Clinical trial NCT03224299
Conditions: Preoperative Skin Preparation
Sponsor: Becton, Dickinson and Company
Start date: 2017-06-21; Completion date: 2017-10-03
Enrollment: 131 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
Objective of this study is to compare the antimicrobial properties of an investigational agent to an active control and a negative control when applied to specific body locations. Testing will be performed according to the procedures outlined in the Food and Drug Administration Tentative Final Monograph (TFM) for Effectiveness Testing of a Patient Preoperative Skin Preparation

Detailed description:

Arms: arm 1: octenidine dihydrochloride in isopropyl alcohol in a single-use applicator - clear arm 2: octenidine dihydrochloride in isopropyl alcohol in a single-use applicator - tinted arm 3: ChloraPrep® - Hi-Lite Orange® applicator arm 4: sterile 0.9% saline applied with single use applicator
Interventions: intervention 1: OCT - clear intervention 2: OCT- tinted intervention 3: ChloraPrep intervention 4: Saline
Intervention descriptions: intervention 1: Investigational Product #1: octenidine dihydrochloride in isopropyl alcohol in a single-use applicator - clear intervention 2: Investigational product #2: octenidine dihydrochloride in isopropyl alcohol in a single-use applicator - tinted intervention 3: Active Control: ChloraPrep® - Hi-Lite Orange® applicator intervention 4: Negative Control: sterile 0.9% saline applied with single use applicator

Protocol documents:
SAP_002.pdf: 
Corporate Clinical Development
Template CCD-6230.T01
Revision 01
Statistical Analysis Plan
Protocol Number/version
MPS-16IPVFT05 Amendment #1 (April 19, 2017)
Protocol Title
Statistical
Analysis
Plan
for
MPS-16IPVFT05:
A
RANDOMIZED,
SINGLE-CENTER, CLINICAL PHASE II EVALUATION OF THE TIME-
DEPENDENT ANTIMICROBIAL EFFECTIVENESS OF OCTENIDINE DI-
HYDROCHLORIDE IN ISOPROPYL ALCOHOL FOR PREOPERATIVE
SKIN PREPARATION
SAP Date
August 2, 2017
Version History
Action
June 15, 2017
Original Release
August 2, 2017
Per Protocol Amendment #3, the order of the comparisons for non-inferiority was changed
per communication with FDA dated June 6, 2017. Instead of using the wording of log
reduction for the primary analysis, the wording of average treatment eﬀect was used.
For product expression volumes, inclusion of data points into Modiﬁed Intent to Treat
(mITT) analysis will be based on whether the matching data point is included for eﬃcacy
for average treatment eﬀect instead of for responder rate. Added a list of abbreviations
and deﬁnitions of terms.
This report is proprietary to Becton Dickinson. No part of it may be distributed or shared with persons other
than BD associates without written permission from Corporate Clinical Development (CCD).
Approval
Signature below indicates approval of the statistical analysis plan as written.
Function
Name
Signature
Statistician
(Author)
Lingzhi Li
This document is signed electronically in the
eTMF system.
Clinical Trial
Manager
Carole Schatz
This document is signed electronically in the
eTMF system.
Statistics
Manager
Julie B´erub´e
This document is signed electronically in the
eTMF system.
MPS-16IPVFT05 Statistical Analysis Plan


Contents
1
List of Abbreviations and Deﬁnitions of Terms
3
2
Executive Summary
3
3
Overall Study Description
3
3.1
Study Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3
3.2
Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3
3.3
Study Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4
3.4
Endpoints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
6
4
Sample Size
6
5
Intended Statistical Software
6
6
Data
7
6.1
Data Sets Analyzed
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
7
6.2
Analysis Population Set(s) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
8
7
Statistical Analysis/Calculations
10
7.1
Derived Variables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.1.1
Log CFU/cm2 of skin
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.1.2
Log Reduction
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.1.3
Responder Rate
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.1.4
Product Expression Volumes
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.2
Handling of Missing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
7.3
Analysis Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
12
7.3.1
Primary Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
12
7.3.2
Secondary Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
13
7.3.3
Exploratory Analyses
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
14
7.3.3.1
Summary Statistics
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
14
7.3.3.2
Log Reduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
14
7.3.3.3
Product Expression Volumes/Weight
. . . . . . . . . . . . . . . . . . . . . . . .
15
7.3.4
Safety Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
15
7.4
Graphs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
17
7.4.1
ATE Per Primary Analysis
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
17
8
Appendix
19
MPS-16IPVFT05 Statistical Analysis Plan
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1
List of Abbreviations and Deﬁnitions of Terms
AC
Active Control
AE
Adverse Event
ATE
Average Treatment Eﬀect
BD
Becton Dickinson and Company
CI
Conﬁdence Interval
Conf Int
Conﬁdence Interval
IP
Investigational Product
ITT
Intent To Treat
LSMean
Least Squares Mean
mITT
Modiﬁed Intent To Treat
NC
Negative Control
SD
Standard Deviation
2
Executive Summary
For average treatment eﬀect / responder rate, the per-protocol data may be diﬀerent from the modiﬁed intent to
treat (mITT) data and analysis will be made separately based on both the per-protocol data and the modiﬁed
intent to treat (mITT) data if two data sets are diﬀerent. Summary statements will be made for each analysis
to show if there are statistically signiﬁcant diﬀerences found, or if Eﬃcacy Standards are met for diﬀerent
comparisons. Links to tables showing analysis results will be provided.
3
Overall Study Description
3.1
Study Background
Pilot clinical studies have been completed by BD in healthy volunteers to assess the safety and eﬃcacy of
topical application at various concentrations of octenidine dihydrochloride in isopropyl alcohol as summarized
in the Investigator’s Brochure (IB). No drug-related adverse events (AEs) have been observed. Additionally, a
clinical study was conducted to measure the systemic absorption of octenidine after topical administration of
octenidine dihydrochloride in isopropyl alcohol under simulated maximal clinical usage to intact and abraded
skin in healthy volunteers. No measurable octenidine in plasma was observed (lower limit of quantiﬁcation was
1 ng/mL).
3.2
Objectives
The primary objective of this study is to compare the immediate antimicrobial properties of octenidine dihy-
drochloride [OCT] in isopropyl alcohol - clear in a single-use applicator and octenidine dihydrochloride [OCT]
in isopropyl alcohol - tinted in a single-use applicator to a Negative Control and an FDA NDA-approved Active
Control. Single use applicators ﬁlled with 0.9% saline will serve as the Negative Control and ChloraPrep® -
Hi-Lite Orange® applicator will serve as the Active Control. In agreement with the Food and Drug Adminis-
tration Tentative Final Monograph (TFM) for Eﬀectiveness Testing of a Patient Preoperative Skin Preparation,
a log10 reduction study (i.e., average treatment eﬀect) will be used to determine antimicrobial eﬃcacy based on
the statistical analyses outlined in published letters from the FDA on January 19, 2017.
The secondary objective is to determine the persistent antimicrobial activity of the two Investigative Products,
Active Control and Negative Control by responder rate. Log10 bacterial counts at 6 hours for each treatment
site will be converted to a binary (yes or no) success measure with success deﬁned as having skin ﬂora counts
that are less than or equal to the baseline skin ﬂora counts.
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3.3
Study Design
This is a single-center and randomized design where each subject receives two diﬀerent treatments out of 4
treatments, one on the left side of the body and one on the right side. This means there are 6 possible combi-
nations of treatments (assuming A, B and C are either Investigational Product #1, Investigational Product #2
or Active Control, D is the Negative Control): A & B, A & C, A & D, B & C, B & D, and C & D.
A minimum of 72 evaluable subjects will be required to achieve a total of at least 144 test sites for groin and
abdominal regions. To achieve 36 evaluable test sites from Investigational Product 1, Investigational Product
2, Active Control and Negative Control arms for groin and abdominal regions, of the minimum 72 evaluable
subjects:
 12 subjects will receive treatments from both A and B.
 12 subjects will receive treatments from both A and C.
 12 subjects will receive treatments from both A and D.
 12 subjects will receive treatments from both B and C.
 12 subjects will receive treatments from both B and D.
 12 subjects will receive treatments from both C and D.
The two investigational products and the active control will be randomly assigned codes of A, B or C. However,
due to visual diﬀerences between each of the test materials, the products cannot be blinded to all study staﬀ.
In order to guard against any bias of the study outcome, technicians who participate in product application
or sample collection from subjects during treatment day procedures will not participate in the processing of
samples. The technicians processing samples and/or counting the resultant plates will be blinded to the study
randomization during the data gathering processes. Plate counts will be reviewed and recorded by separate
BSLI staﬀprior to being entered into a data spreadsheet where product information (unblinding) would occur.
Subjects will be sampled for treatment day baseline microbial populations on randomly assigned sites of the
abdominal and inguinal areas on both sides. Subjects will have samples collected at baseline, 30 seconds (± 5
seconds), 10 minutes (± 30 seconds) and at 6 hours (± 30 minutes) post-treatment application on both abdomen
and inguinal sites. All sampling times will be calculated from the completion of the dry time of each product
following application.
There is a neutralization test before the main treatment study. This test is to assure that the neutralizers used
in the recovery medium quench the antimicrobial activity of the three test materials (The Investigational Prod-
ucts and Active Control), and that the neutralizers are not toxic to the bacteria. Note: this statistical analysis
plan is only for the Main Treatment study and does not include the analysis for the Neutralization study. The
analysis for the Neutralization study will be conducted by the clinical site and included in the ﬁnal clinical report.
The group treatments and number of treatment sites for each group for the main treatment study are shown in
Table 1.
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Table 1: Group treatments and number of treatment sites for the main treatment part
Product Code
Arm
Treatment
Number of evaluable
treatment sites per
anatomical site
A, B or C
Investigational Product #1 (IP1)
Octenidine Dihydrochloride in Isopropyl
Alcohol in a single-use applicator - clear
36
A, B or C
Investigational Product #2 (IP2)
Octenidine Dihydrochloride in Isopropyl
Alcohol in a single-use applicator -tinted
36
A, B or C
Active Control (AC)
ChloraPrep® - Hi-Lite Orange® applicator
36
D
Negative Control (NC)
Sterile 0.9% saline applied with single use
applicator
36
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3.4
Endpoints
 Primary Endpoints
– Log CFU/cm2 of skin for each treatment per body site per time point.
 Secondary Endpoints
– Responder rates will be calculated by converting each log10 CFU/cm2 reduction at 6 hours post-
treatment time point to a binary (yes or no) success measure. Success at 6 hours post-treatment
time point is deﬁned as having skin ﬂora counts that are less than or equal to the baseline skin ﬂora
counts on both body sites.
 Exploratory Endpoints
– Log10 CFU/cm2 reductions from baseline for each treatment per body site per post-application time
point.
– Product expression volumes: the weight of the treatment applied to a test site will be estimated by
subtracting the weight measurement (g) of the treatment after the application from the weight of
the treatment (g) before application.
 Safety Endpoints
– Skin irritation scores (Erythema, Edema, Rash and Dryness scores for each time point)
4
Sample Size
A suﬃcient number of volunteers will be enrolled in the screening phase such that the total numbers of evaluable
samples collected from groin and abdominal regions are not less than 36 evaluable test sites from each treatment
arm (minimum of 144 each for groin and abdominal regions). A minimum of 72 evaluable subjects is required
to achieve a total of at least 144 test sites for groin and abdominal regions. Subjects must satisfy all Screening
Day and Treatment Day Inclusion/Exclusion Criteria prior to screening sample collection and Treatment Day
procedures. If the required numbers of subjects do not qualify from the initial screening group, additional
volunteers will be recruited.
This is a Phase II study to assess the immediate and persistent antimicrobial eﬀect of the Investigational
Products relative to the Negative Control and the Active Control using a log10 reduction study (i.e., average
treatment eﬀect) according to the procedures outlined in the Food and Drug Administration Tentative Final
Monograph (TFM) for Eﬀectiveness Testing of a Patient Preoperative Skin Preparation (FR 59:116, 17 June
94, pp. 31450-31452) and the test criteria outlined in the deferal letters from the FDA published on January
19, 2017. The purpose is to inform design of future eﬃcacy studies, therefore no sample size calculation was
performed to provide statistically-powered evidence of eﬃcacy as part of this study. The sample size is deemed
adequate for these purposes.
5
Intended Statistical Software
R 3.2.0 (2015-04-16) or later version if updated.
MPS-16IPVFT05 Statistical Analysis Plan
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6
Data
6.1
Data Sets Analyzed
Descriptions of the modiﬁed intent to treat (mITT) data, per-protocol data and intent to treat (ITT) data are
shown below. Inclusion for the per-protocol data / mITT data set is evaluated for each body area (left and
right for the groin and abdomen).
 Modiﬁed Intent to Treat (mITT) data: data collected will be included if the treatment-day baseline counts
are within the acceptable range (≥3.00 - ≤5.50 log10 CFU/cm2 on abdominal area, and ≥5.00 - ≤7.50
log10 CFU/cm2 on inguinal area). Data collected will be excluded from mITT data if the treatment-day
baseline counts are outside the acceptable range.
– For responder rate at 6 hours post-treatment time point, missing data at either baseline or a certain
time interval will be imputed as non-responders for that time interval in the mITT data.
– For average treatment eﬀect (ATE) and log10 reduction, missing data will not be imputed for mITT
data.
 Per-Protocol data: data collected will be included if the treatment-day baseline counts are within the
acceptable range (≥3.00 - ≤5.50 log10 CFU/cm2 on abdominal area, and ≥5.00 - ≤7.50 log10 CFU/cm2
on inguinal area). Data collected will be excluded from per-protocol data if the treatment-day baseline
counts are outside the acceptable range.
– Missing data at any time interval will not be imputed for ATE, responder rate and log reduction.
Missing data will be excluded from per-protocol ATE, responder rate and log reduction analysis.
 Intent to Treat (ITT) data: all randomized subjects with available data will be included and used for the
ITT analysis. The full intent to treat (ITT) data set (all randomized subjects) will be used for the safety
analysis.
For ATE, responder rate and log reduction, eﬃcacy analyses will be ﬁrst made on the modiﬁed intent to treat
(mITT) data set. Eﬃcacy analyses will be also conducted on the per-protocol data set as supportive analyses
when per-protocol data are diﬀerent from mITT data.
For product expression volumes, inclusion of data points will be based on whether the matching data point is
included for eﬃcacy for ATE. Analysis will be performed based on the per-protocol data as supportive analyses
when per-protocol data are diﬀerent from mITT data for product expression volumes.
For skin irritation scores, analysis will be performed based on the intent to treat (ITT) data (all randomized
subjects).
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6.2
Analysis Population Set(s)
For each treatment per body site, the number of subjects randomized, the number of subjects available for
mITT analysis [average treatment eﬀect (ATE), responder rate and product expression volumes] and / or for
per-protocol analysis [average treatment eﬀect (ATE), responder rate and product expression volumes] at each
post-treatment time point (30 seconds, 10 minutes and 6 hours) will be provided in tables below (cf. Table
2, Table
3 and Table
4). If the number of subjects randomized is diﬀerent from the number of subjects
randomized with treatments received, the number of subjects randomized with treatment received will be also
summarized.
Table 2: Number of subjects randomized
Treatment
Body Site
N Subjects Randomized with Treatment Received
A
Abdomen
B
Abdomen
C
Abdomen
D
Abdomen
A
Groin
B
Groin
C
Groin
D
Groin
Table 3: Number of subjects available for mITT analysis
Treatment
Body Site
N Subjects
for mITT
ATE at 30
sec
N Subjects
for mITT
ATE at 10
min
N Subjects
for mITT
ATE at 6
hr
N Subjects
for mITT
Responder
Rate at 6
hr
N Subjects
for mITT
Product
Expression
Volumes
A
Abdomen
B
Abdomen
C
Abdomen
D
Abdomen
A
Groin
B
Groin
C
Groin
D
Groin
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Table 4: Number of subjects available for per-protocol analysis
Treatment
Body Site
N Subjects
for Per-
Protocol
ATE at 30
sec
N Subjects
for Per-
Protocol
ATE at 10
min
N Subjects
for Per-
Protocol
ATE at 6
hr
N Subjects
for Per-
Protocol
Responder
Rate at 6
hr
N Subjects
for
Per-Protocol
Product
Expression
Volumes
A
Abdomen
B
Abdomen
C
Abdomen
D
Abdomen
A
Groin
B
Groin
C
Groin
D
Groin
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7
Statistical Analysis/Calculations
7.1
Derived Variables
7.1.1
Log CFU/cm2 of skin
The estimated log10 number of viable microorganisms per cm2 recovered from each sample site will be designated
the “R-value”. To convert the volumetric measure of the sample into the number of colony-forming units per
square centimeter (cm2), the following formula will be employed:
R = log10
"F
 P2
i=1 Ci
n

10-D
A
#
(1)
Where:
R = the average colony-forming unit count in log10 scale per cm2 of sampling surface
F = Total number of mL of sampling solution added to the sampling cylinder (in this study, F = 6 mL for all
samples)
P2
i=1 Ci
n
= average of the duplicate colony counts used for each sample collected (n = 2)
D = Dilution factor of the plates counted
A = Inside area of the sampling cylinder (A= 3.46 cm2 in this study)
Note: The reason that a log10 transformation will be performed on the collected data is to obtain a linear scale.
A linear scale, more appropriately a log10 linear scale, is a basic requirement of the statistical models used in
this study.
If colonies on one of the plates are uncountable, the count from the remaining plate will be used.
In order to avoid potential calculation problems due to taking the logarithm of zero, counts of less than 1
CFU/cm2 are treated as 1 CFU/cm2, such that the log10 transformation is no less than zero.
7.1.2
Log Reduction
Log10 CFU/cm2 reductions from baseline will be calculated separately for each subject, each post-application
time point, and each of the treated anatomical sites by taking the baseline log10 CFU/cm2 values and then
subtracting the log10 CFU/cm2 values for the samples taken after the baseline.
7.1.3
Responder Rate
Responder statuses for each subject will be evaluated separately for each of the four sites and separately at 6
hours post-application time point based on the log10 CFU/cm2 changes from baseline. A subject is considered
a responder at the 6-hour sample time for the abdomen or groin if the log10 CFU/cm2 reduction is equal to or
greater than 0.
7.1.4
Product Expression Volumes
The weight of drug product applied to a test site will be estimated by subtracting the weight measurement of
the test material after the application from the weight of the test material before application.
7.2
Handling of Missing Data
For mITT responder rate analysis, subjects with missing data at either baseline or a certain time interval will
be included as non-responders for that time interval. Responder rate analysis will be performed based on both
MPS-16IPVFT05 Statistical Analysis Plan
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per-protocol data and mITT data if there are any missing data identiﬁed and if per-protocol data and mITT
data are diﬀerent.
For average treatment eﬀect (ATE), log reduction and product expression volumes, missing data will not be
imputed for mITT data. Analysis will be performed based on the per-protocol data as supportive analyses when
per-protocol data are diﬀerent from mITT data.
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7.3
Analysis Methods
For each body site, when per-protocol data are diﬀerent from mITT data for average treatment eﬀect (ATE),
responder rate, log reduction or product expression volumes, per-protocol analysis and mITT analysis will be
performed separately using the same statistical methods. The following analysis methods will be applied to
both per-protocol analysis and mITT analysis, both abdomen and groin.
An alpha level of 5% (two-sided) is used for all analyses.
7.3.1
Primary Analysis
The primary purpose of the study is to compare the immediate antimicrobial activity of the Investigational
Products to the Negative Control and Active Control according to the methods described in the FDA TFM
with the statistical analyses outlined in a deferral letter published January 19, 2017. There are two test criteria
that drive the hypotheses.
 Evaluate the average treatment eﬀect (ATE) of the Active Control (AC) compared to each Investigational
Product (IP) at 30 seconds and 10 minutes for non-inferiority on both abdomen and groin. ATE of the
AC compared to either IP is deﬁned as the contrast of the treatment eﬀect of the IP minus the treatment
eﬀect of the AC (IP - AC) in the linear regression. Statistical hypotheses are:
– Ho: The upper bound 95% conﬁdence interval of the ATE of IP - AC ≥0.5 log10
– Ha: The upper bound 95% conﬁdence interval of the ATE of IP - AC < 0.5 log10
 Evaluate the Average Treatment eﬀect (ATE) of each Investigational Product (IP) compared to the
Negative Control (NC) at 30 seconds and 10 minutes on both abdomen and inguen anatomical area. ATE
of the IP compared to the NC is deﬁned as the contrast of the treatment eﬀect of the NC minus the
treatment eﬀect of the IP (NC - IP) in the linear regression. Statistical hypotheses are:
– Ho: The lower bound 95% conﬁdence interval of the ATE of NC - IP ≤1.2 log10
– Ha: The lower bound 95% conﬁdence interval of the ATE of NC - IP > 1.2 log10
Ho is the null hypothesis and Ha is the alternative hypothesis.
A linear regression model for each body site will be used for primary analysis. In the model, the response
is the post-treatment log10 bacterial counts and predictors are the treatment eﬀect as a ﬁxed eﬀect and the
pre-treatment log10 bacterial counts as a covariate. The interaction between the treatment and pre-treatment
log10 bacterial counts will be also explored. If there is no signiﬁcant interaction detected between treatment and
pre-treatment log10 bacterial counts, the interaction term will be removed from the model and the average dif-
ference between treatments will be estimated from the linear regression model after correcting for pre-treatment
log10 bacterial counts. If there is a signiﬁcant interaction detected between treatment and pre-treatment log10
bacterial counts, the average diﬀerence between treatments will be estimated from the linear regression model
at the average of the covariate. Additional factors (e.g., technician and subject demographics) may be also
included in the model and the eﬀects will be investigated.
Since the study will declare success when and only when both non-inferiority and superiority are demonstrated
for both abdomen and groin, no multiplicity adjustment for multiple primary objectives is required.
An example for results of comparisons above can be found in Tables 5 and 6. All results that do not meet the
targets are highlighted.
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Table 5: Example: ATE for IP1/IP2 - AC for Groin
Comparison
Time Point
N Pairs
Diﬀerence in ATE
Conf Int
Meet Non-inferiority Criteria
IP1 - AC
30 Seconds
36
-0.50
-1.31, 0.30
Yes
IP2 - AC
30 Seconds
36
-0.35
-1.00, 0.31
Yes
IP1 - AC
10 Minutes
36
0.32
-0.52, 1.15
No
IP2 - AC
10 Minutes
36
0.50
-0.39, 1.39
No
Table 6: Example: ATE for NC - IP1/IP2 for Groin
Comparison
Time Point
N Pairs
Diﬀerence in ATE
Conf Int
Meet Superiority Criteria
NC - IP1
30 Seconds
36
1.35
1.01, 1.59
No
NC - IP2
30 Seconds
36
1.23
0.90, 1.50
No
NC - IP1
10 Minutes
36
0.4
0.02, 0.82
No
NC - IP2
10 Minutes
36
1.6
1.28, 1.91
Yes
7.3.2
Secondary Analysis
The secondary purpose of the study is to determine the persistent antimicrobial activity of the two Investiga-
tional Products, Active Control and Negative Control by calculating the responder rate along with 95% lower
conﬁdence bound at 6-hour post-treatment time point for each body site using an Exact method (one-sided).
The goal is to demonstrate a success proportion ≥95% on both the abdomen and the inguen with 95% conﬁ-
dence. However, since this is a Phase II study to inform design of future eﬃcacy studies and is not designed to
provide statistically powered evidence of eﬃcacy, the lower conﬁdence bounds cannot lie entirely above 95%.
An example of comparing responder rates to 95% for each treatment is provided below.
Table 7: Example: Analysis results for responder rate for Groin
Treatment
Time Point
N Non-responder
N Responder
N Total
Responder Rate
95% Lower Conﬁdence Bound
A
6 Hours
1
35
36
97.2%
87.5%
B
6 Hours
0
36
36
100.0%
92.0%
C
6 Hours
2
34
36
94.4%
83.5%
D
6 Hours
1
35
36
97.2%
87.5%
MPS-16IPVFT05 Statistical Analysis Plan
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7.3.3
Exploratory Analyses
7.3.3.1
Summary Statistics
Summary statistics will be provided for log10 CFU/cm2 of skin, log10 CFU/cm2 reduction and product expres-
sion volumes based on mITT data and/or per-protocol data. For continuous variables, data will be summarized
with the following descriptive statistics: number of observations, mean (LSMean), median, standard deviation
and range (minimum - maximum).
See examples for summary statistics Tables
8 and
9 below for log10
CFU/cm2 and log10 CFU/cm2 reduction.
Table 8: Example: summary statistics for log10 CFU/cm2 for Groin
log10 CFU/cm2
Treatment
Time Point
N
Mean
LSMean
Median
SD
Range
log10 CFU/cm2
A
Baseline
36
5.92
5.92
5.82
0.53
5.01 - 7.23
log10 CFU/cm2
A
30 Seconds
36
3.29
3.29
3.44
1.10
0.89 - 5.89
log10 CFU/cm2
A
10 Minutes
36
3.05
3.05
3.09
0.72
1.22 - 4.09
log10 CFU/cm2
A
6 Hours
36
5.78
5.78
5.66
0.54
5.05 - 7.04
log10 CFU/cm2
B
Baseline
36
3.27
3.27
3.43
1.09
0.00 - 4.95
log10 CFU/cm2
B
30 Seconds
36
3.26
3.26
3.39
0.89
0.00 - 4.47
log10 CFU/cm2
B
10 Minutes
36
5.92
5.92
5.82
0.53
5.01 - 7.23
log10 CFU/cm2
B
6 Hours
36
3.27
3.27
3.43
1.09
0.00 - 4.95
log10 CFU/cm2
C
Baseline
36
3.26
3.26
3.39
0.89
0.00 - 4.47
log10 CFU/cm2
C
30 Seconds
36
3.05
3.05
3.09
0.72
1.22 - 4.09
log10 CFU/cm2
C
10 Minutes
36
3.29
3.29
3.44
1.10
0.89 - 5.89
log10 CFU/cm2
C
6 Hours
36
3.05
3.05
3.09
0.72
1.22 - 4.09
log10 CFU/cm2
D
Baseline
36
5.78
5.78
5.66
0.54
5.05 - 7.04
log10 CFU/cm2
D
30 Seconds
36
3.27
3.27
3.43
1.09
0.00 - 4.95
log10 CFU/cm2
D
10 Minutes
36
5.92
5.92
5.82
0.53
5.01 - 7.23
log10 CFU/cm2
D
6 Hours
36
3.29
3.29
3.44
1.10
0.89 - 5.89
Table 9: Example: summary statistics for log10 CFU/cm2 reduction for Groin
log10 CFU/cm2 reduction
Treatment
N
Mean
LSMean
Median
SD
Range
log10 CFU/cm2 reduction
A Baseline - 30 Seconds
36
2.63
2.63
2.57
1.18
0.20 - 4.80
log10 CFU/cm2 reduction
A Baseline - 10 Minutes
36
2.83
2.83
2.83
0.89
1.24 - 4.86
log10 CFU/cm2 reduction
A Baseline - 6 Hours
36
2.52
2.52
2.33
1.09
0.83 - 5.50
log10 CFU/cm2 reduction
B Baseline - 30 Seconds
36
2.53
2.53
2.30
1.11
0.90 - 5.50
log10 CFU/cm2 reduction
B Baseline - 10 Minutes
36
2.63
2.63
2.57
1.18
0.20 - 4.80
log10 CFU/cm2 reduction
B Baseline - 6 Hours
36
2.53
2.53
2.30
1.11
0.90 - 5.50
log10 CFU/cm2 reduction
C Baseline - 30 Seconds
36
2.83
2.83
2.83
0.89
1.24 - 4.86
log10 CFU/cm2 reduction
C Baseline - 10 Minutes
36
2.52
2.52
2.33
1.09
0.83 - 5.50
log10 CFU/cm2 reduction
C Baseline - 6 Hours
36
2.63
2.63
2.57
1.18
0.20 - 4.80
log10 CFU/cm2 reduction
D Baseline - 30 Seconds
36
2.52
2.52
2.33
1.09
0.83 - 5.50
log10 CFU/cm2 reduction
D Baseline - 10 Minutes
36
2.83
2.83
2.83
0.89
1.24 - 4.86
log10 CFU/cm2 reduction
D Baseline - 6 Hours
36
2.53
2.53
2.30
1.11
0.90 - 5.50
7.3.3.2
Log Reduction
Log10 reductions will be compared between test materials by an Analysis of Variance (ANOVA) for each body
site based on mITT data and/or per-protocol data. In the ANOVA model, the response is the log10 reduction
and the predictors are treatment and time point as well as the interaction between treatment and time point.
The average diﬀerence in log reduction between treatments per time point will be estimated. However, all
conclusions for the diﬀerence in the average treatment eﬀect between treatments will be based on the primary
analysis only.
An example for results of comparisons above can be found in Tables 10.
MPS-16IPVFT05 Statistical Analysis Plan
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Table 10: Example: results for product comparisons within time interval (10 minutes) for Groin
Comparison
N Pairs
Diﬀerence
Conf Int
Conclusion
IP1 Baseline-A 10 min vs. AC Baseline-AC 10 min
36
0.13
-0.44, 0.70
no sign. diﬀ.
IP2 Baseline-A 10 min vs. AC Baseline-AC 10 min
36
0.11
-0.65, 0.87
no sign. diﬀ.
IP1 Baseline-A 10 min vs. VC Baseline-VC 10 min
36
0.33
-0.44, 1.09
no sign. diﬀ.
IP2 Baseline-A 10 min vs. VC Baseline-AC 10 min
36
0.33
-0.44, 1.09
no sign. diﬀ.
7.3.3.3
Product Expression Volumes/Weight
An analysis of variance (ANOVA) of the applied volumes/weight will be performed separately for each body
site based on mITT data and/or per-protocol data. In the model, there will be a random subject eﬀect, a ﬁxed
body side (left or right) eﬀect, a ﬁxed treatment eﬀect and the interaction between ﬁxed eﬀects. ANOVA table
will be checked to see whether the treatment has a signiﬁcant eﬀect on product expression volumes as well as
the interaction. If a signiﬁcant diﬀerence is found, subgroup comparisons with Tukey’s method will be made to
determine which pair of comparisons shows signiﬁcant diﬀerence.
7.3.4
Safety Analysis
The ITT data set (all randomized subjects) will be considered evaluable for safety. Skin irritation scores will
be reported for any subject who is scored with a 1 or more at any observation [baseline, post-application/prior
to 30 seconds, 10 minutes, and 6 hours sampling procedures], in any category for any site.
Adverse Events (including post treatment skin irritation scores of 3), will also be summarized. Summary tables
will present incidence rates of adverse events by treatment group for all subjects who enter the treatment period.
Listings of Adverse Events will be provided.
The statistical signiﬁcance of diﬀerences in skin irritation between the Investigational Products, Active Control
and Negative Control will be evaluated using Fisher’s exact test on skin irritation data summarized as follows:
any reaction above zero (no reaction) on the skin irritation rating scale for any category (erythema, edema,
rash, and dryness) will be considered a positive signal for that substance. If Fisher’s exact test shows there
is an overall signiﬁcant treatment eﬀect, Fisher’s exact test or two-proportion test with Score method with
multiplicity adjustment on the alpha level may be used for multiple sub-group comparisons to ﬁnd out which
comparisons show the diﬀerence. Odds ratio (odds of having positive signals for one treatment / odds of having
positive signals for the other treatment) using Fisher’s exact test or the diﬀerence in percentage of positive
signals between treatments using Two-proportion test may be calculated along with the conﬁdence interval
for each category at the certain time interval. The analysis for skin irritation scores will be for informational
purposes only.
Examples for Fisher’s exact test and two-proportion test with Score method are shown below.
MPS-16IPVFT05 Statistical Analysis Plan
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> # overall treatment effect
> irr_tab <- matrix(c(2, 1, 3, 1, 34, 35, 33, 35),
+
nrow = 4,
+
dimnames =
+
list(c("A", "B", "C", "D"),
+
c("Positive (Score > 0)", "Non-positive (Score = 0)")))
> irr_tab
Positive (Score > 0) Non-positive (Score = 0)
A
2
34
B
1
35
C
3
33
D
1
35
> irr <- fisher.test(irr_tab, alternative = "two.sided",
+
conf.level = 0.95)
> print(irr)
Fisher's Exact Test for Count Data
data:
irr_tab
p-value = 0.8361
alternative hypothesis: two.sided
> # subgroup comparisons
> library(PropCIs)
> print(diffscoreci(2, 34, 1, 35, conf.level = 0.972))
data:
97.2 percent confidence interval:
-0.1176006
0.1904725
MPS-16IPVFT05 Statistical Analysis Plan
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7.4
Graphs
7.4.1
ATE Per Primary Analysis
The graph for the ATE of IP1/IP2 - AC with 95% intervals estimated from the regression model (as in Tables
5 and 6) will be provided (see examples in Figure 1 and 2).
Groin
G
G
G
G
−1.0
−0.5
0.0
0.5
1.0
1.5
IP1 − AC 30 Seconds
IP2 − AC 30 Seconds
IP1 − AC 10 Minutes
IP2 − AC 10 Minutes
Treatment per Time Point
Difference in ATE with 95% CI
Example: ATE of IP1/IP2 − AC for Groin
Figure 1: ATE of IP1/IP2 - AC for Groin
MPS-16IPVFT05 Statistical Analysis Plan
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G
G
G
G
0.0
0.5
1.0
1.5
2.0
NC − IP1 30 Seconds
NC − IP2 30 Seconds
NC − IP1 10 Minutes
NC − IP2 10 Minutes
Treatment per Time Point
Difference in ATE with 95% CI
Example:  ATE of NC − IP1/IP2 for Groin
Figure 2: ATE of NC - IP1/IP2 for Groin
MPS-16IPVFT05 Statistical Analysis Plan
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8
Appendix
Subjects not passing the treatment-day baseline criteria will be listed.
Subjects included in mITT analysis, but excluded from per-protocol analysis, if any, will be listed.
MPS-16IPVFT05 Statistical Analysis Plan
Page 19 of 19


Signature Page for MPS-16IPVFT05_Statistical_Analysis_Plan_V2 
Signature Page for MPS-16IPVFT05_Statistical_Analysis_Plan_V2 - System Version N
Approved
Lingzhi Li Statistician, Corporate/Shared Services
Statistician
14-Aug-2017 20:13:12 GMT+0000
Approved
Carole Schatz
Study Manager
14-Aug-2017 20:45:59 GMT+0000
Approved
Julie Berube Manager Statistics, Corporate/Shared
Services
Functional Management
15-Aug-2017 12:28:16 GMT+0000

Prot_001.pdf: