide, the patient’s response will be regarded as invalid, and the 
subscale(s) deficient, in the event that two pain, both stiffness, or more than four physical 
function items are omitted. Further, if one pain, one stiffness, or 1-3 physical function units are 
missing, the average value for the subscale will be substituted in lieu of the missing item 
value(s).  
 


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11 REGULATORY, ETHICAL AND LEGAL OBLIGATIONS 
11.1 Declaration of Helsinki 
The Principal Investigator will ensure that this Study is conducted in accordance with the most 
recent revision of the Declaration of Helsinki. 
11.2 Good Clinical Practice 
The Study will be conducted according to the study protocol and to Standard Operating 
Procedures (SOPs) that meet the guidelines provided by the International Conference on 
Harmonisation (ICH) for Good Clinical Practice in clinical studies. 
11.3 Institutional Review Boards/Ethics Committees 
Before implementing this study, the protocol, the proposed patient informed consent forms and 
other information for the patients, must be reviewed by a properly constituted committee or 
committees responsible for approving clinical studies. The IRB/IEC written, signed approval 
letter/form must contain approval of the designated investigator, the protocol (identifying 
protocol title, date and version number), and of the patient informed consent form (date, version). 
Any change or addition to the protocol can only be made in a written protocol amendment that 
must be approved by the Sponsor, the IRB/IEC and the Health Authorities. 
11.4 Regulatory Authority Approval 
Before this study is implemented, the protocol must be approved by the relevant regulatory 
authority. 
11.5 Informed Consent 
The investigator must fully inform the patient of all pertinent aspects of the trial including the 
written information approved/favorably assessed by the IRB/IEC. 
Prior to the start of the pre-study examination, the written informed consent form must be signed 
and personally dated by the patient and by the physician who conducted the informed consent 
discussion. One copy of the written information and signed consent form must be given to each 
patient and one copy must be retained in the investigator's study records. 
11.6 Patient Confidentiality and Disclosure 
Data on patients collected on eCRFs during the trial will be documented in an anonymous 
fashion and the patient will only be identified by the patient number, and by his/her initials. As 
an exception, if it is necessary for safety or regulatory reasons to identify the patient, all parties 
are bound to keep this information confidential. 


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The investigator will guarantee that all persons involved will respect the confidentiality of any 
information concerning the trial patients. All parties involved in the study will maintain strict 
confidentiality to assure that neither the person nor the family privacy of a patient participating 
in the trial is violated. Likewise, the appropriate measures shall be taken to prevent access of 
non-authorized persons to the trial data. 
11.7 Collection, Monitoring and Auditing Study Documentation, and Data 
Storage 
11.7.1 
Collection of Data and Monitoring Procedures 
This study will use a 21 CFR Part 11 compliant electronic data capture system (eDC). An 
electronic case report form (eCRF) is used for data recording. All data requested on the eCRF 
must be entered and all missing data must be accounted for. 
The data will be checked for completeness and correctness as it is entered by the real-time online 
checks applied by the eDC system. Off-line checks will also be run to perform any additional 
data review required. Discrepancy reports will be generated accordingly and transferred to the 
study center for resolution by the investigator or his/her designee. 
Accurate and reliable data collection will be assured by verification and cross–check of the eCRF 
against the investigator’s records by the study monitor (source document verification), and the 
maintenance of a study drug–dispensing log by the investigator. 
Before study initiation, at a site initiation visit or at an investigator’s meeting, a Sponsor 
representative will review the protocol and case report forms with the investigators and their 
staff. During the study a monitor will visit the site regularly to check the completeness of patient 
records, the accuracy of entries on the case report forms, the adherence to the protocol and to 
Good Clinical Practice, the progress of enrollment. The monitor will ensure during on-site visits 
that study medication is being stored, dispensed and accounted for according to specifications. 
Key trial personnel must be available to assist the monitors during these visits. 
The investigator must give the monitor access to relevant hospital or clinical records, to confirm 
their consistency with the case report form entries. No information in these records about the 
identity of the patients will leave the study center. Monitoring standards require full verification 
for the presence of informed consent, adherence to the inclusion/exclusion criteria, 
documentation of SAEs and the recording of primary efficacy and safety variables.  Additional 
checks of the consistency of the source data with the eCRFs are performed according to the 
study-specific monitoring plan. 
 
 


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11.7.2 
Auditing Procedure 
In addition to the routine monitoring procedures the Sponsor or the regulatory authority can 
conduct an audit or an inspection (during the study or after its completion) to evaluate 
compliance with the protocol and the principles of Good Clinical Practice. 
The investigator agrees that representatives of the Sponsor and Regulatory Authorities will have 
direct access, both during and after the course of this study, to audit and review all study-relevant 
medical records. 
11.7.3 
Retention of Documents 
The investigator must maintain source documents for each patient in the study, consisting of all 
demographic and medical information, including laboratory data, x-ray, etc., and keep a copy of 
the signed informed consent form. All information on case report forms must be traceable to 
these source documents in the patient's file. Data without a written or electronic record will be 
defined before trial start and will be recorded directly on the case report forms, which will be 
documented as being the source data. 
11.8 Disclosure of Information 
All information provided to the investigator by Ampio Pharmaceuticals, Inc. or their designee, 
will be kept strictly confidential. No disclosure shall be made except in accordance with a right 
of publication granted to the investigator. 
No information about this study or its progress will be provided to anyone not involved in the 
study other than to Ampio Pharmaceuticals, Inc., or its authorized representatives, or in 
confidence to the IRB, or similar committee, except if required by law. 
11.9 Discontinuation of the Study 
It is agreed that, for reasonable cause, either the investigator or Ampio Pharmaceuticals, Inc., 
may terminate the investigator’s participation in this study after submission of a written notice. 
Ampio Pharmaceuticals, Inc., may terminate the study at any time upon immediate notice for any 
reason, including the Sponsor’s belief that discontinuation of the study is necessary for the safety 
of patients. 
11.10 Study Report, Publication Policy and Archiving of Study 
Documentation 
11.10.1 Study Report and Publication Policy 
An ICH-compliant integrated clinical and statistical report will be prepared upon completion of 
the study and data analysis. The results of the study will be published in a relevant peer-reviewed 
journal, with authorship status and ranking designated according to the acknowledged 
contributions of participating investigators, institutions and the Sponsor. 


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11.10.2 Study Documents 
The investigator must maintain source documents for each patient in the study, consisting of all 
demographic and medical information, questionnaires, including laboratory data, MRI (as 
applicable), etc., and keep a copy of the signed informed consent form. All information on the e-
case report forms must be traceable to these source documents in the patient's file. Data without a 
written or electronic record will be defined before trial start and will be recorded directly on the 
e-case report forms, which will be documented as being the source data. 
11.10.3 Archiving of Documents 
Essential documents, as listed below, must be retained by the investigator for as long as needed 
to comply with national and international regulations. The Sponsor will notify the 
investigator(s)/institution(s) when the study-related records are no longer required. The 
investigator agrees to adhere to the document retention procedures by signing the protocol.  
Essential documents include: 
• IRB/IEC/REB approvals for the study protocol and all amendments 
• All source documents and laboratory records 
• CRF copies (electronic copies on a CD-ROM) 
• Patients' informed consent forms (with study number and title of trial) 
• FDA form 1572 
• Any other pertinent study document 
 


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12 
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articular injections of hyaluronic acid in knee osteoarthritis: the AMELIA project. Ann 
Rheum Dis. 2011;70:1957-62. 
19. Peterfy C.G., M.D. Ph.D., Guermazi A M.D., S. Zaim M.D., P. F. J. Tirman M.D., Y. Miaux 
M.D., D. White Ph.D., M. Kothari Ph.D., Y. Lu Ph.D., K. Fye M.D., S. Zhao Ph.D. H. K. 
Genant M.D. Whole-Organ Magnetic Resonance Imaging Score (WORMS) of the knee. 
OsteoArthritis and Cartilage (2004) 12, 177–190. 
20. Quinlan GJ, Martin GS, Evans TW. Albumin: biochemical properties and therapeutic 
potential. Hepatology. 2005;41:1211-1219. 
21. Rutjes AW et al, Vicosupplementation for osteoarthritis of the knee: a systematic review and 
meta-analysis Annals of Internal Medicine, 2012; 157(3):280-91 
22. Salottolo K, Stahl E. Minimal clinically important improvement response in patients with 
severe osteoarthritis of the knee: Short report from a survey of clinicians. Journal of 
orthopaedics. 2018 Jun 1;15(2):424-5. 
23. Shimonkevitz R, Thomas GW, Slone DS, et al. A diketopiperazine fragment of human serum 
albumin modulates T-lymphocyte cytokine production through Rap1. J Trauma. 
2008;64:35-41. 
24. Stannus et al. Annals of Rheumatic Disease, Vol. 72, No. 4. 2013 
25. Thomas et al. Anti-inflammatory activity in the low molecular weight fraction of commercial 
human serum albumin (LMWF5A) Journal of Immunoassay and Immunochemistry, 
2016;37:55-67 
26. Thomas et al. The low molecular weight fraction of human serum albumin upregulates 
production of 15d-PGJ2 in Peripheral Blood Mononuclear Cells Biochemical and Biophysical 
Research Communications. 2016b;473(4):1328-1333 


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27. Tubach F, Ravaud P, Baron G, Falissard B, Logeart I, Bellamy N, Bombardier C, Felson D, 
Hochberg M, van der Heijde D, Dougados M. Evaluation of clinically relevant changes in 
patient reported outcomes in knee and hip osteoarthritis: the minimal clinically important 
improvement. Annals of the rheumatic diseases. 2005 Jan 1;64(1):29-33. 
28. Turk DC, Dworkin RH. The initaitive on methods, measurement, and pain assessment in 
clinical trials' IMMPACT'. Pain management: Evidence, outcomes and quality of life. A 
source book. 2nd ed. Edinburgh: Elsevier. 2008:287-304. 
29. Vincent JL, Navickis RJ, Wilkes MM. Morbidity in hospitalized patients receiving human 
albumin: A meta-analysis of randomized, controlled trials* Crit Care Med. 2004;32:2029–
38. 
30. Zhang W, Nuki G, Moskowitz R.W. et al. OARSI recommendations for the management of 
hip and knee osteoarthritis Part III: changes in evidence following systematic cumulative 
update of research published through January 2009. Osteoarthritis and Cartilage. 
2010;18:476-499. 


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13 
CONTACT LIST 
SPONSOR CONTACT: 
Ampio Pharmaceuticals, Inc. 
720-437-6500 
373 Inverness Pkwy #200 
Englewood CO 80112 
 
 

SAP_001.pdf: 
 
 
 
 
A RANDOMIZED, CONTROLLED, DOUBLE‐
BLIND STUDY TO EVALUATE THE EFFICACY 
AND SAFETY OF AN INTRA‐ARTICULAR 
INJECTION OF AMPION™ IN ADULTS WITH 
PAIN DUE TO SEVERE OSTEOARTHRITIS OF 
THE KNEE 
 
STATISTICAL ANALYSIS PLAN 
 
STUDY NUMBER: AP‐013 
 
NCT 03988023 
 
7 JUNE 2019 




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STATISTICAL ANALYSIS PLAN 
A RANDOMIZED, CONTROLLED, DOUBLE-BLIND STUDY TO EVALUATE THE 
EFFICACY AND SAFETY OF AN INTRA-ARTICULAR INJECTION OF 
AMPION™ IN ADULTS WITH PAIN DUE TO SEVERE OSTEOARTHRITIS 
OF THE KNEE  
Protocol Number AP-013 
Version: 
 
1.0 
 
Date: 
7 June 2019 
 
Biostatistician: 
Gary Stevens, PhD  
 
 


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Table of Contents 
1. 
INTRODUCTION ......................................................................................................5 
2. 
OBJECTIVES .............................................................................................................5 
2.1 Primary Objectives .............................................................................................5 
2.2 Secondary and Exploratory ................................................................................5 
2.3 Study Design ......................................................................................................6 
2.4 Sample Size ........................................................................................................6 
3. 
STUDY ENDPOINTS AND COVARIATES ............................................................7 
3.1 Primary Endpoints .............................................................................................7 
3.2 Secondary Endpoints .........................................................................................7 
3.3 Exploratory Endpoints .......................................................................................7 
3.4 Covariates ..........................................................................................................8 
3.5 Subset analyses ..................................................................................................8 
3.6 Sensitivity analyses ............................................................................................8 
4. 
HYPOTHESES ...........................................................................................................8 
4.1 Primary Hypotheses ...........................................................................................8 
5. 
DEFINITIONS ............................................................................................................9 
5.1 Abbreviations .....................................................................................................9 
5.2 Definitions..........................................................................................................9 
6. 
ANALYSIS SUBSETS .............................................................................................12 
6.1 Data Subsets .....................................................................................................12 
6.1.1 
Safety Analysis Set .............................................................................12 
6.1.2 
Intent to Treat (ITT) Analysis Set ......................................................12 
6.1.3 
Per Protocol (PP) Analysis Set ...........................................................12 
7. 
INTERIM ANALYSIS AND EARLY STOPPING GUIDELINES ........................12 
7.1 Data Handling and Electronic Transfer of Data ..............................................13 
7.2 Handling of Missing and Incomplete Data ......................................................13 
7.3 Detection of Bias..............................................................................................14 
7.4 Outliers .............................................................................................................14 
7.5 Testing/Validation Plan ...................................................................................14 
8. 
STATISTICAL METHODS OF ANALYSIS ..........................................................14 
8.1 General Principles ............................................................................................14 
8.2 Patient Accountability ......................................................................................14 
8.3 Demographic and Baseline Characteristics .....................................................15 
8.4 Safety Analyses ................................................................................................15 


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8.5 Adverse Events ................................................................................................15 
8.5.1 
Concomitant Medications ...................................................................15 
8.5.2 
Vital Signs ..........................................................................................15 
8.5.3 
Physical Examinations........................................................................15 
8.6 Efficacy Analyses ............................................................................................15 
8.6.1 
Primary Effectiveness Endpoints .......................................................16 
8.6.2 
Secondary Effectiveness Endpoints ...................................................17 
8.6.3 
Subset analyses ...................................................................................17 
8.6.4 
Sensitivity analyses ............................................................................17 
8.6.5 
Exploratory Endpoints ........................................................................17 
8.6.6 
Post Hoc/Ad Hoc Analyses ................................................................18 
8.6.7 
Multiplicity Adjustment .....................................................................18 
9. 
CHANGES FROM PROTOCOL-SPECIFIED ANALYSES ..................................19 
10. LIST OF PLANNED TABLES, FIGURES, AND LISTINGS ................................19 
10.1 Tables ...............................................................................................................19 
10.2 Listings .............................................................................................................21 
10.3 Figures..............................................................................................................22 
11. LITERATURE CITATIONS  REFERENCES........................................................22 
12. HANDLING OF MISSING OR INCOMPLETE DATES FOR ADVERSE 
EVENTS AND CONCOMITANT MEDICATIONS ..............................................22 
12.1 Imputation Rules for Partial or Missing Stop Dates ........................................22 
13. Schedule of Assessments and Procedures .................................................................24 


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1. 
INTRODUCTION 
This statistical analysis plan (SAP) outlines the proposed statistical methods to be 
implemented during the review of data to ensure that it confirms with categories 
determined by the CRF or the anticipated ranges for continuous variables and analysis of 
data collected within the scope of Ampion Protocol AP-013, “A Randomized, Controlled, 
Double-Blind Study To Evaluate The Efficacy And Safety Of An Intra-Articular Injection 
Of Ampion™ In Adults With Pain Due To Severe Osteoarthritis Of The Knee,” dated 7 
June 2019. 
It is not intended that each and every table, listing, or graph will be included in the clinical 
study report (CSR). It is also possible that additional analyses will be conducted after 
review of the data. Any analyses or summaries not specified in the SAP, but performed 
after review of the data, will be identified in the CSR as post hoc. 
2. 
OBJECTIVES 
2.1 Primary Objectives 
The co-primary trial objectives are to evaluate the greater efficacy for pain improvement 
and functional improvement of 4 mL Ampion versus saline IA injection when applied to 
patients suffering from severe OAK. Efficacy will primarily be assessed with WOMAC A 
pain and WOMAC C function scores. 
Mean change in the WOMAC A weekly pain scores from Baseline to Week 12 will be 
compared between Ampion and saline control.  Mean change in WOMAC C function score 
from Baseline to Week 12 will be compared between Ampion and saline control. This will 
ensure that Ampion is superior to saline in improving pain and function. 
2.2 Secondary and Exploratory  
The secondary trial objectives include the following evaluations of an intra-articular 
injection of Ampion, as compared to saline, when administered to patients suffering from 
severe OAK: assessment of safety; improvement in the Patient Global Assessment; 
improvement in pain utilizing a 100 mm VAS scale; the amount of rescue analgesia.  
Primary and secondary objectives will be reported by age subsets, race, and sex. 
Exploratory objectives include analyzing the effect, if any, of the efficacy of Ampion, as 
compared to saline, on stiffness; Outcome Measures in Rheumatology Clinical Trials and 
Osteoarthritis Research Society International (OMERACT-OARSI) responder status; 
proportion of treatment failures; and change in pain and function as a cumulative 
distribution function (CDF) plot. 
Analysis with biomarkers, such as those for analgesic use and chemokines and cytokines 
associated with inflammation and cartilage regeneration, may be analyzed to compare 
differences between treatment groups using urine samples collected at Week 12.  
Additionally, a Bayesian meta-analysis will be performed for WOMAC A pain sub score 
using all KL 4 patients in studies AP-003A, AP-003-B, AP-003-C, AP-004, and AP-013.  


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2.3 Study Design 
This is a randomized, saline-controlled, double-blind study with a 28-day screening period 
for each patient followed by a 24-week participation period. This trial will use an adaptive 
design with promising zone method for sample size re-estimation at the interim analysis.  
The clinical effects of treatment on severe OAK will be evaluated at baseline 2, 4, 6, 8, 10, 
12, and 24 weeks, using the Western Ontario and McMaster Universities Arthritis Index 
(WOMAC®) osteoarthritis Index 3.1, and the Patient’s Global Assessment of disease 
severity (PGA).  
Daily assessments of WOMAC A score using a subject diary will be assessed for seven 
days beginning one week prior to the clinic visits at Week 0 (baseline) and Week 12 to 
allow for calculation of a weekly average of WOMAC A Daily Pain scores, which will be 
used for conducting the primary efficacy analysis. 
The WOMAC® is a validated pain scoring system and sets the standard for the patient 
response. In order not to bias the collection of data, only questions from the validated 
WOMAC pain scale will be asked of patients. Clinical meaningfulness will be determined 
by the end results of this trial, specifically by the apparent clinical benefit versus any 
adverse events or any increased apparent risk. Safety will be assessed by recording adverse 
events (through 24 hours post-dose and at all follow up contacts) and physical examination 
and vitals (Baseline, Weeks 6, 12 and 24). 
2.4 Sample Size  
This trial will use an adaptive design with promising zone method for sample size re-
estimation. 
This trial is designed for 1,034 patients, 517 per study arm, randomized 1:1 for Ampion 
and saline IA injection. There will be an interim analysis when at 724 patients, 362 per 
study arm, have completed their Week 12 visit. 
A promising zone analysis will be performed at the interim.  Following the interim analysis, 
enrollment will either a) continue up to a total of 1034 patients, 517 patients per study arm, 
randomized 1:1 for Ampion and saline IA injection; b) allow for sample size re-estimation 
up to a maximum of 1.5 times the original trial size for a total of 1,552 patients, 776 per 
study arm, randomized 1:1 for Ampion and saline IA injection.  
The Ampion effect was estimated as the average change in pain from Baseline to Week 12 
for KL 4 subjects receiving Ampion from all prior single injection studies randomized 1:1 
with saline control. Saline effect was estimated as the average change in pain from Baseline 
to Week 12 for KL 4 subjects receiving saline from all prior single injection studies 
randomized 1:1 for Ampion and saline.  The sample size for assessing the WOMAC A 
Pain score alone would be 690 total patients for 90% power.   
The Ampion effect was estimated as the average change in function from Baseline to Week 
12 for KL 4 subjects receiving Ampion from all prior single injection studies randomized 
1:1 with saline control. Saline effect was estimated as the average change in function from 
Baseline to Week 12 for KL 4 subjects receiving saline from all prior single injection 


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studies randomized 1:1 for Ampion and saline.  The sample size for assessing the WOMAC 
C Function score alone would be 984 patients for 90% power.   
The total sample size is chosen as the larger of the two values. To be conservative sample 
sizes were estimated with a 5% dropout so 1,034 patients will be enrolled to achieve 984 
total patients.  All ITT patients will be included in the statistical analysis. At the assumed 
effect size there is a 90% probability of success at 1,034 patients. 
The study design of 1,034 total patients achieves 97% power assuming a WOMAC A pain 
score with mean pain decreases of -0.83 for Ampion and -0.62 for saline and a standard 
deviation of 0.85. 
3. 
STUDY ENDPOINTS AND COVARIATES 
3.1 Primary Endpoints  
The trial will test whether Ampion can produce a superior improvement in pain vs. saline 
control and a superior improvement in function vs. saline. This goal is achieved using two 
co-primary endpoints: 
Change in the WOMAC A weekly pain sub score between Baseline and Week 12  
Change in WOMAC C physical function sub score between Baseline and Week 12 
An average of the daily WOMAC A pain assessments will be used to calculate the weekly 
pain score, at baseline and at Week 12. 
Mean 12-week change in the WOMAC A pain score will be compared between Ampion 
and saline control. Mean 12-week change in WOMAC C physical function score will be 
compared between Ampion and saline control.  
3.2 Secondary Endpoints  
 Change in PGA between Baseline and Week 12  
 Change in 100mm VAS Pain scale between Baseline and Week 12 
 Use of rescue analgesia (amount of acetaminophen used between Baseline and 
Week 12) 
 Incidence and severity of treatment-emergent adverse events (TEAEs) 
 
3.3 Exploratory Endpoints  
 Change in WOMAC B stiffness sub score between Baseline and Weeks 2, 4, 6, 8, 
10, 12, and 24 
 Response status based on the OMERACT-OARSI criteria at Weeks 2, 4, 6, 8, 10, 
12, and 24 
 Proportion of treatment failures at Week 12 
 Change in pain and function analysis as a cumulative distribution function (CDF) 
plot 
 A Bayesian meta-analysis for WOMAC A pain sub score including KL 4 patients 
from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013 


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 Change in WOMAC A pain sub score between Baseline and Weeks 2, 4, 6, 8, 10, 
and 24  
 Change in WOMAC C function sub score between Baseline and Weeks 2, 4, 6, 8, 
10, and 24 
 Change in WOMAC A pain sub score between Baseline and Weeks 2 through 12 
and Weeks 2 through 24  
 Change in WOMAC C function sub score between Baseline and Weeks 2 through 
12 and Weeks 2 through 24  
 Change in PGA between Baseline and Weeks 2, 4, 6, 8, 10, and 24 
 Change in WOMAC C physical function sub score between Baseline and Weeks 2, 
4, 6, 8, 10, 12, and 24 
 Difference in urine analysis between treatment groups at Week 12 
 
3.4 Covariates 
Analyses of change will include treatment assignment, Week, and the Baseline measure of 
the variable being analyzed as a covariate in the model.  
3.5 Subset analyses 
The two co-primary endpoints and three efficacy secondary endpoints will be re-analyzed 
separately for (a) ages <60, [60, 70), and ൒70, (b) men and women, (c) white and other, 
(d) Non-Hispanic and Hispanic. 
3.6 Sensitivity analyses 
The two co-primary endpoints and three efficacy secondary endpoints will be re-analyzed 
separately in the per-protocol population to exclude treatment failures.  
4. 
HYPOTHESES 
4.1 Primary Hypotheses 
The trial has co-primary hypotheses – requiring success on both for a successful trial.  First, 
pain reduction must be demonstrated for Ampion vs. saline control.  Next a function 
improvement must be shown on Ampion vs. saline control. 
Reduction in Pain, as measured by a change in the WOMAC A pain subscore as measured 
by the 5-point Likert scale between Baseline and Week 12, will be greater in patients 
treated with Ampion than with saline: 
Null Hypothesis: 
 
 
μA = μS 
Alternate Hypothesis: 
 
μA < μS 
 
Where μA = mean pain reduction in Ampion arm; μS = mean pain reduction in saline arm. 
Lower mean change scores indicate greater pain reduction. 
Improvement in function, as measured by a reduction in the WOMAC C function sub score 
as measured by the 5-point Likert scale between Baseline and Week 12, will be greater in 
patients treated with Ampion than with saline: 


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Null Hypothesis: 
 
 
μA = μS  
Alternate Hypothesis: 
 
μA < μS  
Where μA = mean function reduction in Ampion arm; μS = mean function reduction in 
saline arm. Lower mean change scores indicate greater improvement in function. 
5. 
DEFINITIONS 
5.1 Abbreviations 
Abbreviation  
Definition 
ATC 
Anatomical Therapeutic Chemical Classification System 
ITT 
Intent to Treat Population 
MedDRA® 
Medical Dictionary for Regulatory Activities 
mL 
Milliliter 
mmHg 
Millimeters of mercury 
MMRM 
Mixed-effects model with repeated measures 
NA 
Not applicable 
OA 
Osteoarthritis 
OARSI 
Osteoarthritis Research Society International 
OMERACT 
Outcome Measures in Rheumatology 
PGA 
Patient’s global assessment of disease severity 
PP 
Per protocol population 
SAE 
Serious adverse event 
SAS 
Statistical Software from SAS Institute 
SD 
Standard deviation 
SEM 
Standard error of the mean 
TEAE 
Treatment-emergent adverse event 
WOMAC® 
Western Ontario and McMaster Universities Arthritis Index 
 
5.2 Definitions 
Adverse Event (AE) 
An adverse event (AE) is defined as any undesired medical occurrence in a patient or 
clinical investigation patient receiving a pharmaceutical product and which does not 
necessarily have a causal relationship with this treatment. Therefore, an AE can be any 
unfavorable sign and unintended sign (including an abnormal laboratory finding), 


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symptom, or disease temporarily associated with the use of a study drug, whether or not 
related to the study drug. 
AEs will be graded for severity using the following categories. Missing grade will be 
assigned a grade of 3 (severe) in tabulations: 
Grade 1 (MILD): The symptom is barely noticeable to the study patient and does not 
influence performance or functioning. Concomitant medication is not ordinarily indicated 
for relief of mild AEs. 
Grade 2 (MODERATE): The symptom is of sufficient severity to make the study patient 
uncomfortable and to influence performance of daily activities. Concomitant medication 
may be indicated for relief of moderate AEs. 
Grade 3 (SEVERE): The symptom causes severe discomfort, sometimes of such severity 
that the study patient cannot continue in the study. Daily activities are significantly 
impaired or prevented by the symptom. Concomitant medication may be indicated for relief 
of severe AEs. 
Relationship to study drug will be coded using the following categories. Missing 
relatedness will be assigned to ‘Related’ in tabulations: 
Unrelated: The adverse event is unlikely to have been caused by study drug. 
Possibly related: It is unclear whether the adverse event may have been caused by study 
drug. 
Related: The adverse event is likely to have been caused by study drug.  
Relationship to procedure will be coded using the following categories. Missing 
relatedness will be assigned to ‘Related’ in tabulations: 
Unrelated: The adverse event is unlikely to have been caused by procedure. 
Possibly related: It is unclear whether the adverse event may have been caused by 
procedure. 
Related: The adverse event is likely to have been caused by procedure.  
Treatment-Emergent AE: 
A treatment-emergent AE (TEAE) is any AE that begins or increases in severity after the 
initial dose of study drug. 
Serious Adverse Event: 
 A serious adverse event (SAE) isany untoward medical occurrence that occurs at any dose 
that: 
 Results in death 


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 Is life-threatening (patient is at immediate risk of death from the event as it occurred) 
 Requires in-patient hospitalization (formal admission to a hospital for medical 
reasons) or prolongation of existing hospitalization 
 Results in persistent or significant disability/incapacity 
 Results in a congenital anomaly/birth defect 
Hospitalizations for elective surgery or other medical procedures that are not related to a 
treatment-emergent AE are not considered SAEs. 
Age 
Patient’s age is defined as its integer value in years at enrollment.  
Baseline 
For any variable, unless otherwise defined, Baseline at Day 0, Visit 1. If data are not 
available from Visit 1, then the last measurement recorded prior to Day 0 will be selected 
from the screening period. 
Change from Baseline: 
The arithmetic difference between a post-Baseline value and the Baseline value: 
Change from Baseline = (Post-Baseline Value – Baseline Value) 
Percentage Change from Baseline = [(Post-Baseline Value – Baseline Value) / 
Baseline Value] x 100 
End of Study 
End of study is at Visit 10 (Week 24), unless terminated early. 
Enrollment date 
Enrollment date is the same as the randomization date and is designated Day 0. 
Study drug 
Study drug in this study is 4 mL Ampion injection or 4mL saline. 
Randomization date 
Randomization date is the day the patient is assigned a randomization number on study 
Day 0. 
Study Day 0 
Day 0 is defined as the first day that study drug is administered to the patient.  
Study Day 
Day of treatment: study day = (visit date - date of Study Day 0).  


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Treatment failure 
Treatment failures are defined as patients who receive additional medications or surgical 
procedures in order to treat worsened symptoms in the treated knee during the study; 
specifically, treatment failures are any patients who:  
 Increase their corticosteroid dose of ≤ 10 mg above their starting dose of 
corticosteroid during the study to treat worsened symptoms in the treated knee;  
 Increase use of NSAID pain medications by a clinically significant amount during 
the study to treat worsened symptoms in the treated knee; 
 Begin opioid treatment during the study to treat worsened symptoms in the treated 
knee; 
 Receive intra-articular injection of pain medicine in the treated knee during the 
study to treat worsened symptoms in the treated knee; 
 Undergo total knee replacement or other surgical procedure in the treated knee 
prior to assessment of the primary efficacy endpoint. 
 
6. 
ANALYSIS SUBSETS 
6.1 Data Subsets  
6.1.1 Safety Analysis Set 
The safety analysis population is defined as all patients who are randomized and receive 
study medication (Ampion or saline). Patients will be analyzed as treated. Summaries of 
data will include all collected data. 
6.1.2 Intent to Treat (ITT) Analysis Set 
The intent-to-treat (ITT) analysis population is defined as all patients who are randomized. 
All efficacy analyses will be performed in the ITT population. Patients will be analyzed as 
randomized. 
The ITT analysis set is the primary effectiveness analysis population. For the primary 
effectiveness analysis at Week 12, multiple imputation will be used to assign a value to 
those cases with missing data. The full conditional specification method with predictive 
means matching as described in Berglund & Heeringa (2014) will be used.  This method 
uses all of an individual’s known primary outcome measures at Baseline, 2, 4, 6, 8, 10, 
and/or 12 weeks to impute any missing values. 
6.1.3 Per Protocol (PP) Analysis Set 
The per protocol (PP) analysis population is defined as all patients who met all entry criteria 
and had no major protocol violations. All efficacy analyses will be repeated in the PP 
population. These analyses will be supportive of the ITT analysis. Patients will be analyzed 
as treated. No imputations will be performed. 
7. 
INTERIM ANALYSIS AND EARLY STOPPING GUIDELINES  


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There will be one interim analysis once 724 subjects have reached final assessment at Week 
12.  
At the interim analysis, the conditional probability based upon the observed treatment 
effect on the WOMAC C function score will be calculated.  The minimal conditional 
probability that allows a sample size increase without inflating Type I error under the 
promising zone for a sample size increase up to 150% is 0.391.  Any conditional power < 
0.391 results in going to 1,034 patients, randomized 1:1 for Ampion and saline IA injection.  
Sample size re-estimation according to the promising zone is done for conditional powers 
between 0.391 and 0.90 with a sample size increase capped to 1,552 total patients, 
randomized 1:1 for Ampion and saline IA injection. 
The final analysis, defined by a sample size established using the promising zone, will use 
the critical value with one-sided = 0.025 for WOMAC A (Ampion vs. saline control) and 
WOMAC C (Ampion vs. saline control).  
7.1 Data Handling and Electronic Transfer of Data  
See Data Management Plan (DMP). 
7.2 Handling of Missing and Incomplete Data 
All data collected under this study protocol will be included in the assessment of patient 
safety. Missing or incomplete AE data will assume greatest relationship to study drug 
and/or severity. Incomplete adverse event and concomitant medication dates will be 
imputed as described in Section 12. If imputed dates are used, then they will be identified 
as such in the final study report. If an AE start date is missing or partially missing and no 
additional information is available from the site in order to establish whether the event 
started before or after the dose of study drug, the event will be considered to have started 
after dose. Partially missing dates where the month and year is prior to Day 0 will be 
classified as pre-dose. 
For the co-primary effectiveness analysis of WOMAC pain change and WOMAC C 
function change, missing change scores will be imputed when the ITT analysis population 
is used. Missing data will be imputed by the fully conditional specification predictive mean 
matching multiple imputation approach of Berglund and Heeringa (2014). A sensitivity 
analysis will be conducted in which the missing 12-week endpoint is replaced by the 
Baseline WOMAC pain score. 
For the secondary endpoints of PGA, 100mm VAS, and number of acetaminophen pills 
used, missing values will be imputed when the ITT analysis population is used. Imputation 
methods employed will be multiple imputation (SAS PROC MI and SAS PROC 
MIANALYZE) and single imputation methods (i.e., regression-based estimates, Baseline 
imputation, LOCF, and a worst/best case imputation).  
Per the WOMAC User Guide, the patient’s response will be regarded as invalid, and the 
subscale(s) deficient, in the event that two pain, both stiffness, or more than four physical 
function items are omitted. Further, if one pain, one stiffness, or 1-3 physical function units 


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are missing, the average value for the subscale will be substituted in lieu of the missing 
item value(s).  
7.3 Detection of Bias 
Any breaking of the blind for individual patients prior to formal unblinding will be 
documented in the clinical study report. Data collected after unblinding will be noted.  
7.4 Outliers 
No formal outlier tests are planned. If necessary, values outside the pre-defined range, such 
as a WOMAC score not between zero and four, or a PGA score not between one and five, 
would be queried and excluded prior to database lock. 
7.5 Testing/Validation Plan 
All statistical analyses will be programmed using SAS® software version 9.3, or later. 
Graphic displays may be produced using R, version 3.0.0, or later. Standard macros will 
be used in programming when possible. Testing and validation plans for all programs will 
be developed in accordance with contract research organization guidelines and will include 
independent programming of tables and analyses. 
8. 
STATISTICAL METHODS OF ANALYSIS 
8.1 General Principles 
Data will be summarized by saline and Ampion arms. Descriptive statistics on continuous 
variables will include mean, standard deviation, median, 25th and 75th percentiles, and 
range. Change from Baseline will include a 95% confidence interval. Categorical variables 
will be summarized using frequency counts and percentages. Data listings of individual 
patient’s data will be provided.  
Statistical testing for the primary endpoints of pain and function will be performed using a 
one-sided p-value compared to =0.025.   
Other efficacy analyses will be presented with one-sided p-values. Safety analyses will be 
presented as two-sided.  
8.2 Patient Accountability 
The number of patients who are randomized, receive study drug, and complete the study 
will be summarized. The number of patients included in the safety, ITT, and PP analysis 
sets will be included in the table. Attendance at each visit, including missed visits, 
discontinuations, lost to follow-up, and percentage accountability will be summarized. A 
list of patients who withdraw early will be provided. It will include the reason and timing 
of the withdrawal. Similarly, the reason any patient is excluded from an analysis set will 
also be provided. In addition, significant known protocol deviations will be noted for 
individual patients; a summary table may also be provided. 


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8.3 Demographic and Baseline Characteristics 
Age, race, ethnicity, sex, height, weight, and Baseline measures of WOMAC and PGA will 
be summarized by treatment arm for all patients randomized, using descriptive statistics.  
8.4 Safety Analyses 
The safety profile will be based on adverse events, concomitant medications, vital signs, 
and results of physical examination. All treated patients will be included in the safety 
analyses. 
8.5 Adverse Events 
Adverse events will be grouped by system organ class and by preferred term within system 
organ class according to the latest version of the MedDRA coding dictionary. The number 
of patients reporting at least one adverse event and each adverse event will be summarized 
treatment group. Tables and/or narratives of any on-study death, serious or significant 
adverse events, including early withdrawals because of adverse events, will be provided 
should they occur. 
8.5.1 Concomitant Medications 
The number and percent of patients receiving concomitant medications or treatments prior 
to and during the study and at the final visit will be tabulated and presented overall and by 
treatment group for the Safety analysis dataset. Concomitant medications, using the generic 
medicine name, and treatments will be summarized using descriptive statistics and will be 
presented by type of drug (WHO drug classification preferred term and Anatomical 
Therapeutic Chemical Classification [ATC] Level 1) by treatment group. 
8.5.2 Vital Signs 
Vital signs will be listed for each patient. These will include temperature, respiration, pulse, 
and blood pressure. Summaries over time and changes from Baseline will be provided. 
8.5.3 Physical Examinations 
Any new or abnormal findings will be recorded as adverse events. Status of each body 
system will be summarized at each visit. 
8.6 Efficacy Analyses 
All efficacy variables will be assessed at Baseline (Day 0), Week 2, Week 4, Week 6, Week 
8, Week 10, Week 12, and Week 24. Differences between treatments (Ampion and saline 
arms) will be presented using mean differences and 95% confidence intervals. Endpoints 
will be compared between Ampion and saline. 
For summaries based on the PP, missing data will not be estimated or carried forward in 
any of the descriptive analyses. For summaries based on the ITT population, estimates from 
the MMRM will be shown incorporating imputed missing data via the full conditional 
specification method with predictive mean matching.  


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8.6.1 Primary Effectiveness Endpoints 
Change in WOMAC A pain score as measured by a 5-point Likert scale, from Baseline to 
Week 12, will be analyzed by including change from Baseline at 2, 4, 6, 8, 10, and 12 
weeks in a mixed-effects model with repeated measures (MMRM). Treatment assignment 
Week, the treatment*week interaction and Baseline WOMAC pain score will be included 
as fixed effects. Patient ID will be included to denote repeated measures and a compound 
symmetric covariance matrix will be used for residuals. Least squares means and treatment 
difference with 95% confidence intervals will be presented. 
SAS code used will follow the following format: 
Proc mixed; 
Class PtID treatment week; 
Model delta_pain = treatment baseline_pain week treatment*week; 
Repeated week / subject=newID type = CS; 
Lsmeans active*week / SLiCE=week diff; 
run; 
 
The primary analysis will include the repeated measures of WOMAC C function sub score 
from Baseline to Weeks 2, 4, 6, 8, 10, and 12 and will use a mixed-effects model with 
repeated measures MMRM model, with Week as a fixed effect, and including patient ID 
as a repeated measure with a compound symmetric covariance matrix for residuals.  
The primary effectiveness endpoints will be analyzed using the ITT analysis population. 
In order to preserve the randomization, any missing values at 12 weeks will be imputed. 
Week 12 visit data collected more than 14 days after the target day of 84 will be used and 
not imputed. The primary method of imputation will the full conditional specification 
method with predictive means matching (Berglund & Heeringa 2014). This imputation 
method uses all observed values of the primary outcome at Baseline, 2, 4, 6, 8, 10, and 12 
weeks to impute an individual’s missing values.  To determine the possible effect of 
missing data on the primary analysis, the following sensitivity analyses will be performed: 
1 – multiple imputation (SAS PROC MI, using the default method MCMC along with 
PROC MIANALYZE); 2 – last observation carried forward (LOCF) 3 – a regression-based 
prediction using individual’s prior data and time of prior data will be used to impute Week 
12 data; and 4 – a conservative imputation approach will be performed where the greatest 
pain reduction observed in the study will be imputed for the saline arm and the worst 
change in pain score observed in the study will be used to impute missing 12-week values 
in the Ampion arm. Any imputed value that results in more than one decimal place will be 
rounded to one decimal place. 
An analysis of site effect will be performed. If the main effect has a p-value of 0.15 or 
smaller in a secondary analysis, site will be included in the model and summaries of the 
primary effectiveness endpoint will be produced by site as well as pooled over sites. 
WOMAC A Pain scores and the WOMAC C functions scores and their respective change 
and percent change from Baseline will be summarized by visit. The Change from Baseline 
scores will include a 95% CI.   For the WOMAC A Pain subscore and WOMAC C Function 
subscore, a test for the main effect for Ampion vs. saline will be included on the summary 
tables at weeks indicated as primary or secondary endpoints.    


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8.6.2 Secondary Effectiveness Endpoints 
Secondary effectiveness endpoints will be analyzed using the ITT and the PP analysis 
populations. All secondary efficacy tests will be performed and reported with one-sided p-
values for superiority of Ampion. 
Change from Baseline to 2, 4, 6, 8, 10, 12, and 24 weeks for the PGA will be analyzed 
using a MMRM model. Treatment assignment, Week, and Baseline PGA will be included 
as fixed effects and a compound symmetric covariance matrix for residuals to reflect the 
repeated measures.  
Change from Baseline to 12 weeks for the 100mm VAS Pain Score will be analyzed using 
a MMRM model. Treatment assignment, and Baseline PGA will be included as fixed 
effects and a compound symmetric covariance matrix for residuals to reflect the repeated 
measures.  
Use of rescue analgesia (amount of acetaminophen used) will be compared by treatment 
arm. A Wilcoxon rank-sum test will be used to compare overall pill count use over the 
study between saline and Ampion. 
8.6.3 Subset analyses 
The two co-primary endpoints and three secondary effectiveness endpoints will be 
analyzed by subsets for: 
(a) ages <60, [60, 70), and ൒70,  
(b) men and women,  
(c) white and other,  
(d) Non-Hispanic and Hispanic 
 
8.6.4 Sensitivity analyses 
The two co-primary endpoints and three efficacy secondary endpoints will be re-
analyzed separately in the per-protocol population to exclude treatment failures.  
 
8.6.5 Exploratory Endpoints 
A Bayesian meta-analysis for Likert WOMAC A pain subscore including KL4 patients 
from studies AP-003-A, AP-003-B, AP-003-C, AP-004, and AP-013. The model is 
constructed for WOMAC A change from baseline for patient i in study j 
Yi,j ~ N(μi,j, σ2) 
μi,j = αj + δj Ti +β BLi 
αj ~N(α, σα2) 
δj ~N(δ, σδ2) 
α ~N(0,100) 


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δ ~N(0,100) 
σα ~U(0,10) 
σδ ~ U(0,10) 
σ ~ U(0,10) 
Where Ti is the treatment assignment (0 saline or control; 1=Ampion) and BLi is the 
baseline WOMAC A score.  j’s represent the control means across studies. j’s represent 
the treatment effects across studies.  σα and σα represent the heterogeneity of control rates 
and treatment effects, respectively. Sensitivity analyses will be reported that increase the 
priors on the variance parameters from U(0,10) to U(0,500).  The posterior means and 95% 
credible intervals will be reported for each treatment by study and treatment effect by study. 
Change in WOMAC A pain sub score between Baseline and Weeks 2, 4, 6, 8, 10, and 24 
will be analyzed using the MMRM model described for the primary analysis. 
Change in WOMAC B stiffness sub score between Baseline and Weeks 2, 4, 6, 8, 10, 12 
and 24 will be analyzed using the MMRM model described for the primary analysis. 
Change in WOMAC C physical function sub score between Baseline and Weeks 2, 4, 6, 8, 
10, and 24 will be analyzed using the MMRM model described for the primary analysis. 
Change in PGA between Baseline and Weeks 2, 4, 6, 8, 10, and 24 will be analyzed using 
the MMRM model described for the primary analysis. 
OMERACT-OARSI responder status, defined using Scenario D criteria, will be analyzed 
using separate logistic regression models for each visit. 
Treatment failures will be analyzed with a Fisher’s exact test. 
Cumulative distribution function (CDF) plots will be used to compare WOMAC A pain 
and WOMAC C function endpoints for Ampion vs. saline arms. 
If collected, the differences in urine analysis will be evaluated between treatment groups 
at Week 12. 
Continuous secondary endpoints and their change from Baseline will be summarized by 
visit and treatment arm. Change from Baseline will include a 95% CI.  
8.6.6 Post Hoc/Ad Hoc Analyses 
Any analysis not described in this plan will be considered post-hoc and identified as such 
in the CSR. 
8.6.7 Multiplicity Adjustment 
Hierarchical testing for the ITT analysis population will be applied to the analysis of the 
primary endpoints.  It will also be utilized for the secondary endpoints, assuming 
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for the primary effectiveness tests (WOMAC A and WOMAC C (as specified)), then and 
only then would PGA score be tested at the one-sided alpha of 0.025; if the PGA score is 
significant then and only then, will the 100 mm VAS hypotheses be tested at a one-sided 
alpha of 0.025; if the 100 mm VAS hypothesis is significant then, and only then, will the 
amount of rescue analgesia through Week 12 be tested at a one-sided alpha 0.025.  
Multiplicity adjustment will not be applied to exploratory endpoints. 
9. 
CHANGES FROM PROTOCOL-SPECIFIED ANALYSES  
Modifications to the planned statistical analyses should be minimized. Nonetheless, the 
data obtained from the study may indicate that the planned analyses are inappropriate, that 
additional analyses need to be performed, or that the design of the study needs to be 
modified, due to factors such as the distribution of the data or imbalance in important 
covariates. The study report will provide a detailed explanation for any deviations from the 
planned analyses. 
10. 
LIST OF PLANNED TABLES, FIGURES, AND LISTINGS 
Tables are categorized and numbered in accordance with ICH E3 guidelines. Each table, 
figure and listing is presented by treatment arm.  
Accountability tables listed below will also include an overall column. Listings will be 
sorted by treatment, patient ID, and by visit, if multiple visits exist. 
10.1 Tables 
14.1 Accountability 
14.1.1 
Accountability (Analysis population: All Enrolled) 
14.1.2 
Enrollment by Site (Analysis population: All Enrolled) 
14.1.3 
Analysis Populations (Analysis population: All Enrolled) 
14.1.4 
Patient Disposition (All Screened) 
14.1.5 
Major Protocol Deviations (Analysis population: All Enrolled) 
14.1.6.1 
Demographics and Baseline Characteristics (Analysis population: ITT) 
14.1.6.2 
Demographics and Baseline Characteristics (Analysis population: PP) 
The following efficacy tables will be provided both for ITT and PP. ITT tables will have a 
.1 suffix and PP tables will have a .2 suffix added to the table numbers (ex. 14.2.1.1 = ITT 
and 14.2.1.2 = PP) 
14.2 Efficacy 
14.2.1 
Summary of WOMAC A Pain Subscale 
14.2.2 
Summary of WOMAC B Stiffness Subscale  
14.2.3 
Summary of WOMAC C Physical Function Subscale 
14.2.4 
Summary of 100mm VAS Pain Score 
14.2.5 
Summary of PGA 
14.2.6 
Summary of Rescue Analgesia Use 
14.2.7 
Summary of OMERACT-OARSI responder rates 
14.2.8 
Summary of WOMAC A Pain Subscale by Age 


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14.2.9 
Summary of WOMAC A Pain Subscale by Sex 
14.2.10 
Summary of WOMAC A Pain Subscale by Race 
14.2.11 
Summary of WOMAC A Pain Subscale by Ethnicity 
14.2.12 
Summary of WOMAC C Pain Subscale by Age 
14.2.13 
Summary of WOMAC C Pain Subscale by Sex 
14.2.14 
Summary of WOMAC C Pain Subscale by Race 
14.2.15 
Summary of WOMAC C Pain Subscale by Ethnicity 
14.2.16 
Summary of 100mm VAS Pain Score by Age 
14.2.17 
Summary of 100mm VAS Pain Score by Sex 
14.2.18 
Summary of 100mm VAS Pain Score by Race 
14.2.19 
Summary of 100mm VAS Pain Score by Ethnicity 
14.2.20 
Summary of PGA by Age 
14.2.21 
Summary of PGA by Sex 
14.2.22 
Summary of PGA by Race 
14.2.23 
Summary of PGA by Ethnicity 
14.2.24 
Summary of Rescue Analgesia Use by Age 
14.2.25 
Summary of Rescue Analgesia Use by Sex 
14.2.26 
Summary of Rescue Analgesia Use by Race 
14.2.27 
Summary of Rescue Analgesia Use by Ethnicity 
14.2.28 
Summary of treatment failure rates (%) 
 
14.3 Safety 
14.3.1 
Overall Summary of Treatment-Emergent Adverse Events (Analysis 
population: Safety) 
14.3.2 
Incidence of Treatment-Emergent Adverse Events by System Organ Class 
and Preferred Term (Analysis population: Safety) 
14.3.3 
Incidence of Treatment-Emergent Adverse Events by System Organ Class 
and Preferred Term by Maximum Severity (Analysis population: Safety) 
14.3.4 
Incidence of Treatment-Emergent Related Adverse Events by System 
Organ Class and Preferred Term (Analysis population: Safety) 
14.3.5 
Incidence of Treatment-Emergent Related Adverse Events by System 
Organ Class and Preferred Term by Maximum Severity (Analysis 
population: Safety) 
14.3.6 
Incidence of Treatment-Emergent Serious Adverse Events by System 
Organ Class and Preferred Term (Analysis population: Safety) 
14.3.7 
Incidence of Treatment-Emergent Adverse Events by Preferred Term in 
Descending Order of Frequency (Analysis population: Safety) 
14.3.8 
Incidence of Treatment-Emergent Related Adverse Events by Preferred 
Term in Descending Order of Frequency (Analysis population: Safety) 
14.3.9.1 
Summary of Vital Signs and Change from Baseline over Time for Pulse 
(bpm) (Analysis population: Safety) 
14.3.9.2 
Summary of Vital Signs and Change from Baseline over Time for 
Temperature (F) (Analysis population: Safety) 
14.3.9.3 
Summary of Vital Signs and Change from Baseline over Time for Systolic 
BP (mmHg) (Analysis population: Safety) 


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14.3.9.4 
Summary of Vital Signs and Change from Baseline over Time for 
Diastolic BP (mmHg) (Analysis population: Safety) 
14.3.9.5 
Concomitant Medication Use by ATC Level 1 and WHO Preferred Term 
(Analysis population: Safety) 
14.3.9.6 
Medication Started on Study by ATC Level 1 and WHO Preferred Term 
(Analysis population: Safety) 
14.3.9.7 
Concomitant Medications by WHO Preferred Term in Descending Order 
of Use (Analysis population: Safety) 
14.3.9.8 
Medication Started on Study by WHO Preferred Term in Descending 
Order of Use (Analysis population: Safety) 
 
10.2 Listings 
Subject Accountability 
1. Randomization List (including: patient ID, randomization number, 
randomized treatment and treatment administered, and date of treatment) 
2. Inclusion and Exclusion Criteria 
3. Protocol Deviations 
4. Patients Withdrawing from the Study Prematurely (date and reason) 
5. Analysis Populations with Reason for Exclusion (if populations differ) 
Demographics and Baseline Characteristics 
6. Demographics and Baseline Characteristics (age, sex, race, ethnicity, weight, 
height, Baseline WOMAC and PGA scores) 
7. Medical History 
8. Baseline Medication Use 
Efficacy  
9. WOMAC A, B, and C Scores (includes all subscales) 
10. PGA Scores 
11. 100mm VAS Pain Scores 
12. Rescue Analgesia 
13. OMERACT-OARSI responder rates 
14. Treatment failures 
Safety  
15. All Adverse Events [with indication of TEAE] 
16. Physical Exams 


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17. Vital Signs Data [systolic and diastolic blood pressure, pulse rate, respiration, 
and temperature] 
10.3 Figures 
Note that all figures, unless otherwise stated, will be line plots showing mean ± SEM at 
each visit for each treatment arm. 
1. WOMAC A Pain Subscale at Baseline and Week 12 [Boxplot] 
2. Summary of WOMAC A Pain Subscale over Time 
3. Summary of WOMAC B Stiffness Subscale over Time 
4. Summary of WOMAC C Physical Function Subscale over Time 
5. Summary of PGA over Time 
6. Summary of Rescue Analgesia Used [stacked histogram] 
7. Summary of OMERACT-OARSI responder rates over time [Bar chart] 
8. Change in WOMAC A Pain Subscale from Baseline to Week 12 for subgroups 
[Forest plot] 
9. Change in WOMAC C Function Subscale from Baseline to Week 12 for 
subgroups [Forest plot] 
11. 
LITERATURE CITATIONS  REFERENCES 
SAS Institute Inc. SAS Language: version 8 first edition. SAS Institute, Inc, Cary, NC, 
USA, 1990. 
R Core Team (2012). R: A language and environment for statistical computing. R 
Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, URL: 
http://www.R-project.org/. 
12. 
HANDLING OF MISSING OR INCOMPLETE DATES FOR 
ADVERSE EVENTS AND CONCOMITANT MEDICATIONS 
12.1 Imputation Rules for Partial or Missing Stop Dates 
If the month and year are present, impute the last day of the month. If only the year is 
present, impute December 31 of that year. If the stop date is entirely missing, assume the 
event or medication is ongoing. If a partial or complete stop date is present and the 
‘ongoing’ or ‘continuing’ box is checked, then it will be assumed that the AE or 
concomitant medication stopped and the stop date will be imputed, if partial.  


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Stop Date 
 
 
Complete: 
yyyymmdd 
Partial: yyyymm 
Partial: yyyy Missing 
Start 
Date 
 
<1st 
Dose 
1st 
Dose 
<1st 
Dose 
yyyymm 
1st 
Dose 
yyyymm 
<1st 
Dose 
yyyy 
1st 
Dose 
yyyy 
Partial: 
yyyymm 
=1st 
Dose 
yyyymm 
2 
1 
2 
1 
N/A 
1 
1 
 1st 
Dose 
yyyymm 
2 
2 
2 
2 
2 
Partial: 
yyyy 
=1st 
Dose 
yyyy 
3 
1 
3 
1 
N/A 
1 
1 
 1st 
Dose 
yyyy 
3 
3 
3 
3 
3 
Missing 
4 
1 
4 
1 
4 
1 
1 
1 = Impute the date of first dose 
2 = Impute the first of the month 
3 = Impute January 1 of the year 
4 = Impute January 1 of the stop year 
Note: For patients who were never treated (first dose date is missing), partial start dates 
will be set to the first day of the partial month. 
Note: If the start date imputation leads to a start date that is after the stop date, then do not 
impute the start date.


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13. 
SCHEDULE OF ASSESSMENTS AND PROCEDURES 
 
Screening 
Pre-
Baseline 
(phone) 
Baseline 
24-hours 
(phone) 
Week 2 
(phone) 
Week 4 
(phone) 
 
Week 6 
 
Week 8 
(phone) 
Week 10 
(phone) 
Pre-Week 
12 
(phone) 
Week 12 
 
Week 24 
 
Early 
Term. 
Assessments 
& Procedures 
Visit -1 
Day-28 to 0 
Visit 0 
Day -6 to 0 
Visit 1 
Day 0 
Visit 2 
Day 1 
Visit 3 
Day 14 ± 7 
Visit 4 
Day 28 ± 7 
Visit 5 
Day 42 ± 7 
Visit 6 
Day 56 ± 7 
Visit 7 
Day 70 ±7 
Visit 8 
Day 77 ±7 
Visit 9 
Day 84 ± 7 
Visit 10 
Day 174 ± 7 
 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/ 
exclusion criteria 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
Medical history/prior 
medications 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
Concomitant 
medications 
X 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Urine sample 
 
 
X** 
 
 
 
 
 
 
 
X 
 
 
Physical examination 
X 
 
X 
 
 
 
X 
 
 
 
X 
X 
X 
Vital Signs 
X 
 
X 
 
 
 
X 
 
 
 
X 
X 
X 
Randomization 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Review Diary 
Completion Instructions 
X 
 
 
 
 
 
 
 
X 
 
 
 
 
Phone Contact Daily 
Collection Reminder  
 
X 
 
 
 
 
 
 
 
X 
 
 
 
WOMAC A (Likert) 
X 
X* 
X*** 
 
X 
X 
X 
X 
X 
X* 
X*** 
X 
X 
WOMAC A (VAS) 
X 
X* 
X*** 
 
 
 
 
 
 
X* 
X*** 
 
X 
WOMAC B and C 
(Likert) 
X 
 
X 
 
X 
X 
X 
X 
X 
 
X 
X 
X 
PGA 
X 
 
X 
 
X 
X 
X 
X 
X 
 
X 
X 
X 
X-ray1 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Study treatment 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Dispense Rescue 
medication 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Review Rescue 
medication 
 
 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Adverse Events 
 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Visits are in clinic except for Day 1 and Weeks 2, 4, 8 and 10 when patients will be contacted by telephone 
1X-ray may be acquired at Screening to satisfy inclusion criteria, “Index knee must be symptomatic for greater than six months with a clinical diagnosis of OA and supported by radiological evidence 
(KL 4) that is not older than six months prior to the date of screening”.  
*Patients will complete 7 daily pain score assessments up to and including the Baseline and Week 12 visits to allow calculation of WOMAC A weekly pain scores. The Screening and first Pre-
Baseline daily pain score collection may occur on the same day, in office or at home, as appropriate. 
**Urine will only be collected at Baseline if necessary to conduct a pregnancy test; collection to occur as applicable 
***The WOMAC may occur in office or at home, and may serve as the 7th WOMAC Daily Pain collection