Clinical trial NCT06719986
Conditions: Opioid Overdose
Sponsor: Purdue Pharma LP
Start date: 2022-10-03; Completion date: 2022-12-23
Enrollment: 24 participants; Allocation: RANDOMIZED; Intervention model: CROSSOVER

Brief summary:
The purpose of this study is to determine the pharmacodynamics (change in minute ventilation) of nalmefene when given as an autoinjector intramuscularly (IM; into the thigh) compared to naloxone when given intranasally (IN; into the nose) to healthy subjects with prior opioid exposure under steady state fentanyl concentrations (opioid agonism).

Detailed description:

Arms: arm 1: Nalmefene Hydrochloride injection 1.5 mg for intramuscular (IM) administration arm 2: Naloxone 4 mg for intranasal (IN) administration
Interventions: intervention 1: Nalmefene HCl injection intervention 2: Naloxone HCl intranasal
Intervention descriptions: intervention 1: Nalmefene Hydrochloride injection 1.5 mg for intramuscular (IM) administration intervention 2: Naloxone 4 mg for intranasal (IN) administration

Protocol documents:
Prot_000.pdf: 
Official Title: A Study to Characterize the Time Course of Reversal of Opioid 
(Fentanyl)-Induced Respiratory Depression Following Administration of 
Nalmefene Autoinjector 1.5 mg (0.94% MgCl2) Intramuscular·and Narcan® 4 mg 
Intranasal in Healthy Subjects 
 
NCT #: NCT06719986 
 
Date of Document: October 12, 2022 



























SAP_001.pdf: 
Official Title: A Study to Characterize the Time Course of Reversal of Opioid 
(Fentanyl)-Induced Respiratory Depression Following Administration of 
Nalmefene Autoinjector 1.5 mg (0.94% MgCl2) Intramuscular·and Narcan® 4 mg 
Intranasal in Healthy Subjects 
 
NCT #: NCT06719986 
 
Date of Document: August 7, 2022 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Confidential 
Page 1 | 28 
STATISTICAL ANALYSIS PLAN 
A Study to Characterize the Time Course of Reversal of Opioid (Fentanyl)-
Induced Respiratory Depression Following Administration of Nalmefene 
Autoinjector 1.5 mg (0.94% MgCl2) Intramuscular and Narcan® 4 mg 
Intranasal in Healthy Subjects 
Protocol Number: 
NAL1004 
Protocol Version and Date: 
Amendment 1: 28-JULY-2022 
Study Drug Name: 
Fentanyl, Nalmefene Hydrochloride, Naloxone 
Hydrochloride 
Phase: 
Phase 1 
Sponsor: 
Purdue Pharma, LP  
One Stamford Forum  
Stamford, CT 06901-3431  
USA 
Analysis Plan Date: 
August 7, 2022 
Analysis Plan Version: 
Version 1.0 




Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 3 of 28 
TABLE OF CONTENTS 
STATISTICAL ANALYSIS PLAN ............................................................................................................. 1 
SPONSOR SIGNATURE PAGE.................................................................................................................. 2 
TABLE OF CONTENTS .............................................................................................................................. 3 
LIST OF TABLES INCLUDED IN THE TEXT ......................................................................................... 5 
1 
Introduction .......................................................................................................................................... 9 
2 
Study Design Overview ....................................................................................................................... 9 
 
 
3 
Study Objectives ................................................................................................................................ 17 
 
 
 
 
 
 
4 
Study Endpoints and Evaluations....................................................................................................... 18 
 
 
 
 
 
 
 
 
5 
Analysis Populations and Applications .............................................................................................. 19 
 
 
 
 
 
 
 
 
6 
Statistical Considerations ................................................................................................................... 20 
 
 
6.2.1 
Multiplicity Adjustment .................................................................................................. 22 
 
 
 
 
7 
Efficacy Analysis ............................................................................................................................... 23 
8 
Safety Analysis .................................................................................................................................. 23 
 
 
  
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
  
 
   
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 4 of 28 
9 
Pharmacodynamic Analysis (Part 2 Only) ......................................................................................... 24 
10 
Definitions and Conventions for Data Handling ................................................................................ 26 
11 
REFERENCES................................................................................................................................... 28 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 5 of 28 
LIST OF TABLES INCLUDED IN THE TEXT
Table 1 Part 1 Study Flow Chart - Overall Procedures……………………………………………………11 
Table 2 Part 2 Study Flow Chart - Overall Procedures…………………………………………………….14 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 6 of 28 
ABBREVIATIONS 
® 
registered trademark  
AE 
adverse event  
ALT 
alanine aminotransferase (alanine transaminase; also SGPT)  
AST 
aspartate aminotransferase (aspartate transaminase; also SGOT) 
bpm 
beats per minute  
BMI 
body mass index  
BUN 
blood urea nitrogen  
CO2 
carbon dioxide  
CRF 
case report form  
DO 
DoR 
Doctor of Osteopathic Medicine 
duration of reversal  
dL 
deciliter(s)  
ECG 
EOQ 
electrocardiogram 
end of qualification  
EOS 
end of study  
ET 
end-tidal  
fax 
facsimile transmission  
FDA 
Food and Drug Administration  
FSH 
follicle stimulating hormone  
GCP(s) 
Good Clinical Practice(s)  
gm or g 
gram(s)  
HBsAg 
hepatitis B surface antigen  
HCV 
hepatitis C virus (antibody)  
HIV 
human immunodeficiency virus  
hr or h 
hour  
HS 
healthy subjects  
ICF 
informed consent form  
ICH 
International Conference on Harmonisation  
IM 
intramuscular  
IRB 
Institutional Review Board  
IV 
intravenous  
K2EDTA 
dipotassium ethylenediaminetetraacetic acid 
kg 
kilogram(s)  
L 
liter(s)  
ln 
natural logarithm  
L.P.
Limited Partnership  
LDH
lactate dehydrogenase  


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 7 of 28 
LFT 
liver function test  
LLT 
lower level term(s)  
mcg 
microgram  
MD 
Medical Doctor  
MedDRA 
Medical Dictionary for Regulatory Activities  
mEq 
milliequivalent(s)  
mg 
milligram(s)  
min 
minute(s)  
mL or ml 
milliliter(s)  
mm 
millimeter(s)  
mm Hg 
millimeters of mercury  
mmol 
millimole(s)  
MV 
minute volume  
N/A or NA 
not applicable  
OIRD 
opioid-induced respiratory depression  
OOWS 
Objective Opioid Withdrawal Scale  
PaCO2 
arterial partial pressure of carbon dioxide  
PCO2 
partial pressure of CO2 in mm Hg (transcutaneous CO2)  
PD 
pharmacodynamics(s)  
pH 
negative log of hydrogen ion concentration  
PI 
principal investigator  
PK 
pharmacokinetic(s)  
PO2 
Partial pressure of oxygen  
PPLP 
Purdue Pharma Limited Partnership  
PT 
preferred term(s)  
RBC 
red blood cell (count)  
ROU 
healthy recreational opioid users  
RR 
respiratory rate  
RVM 
respiratory volume monitor(ing)  
SAP 
Statistical Analysis Plan 
SAS 
Statistical Analysis System  
SC 
subcutaneous  
SGOT 
serum glutamic-oxaloacetic transaminase (also AST)  
SGPT 
serum glutamate pyruvate transaminase (also ALT)  
SoA 
Schedule of Assessments  
SOC 
System Organ Class  
SOP 
Standard Operation Practices  
SpO2 
hemoglobin-O2 saturation determined by pulse oximetry 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 8 of 28 
TCO2 
THC 
transcutaneous CO2 
9-delta-tetrahydrocannabinol
TTO 
time to onset
TV 
tidal volume
uL 
microliter(s)
ULN 
upper limit of normal
US 
United States
VAS 
visual analog scale
WBC 
white blood cell (count)
WHO 
World Health Organization


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 9 of 28 
1 Introduction 
This statistical analysis plan (SAP) is designed to outline the statistical methods for the NAL1004 
study regarding test drug of fentanyl, naloxone, nalmefene and to assess the change in minute 
ventilation at 5 minutes of intramuscular nalmefene and intranasal naloxone reversal from the 
opioid induced nadir. 
This document has been prepared based on protocol Amendment 1 dated July 28, 2022. Details 
will be described in this analysis plan to ensure that the data listings, summary tables and figures 
which will be produced, and the statistical methodologies that will be used, are complete and 
appropriate to allow valid conclusions regarding the study objectives. 
This analysis plan was finalized and approved by the sponsor 
 prior 
to the planned database lock. 
This SAP supersedes the statistical considerations identified in the protocol; where considerations 
are substantially different, they will be identified, and justification will be provided. If additional 
analyses are required to supplement the planned analyses described in the SAP, they may be 
performed and will be identified in the clinical study report (CSR). 
2  Study Design Overview 
2.1 
Overall Study Design 
This will be a 2-part study. Part 1 is a Qualification Phase prior to Part 2 which will be conducted 
to identify subjects eligible to participate in the Treatment Phase. This will be based upon 
satisfying specified qualification criteria following completion of procedures and/or study drug 
administrations. Qualification may not be required if a subject has participated in Clinical Study 
NAL1003. Each Qualification cohort will have a screening, treatment, EOS and Follow-up.  The 
qualification treatment phase will consist of 1 period and evaluate the pharmacodynamics (minute 
ventilation) of OIRD.  
Part 2 will be a single-center, randomized, 4 period crossover, 2 treatment replicate study to 
evaluate the pharmacodynamic effects (change in minute ventilation from opioid induced nadir 
(OIRD)) of nalmefene (1.5 mg) when given as an autoinjector intramuscularly (IM; into the thigh) 
compared to naloxone (Narcan 4 mg) when given intranasally (IN; into the nose) to reverse opioid 
(fentanyl) -induced respiratory depression (OIRD) in healthy subjects with prior opioid exposure. 
The Treatment Phase will have a screening, treatment, EOS and Follow-up. 
The two treatment sequences for first 2 periods: 
•
Sequence 1: Narcan 4 mg, IN - Nalmefene Autoinjector 1.5 mg, IM
•
Sequence 2: Nalmefene Autoinjector 1.5 mg, IM - Narcan 4 mg, IN
Table 1 and Table 2 below present the complete schedule of activities for NAL1004. 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 10 of 28 
Table 1 
Part 1 Study Flow Chart - Overall Procedures 
Phase 
Screening 
Qualification 
F/U 
Notes 
Period 
Screening 
Check-In 
1 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
2 
8-11
Informed consent 
X 
Medical history 
X 
Including drug and alcohol use history 
Demography 
X 
Including weight, height, and BMI 
Physical exam 
X 
X 
X 
At check-in:  Symptom-directed, at discretion of investigator or designee 
Inclusion/exclusion 
X 
X 
DSM-IV-TR (dependence) 
X 
Clinical laboratory 
X 
X 
X 
Chemistry, hematology and urinalysis 
At check-in:  chemistry and hematology testing only 
Vital signs and SpO2 with 
HDYF? 
X 
X 
X 
X 
Taken after 5 minutes seated (or during infusions when supine, seated, or 
semirecumbent):  heart rate, blood pressure, respiratory rate, SpO2  
Screening:  Additional standing (for ~2 minutes) vital signs (heart rate and blood 
pressure)  
Obtained predose before saline infusion  
During fentanyl infusion: Predose, 30, and 60 minutes 
End of fentanyl infusion:   end of infusion, 2 and 4 hrs.   
Time points may be added, omitted, or adjusted.  
Vital signs also taken pre- and post-naloxone challenge test. 
Oral temperature 
X 
X 
X 
X 
For safety purposes, forehead temperature may be substituted for oral. 
Days 1:   
Predose, and 8-12 hrs post infusion 
Serum pregnancy test (females 
only) 
X 
X 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 11 of 28 
Phase 
Screening 
Qualification 
F/U 
Notes 
Period 
Screening 
Check-In 
1 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
2 
8-11
Serum FSH (post-menopausal 
females only) 
X 
Urine pregnancy test (females 
only) 
X 
Conventional 12-lead ECG 
X 
X 
X 
Drug/alcohol screen 
X 
X 
Serology (HBsAg and HCV) 
X 
Naloxone challenge test 
X 
Performed at least approximately 12 hours prior to first dose in Period 1. 
Includes Objective Opioid Withdrawal Scale (OOWS). 
Does not need to be performed if conducted during a previous check-in visit 
within 60 days. 
RVM/capnography/ 
PD training and practice session 
X 
X 
May be performed at screening or Check-in day -1 or both. 
Study drug dosing 
X 
Time = 0, relative to other procedures.  
 
 
Respiratory volume monitoring 
and capnography 
X 
Procedure will be performed pre (Baseline) and post IV study drug 
administration. 
Predose (baseline) and continuous RVM throughout dosing and for 15 minutes 
after the end of the infusion for remifentanil and for 1 hr after the end of the 
infusion for fentanyl. 
Time interval may be adjusted or separate time points may be examined. 
Transcutaneous 
CO2 
monitoring 
X 
Obtained prior to the start of the saline infusion, every 15 minutes during saline 
infusion and at the end of the saline infusion, every 5 minutes during the fentanyl 
Infusion, and at the end of the fentanyl Infusion. Time points may be added and 
adjusted. 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 12 of 28 
Phase 
Screening 
Qualification 
F/U 
Notes 
Period 
Screening 
Check-In 
1 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
2 
8-11
Continuous SpO2 monitoring 
X 
Relative to IV dosing:  Predose to 8-12 hours for fentanyl 
Note: For fentanyl dosings, subjects can be taken off the pulse oximetry 
monitoring for up to a half hour each day to shower 
Follow-up phone call 
X 
A follow-up phone call will be made to (or received from) each subject 7 to 10 
days after Day 1 or after early withdrawal from the study. 
Confined to study unit 
◄---------------------►
Concomitant therapies 
◄--------------------------------------►
Adverse events 
◄--------------------------------------►
EOQ: End of Qualification; F/U: Follow-up; HYDF?=How do you feel question; IV=Intravenous; RVM= respiratory volume monitor(ing); 
SpO2=Hemoglobin-O2 saturation determined by pulse oximetry


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 13 of 28 
Table 2 
Part 2 Study Flow Chart - Overall Procedures 
Phase 
Screen-
ing 
Treatment 
F/U 
Notes 
Period 
Screen-
ing 
Check-
In 
1 
2 
3 
4 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
4 
7 
10 
11 
18-21 
Informed consent 
X 
Medical history 
X 
Including drug and alcohol use history 
Demography 
X 
Including weight, height, and BMI 
Physical exam 
X 
X 
X 
At check-in:  Symptom-directed, at discretion of investigator or designee 
Inclusion/exclusion 
X 
X 
DSM-IV-TR 
(dependence) 
X 
Clinical laboratory 
X 
X 
X 
Chemistry, hematology and urinalysis 
At check-in:  chemistry and hematology testing only 
Testing for COVID-19 may also be done at any time 
Serum pregnancy test 
(females only) 
X 
X 
Serum FSH (post-
menopausal females 
only) 
X 
Urine pregnancy test 
(females only) 
X 
Conventional 12-lead 
ECG 
X 
X 
X 
Drug/alcohol screen 
X 
X 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 14 of 28 
Phase 
Screen-
ing 
Treatment 
F/U 
Notes 
Period 
Screen-
ing 
Check-
In 
1 
2 
3 
4 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
4 
7 
10 
11 
18-21 
Serology (HBsAg 
 and HCV) 
X 
Naloxone challenge 
test 
X 
Performed at least approximately 12 hours prior to first dose in Period 1. 
Includes Objective Opioid Withdrawal Scale (OOWS).
Does not need to be performed if conducted during Part 1 and subject enters 
Part 2 within 60 days 
Vital signs and SpO2 
with HDYF? 
X 
X 
X 
X 
X 
X 
X 
Taken after 5 minutes seated (or during infusions when supine, seated, or 
semirecumbent):  heart rate, blood pressure, respiratory rate, SpO2  
Screening:  Additional standing (for ~2 minutes) vital signs (heart rate and 
blood pressure)  
Days 1, 4, 7, 10:  During fentanyl infusion: Predose (-15 minutes), end of 
infusion, 2 and 4 hrs post end of infusion.   
Time points may be added, omitted, or adjusted.  
Vital signs also taken pre- and post-naloxone challenge test. 
Oral temperature 
X 
X 
X 
X 
X 
X 
X 
For safety purposes, forehead temperature may be substituted for oral. 
Days 1, 4, 7, 10:  Predose, and 8-12 hrs post infusion.  Time points may be 
added, omitted, or adjusted. 
Respiratory 
monitoring/PD 
training and practice 
session 
X 
X 
May be performed at screening or Check-in Day -1 or both. 
Randomization 
X 
Prior to study drug dosing in Period 1 only 


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Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 15 of 28 
Phase 
Screen-
ing 
Treatment 
F/U 
Notes 
Period 
Screen-
ing 
Check-
In 
1 
2 
3 
4 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
4 
7 
10 
11 
18-21 
Fentanyl 
Infusion 
administration 
X 
X 
X 
X 
Time is relative to start of IV infusion. 3-step fentanyl infusion:  first 
infusion at an initial rate of 5.0 mcg/min, a second infusion for 20 min and 
a third infusion up to 90 min, see Table 7. 
Nalmefene 
or 
Naloxone Study drug 
dosing 
X 
X 
X 
X 
  Nalmefene or Naloxone will be administered approximately 10 min 
following the start of Fentanyl Infusion #2. 
Time = 0, relative to other procedures. 
Local Tolerability 
X 
X 
X 
X 
X 
Nalmefene Treatment only, Days 1, 4, 7, 10: Prior to the start of IV infusion 
Relative to Nalmefene administration:  15, 30 minutes, 1 hr and 24 hr after 
nalmefene administration. 
Blood 
sample 
for 
drug concentration  
X 
X 
X 
X 
Relative to Nalmefene/Naloxone administration:  2.5, 5, 10, 15, 20, and 30 
minutes and at 1, 2, 4, 8, 12,  and 24 hours after nalmefene/naloxone 
administration. 
Respiratory volume 
monitoring 
and 
capnography 
X 
X 
X 
X 
Relative to fentanyl infusion, Predose (baseline) and continuous RVM 
throughout dosing and for 1 hr after the end of the infusion for fentanyl. 
Time interval may be adjusted, or specific time points or multiple shorter 
intervals may be selected. 
Transcutaneous CO2 
monitoring 
X 
X 
X 
X 
Relative to fentanyl infusion, Predose (baseline) and continuous throughout 
dosing and for 1 hr after the end of the infusion for fentanyl. 
Time interval may be adjusted, or specific time points or multiple shorter 
intervals may be selected. 


Purdue Pharma, LP 
Protocol No: NAL1004 
 Statistical Analysis Plan 
Page 16 of 28 
Phase 
Screen-
ing 
Treatment 
F/U 
Notes 
Period 
Screen-
ing 
Check-
In 
1 
2 
3 
4 
EOS 
F/U 
Study Day 
Study Procedures 
-29 - -2
-1
1 
4 
7 
10 
11 
18-21 
Continuous SpO2 
monitoring
◄-------------------►
Relative to fentanyl infusion:  Predose to 8-12 hrs post dose 
Time interval may be adjusted, or specific time points or multiple shorter 
intervals may be selected dependent on the agonist administered. 
Note: Subjects can be taken off the pulse oximetry monitoring for up to a 
half hour each day to shower. 
Device deficiencies 
◄-------------------►
Follow-up phone call 
X 
A follow-up phone call will be made to (or received from) each subject 7 
to 10 days after EOS or after early withdrawal from the study. 
Confined to study 
unit 
◄---------------------------------------►
Concomitant 
therapies 
◄-------------------------------------------------------------►
Adverse events 
◄-------------------------------------------------------------►
EOS: End of study; F/U: Follow-up; HYDF?=How do you feel question; IV=Intravenous; RVM=respiratory volume monitor(ing); 
SpO2=Hemoglobin-O2 saturation determined by pulse oximetry


Purdue Pharma, LP 
Statistical Analysis Plan 
Protocol No: NAL1004 
Dated: August 7, 2022, Version 1.0 
Confidential 
Page 17 | 28 
2.2 
Sample Size Justification 
Part 1:  A sufficient number of subjects will be screened and entered into Part 1 to provide the 
number of subjects required in Part 2.  
Part 2:  A total of up to 22 subjects will be randomized to complete approximately 20.  
In a non-inferiority test on MV at 5min from a 4 period, 2 treatment replicate cross-over design, a 
total sample size of 20 would achieve >90% power at a 2.5% significance level (1-sided) when the 
true difference between the means is 0.8L/min, the non-inferiority margin is -0.5L/min, and the 
within-subject standard deviation is 1.2L/min.  
In a superiority test on MV at 5min from a 4 period, 2 treatment replicate cross-over design, a total 
sample size of 20 would achieve 83% power at a 2.5% significance level (1-sided) when the true 
difference between the means is 0.8L/min and the within-subject standard deviation is 1.2L/min. 
The planned sample size is adequate for sequential testing of noninferiority followed by 
superiority.  
3 Study Objectives 
3.1 
Primary Objectives 
•
To assess the change in minute ventilation at 5 minutes of intramuscular nalmefene and
intranasal naloxone reversal from the opioid induced nadir
3.2 
Secondary 
•
To assess the time course of changes in minute ventilation of intramuscular nalmefene
and intranasal naloxone reversal from the opioid induced nadir
•
To evaluate the pharmacokinetics of nalmefene and naloxone following various routes
of administration
•
To assess the safety and tolerability of fentanyl when co-administered with and without
nalmefene or naloxone


Purdue Pharma, LP 
Statistical Analysis Plan 
Protocol No: NAL1004 
Dated: August 7, 2022, Version 1.0 
Confidential 
Page 18 | 28 
3.3 
Exploratory 
•
To characterize the time course (Magnitude of change in minute ventilation) of
intramuscular nalmefene and intranasal naloxone reversal from the opioid induced
nadir
4 Study Endpoints and Evaluations 
4.1 
Pharmacokinetic Endpoints and Evaluations 
Pharmacokinetic concentration and endpoint will be included in a separate data analysis plan 
(DAP) and will not be included in this SAP. 
4.2 
Pharmacodynamics Evaluations 
Pharmacodynamic (PD) evaluations will be performed for the following time frames: prior to 
fentanyl, after fentanyl and before any antagonists, and after antagonist administration (nalmefene, 
naloxone). The PD variables include: 
•
Minute Ventilation (MV) in L/min - calculated as the product of tidal volume and respiratory
rate, determined in a continuous fashion by noninvasive respiratory volume monitor
•
Transcutaneous CO2 (TCO2) in mmHg – a lagging measure of respiratory status in OIRD, it is
an exploratory supplement endpoint to MV
4.3 
Safety Endpoints and Evaluations 
Safety will be assessed using 
•
Treatment emergent adverse events (TEAEs)
•
Clinical laboratory test results
•
Vital signs, including SpO2
•
Physical examinations
•
Conventional 12-lead ECGs
•
Local tolerability
4.4 
Other Evaluations 
Other evaluations include: 
•
Demographics and baseline characteristics
•
Disposition
•
Study drug dosing


Purdue Pharma, LP 
Statistical Analysis Plan 
Protocol No: NAL1004 
Dated: August 7, 2022, Version 1.0 
Confidential 
Page 19 | 28 
•
Medical history
•
Drug screen
•
Pregnancy test
•
Medications (prior and concomitant)
5 Analysis Populations and Applications 
For purposes of analysis, the following analysis populations are defined: 
5.1 
Enrolled Population  
Enrolled population includes all subjects who signed informed consent. 
5.2 
Randomized Safety Population 
Randomized safety population, also known as the safety population, includes subjects who are 
randomized and receive at least 1 dose of the study drug. 
5.3 
Eligible Analysis Set (EAS) 
Twenty subjects will be expected to complete the study, each of whom will receive nalmefene or 
naloxone in different order after fentanyl -induced respiratory depression (OIRD).  
For each period, eligible analysis set (EAS) include subjects/periods who are randomized, receive 
study drug, have at least 1 valid PD measurement, and their OIRDs are worse than baseline for 
MV and TCO2. Note: baseline and OIRD for MV and TCO2 may be different. Therefore, eligible 
analysis set for each PD parameter and period may be different.  
To evaluate the treatment difference between nalmefene and naloxone, the same treatment under 
different periods will be pooled by averaging PD parameters from eligible data for each subject.  
This analysis set will be used as primary analysis.  
5.4 
Full Analysis Set (FAS) 
Full analysis set (FAS) includes subjects/periods who are randomized, receive study drug, and 
have at least 1 valid PD measurement for one individual period.  
To evaluate the treatment difference between nalmefene and naloxone, the same treatment under 
different periods will be pooled by averaging PD parameters from eligible data for each subject.  
This set will be used as sensitivity analyses. However, if it is determined that EAS is very similar 
to FAS, only EAS will be performed. 


Purdue Pharma, LP 
Statistical Analysis Plan 
Protocol No: NAL1004 
Dated: August 7, 2022, Version 1.0 
Confidential 
Page 20 | 28 
6 Statistical Considerations 
All summaries and statistical analysis will be performed by SAS version 9.4 or later. 
Following each administration of Nalmefene Autoinjector or Narcan, the reversal time course of 
the fentanyl induced respiratory depression for MV and TCO2 will be evaluated. 
The following data will be provided in the summary tables: 
•
Disposition
•
Demographics and baseline characteristics
•
Medical history
•
Concomitant medications
•
Treatment exposure
•
Treatment emergent adverse events
•
Derived pharmacodynamic (PD) parameters based on PD assessment data
o
Change in MV from opioid induced nadir (Opioid Induced Respiratory Depression
(OIRD))
o
Maximum of reversal in MV
o
Time to maximum of reversal in MV
The summary figures/tables for PD assessment will include 
o
Individual profiles
o
Treatment mean profiles
o
Change from baseline and percent change from baseline in MV and TCO2 from
opioid induced nadir
o
Change in MV from opioid induced nadir (Opioid Induced Respiratory Depression
(OIRD))
o
Maximum of reversal in MV
o
Time to maximum of reversal in MV
o
Time to onset
•
Vital signs
•
Physical examinations
The following data will be provided in data listings: 
•
Demographics and baseline characteristics
•
Treatment exposure
•
Medical history
•
Prior and concomitant medication/therapy


Purdue Pharma, LP 
Statistical Analysis Plan 
Protocol No: NAL1004 
Dated: August 7, 2022, Version 1.0 
Confidential 
Page 21 | 28 
•
Treatment emergent adverse events
•
Derived pharmacodynamic (PD) parameters
•
Vital signs
•
Physical examinations
•
Electrocardiogram
6.1 
General Statistical Procedures 
Frequency distributions for categorical variables will be provided as number and percentage of 
subjects with a response in the category. Unless otherwise specified, percentages will be based on 
number of subjects in the given population as noted. Percentages will be reported to one decimal 
place. 
The descriptive statistics for continuous variables will be number of subjects, mean, standard 
deviation (SD), median, minimum and maximum. Mean and median will be reported to 1 more 
decimal place than the raw data, while the SD will be reported to 2 more decimal places than the 
raw data. Minimum and maximum will be reported the same as the original data.  
Unless otherwise stated, all statistical summary will be performed separately for each treatment in 
each part.  
In general, all listings will be ordered by Part, subject ID and visit for available data unless 
otherwise specified in the footnote. 
6.2 
Statistical Hypotheses 
Hypotheses #1: Non-inferiority Test 
The null hypothesis is that Nalmefene (μT) is inferior to reference treatment (μR). 
The alternative hypothesis is that Nalmefene is not inferior to reference treatment. 
H0: μT - μR ≤ -0.5   vs.   H1: μT - μR > -0.5 
Hypothesis #2: Superiority Test 
The null hypothesis is that Nalmefene (μT) is not superior to reference treatment (μR). 
The alternative hypothesis is that Nalmefene is superior to reference treatment. 
H0: μT - μR ≤ 0   vs.   H1: μT - μR > 0 
Statistical analysis will be performed for hypothesis #1 first, if noninferiority 
is met the analysis will proceed to hypothesis #2. 


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6.2.1 Multiplicity Adjustment  
For the primary endpoint, the non-inferiority test will be performed first. If non-inferiority is 
achieved, superiority will be tested sequentially. Using this hierarchical test procedure, the 
comparison of the two treatments will be tested based on a one-sided alpha = 0.025. 
The secondary endpoints will not be adjusted for multiplicity. 
6.3 
Subject Enrollment and Disposition 
Number of subjects will be provided overall for Enrolled population for both Qualification Phase 
and Treatment Phase as follows: 
Number of subjects enrolled 
Number of subjects in randomized safety population 
Number of subjects in full analysis population  
Number of subjects completed study 
Number of subjects completed treatment phase 
Number of subjects discontinued (including screen failures) 
Reasons for discontinuation 
For the Randomized Safety population, the denominators for percentages are based on number of 
Enrolled population. 
Enrollment information and disposition will be provided in a data listing. 
6.4 
Demographic and Other Baseline Characteristics 
The following variables will be summarized overall using Enrolled population and randomized 
safety population.  
•
Age (years) at screening. If not reported, calculated as integer of (date of informed consent
signed – date of birth)/365.25.
•
Age category (18 ≤ age < 35, 35 ≤ age < 50, 50 ≤ age < 55)
•
Sex
•
Race
•
Ethnicity
•
Height (cm)
•
Weight (kg)
•
BMI (body mass index, kg/m2, calculated as Weight (kg) / (Height (m)2)
•
Child-bearing potential (Yes, No, Not Applicable: Subject is Male)
Conversions for height and weight are as follows: 


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Height (cm) = Height (inches) × 2.54 
Weight (kg) = Weight (lb) × 0.4536 
The above demographics and baseline characteristics will be summarized for the Qualification 
Phase using the enrolled population and the Treatment Phase using randomized safety population. 
Demographic characteristics will also be listed. 
7 Efficacy Analysis 
Not applicable. 
8 Safety Analysis 
8.1 
Adverse Events 
Adverse events (AEs) will be presented separately by treatment assignment. AEs will be coded 
using the most current version of MedDRA and will be classified by SOC and PT of MedDRA. 
Severity of AEs will be assessed by investigators according to CTCAE. 
Treatment-emergent adverse events are defined as AEs that: 
•
Emerge during treatment, having been absent at pretreatment; or
•
Reemerge during treatment, having been present at pre-treatment but stopped prior to
treatment; or
•
Worsen in intensity during treatment relative to the pre-treatment state, when the AE is
continuous.
An AE is treatment emergent if the start date/time of the event occurs any time on or after the first 
dose received on Day 1. 
Adverse events that start within seven days after the last dose of study drug will be considered 
TEAEs. 
The following types of summary will be provided by treatment using Randomized Safety 
population for each treatment phase separately: 
1. Overview of TEAE
2. TEAEs by SOC, PT, and Maximum Severity
3. TEAEs by SOC and PT
4. Drug-Related TEAEs by SOC and PT
5. Drug-Related TEAEs by SOC, PT and Maximum Severity
6. TEAEs by Preferred Term in Descending Order
If a PT or SOC was reported more than once for a subject, the subject would only be counted once 
in the incidence for that PT or SOC.  
In tabulation by severity grade, 


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•
For a given SOC, only the most severe SOC for each subject will be included.
•
For a given PT, only the most severe PT for each subject will be included.
In tabulations by relationship to study drug,  
•
For a given SOC, only the most related SOC for each subject will be included.
•
For a given PT, only the most related PT for each subject will be included.
All AEs, TEAEs, AEs leading to treatment discontinuation will be provided in listings.  
8.2 
Vital signs 
For each continuous vital sign parameter, descriptive summary statistics for values observed at 
Baseline and at each scheduled timepoint, and for the change from Baseline will be presented. For 
this summary, the Baseline is the last non-missing value prior to the start of Fentanyl treatment. 
This summary will be presented in the randomized safety population. 
Vital sign data will be provided in a listing for subjects in Randomized Safety population. 
8.3 
Electrocardiogram (ECG) 
For each continuous ECG parameter, descriptive summary statistics for values observed at 
Baseline and at each scheduled timepoint, and for the change from Baseline will be presented. For 
this summary, the Baseline is the last non-missing value prior to the start of Fentanyl treatment. 
This summary will be presented in the randomized safety population. 
ECG data will be provided in a data listing for subjects in Randomized Safety population. 
9 Pharmacodynamic Analysis (Part 2 Only) 
9.1 
Pharmacodynamics Over Time Profiles 
The mean profile of smoothed data of MV and raw data of TCO2 for each treatment (i.e., each pooled 
period) will be plotted with overlay of individual subject’s replicated treatments. In addition, mean profiles 
for each treatment will be overlaid in one figure.  
Individual and mean profiles will also be provided for change from baseline and % Change from Baseline 
based on the smoothed data of MV and raw data of TCO2. 
Minute Ventilation (MV): Individual time profiles of MV raw data are generally highly variable. To better 
characterize the magnitude and time course of opioid agonist/antagonist effects on ventilation, medians will 
be moved over a 4-minute centered window continuously to smooth the raw data. In the earlier NAL1003 
study, this 4-minute window was determined to be the shortest window that adequately reduced the 
moment-moment variation while accurately preserving time course differences. 
To avoid any carryover effects from the previous drug, the smoothing algorithm will be applied in a piece-
wise fashion, as follows: 


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•
Data before any fentanyl (without any opioid agonist) administration
•
Data from beginning of fentanyl administration to antagonist administration (only the
effect from fentanyl)
•
Data on and after antagonist administration with continuing fentanyl administration (the
effect of naloxone and any antagonist)
Transcutaneous CO2: Individual time profiles of TCO2 raw data will be used as is. 
Parameters (Prior to Antagonist Administration Phase) 
For MV and TCO2, the following parameters will be calculated to evaluate the action of OIRD for each 
subject and each period, during the period of fentanyl only administration: 
•
Baseline (Pre-treatment value) - Median value of the PD data during the 10 minutes
before the start of Fentanyl
•
OIRD value - Median value of the PD data between -10 and -5 minutes before the start of
the antagonist
9.2 
 Primary Endpoint(s) Analysis 
Change in MV from opioid induced nadir (Opioid Induced Respiratory Depression (OIRD))  
•
Pre-specified Time Point: 5 min after administration of antagonist
The change in MV from OIRD at 5 min after administration of antagonist will be provided by 
treatment in tables and figures with 95% confidence interval presented. 
9.3 
Secondary Endpoint(s) Analysis 
Change in MV from opioid induced nadir (Opioid Induced Respiratory Depression (OIRD)  
•
Pre-specified Time Point: 2.5 min, 10 min, 15 min, 20 min, 90 min after administration of
antagonist
The change in MV from OIRD will be provided by treatment in tables and figures with 95% 
confidence interval presented. 
Maximum of Reversal in MV - The highest MV achieved after administration of antagonist   
•
Time frames: 0-2.5 min, 0-5 min, 0-10 min, 0-15 min, 0-20 min, 0-90 min from
administration of antagonist
The maximum reversal on MV (L/min) will be summarized by treatment in tables with 95% 
confidence interval provided. 


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Time to Maximum of Reversal in MV - Time from administration of antagonist to Maximum 
reversal (change in MV)   
•
Time frames: 0-2.5 min, 0-5 min, 0-10 min, 0-15 min, 0-20 min, 0-90 min from
administration of antagonist
The time to maximum reversal on MV (L/min) will be summarized by treatment in tables with 
95% confidence interval provided.   
To reduce the moment-to-moment variation, additional smoothing windows may be applied to 
assess the robustness of PD results. For example:   
Approach 1: Based on 1-minute window smoothed MV data (Smooth every 1 min of the raw 
data with center smoothing).  
Approach 2: Based on 4-minutes window smoothed MV data (Smooth every 4 mins of the raw 
data with center smoothing)  
9.4 
Exploratory Endpoint(s) Analysis 
Time to Onset (TTO) - onset of reversal 
In the setting of opioid overdose, onset of reversal is expected within minutes following antagonist 
administration. A 30-minute window was selected to ensure the initial onset is fully captured. If 
TTO for X% of reversal is not achieved, then it will be considered a reversal failure and will be 
imputed to 30 minutes. 
Onset of reversal for each treatment will include: 
•
TTO: Time from administration of antagonist (Nalmefene, Narcan) to X% of baseline
reversal, where baseline is defined as median value of the PD of interest during the 10
minutes before the start of Fentanyl
Thresholds (X%) of reversal is defined as X% of baseline reversal and calculated as X %) * 
(Baseline – OIRD) + OIRD, where X are 25, 33, 50, 67, 75, 90, and 100.From the onset of 
reversal from individual periods of each treatment, observed onset of reversal for a pooled period 
is calculated based on: 
•
Eligible analysis set - the observed onset of reversal for a pooled treatment period will be
the average of onset from eligible periods
Bar charts, scatter plots and overlay boxplots of time to onset of reversal will be plotted by 
treatment and pooled treatment periods if data warrants. 
10 Definitions and Conventions for Data Handling 


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10.1 Definition of Baseline 
For all evaluations unless otherwise noted, baseline is defined as the most recent non-missing 
measurement prior to or on the first administration of Fentanyl for qualification phase or each 
period during treatment phase, including unscheduled measurements. Baseline can be the same 
date as first dose, given the measurement is expected prior to first dose when only date information 
is available. 
10.2 Definition of Study Days 
Unless otherwise noted, study days of an evaluation are defined as number of days relative to the 
first dosing date of Fentanyl of qualification or treatment phase which is designated as Day 1, and 
the preceding day is Day -1, the day before that is Day -2, etc.  
•
If evaluation date is on or after first dose date, then study days are calculated as
−
Evaluation date minus - first dose date of study drug + 1
•
If evaluation date is before first dose date, then relative study days are calculated as
−
Evaluation date - first dose date of study drug
10.3 First Dose Date of Study Treatment 
The date of first dose of study treatment is defined as the earliest dose date of study drugs in the 
treatment period. 


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11 REFERENCES 
None.