Clinical trial NCT03616223
Conditions: Sensorineural Hearing Loss
Keywords: Intratympanic Administration Hearing Restoration
Sponsor: Frequency Therapeutics
Start date: 2018-07-03; Completion date: 2018-12-18
Enrollment: 23 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This is a phase 1/2 study of FX-322 at two dose levels compared to placebo in male and female adults otherwise healthy with stable sensorineural hearing loss.

Detailed description:
Sensorineural hearing loss (SNHL) accounts for about 90% of all cases of hearing loss. The study will assess the safety of FX-322 (laduviglusib and sodium valproate) given as a single intratympanic injection in subjects with a medical history of sensorineural hearing loss that is associated with noise exposure or sudden hearing loss. Safety will be evaluated both systemically (lab and clinical monitoring) and locally (otoscopy and audiometry) in 24 subjects, and a blood PK profile of FX-322 will also be determined.

Arms: arm 1: Cohort of 8. Single intratympanic injection arm 2: Cohort of 8. Single intratympanic injection arm 3: Cohort of 4. Single intratympanic injection arm 4: Cohort of 4. Single intratympanic injection
Interventions: intervention 1: FX-322 intervention 2: Placebo
Intervention descriptions: intervention 1: Intratympanic injection of FX-322 consists of laduviglusib and sodium valproate intervention 2: Intratympanic injection

Protocol documents:
Prot_000.pdf: 
Frequency Therapeutics  
Page 1 of 42 
Version 3.0 02Aug2018  
Page 1 of 42 
CLINICAL STUDY PROTOCOL 
A Phase 1/2 Randomized, Double-blind, Placebo-controlled Single Dose 
Study at Two Dose Levels of FX-322 Administered by Intratympanic 
Injection in Adults with Stable Sensorineural Hearing Loss 
Protocol Number: 
FX-322-201 
EudraCT Number: 
N/A 
Investigational Product: 
FX-322 
Phase: 
Phase 1/2 
Sponsor: 
Frequency Therapeutics 
19 Presidential Way 
Woburn. MA 01801 
USA 
Protocol Date: 
02 Aug  2018 
Protocol Version: 
Version 3.0 
CONFIDENTIAL 
This protocol may not be reproduced or communicated to a third party without the 
written permission of Frequency Therapeutics. 




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CONFIDENTIAL 
2 
SYNOPSIS 
Protocol Number: 
FX-322-201 
Title: 
A Phase 1/2 Randomized, Double-blind, Placebo-controlled Single Dose Study at Two Dose 
Levels of FX-322 Administered by Intratympanic Injection in Adults with Stable 
Sensorineural Hearing Loss  
Investigational Product: 
FX-322 
Study Centers: 
2-5 centers in the US
Phase: 
Phase 1/2 
Objectives: 
Primary objectives: 
•
To assess the systemic safety of FX-322
•
To assess the plasma pharmacokinetic (PK) profile to determine the systemic
exposure to the active pharmaceutical ingredients
•
To assess the effect of FX-322 on otologic and audiologic measures
Study Design: 
This is a Phase 1/2 randomized, double-blind, placebo-controlled, single dose study at two 
dose levels of FX-322 compared to placebo in adults with stable sensorineural hearing loss 
(no changes over 6 months of 10 dB or more in any frequency).  The 2 dose levels proposed 
in this study, FX-322L and FX-322H will be dosed concurrently.   
The study will have 3 phases: Screening, Treatment with Observation, and Follow-up. 
Screening:  Can occur up to 30 days prior to study drug administration (Day 1). All subjects 
will be screened to determine study eligibility. 
Double Blind Treatment with Observation: Approximately 24 subjects are planned to be 
randomized to one of four treatment groups to receive a single dose of either FX-322L, 
placeboL, FX-322H, or placeboH.  Randomized subjects will be allocated 1:1 to one of 2 
cohorts (12 in each cohort) and 2:1 allocation ratio to study drug (8 FX-322:4 placebo) within 
each cohort.  The otolaryngologist will be blinded to whether a patient is randomized to drug 
or placebo within a cohort, however, may not be blinded to the cohort (high dose or low 
dose) because low and high dose cohorts have different injection volumes. 
The study treatment will be administered on Day 1 by an intratympanic injection. 
The investigational drug, FX-322, is a fixed ratio dose combination of two small molecules: a 
glycogen synthase kinase (GSK) inhibitor, FX03, and Valproate Sodium (FX00) (a histone 
deacetylase inhibitor; HDAC inhibitor).  There will be a matched placebo, identical in 
consistency and color to FX-322.   


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CONFIDENTIAL 
Once a subject has been identified, as meeting all inclusion/exclusion criteria, the ear with the 
worse hearing will be selected, unless a rationale exists to treat the other ear.  The 
Investigator will need to discuss that rationale with the Sponsor prior to randomization.   
Each subject will be placed in the supine position. EMLA® topical anesthetic cream will be 
administered to the tympanic membrane using the provided applicator kit. The cream will 
cover the eardrum and remain in place for 2-3 minutes then suctioned clear.  Under a 
microscope, a 25-gauge needle will be used to inject FX-322 or placebo into the middle ear  
at the junction of the posterior inferior and posterior superior quadrant with the needle tip 
directed towards the round window. The posterior superior quadrant should be avoided to 
prevent injury to the ossicles.  
After injection, the subject will continue to lie with the injected ear facing up for 10-15 
minutes. The injections will be performed at an appropriate location designated by a Board 
Certified Otolaryngologist trained and experienced in performing intratympanic injections. 
The 24 subjects will be required to stay in the Phase 1 Unit overnight for observation for up 
to 28 hours post injection.  
Blood samples for plasma pharmacokinetic (PK) will be obtained at pre-dose, 0.5 hour (+/- 
10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8 hour (+/- 15 min), 
and 24 hours (+4 hours) post injection. 
Safety monitoring will include recording of adverse events (AEs) and 12-lead 
electrocardiogram (ECG) findings at baseline, 1 hour (+/-30 min), and 24 hours post 
injection. 
Safety labs and urine testing will be collected per the Schedule of Events. 
In addition, a blood sample will be taken for exploratory biomarker analysis prior to, 24 
hours post, 15 days post and 3 months post injection. 
Follow up: Subject will be required to return for safety, otologic, and audiologic assessments 
2 weeks post injection.  Subjects will return at 1, 2, and 3 months for otoscopic and 
audiologic assessments. 
Number of Subjects:  
Approximately 24 evaluable subjects 
Study Duration: 
Screening phase: up to 30 days before study drug administration. 
Treatment with observation phase: 1 day (up to 28 hours) 
Follow-up phase: 3 months 
Study Population: 
Male and female adults, 18 to 65 years, inclusive, otherwise healthy with stable sensorineural 
hearing loss  
Primary Endpoint(s): 
•
To assess the systemic safety of FX-322
•
To assess the plasma pharmacokinetic (PK) profile to determine the systemic
exposure to the active pharmaceutical ingredients
•
To assess the effect of FX-322 on otologic and audiologic measures
Pharmacokinetics: 
Blood samples for determination of plasma concentrations will be drawn pre-dose and 0.5 
hour (+/- 10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8 hour (+/- 


Frequency Therapeutics  
Page 5 of 42 
CONFIDENTIAL 
15 min), and 24 hours (+4 hours) post injection.  Calculated PK parameters include Cmax, 
AUC0-inf, CL, Vss, T1/2, and Tmax. 
Safety: 
Safety assessments including adverse events, laboratory parameters, vital signs, physical 
examination, ECG, audiometry, and otoscopy. 
Statistical Analysis: 
Sample Size:  Since, the study is aimed at demonstrating safety of FX-322, the selected 
sample size was considered adequate for an initial assessment of safety and tolerability and 
was not based on formal statistical considerations such as power.  As such, the statistical 
analyses will consist of descriptive statistics including the mean, standard deviation (SD), 
median, minimum and maximum statistics for continuous endpoints and numbers and percent 
for categorical endpoints.  Where relevant, 95% confidence intervals will also be provided.   
Statistical Methods:  Subjects will be randomized to one of four treatment groups:  FX-
322L, PlaceboL, FX-322H, or PlaceboH using a 1:1 allocation ratio for dose cohort (12 per 
cohort) and within each cohort a 2:1 allocation ratio for study drug (8 FX-322: 4 placebo).   
Since the volume in the syringe may be potentially unblinding to the otolaryngologist, for this 
study, cohort assignment may not be blinded to the otolaryngologist (all subjects and study 
staff will remain blinded), and randomization to FX-322 or placebo will remain double-blind 
to all study staff (including the otolaryngologist) and subjects. Patient disposition, 
demographic and medical history will be tabulated.  Summary tabulations for treatment-
emergent adverse events will be presented by preferred term and body system using the 
Medical Dictionary for Regulatory Activities (MedDRA®).  Tabulations will be provided by 
seriousness, severity, and relationship to study drug.  Laboratory data will be presented as 
summary statistics at each timepoint measured as well as changes from baseline and/or shift 
tables.  Otologic and audiologic endpoints will be summarized in a similar manner. 
Pharmacokinetics will summarize the PK parameters for relevant endpoints and present 
individual as well as summary FX-322 concentration-time profiles. All subjects exposed to 
study drug will be included in the Safety Analysis Set and included in the safety analyses 
according to the treatment received regardless of the randomized assignment.  While 
statistical tests may be conducted, given the lack of formal consideration for statistical power, 
these will be viewed as exploratory analyses rather than confirmatory.  Details of the planned 
statistical analyses and methods will be provided in the Statistical Analysis Plan (SAP). 


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CONFIDENTIAL 
PROTOCOL Version 3.0 Amendments 
Item No. 
Change 
Section and Page Number(s) 
1 
Updated date and version of protocol. Title Page, pg.1 
Footer, every page 
2 
Updated Day 15 Visit window to +/- 
3 days. 
Table 1. pg. XX 
Section 8.1.4. Day 15 Visit, pg. 
3 
Updated Vitals to sitting and 
removed supine 
Section 8.1.1.  Screening Visit, pg. 12 
4 
Added coagulation to the testing to be 
performed. 
Section 8.1.2. Baseline/Treatment 
Visit, pg. 21 
5 
Updated drug testing to naming of the 
tests performed at site. 
Section 8.1.1.  Screening Visit, pg. 17 
Section 9.1.2.  Clinical Laboratory 
Evaluation, pg. 21 
6 
Updated the Day 50 Visit to Day 60 
to correct error. 
Section 9.2.3.Word Recognition. pg. 
21 
7 
Added “or back up designee” to 
ensure coverage if Medical Monitor 
is not available.. 
Section 8.1.1.Screening Visit. pg. 21 


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CONFIDENTIAL 
Item No. 
Change 
Section and Page Number(s) 
1 
Updated date and version of protocol. Title Page, pg.1 
Footer, every page 
2 
Updated Sponsor Signatory. 
Name and contact information, pg. 2 
3 
Clarified type of topical anesthetic to 
EMLA. 
Synopsis, pg. 4 
Section 7.1 Overall Study Design and 
Plan: Description, pg. 12 
Section 7.4.1 Investigational Products 
Administered, pg. 17 
Section 7.6.5 Selection and Timing of 
Dose for Each Subject, pg. 21 
4 
Clarified location of the 
intratympanic injection. 
Synopsis, pg. 4 
Section 7.1 Overall Study Design and 
Plan: Description, pg. 12 
Section 7.4.1 Investigational Products 
Administered, pg. 17 
Section 7.6.5 Selection and Timing of 
Dose for Each Subject, pg. 21 
5 
Updated Table of Contents. 
Table of Contents, pg. 7-9 
6 
Updated list of abbreviations. 
Abbreviations, pg. 11 
7 
Updated Table 1: Schedule of 
Assessments. 
Table 1: Schedule of Assessments, 
pg. 14 
8 
Added additional exclusion criteria 
Section 7.3.3 Exclusion Criteria pg.15 
9 
Removed “use of insulin (by any 
route)” . 
Section 7.6.7 Prohibited 
Medication/Therapy, pg.21 
10 
Added additional Words-In-Noise 
(WIN) testing to applicable visits. 
Section 8.1.1 Screening Visit, pg. 22 
Section 8.1.4 Follow-up Visit 4, pg. 
23 
Section 8.1.5 Follow-up Visits 5, 6, 
and 7, pg. 23 
11 
Added description of Words-In-Noise 
(WIN) testing. 
Section 9.2.4, pg. 27 
12 
Added reference for WIN testing 
Section 15 References, pg. 38 
PROTOCOL 2.0 


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CONFIDENTIAL 
3 
TABLE OF CONTENTS 
1 
Protocol Approval Signatures .................................................................................. 2 
2 
Synopsis ...................................................................................................................... 3 
3 
Table of Contents ....................................................................................................... 8 
4 
List of Abbreviations and Definition of Terms ..................................................... 11 
5 
Introduction.............................................................................................................. 12 
6 
Study Objectives ...................................................................................................... 13 
6.1 
Primary Objective(s) ............................................................................................... 13 
7 
Investigational Plan ................................................................................................. 13 
7.1 
Overall Study Design and Plan: Description ......................................................... 13 
7.1.1 
Study Design ............................................................................................. 14 
7.1.2 
Schedule of Assessments .......................................................................... 14 
7.2 
Discussion of Study Design ..................................................................................... 16 
7.3 
Selection of Study Population ................................................................................. 16 
7.3.1 
Number of Planned Subjects .................................................................... 16 
7.3.2 
Inclusion Criteria ...................................................................................... 16 
7.3.3 
Exclusion Criteria ..................................................................................... 16 
7.3.4 
Withdrawal of Subjects From Assessments ............................................. 17 
7.4 
Investigational Products .......................................................................................... 18 
7.4.1 
Investigational Products Administered ..................................................... 18 
7.4.2 
Identity of Investigational Products .......................................................... 18 
7.5 
Preparation and Dispensing .................................................................................... 19 
7.5.1 
Packaging and Labeling ............................................................................ 20 
7.5.2 
Supply, Storage and Handling .................................................................. 20 
7.5.3 
Compliance and Drug Accountability ...................................................... 20 
7.5.4 
Disposal, Return or Retention of Study Drug ........................................... 20 
7.6 
Blinding and Un-blinding ....................................................................................... 20 
7.6.1 
Blinding .................................................................................................... 20 
7.6.2 
Un-blinding ............................................................................................... 21 
7.6.3 
Method of Assigning Subjects to Treatment Groups ............................... 21 
7.6.4 
Selection of Doses in the Study ................................................................ 21 
7.6.5 
Selection and Timing of Dose for Each Subject ....................................... 22 
7.6.6 
Prior and Concomitant Therapy ................................................................ 22 
7.6.7 
Prohibited Medication/Therapy ................................................................ 22 
7.6.8 
Rescue Medication .................................................................................... 22 
7.6.9 
Treatment Compliance .............................................................................. 23 
8 
Timing of Study Procedures ................................................................................... 23 


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CONFIDENTIAL 
8.1 
Pre-treatment ........................................................................................................... 23 
8.1.1 
Screening Visit (Visit 1; Day -30 to 0) ..................................................... 23 
8.1.2 
Baseline/Treatment Visit (Visit 2, Day 1) ................................................ 23 
8.1.3 
Observation Visit (Visit 3, Day 2) ............................................................ 24 
8.1.4 
Follow-up (Visit 4, [Day 15 +3 days]) ..................................................... 24 
8.1.5 
Follow-up (Visit 5 [Day 30±5 days], Visit 6 [Day 60±5 days], 
Visit 7 [Day 90±5 days]) .......................................................................... 24 
8.2 
Duration of Treatment ............................................................................................ 25 
9 
Safety ASSESSMENTS ........................................................................................... 25 
9.1 
Safety Measurements Assessed ............................................................................... 25 
9.1.1 
Concomitant Medication .......................................................................... 25 
9.1.2 
Clinical Laboratory Evaluation ................................................................. 25 
9.1.3 
Vital Signs ................................................................................................ 26 
9.1.4 
Electrocardiogram ..................................................................................... 27 
9.1.5 
Physical Exam (PE) .................................................................................. 27 
9.1.6 
Body Mass Index (BMI) ........................................................................... 27 
9.1.7 
Noise Exposure ......................................................................................... 27 
9.1.8 
Otoscopy ................................................................................................... 27 
9.2 
Audiometry ............................................................................................................... 27 
9.2.1 
Comprehensive audiogram ....................................................................... 27 
9.2.2 
Tympanometry .......................................................................................... 28 
9.2.3 
Word Recognition (quiet) ......................................................................... 28 
9.2.4 
Words-In-Noise (WIN) ............................................................................. 28 
9.3 
Pharmacokinetic Assessment .................................................................................. 28 
10 
Adverse Events ......................................................................................................... 29 
10.1 Serious Adverse Events ........................................................................................... 31 
10.2 Unexpected Adverse Reactions ............................................................................... 32 
11 
Statistical Methods ................................................................................................... 32 
11.1 Statistical and Analytical Plans .............................................................................. 32 
11.1.1 
Descriptive Statistics ................................................................................ 32 
11.1.2 
Sample Size .............................................................................................. 34 
11.1.3 
Analysis Sets ............................................................................................. 34 
11.1.4 
Subject Disposition ................................................................................... 34 
11.1.5 
Description of Subgroups to be Analyzed ................................................ 35 
11.1.6 
Subject Demographics and Baseline Characteristics ................................ 35 
11.1.7 
Safety Analyses ........................................................................................ 35 
11.1.7.1 Adverse Events ......................................................................................... 35 
11.1.7.2 Clinical Laboratory Evaluations ............................................................... 35 
11.1.7.3 Vital Signs ................................................................................................ 35 
11.1.7.4 Physical Examination and Electrocardiogram .......................................... 35 
11.1.7.5 Prior and Concomitant Medications ......................................................... 36 
11.1.7.6 Tympanometry and Otoscopy ................................................................... 36 


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CONFIDENTIAL 
11.1.7.7 Comprehensive Audiometry ..................................................................... 36 
11.2 Handling of Missing Data and Subject Withdrawals ........................................... 36 
11.3 Protocol Deviations .................................................................................................. 36 
12 
Quality Assurance and Quality Control ................................................................ 36 
12.1 Audit and Inspection ............................................................................................... 36 
12.2 Monitoring ................................................................................................................ 36 
12.3 Data Management and Coding ............................................................................... 37 
13 
Records and Supplies .............................................................................................. 37 
13.1 Drug Accountability ................................................................................................ 37 
13.2 Financing and Insurance ......................................................................................... 37 
14 
Ethics ......................................................................................................................... 38 
14.1 Independent Ethics Committee or Institutional Review Board .......................... 38 
14.2 Regulatory Authorities ............................................................................................ 38 
14.3 Ethical Conduct of the Study .................................................................................. 38 
14.4 Informed Consent .................................................................................................... 38 
14.5 Subject Confidentiality ............................................................................................ 39 
15 
References ................................................................................................................. 40 
16 
Appendix 1 ................................................................................................................ 41 
Word Recognition Scoring Chart .............................................................................. 41 
17 
Investigator Signature Page .................................................................................... 42 


Frequency Therapeutics                                                                           Page 11 of 42 
 
 
CONFIDENTIAL 
4 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
AE 
Adverse event 
ASHA 
American Speech-Language-Hearing Association 
BMI 
Body mass index 
BPPV 
Benign Paroxysmal Positional Vertigo 
CNIHL 
Chronic noise induced hearing loss 
CL 
Total clearance 
Cmax 
Maximal plasma concentration 
CRO 
Contract Research Organization 
dB 
decibel 
DMSO 
Dimethyl sulfoxide 
DPOAE 
Distortion product otoacoustic emissions 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
EDC 
Electronic data capture 
GCP 
Good Clinical Practice 
GSK 
Glycogen Synthase Kinase 
HDAC 
Histone deacetylase 
HCG 
Human chorionic gonadotrophin 
IB 
Investigator’s Brochure 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IEC 
Independent Ethics Committee 
IRB 
Institutional Review Board 
IUD 
Intrauterine device 
LGR5+ 
Leucine-rich repeat-containing G-protein coupled receptor 5 
MedDRA  
Medical Dictionary for Regulatory Activities 
PE 
Physical exam 
PI 
Principal Investigator 
PK 
Pharmacokinetics 
QTcF 
QT interval data corrected using Fridericia’s formula 
SAE 
Serious Adverse Event 
SD 
Standard deviation 
SNHL 
Sensorineural hearing loss  
TEAE 
Treatment-emergent adverse event 
Tmax 
Time to maximal concentration 
TM 
Tympanic membrane 
Vss 
Apparent volume of distribution at steady state 
WHO 
World Health Organization 
WIN 
Words-In-Noise Testing 
 
 
 
 


Frequency Therapeutics                                                                           Page 12 of 42 
 
 
CONFIDENTIAL 
5 
INTRODUCTION 
Worldwide, an estimated 1.1 billion people are at risk for hearing loss due to exposure to 
damaging levels of sound (WHO, 2015). Excessive sound levels or loud sounds for extended 
periods time hyperstimulate cochlear hair cells, which can lead to increased production of 
reactive oxygen species and oxidative cell death. Structural damage to hair cells results in 
sensorineural hearing loss (SNHL), which can be characterized by an attenuation and 
distortion of response to incoming auditory stimuli. Presently, no curative treatments exist, 
but rather assistive devices such as hearing aids and cochlear implants are used to address the 
symptoms of SNHL. These options, however, do not address the cause of SNHL and are not 
sufficient for normal quality of life in individuals with SNHL. 
Sensorineural hearing loss (SNHL) accounts for about 90% of all cases of hearing loss (Li et 
al, 2017). Leading causes include noise exposure, ototoxic medications, advanced age, 
inherited, and autoimmune disorders. SNHL is usually irreversible and managed with hearing 
aids or cochlear implants. Noise is a major occupational and environmental hazard, causing 
hearing loss, sleep disturbance, fatigue, and hypertension (Hong et al, 2013). SNHL has long 
been recognized as the primary and direct health effect of excessive noise exposure (Basner 
et al, 2015). In the United States, an estimated 48 million people or 20.3% of the population 
12 years or older has hearing loss in one or both ears (Lin et al, 2011). The World Health 
Organization reported that 16% of the disabling hearing loss in adults is attributable to 
occupational noise exposure (Nelson et al, 2005). Hearing loss is associated with increased 
dementia, depression, and other mental health disorders, especially in the elderly (Choi et al, 
2016; McGilton et al, 2016; Deal et al, 2016). 
FX-322 is a fixed ratio dose combination of two small molecules: a glycogen synthase kinase 
(GSK) inhibitor, FX03, and Valproate Sodium (an HDAC inhibitor approved in multiple 
countries including the US for human use as Depacon®). 
The active ingredients in FX-322 target two cellular mechanisms, histone deacetylase 
(HDAC) and GSK inhibition, which act synergistically to induce a regenerative response in 
cochlear tissue and thus may restore hearing function in subjects with hearing loss including 
CNIHL. Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5+) progenitor 
cells have the capability of asymmetric division to create a copy of themselves and a daughter 
cell that can become a hair cell.  
In preclinical experiments, a fixed ratio dose combination of FX03 and Valproate Sodium 
demonstrates: 
• Expansion of the Lgr5+ progenitor cells and formation of new hair cells in cell culture 
studies in newborn and adult mice. Substantial Lgr5+ expansion occurs with the 
combination of agents but not with the individual agents alone.  
• Expansion of progenitor cells in adult primate and adult human inner ear tissue. 
• Formation of new hair cells in explanted mouse cochlea cultures. 
• Significant improvement in hearing and increase in hair cell count in an adult mouse 
model of noise-induced hearing loss maintained for at least 3 months after a single 
treatment. 
Based on these findings, FX-322 is formulated as a fixed ratio dose combination, 
 
 
 


Frequency Therapeutics                                                                           Page 13 of 42 
 
 
CONFIDENTIAL 
 
r. FX-322 will be administered by an 
intra-tympanic injection into the middle ear onto the round window membrane area, for the 
active ingredients to be locally absorbed into the cochlea. Systemic exposure from local 
deposition of the drug in the gel is expected to be negligible. There has been one clinical 
study performed with FX-322, study FX-322-103.  Nine patients undergoing cochlear 
implantation surgery received a single unilateral intra-tympanic injection of FX-322 (6 
patients) or placebo (3 patients) to study the uptake in the cochlea.  The results demonstrated 
acceptable local and systemic acute safety and tolerability and very limited systemic exposure 
after the local intra-tympanic injection which was unmeasurable within 24 hours post 
injection.  In addition, the cochlear pharmacokinetic (PK) data demonstrated the drug was 
present at 4 h post-IT injection in one patient but unmeasurable after 24 hours in the 
perilymph in two subjects. 
 
6 
STUDY OBJECTIVES 
6.1 
Primary Objective(s) 
• To assess the systemic safety of FX-322  
• To assess the plasma pharmacokinetic (PK) profile to determine the systemic 
exposure to the active pharmaceutical ingredients 
• To assess the effect of FX-322 on otologic and audiologic measures  
 
 
7 
INVESTIGATIONAL PLAN 
7.1 
Overall Study Design and Plan: Description 
This is a Phase 1/2 randomized, double-blind, placebo-controlled, single dose study to 
evaluate the safety at two dose levels of FX-322L and FX-322H compared to placebo in 
adults with stable sensorineural hearing loss (no changes over 6 months of 10 dB or more in 
any frequency). The trial will have three phases: Screening, Treatment with Observation, and 
Follow up.  
 
Screening:  Can occur up to 30 days prior to study drug administration (Day 1). All subjects 
will be screened to determine study eligibility. 
 
Double Blind Treatment: Treatment with Observation: Approximately 24 subjects are 
planned to be randomized using a 1:1 allocation ratio to cohort (12 in each cohort) and a 2:1 
 allocation ratio to study drug (8 FX-322:4 placebo) within each cohort.  Subjects will be 
randomized to one of four treatment groups to receive a single dose of either FX-322L, 
placeboL, FX-322H, or placeboH.   
Each subject will be placed in the supine position. EMLA® topical  anesthetic cream will be 
administered to the tympanic membrane using the provided applicator kit.  The cream will 
cover the eardrum and remain in place for 2-3 minutes then suctioned clear.  Under a 
microscope, a 25-gauge needle will be used to inject of FX-322 or placebo into the middle 
ear at the junction of the posterior inferior and posterior superior quadrant with the 
needle tip directed towards the round window. The posterior superior quadrant should be 
avoided to prevent injury to the ossicles.  


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CONFIDENTIAL 
After injection, the subject will continue to lie with the injected ear facing up for 10-15 
minutes. The injections will be performed at an appropriate location designated by a Board 
Certified Otolaryngologist trained and experienced in performing intratympanic injections.  
The 24 subjects will be required to stay in the Phase 1 Unit overnight for observation for up 
to 28 hours post injection.  
Blood samples for plasma pharmacokinetic (PK) will be obtained pre-dose and at 0.5 hour 
(+/- 10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8 hour (+/- 15 
min), and 24 hours (+4 hours) post injection. 
Safety monitoring will include recording of adverse events (AEs) and 12-lead 
electrocardiogram (ECG) findings at baseline (pre-dose), 1 hour (+/- 30 mins), and 24 hours 
post injection.  Safety labs and urine testing will be collected per the Schedule of Events. 
In addition, a blood sample will be taken for exploratory biomarker analysis prior to and 24 
hours post, 15 days post, and 3 months post injection. 
Follow up: Subject will be required to return for safety, otologic, and audiologic assessments 
2 weeks post injection.  Subjects will return at 1, 2, and 3 months for otoscopic and 
audiologic assessments. 
7.1.1 Study Design 
7.1.2 Schedule of Assessments 
The schedule of planned study assessments is shown in Table 1. 


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CONFIDENTIAL 
Table 1: 
Schedule of Assessments
Subjects will be given the option of staying overnight the day before dosing (Day 1) 
a- Assessment performed prior to injection.
b- Assessment performed up to 24 hr. prior to injection with a standard audiogram
c- 12-lead ECG to be performed in a supine position prior to injection, 1 hour post injection (+/-30 mins), and 24 hours
d- Should there be a positive result from the battery of drug screen tests, and the Investigator feels the subject should not be
excluded, the Sponsor will be contacted to determine final eligibility.
e- Assessments performed after injection
f -Time points: prior to injection and 0.5 hour (+/- 10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8
hour (+/- 15 min), and 24 hours (+4 hours) post injection.
g- Interval noise history will be collected by querying the subject about noise exposure since receiving study treatment
h- Visit windows: +/- 5 day for all visits
i- Perform only at the Follow Up Day 90 Visit
j-Randomization will occur on Day 1 prior to study drug administration.
Visit 
Screening 
Treatment 
Observation 
Follow-up 
Assessment/Day 
-30 to 0
1 
2 
15 (+/-3 
days) 
30,60,90 h 
Informed Consent 
X 
Inclusion/ Exclusion 
Criteria 
X 
X 
Demographics 
X 
Medical History 
X 
X 
Interval noise history 
X g 
X g 
Concomitant Medication 
X 
Xa 
X 
X 
X 
Physical Examination 
including weight and height 
X 
Vital Signs (body 
temperature, pulse rate, bp) 
X 
Xa 
X 
X 
X 
ECG (12–lead) 
X 
Xc 
Xc 
Tympanometry 
X 
X 
X 
Comprehensive audiogram 
X 
Xb 
X 
X 
Word Recognition, quiet 
X 
X 
X 
Words-In-Noise Testing 
(WIN) 
X 
X 
X 
Otoscopy 
X 
Xa 
X 
X 
Safety blood samples 
X 
Xa 
X 
X 
Hepatitis B and C testing 
X 
Urinalysis 
X 
Xa 
X 
X 
Urine Pregnancy Test 
(women of child bearing 
potential only) 
X 
Xa 
X 
X 
Drug Screen 
Xd 
PK: Plasma FX-322 
XXXXXXf 
X 
Blood sample for biomarker 
analysis  
Xa 
X 
X 
Xi 
Randomization 
Xj 
Study Medication (FX-322 
or Placebo injection) 
X 
Adverse Events 
Xe 
X 
X 
X 


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CONFIDENTIAL 
7.2 
Discussion of Study Design 
This is a Phase 1/2 randomized, double-blind, placebo-controlled, single dose study to 
evaluate the safety at two dose levels of FX-322 (FX-322L and FX-322H) compared to 
placebo in adults with stable sensorineural hearing loss (no changes of 10 dB or more in any 
frequency at least 6 months prior to screening).  
7.3 
Selection of Study Population 
7.3.1 Number of Planned Subjects 
Approximately 24 subjects are planned to enter the study, and 24 subjects are expected to 
complete the study.  Additional subjects will be enrolled to account for early 
termination/withdrawal subjects, unless the reason for discontinuation was a treatment 
emergent adverse event. 
7.3.2 Inclusion Criteria 
To be eligible for study entry subjects must satisfy all of the following criteria: 
1. Subject has read and voluntarily signed the Informed Consent Form (ICF) after all
questions have been answered and prior to any study-mandated procedure.
2. Adult aged 18-65 years.
3. Established diagnosis of stable sensorineural hearing loss (no changes of 10 dB or
more in any frequency) by standard audiometric measures for >6 months.
4. Documented medical history consistent with hearing loss being caused by noise
exposure or sudden sensorineural hearing loss (documented audiogram at least 6
months prior to screening required).
5. Ability to communicate well with the Investigator and is willing to comply with and
complete all the study procedures.
6. Female subjects must be of non-childbearing potential or will need to utilize two
methods of highly effective contraception during the study participation (e.g.
hormonal contraception or an intrauterine device and condoms) or remain abstinent.
Male subjects should use condoms with spermicide during the course of the study or
remain abstinent.  Subjects should not donate sperm or ova during the study period.
7.3.3 Exclusion Criteria 
Subjects will be excluded from the study if one or more of the following criterion are 
applicable: 
1. Subjects currently on any medication consisting of valproic acid, valproate sodium, or
divalproex sodium (e.g. Depacon®, Depakote®, Depakene, Epival®, Valpro, and
Epilim).
2. Perforation of tympanic membrane or other tympanic membrane disorders that would
interfere with the delivery and safety assessment of an intratympanic medication or
reasonably be suspected to affect tympanic membrane healing after injection in either
ear.  This includes a current tympanostomy tubes.
3. Any conductive hearing loss of 10 dB or more at two or more frequencies in either
ear.


Frequency Therapeutics                                                                           Page 17 of 42 
 
 
CONFIDENTIAL 
4. A pure tone average of 70 dB or greater at 500Hz, 1000Hz, 2000Hz, and 4000Hz in 
the ear to be injected. 
5. Active chronic middle ear disease or a history of major middle ear surgery, as an 
adult, in the ear to be injected. 
6. Subject has had an intratympanic injection in either ear within 6 months of the 
screening visit. 
7. History of clinically significant vestibular symptoms at the discretion of the 
investigator.  For example, BPPV may be considered acceptable whereas Ménière’s 
would not.   
8. History of clinically significant systemic autoimmune disease (e.g. rheumatoid 
arthritis, Sjogren's syndrome, multiple sclerosis, psoriasis). 
9. History of head or neck radiation treatment or exposure. 
10. Exposure to another investigational drug within 28 days prior to injection of study 
drug. 
11. Evidence of any active or chronic disease, or condition that could interfere with, or for 
which the treatment of might interfere with, the conduct of the study, or that would 
pose an unacceptable risk to the subject in the opinion of the investigator (following a 
detailed medical history, physical examination, vital signs (systolic and diastolic 
blood pressure, pulse rate, body temperature), 12-lead ECG, and clinical laboratory 
parameters (hematology, blood chemistry, and urinalysis)). Deviations of laboratory 
values from the normal range may be accepted, if judged by the Investigator to have 
no clinical relevance. In the case of uncertain or questionable results, tests performed 
during screening may be repeated before randomization to confirm eligibility or 
judged to be clinically irrelevant for healthy subjects in the opinion of the 
Investigator. 
12. History of substance abuse within 2 years of the Screening Visit. 
13. Positive test for drugs of abuse at screening. In cases when the Investigator feels the 
subject should not be excluded for a positive drug test, the Sponsor will be consulted 
and determine final eligibility. 
14. Positive urine pregnancy test or breast-feeding. 
15. Any known factor, condition or disease that, in the view of the investigator, might 
interfere with treatment compliance, study conduct or interpretation of the results such 
as psychiatric disease or suicidal tendencies. 
 
7.3.4 Withdrawal of Subjects From Assessments 
Subjects may stop the study for any of the following reasons: 
• 
Subject request 
• 
Use of non-permitted concurrent therapy 
• 
Non-compliance with the study schedule 
• 
Lost to follow-up 
• 
Occurrence of adverse events (AEs) not compatible with the continuation of subject 
participation in the study, in the investigator’s opinion, or unacceptable to the subject to 
continue 
• 
Investigator request 
• 
Intercurrent illness 
• 
Sponsor request 
• 
Pregnancy 


Frequency Therapeutics  
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CONFIDENTIAL 
Subjects who do not comply with the protocol or who withdraw consent may be replaced. 
Subjects are free to withdraw from the study at any time without providing reason(s) for 
withdrawal and without prejudice to further treatment. The reason(s) for withdrawal will be 
documented in the case report form (CRF).  Any subjects that withdraw due to a treatment 
emergent adverse event (TEAE) will not be replaced. 
Subjects withdrawing from the study will be encouraged to complete the same final 
evaluations as subjects completing the study according to this protocol, particularly safety 
evaluations. The aim is to record data in the same way as for subjects who completed the 
study. 
Reasonable efforts will be made to contact subjects who are lost to follow-up. These efforts 
must be documented in the subject’s file.  
Pregnancy 
Subjects will be instructed that known or suspected pregnancy occurring during the study, in 
subjects or female partners of male subjects, should be confirmed and reported to the 
investigator.  All subjects will be followed until the end of the study, completing study 
assessments as appropriate during pregnancy. The investigator should also be notified of 
pregnancy occurring during the study but confirmed after completion of the study. In the 
event that a subject is subsequently found to be pregnant after inclusion in the study, any 
pregnancy will be followed to term, and the status of mother and child will be reported to the 
sponsor after delivery. 
Full details will be recorded on the withdrawal page of the CRF, or an SAE report will be 
completed if the subject has completed the study. 
7.4 
Investigational Products 
7.4.1 Investigational Products Administered 
A single injection of FX-322 or placebo will be administered on Day 1 by intratympanic 
injection. Subjects will be randomized to one of four treatment groups to receive a single 
dose of either FX-322L, placeboL, FX-322H, or placeboH. Randomized subjects are to be 
allocated 1:1 to each of 2 cohorts (12 in each cohort) and allocated 2:1  to study drug (8 FX-
322:4 placebo) within each cohort.  EMLA® topical anesthetic cream will be administered to 
the tympanic membrane using the provided applicator kit. The cream will cover the eardrum 
and remain in place for 2-3 minutes then suctioned clear.  Under a microscope, a 25-gauge 
needle will be used to inject of FX-322 or placebo into the middle ear at the junction of the 
posterior inferior and posterior superior quadrant with the needle tip directed towards the 
round window. The posterior superior quadrant should be avoided to prevent injury to the 
ossicles.  
After injection, the subject will continue to lie with the injected ear facing up for 10-15 
minutes. The injections will be performed at an appropriate location designated by a Board 
Certified Otolaryngologist trained and experienced in performing intratympanic injections.   
7.4.2 Identity of Investigational Products 
The investigational drug is FX-322 which is a fixed ratio dose combination of 2 small 
molecules: a glycogen synthase kinase (GSK) inhibitor (FX03) and valproate sodium (FX00), 


Frequency Therapeutics  
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CONFIDENTIAL 
a histone deacetylase (HDAC) inhibitor. FX-322 is in the form of a 
. 
7.5 
Preparation and Dispensing 
The FX-322 investigational product is a sterile liquid for intratympanic injection only. FX-
322 is prepared by mixing a diluent vial with a lyophilized active vial, which is stored 
refrigerated until use, within 6 hours of mixing. 
An osmolarity matched placebo for FX-322 is a sterile liquid for intratympanic injection 
only. The placebo is prepared by mixing a diluent vial with a lyophilized vial, which is stored 
refrigerated until use, within 6 hours of mixing. 
The components in the study drug and placebo are shown in the following tables: 
Table FX-322 Study Drug: 
Active Ingredients 
 
FX03 
 
Valproate Sodium 
 
 
 
 
 
 
 
The matched placebo has been developed with similar pH, osmolality and gelation as FX-
322. In a 0.2 mL injection, placeboH contains 
  
Proposed Amounts of Active Agents to be Injected in Humans 
FX-322L 
Total 
(in a 0.05 mL injection) 
FX-322H 
Total 
(in a 0.20 mL injection) 
FX03 
 
 
FX00 
 
 
Frequency Therapeutics will supply sterile FX-322 and placebo, as a sealed sterile vial 
containing lyophilized ingredients and another sterile vial containing a diluent. The placebo 
or FX-322 is prepared by mixing a diluent vial with a lyophilized vial. Mixing is achieved by 
tapping the vial on the palm of the hand to wet the lyophilized cake. FX-322 and placebo are 
stored refrigerated until use, within 6 hours of mixing. Detailed instructions will be included 
with the vials and in the Pharmacy Manual. 


Frequency Therapeutics  
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CONFIDENTIAL 
7.5.1 Packaging and Labeling 
For the injection, the study drug or placebo will be packaged in a sterile 1 mL tuberculin 
syringe with a sterile cap.  The syringe will be labelled clearly with the details for each 
randomized patient.  Two syringes per patient will be provided in case a back-up syringe is 
needed for injection.  The low dose (0.05 mL) syringe will be filled to the 0.1 mL mark.  The 
high dose (0.2 mL) will be filled to the 0.25 mL mark.  The FX-322 and placebo preparations 
drawn into the syringes must be warmed in a heating unit. Refer to the Pharmacy Manual for 
further instructions.  
7.5.2 Supply, Storage and Handling 
The vials must be stored as indicated in the Pharmacy Manual. Empty vials and containers 
may be destroyed at the conclusion of the study, after the site monitor has performed 
accountability and in compliance with the site’s SOP.  
7.5.3 Compliance and Drug Accountability 
Accountability for the study drug at the study site is the responsibility of the Investigator. The 
investigator will ensure that the study drug is used only in accordance with this protocol. The 
otolaryngologist will complete a form post injection to note any complications or if any of the 
study treatment was not able to be administered.  Where allowed, the Investigator may 
choose to assign some of the drug accountability responsibilities to a pharmacist or other 
appropriate individual. Drug accountability records indicating the drug’s delivery date to the 
site, inventory at the site, use by each subject, and return to Frequency Therapeutics (or 
destruction, if approved by Frequency Therapeutics) will be maintained by the clinical site. 
These records will adequately document that the subjects were provided the doses as 
specified in the protocol and should reconcile all study drug received from Frequency 
Therapeutics or its designee. Accountability records will include dates, quantities, 
batch/serial numbers, and subject identification numbers. The site monitor will review drug 
accountability at the site on an ongoing basis during monitoring visits. 
7.5.4 Disposal, Return or Retention of Study Drug 
All unused and used study drug will be retained at the site until inventoried by the site 
monitor. All used, unused or expired study drug will be returned to Frequency Therapeutics 
or if authorized, disposed of at the study site in accordance with governing regulations and 
documented.  
7.6 
Blinding and Un-blinding 
7.6.1 Blinding 
The subjects, all Frequency Therapeutics staff and representatives, investigators and site 
personnel will be blinded to the study drug assignment.  The otolaryngologist may not be 
blinded to cohort assignment, but will be blinded to randomized treatment (FX-322 or 
placebo). The pharmacy staff who prepare the study drug will be unblinded and will not 
perform any other roles on the study other than drug preparation and accountability.  The 
independent statistician and/or independent statistical programmer who is not otherwise 
involved with the study will generate the randomization schedule and will be un-blinded. 


Frequency Therapeutics                                                                           Page 21 of 42 
 
 
CONFIDENTIAL 
7.6.2 Un-blinding 
Only in the case of emergency, when knowledge of the study drug administered is essential 
for the clinical management or welfare of the subject, may the Investigator un-blind a 
subject’s treatment assignment. Under such conditions, the identity of the study drug will be 
obtained by contacting the CRO. 
If possible, the Medical Monitor and Frequency Therapeutics should be consulted prior to 
breaking the blind. If the blind is broken for any reason, the Investigator must notify 
Frequency Therapeutics immediately of the un-blinding incident without revealing the 
subject’s study treatment assignment to Frequency Therapeutics.  
In the event that the treatment assignment is broken, the date, the signature of the person who 
broke the code and the reason for breaking the code must be recorded on the sourced 
documents.  Any code-breaks that occur must be immediately reported to the Medical 
Monitor.    
Any subject whose treatment assignment has been un-blinded will be followed up for safety 
purposes.   
Each box will be labelled Study drug in compliance with applicable local regulations. FX-
322, Placebo, and Diluent vials will be packaged in separate boxes.  
 
Instructions on compounding the study drug will be provided in the pharmacy manual.  
 
All study drug will be transported, received, stored and handled strictly in accordance with 
the product label, the instructions provided to the investigational sites, the sites standard 
operation procedures and applicable regulations.   
 
7.6.3 Method of Assigning Subjects to Treatment Groups 
Twenty-four subjects will be randomized to one of four treatment groups:  FX-322L, 
placeboL, FX-322H, or placeboH.  The allocation ratio to dose cohort will be 1:1, 12 subjects 
per dose cohort, and within each dose cohort the allocation ratio will be 2:1 (8 FX-322: 4 
placebo).  A permuted randomized block algorithm will be developed by an independent 
statistician and/or statistical programmer not otherwise involved in the study.   
There will be no randomized stratification in this study. 
7.6.4 Selection of Doses in the Study 
The 2 doses proposed in this study, FX-322L and FX-322H will be dosed concurrently.  In a 
previous human study, Study FX-322-103, we have demonstrated that a single dose FX-322H 
has been well tolerated in patients with hearing loss with no serious drug related adverse 
events.  The only treatment-emergent adverse events occurring after intratympanic injection 
of study drug or placebo, but before cochlear implant surgery in Study FX-322-103 were 
transient ear pain, reported in both the active and placebo subjects, were most likely related to 
the intratympanic injection procedure.  The systemic exposure to the drug in these patients 
was also very limited.  Safety margins for plasma clinical maximal concentrations (Cmax) for 
FX03 determined in this study range from 31 to > 4000-fold based on nonclinical systemic 
exposure in toxicity studies with Guinea Pig, Rat and Dog, while safety margins for plasma 
clinical Cmax for FX00 were > 30-fold. FX00 is also approved in the US for the treatment of 
epilepsy and has been extensively studied in healthy volunteers and patients as both oral and 
intravenous formulations and has a well-characterized safety profile since its first approval in 
1996. Further, valproate sodium Cmax in the clinical PK study was 113 to 226-fold lower than 


Frequency Therapeutics  
Page 22 of 42 
CONFIDENTIAL 
the accepted therapeutic range following systemic treatment for epilepsy (50-100 µg/mL; 
Depacon Prescribing Information, 1996).    
While the highest proposed dose of FX-322 (with FX03 and FX00 as active ingredients) is 
considered well characterized for safety and tolerability with limited systemic exposure, we 
plan to also assess the safety and efficacy of a lower dose 
 
. Based on these data, it is considered that assessing 
the lower dose concurrent to the higher dose is well justified and does not pose any additional 
risk.  
7.6.5 Selection and Timing of Dose for Each Subject 
Once a subject has been identified who meets all inclusion/exclusion criteria, the ear with the 
worse hearing will be selected, unless a rationale exists to treat the other ear.  The 
Investigator will need to discuss that rationale with the Sponsor prior to randomization.   
Each subject will be placed in the supine position. EMLA® topical anesthetic cream will be 
administered to the tympanic membrane using the provided applicator kit. The cream will 
cover the eardrum and remain in place for 2-3 minutes then suctioned clear.  Under a 
microscope, a 25-gauge needle will be used to inject FX-322 or placebo into the middle ear at 
the junction of the posterior inferior and posterior superior quadrant with the needle tip 
directed towards the round window. The posterior superior quadrant should be 
avoided to prevent injury to the ossicles.  
After injection, the subject will continue to lie with the injected ear facing up for 10-15 
minutes. The injections will be performed at an appropriate location designated by a Board 
Certified Otolaryngologist trained and experienced in administering intratympanic injections. 
Subjects will be required to stay in the Phase 1 research unit overnight for observation, up to 
28 hours post injection. Subjects will be given a choice of staying at the Phase 1 research unit 
the night before study drug injection. 
7.6.6 Prior and Concomitant Therapy 
At each study visit the study personal will question the subject about any medication taken, 
including vitamin supplements and herbal remedies. Any concurrent medications will be 
recorded in the subject's records and the eCRF. Any changes in doses or introduction of a 
new medication during the course of the study will be recorded. 
7.6.7 Prohibited Medication/Therapy 
Concomitant medications that are not allowed during the study are valproic acid (brand name 
(e.g. Depacon®, Depakote®, Depakene, Epival®, Valpro, and Epilim) or derivatives. 
Intratympanic injections of anything other than study medication are prohibited during the 
course of the study. 
7.6.8 Rescue Medication 
No rescue medication will be provided as no FDA approved treatment for sensorineural 
hearing loss exists.  The Investigator should treat any adverse events as medically appropriate 
and per their standard of care.  Investigators will discuss the need for oral steroids on an 
individual subject’s basis with the Medical Monitor prior to prescribing the medication. 


Frequency Therapeutics  
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CONFIDENTIAL 
7.6.9 Treatment Compliance 
Treatment will be given one time only by the investigator so subject treatment compliance 
will not be assessed. 
8 
TIMING OF STUDY PROCEDURES 
Subjects will provide written informed consent before any study related procedures are 
performed. 
The planned study assessments are in Section 7.1.2. 
8.1 
Pre-treatment 
8.1.1 Screening Visit (Visit 1; Day -30 to 0) 
•
Obtain signed informed consent.
•
Assess for eligibility (against the inclusion and exclusion criteria).
•
Collect full medical history, including concomitant illnesses/diseases and concomitant
medications.
•
Record demographic data, such as ethnic origin, date of birth, and sex.
•
Record historical disease data and diagnostic information (including prior audiogram
for inclusion determination).
•
Perform a physical examination, including body weight and height.
•
Record vital signs (sitting blood pressure, body temperature, and heart rate).
•
Perform a 12-lead ECG.
•
Collect samples for hematology, coagulation, clinical chemistry, and urinalysis tests.
•
Collect samples for hepatitis B surface antigen and hepatitis C antibody.
•
Collect urine sample for pregnancy test, if applicable.
•
Collect urine sample for drug screening (alcohol, cocaine, opiates, benzodiazepines,
cannabinoids, barbiturates, and amphetamines).
•
Perform otoscopy, comprehensive audiogram, word recognition testing, WIN testing,
and tympanometry
•
Each subject’s audiogram will be sent to the Medical Monitor or back up designee for
review and determination of final eligibility.
8.1.2 Baseline/Treatment Visit (Visit 2, Day 1) 
The following procedures will be performed at the Baseline/Treatment Visit: 
•
Before injection of study drug:
o
Reassess for eligibility against the inclusion and exclusion criteria.
o
Up to 24 hours before study drug injection, perform a simple audiogram.
o
At least 30 minutes before study drug injection, 12 lead ECG will be recorded
o
Collect samples for hematology, coagulation, clinical chemistry, and
urinalysis tests, and baseline serum sample for PK testing.
o
Collect concomitant medications.
o
Record vital signs.
o
Perform a urine pregnancy test, if applicable.
o
Perform otoscopy.


Frequency Therapeutics  
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CONFIDENTIAL 
o
Record any changes in medical history that have occurred since the previous
visit.
•
When all the baseline procedures have been performed and the investigator has
confirmed the subject’s eligibility for the study, the subject will be randomized.
•
Randomization will take place for eligible subjects, and each will receive an unique
subject number.
•
Inject FX-322 or placebo via intratympanic injection to the study ear.
•
After study drug injection:
o
Collect serum samples for PK testing at the following time points: 0.5 hour
(+/- 10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8
hour (+/- 15 min), and 24 hours (±4 hours) post injection.
o
Record any adverse events that have occurred since study treatment.
8.1.3 Observation Visit (Visit 3, Day 2) 
The Observation Visit (Visit 3) will take place 1 day after the Baseline/Treatment visit. The 
following procedures will be performed at the Observation Visit: 
•
Record any AEs that have occurred since the previous visit and any changes in
concomitant medication.
•
Record vital signs.
•
Perform 12-lead ECG.
•
Collect samples for hematology, coagulation, clinical chemistry, and urinalysis tests.
•
Collect blood sample for biomarker analysis.
•
Collect serum sample for PK testing at 24 hours (±4 hours) post injection.
8.1.4 Follow-up (Visit 4, [Day 15 ±3 days]) 
The Follow-up Visit (Visit 4) will take place at Day 15 (± 3 days). The following procedures 
will be performed at the Follow-up Visit: 
•
Record any AEs that have occurred since the last visit and any changes in concomitant
medication.
•
Perform interval noise history.
•
Perform otoscopy, comprehensive audiogram, word recognition testing, WIN testing,
and tympanometry.
•
Record vital signs.
•
Collect samples for hematology, coagulation, clinical chemistry, and urinalysis tests.
•
Collect blood sample for biomarker analysis.
•
Perform a urine pregnancy test, if applicable.
8.1.5 Follow-up (Visit 5 [Day 30±5 days], Visit 6 [Day 60±5 days], Visit 7 [Day 90±5 
days]) 
The Follow-up Visits (Visits 5, 6, and 7) will take place 30, 60, and 90 ± 5 days after the 
Baseline/Treatment visit. The following procedures will be performed at the Follow-up 
Visits: 


Frequency Therapeutics                                                                           Page 25 of 42 
 
 
CONFIDENTIAL 
• 
Record any AEs that have occurred since the last visit and any changes in concomitant 
medication. 
• 
Obtain interval noise history. 
• 
Perform otoscopy, comprehensive audiogram, word recognition testing, WIN testing, 
and tympanometry. 
• 
Record vital signs. 
• 
Perform a urine pregnancy test, if applicable. 
• 
Collect blood sample for biomarker analysis (at the Day 90 visit only) 
 
8.2 
Duration of Treatment 
The duration of treatment will be 3 months plus 1 day.  
9 
SAFETY ASSESSMENTS 
The planned schedule of assessments is in Section 7.1.2. 
9.1 
Safety Measurements Assessed  
9.1.1 Concomitant Medication 
Subjects will be asked about concomitant medications at time points outlined in the Table of 
Assessments.   All concomitant medication information will be recorded in the CRF. 
9.1.2 Clinical Laboratory Evaluation 
The hematology and clinical chemistry laboratory analyses will be performed and used by the 
investigator to assess the laboratory data for clinical significance and pathological changes. 
The following laboratory safety tests will be performed at time points: 
Hematology 
• HEMOGLOBIN 
• MEAN CELL VOLUME (MCV) 
• HEMATOCRIT 
• MEAN CELL HEMOGLOBIN 
(MCH) 
• WHITE BLOOD CELL 
(WBC) COUNT (TOTAL 
AND DIFFERENTIAL) 
• PLATELET COUNT 
• RED BLOOD CELL (RBC) 
COUNT 
• MCH CONCENTRATION (MCHC) 
Coagulation 
• PT 
• APTT 
 
 
 


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Clinical Chemistry 
•
CREATININE
•
SODIUM
•
UREA (OR BLOOD UREA
NITROGEN [BUN])
•
POTASSIUM
•
ASPARTATE
TRANSFERASE (AST)
•
CHLORIDE
•
ALANINE TRANSFERASE
(ALT)
•
GLUCOSE
•
GAMMA GLUTAMYL
TRANSFERASE (GGT)
•
URIC ACID
•
ALKALINE PHOSPHATASE
•
TOTAL
CHOLESTEROL
•
LACTATE
DEHYDROGENASE (LDH)
•
TRIGLYCERIDES
•
TOTAL BILIRUBIN
•
CALCIUM
•
ALBUMIN
•
PHOSPHORUS
•
TOTAL PROTEIN
Urinalysis (random) 
•
PH
•
BLOOD
•
GLUCOSE
•
PROTEIN
•
KETONES
•
MICROSCOPY
Urine drug screen (alcohol, cocaine, opiates, benzodiazepines, cannabinoids, barbiturates, 
and amphetamines) will be performed at Screening only. 
Screening for hepatitis B surface antigen and hepatitis C antibody at Screening only. 
Screening for pregnancy will be performed (urine β-HCG) as outlined in Table 1. 
9.1.3 Vital Signs 
Vital signs (temperature, blood pressure and heart rate) will be recorded at time points 
outlined in Table 1 and will be performed in a standardized manner, i.e., after the subject has 
rested in the sitting position for 5 minutes. 


Frequency Therapeutics                                                                           Page 27 of 42 
 
 
CONFIDENTIAL 
 
9.1.4 Electrocardiogram 
At Screening, prior to treatment at Day 1, and at Day 2, a 12-lead ECG will be performed. 
ECG tracings will be retained and labeled as per the standard procedures applicable to the 
clinical study site. 
9.1.5 Physical Exam (PE) 
A complete physical examination will be performed by a licensed physician at Screening. 
Complete physical examinations include: general appearance, head, ears, eyes, nose, throat, 
dentition, thyroid, chest (heart, lungs), abdomen, skin, neurological, extremities, back, neck, 
musculoskeletal, and lymph nodes and any pertinent system based on any prior findings. 
Physical examinations may be performed at various unscheduled time points, if deemed 
necessary by the Investigator. 
9.1.6 Body Mass Index (BMI) 
BMI will be calculated by dividing the subject’s body weight in kilograms by the subject’s 
height in meters squared (BMI = kg/m2). Body height (centimeter) and body weight 
(kilograms) will be measured at the time points delineated in the Schedule of Assessments. 
Body weight and height will be obtained with the subject’s shoes and jacket or coat removed. 
9.1.7 Noise Exposure 
The subject will be queried about noise exposure since the last visit (Interval noise history) at 
Day 15, Day 30, Day 60, and Day 90 visits. 
9.1.8 
Otoscopy 
Microscopic otoscopy will be included to specifically record any abnormalities of the 
external ear canal, tympanic membrane and middle ear and will be performed at Screening, 
prior to injection on Day 1 and at the Day 15, Day 30, Day 60, and 90 visits. 
9.2 
Audiometry 
Refer to the Schedule of Assessments for timing and order of assessments. 
9.2.1 Comprehensive audiogram 
A comprehensive audiometric exam (a graphical representation) to determine how well a 
subject can hear as well as a person’s ability to understand speech. will be performed at the 
Screening visit, prior to baseline, and Day 15, 30, 60, and 90.  This will be performed on a 
calibrated audiometer by a licensed audiologist.  At the is following different frequencies will 
be obtained:  Air: 250 Hz, 500 Hz, 1 kHz, 2 kHz, 3 kHz, 4 kHz, 6 kHz, and 8 kHz; Bone: 500 
Hz, 1 kHz, 2 kHz, and 4 kHz. 


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9.2.2 Tympanometry 
Tympanometry (an objective test of middle-ear function is an examination used to test the 
condition of the middle ear and mobility of the eardrum (tympanic membrane) and the 
conduction bones, by creating variations of air pressure in the ear canal.  Middle ear 
compliance, peak pressure, and tympanogram type will be recorded.  Tympanometry will be 
performed at the Screening, Day 15, 30, 60, and 90 Visits.  The assessment will be performed 
by a licensed audiologist on a calibrated tympanometer.  A print-out containing the 
tympanometry from the machine will be collected and store in the source document.   
9.2.3 Word Recognition (quiet) 
Word recognition in quiet (AB words) will be measured with recorded CNC word lists.  The 
test will consist of 50 words presented to the subject and the percentage of words correctly 
identified will be recorded.  Word recognition will be performed at Screening, Day 15, 30, 
60, and 90 Visits.  Testing will be performed at 40dB above speech reception threshold. 
9.2.4 Words-In-Noise (WIN) 
The Words-in-Noise Test (WIN) was developed as an instrument to quantify the ability of 
listeners to understand monosyllabic words in background noise using multitalker babble 
(Wilson, 2003). Materials are recorded at 7 signal-to-noise ratios (0, 4, 8, 12, 16, 20, 24 dB) 
that are presented in a descending manner.  The purpose of this test is to mimic the real world 
in which background noises are maintained at consistent levels for different listening 
situations.  This test is conducted by presenting 2 lists of 35 recorded words to each ear.  
Overall accuracy will be recorded as the number of words identified correctly.  This test will 
occur at Screening, Day 15, 30, 60, and 90.  Testing will be performed at 40dB above speech 
reception threshold. 
9.3 
Pharmacokinetic Assessment 
The systemic exposure of the active pharmaceutical ingredients will be assessed.  Plasma 
levels of FX00 and FX03 will be measured from plasma samples obtained at time points 
according to Table 1.  Calculated PK parameters include Cmax, AUC0inf, CL, Vss, T1/2, and 
Tmax. 


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10 ADVERSE EVENTS 
Adverse Event Definition 
An AE is defined as any untoward medical occurrence in a clinical study subject 
administered a medicinal product which does not necessarily have a causal relationship with 
this treatment. An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease temporally associated with the use of a 
medicinal (investigational) product, whether or not it is related to the medicinal 
(investigational) product. This includes an exacerbation of pre-existing conditions or events, 
intercurrent illnesses, drug interaction or the significant worsening of the indication under 
investigation that is not recorded elsewhere in the CRF under specific efficacy assessments. 
Anticipated fluctuations of pre-existing conditions, including the disease under study that do 
not represent a clinically significant exacerbation or worsening need not be considered AEs. 
It is the responsibility of the investigator to document all AEs that occur during the study. 
AEs will be elicited by asking the subject a nonleading question, for example, “Have you 
experienced any new or changed symptoms since we last asked/since your last visit?”. AEs 
should be reported on the appropriate page of the CRF. 
Assessment of Severity 
Each AE will be assigned a category by the investigator as follows: 
Mild: 
An AE that is easily tolerated by the subject, causes minimal discomfort 
and does not interfere with everyday activities. 
Moderate: 
An AE that is sufficiently discomforting to interfere with normal 
everyday activities; intervention may be needed. 
Severe: 
An AE that prevents normal everyday activities; treatment or other 
intervention usually needed. 
If there is a change in severity of an AE, it must be recorded as a separate event. 
Assessment of Causality 
Every effort will be made by the investigator to assess the relationship of the AE, if any, to 
the study drug. Causality should be assessed using the categories presented in the following 
table: 
Unrelated: 
Clinical event with an incompatible time relationship to study 
drug administration, and that could be explained by underlying 
disease or other drugs or chemicals or is incontrovertibly not 
related to the study drug. 
Unlikely: 
Clinical event whose time relationship to study drug 
administration makes a causal connection improbable, but that 
could plausibly be explained by underlying disease or other drugs 
or chemicals. 
Possible: 
Clinical event with a reasonable time relationship to study drug 
administration, but that could also be explained by concurrent 
disease or other drugs or chemicals. 


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Probable: 
Clinical event with a reasonable time relationship to study drug 
administration and is unlikely to be attributed to concurrent 
disease or other drugs or chemicals. 
Treatment Emergent Adverse Events (Audiometric and Otoscopic) 
If any of the following criteria are met, the event will be recorded as an treatment emergent 
adverse event (TEAE) per ASHA guidelines (ASHA 1994): 
• Asymmetric loss of hearing greater than 20 dB at any one frequency in the treated ear. 
• Asymmetric loss of hearing greater than 10 dB at two adjacent frequencies in the 
treated ear. 
• Asymmetric loss of response at three consecutive test frequencies where responses 
were previously obtained in the treated ear. 
For word recognition testing, a follow up visit score that falls below the lower limit of the 
95% confidence interval for the baseline word recognition score will be considered a TEAE 
(Thorton and Raffin 1978). 
Additionally, if the subject experiences a perforation greater than 25% of the tympanic 
membrane in the treated ear this will be recorded as a TEAE.  
If two subjects experience the same TEAE, randomization and dosing will be paused, data 
will be evaluated, and a determination will be made about dosing remaining subjects in the 
study. 
Given that study participants are to be diagnosed with stable SNHL, there is the possibility 
that patients could be exposed to loud noise during the observation period of the study and 
before their scheduled audiometric testing. This noise exposure during could potentially 
contribute to the incidence of audiometric TEAEs and may complicate the interpretation of 
causality. For this reason, confirmation of audiometric TEAEs if observed, will be done at the 
subject’s next study visit.  Based on the repeat audiometric testing, the final interpretation of 
causality will be determined. 
 
Action Taken 
The investigator will describe the action taken in the appropriate section of the CRF, as 
follows: 
• 
None 
• 
Concomitant medication 
• 
Other, specify. 
Follow-up of Adverse Events 
All investigators should follow up subjects with AEs until the event is resolved or until, in the 
opinion of the investigator, the event is stabilized or determined to be chronic. Details of AE 
resolution must be documented in the CRF. 


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Documentation and Reporting of Adverse Events 
AEs should be reported and documented in accordance with the procedures outlined below. 
All AEs occurring during the study must be documented on the relevant CRF pages. The 
following data should be documented for each AE: 
•
Description of the symptom event
•
Classification of ‘serious’ or ‘not serious’
•
Severity
•
Date of first occurrence and date of resolution (if applicable)
•
Action taken
•
Causal relationship
•
Outcome of event (unknown, recovered, not yet recovered, recovered with sequelae,
death [with date and cause reported])
10.1 Serious Adverse Events 
Serious Adverse Event Definition 
An SAE is any untoward medical occurrence or effect that, at any dose, 
•
Results in death.
•
Is life-threatening (an AE is life-threatening if the subject was at immediate risk of
death from the event as it occurred, i.e., it does not include a reaction that might have
caused death if it had occurred in a more serious form).
•
Requires or prolongs inpatient hospitalization. (Complications occurring during
hospitalization are AEs and are SAEs if they cause prolongation of the current
hospitalization. Hospitalization for elective treatment of a pre-existing non--worsening
condition is not, however, considered an AE. The details of such hospitalizations must
be recorded on the medical history or physical examination page of the CRF).
•
Results in persistent or significant disability/incapacity. (An AE is incapacitating or
disabling if it results in a substantial and/or permanent disruption of the subject’s ability
to carry out normal life functions).
•
Results in a congenital anomaly/birth defect.
In addition, medical and scientific judgement is required to decide if prompt notification is 
required in situations other than those defined for SAEs above. This may include any event 
that the Investigator regards as serious that did not strictly meet the criteria above but may 
have jeopardized the subject or required intervention to prevent one of the outcomes listed 
above, or that would suggest any significant hazard, contraindication, side effect, or 
precaution that may be associated with the use of the investigational product. 
Reporting of Serious Adverse Events 
Any SAE must be reported by the Investigator if it occurs during the clinical study, whether 
or not the SAE is considered to be related to the investigational product. An SAE report 
consists of the SAE form, the AE form, and the concomitant medication form. A copy of 
these forms must be emailed within 24 hours for the attention of the Sponsor at 
 .  Additional contact details of the Medical Monitor can 
be found in the Medical Monitoring Plan. 


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The Investigator should not wait to receive additional information to document fully the 
event before notification of a SAE, though additional information may be requested. Where 
applicable, information from relevant laboratory results, hospital case records, and autopsy 
reports should be obtained. 
Instances of death, congenital abnormality, or an event that is of such clinical concern as to 
influence the overall assessment of safety, if brought to the attention of the investigator at any 
time after cessation of study drug administration and linked by the investigator to this study, 
should be reported to the study monitor. 
The sponsor and/or CRO will promptly notify all relevant investigators and the regulatory 
authorities of findings that could adversely affect the safety of subjects, impact on the 
conduct of the study or alter the independent ethics committee (IEC)/institutional review 
board (IRB) approval/favorable opinion of the study. In addition, CRO on behalf of the 
sponsor, will expedite the reporting to all concerned investigators, to the IRBs, where 
required, and to the regulatory authorities of all adverse reactions that are both serious and 
unexpected. 
Details of the procedures to be followed if a pregnancy occurs are provided in Section 7.3.4. 
10.2 Unexpected Adverse Reactions 
Unexpected Adverse Reaction Definition 
An unexpected adverse reaction is any untoward and unintended response that is related to 
the administration of the study drug at any dose that is not consistent with the applicable 
product information (e.g. investigators brochure for an unauthorized investigational 
medicinal product or summary of product characteristics for an authorized product). 
All suspected unexpected serious adverse reactions (SUSARs) will be the subject of 
expedited reporting. The sponsor and/CRO shall ensure that all relevant information about a 
SUSAR that is fatal or life-threatening is reported to the relevant competent authorities and 
IRB within 7 days after knowledge by the sponsor of such a case and that relevant follow up 
information is communicated within an additional 8 days. All other SUSARs will be reported 
to the relevant competent authorities and IRB within 15 days after knowledge by the sponsor 
of such a case. All investigators should follow up SUSARs until the event is resolved or until, 
in the opinion of the investigator, the event is stabilized or determined to be chronic. Post 
study SUSARs that occur after the subject has completed the clinical study must be reported 
by the investigator to the sponsor. 
11 STATISTICAL METHODS 
11.1 Statistical and Analytical Plans 
11.1.1 Descriptive Statistics 
The study is a phase 1 / 2, randomized, double-blind, placebo-controlled single dose study to 
evaluate the safety at two dose levels of FX 322 (FX-322L and FX-322H) compared to 
placebo in adults with stable sensorineural hearing loss (no changes over 6 months of 10dB 
or more in any frequency).  Approximately 24 subjects will participate in the trial.  Subjects 
will be randomized to one of four treatment groups:  FX-322L, placeboL, FX-322H, or 


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placeboH using a 1:1 allocation ratio for dose cohort (12 per cohort) and within each cohort a 
2:1 allocation ratio for study drug (8 FX-322: 4 Placebo).  Since the volume in the syringe 
may be potentially unblinding to the investigator, for this study cohort assignment may not be 
blinded to the otolaryngologist; however, randomization to FX-322 or placebo will remain 
double-blind.  Following randomization subjects will receive a single intratympanic injection 
of blinded study drug into the “treated” or “study” ear.  The contralateral ear will be 
designated as the “untreated” ear. 
 
The primary objective(s) for this study is to assess the systemic safety of FX-322, the effect 
of FX-322 on otologic and audiologic measures, and the plasma PK profile to determine the 
systemic exposure to the active pharmaceutical ingredients.  The statistical analyses will 
consist of descriptive statistics including the mean, standard deviation (SD), median, 
minimum and maximum statistics for continuous endpoints and numbers and percent for 
categorical endpoints.  Statistics will be presented by dose cohort, treatment group, and 
overall.  Where relevant, 95% confidence intervals for summary statistics will also be 
provided.  Figures describing individual subject data may also be provided.  The general 
statistical methods are outlined below; however, more detailed descriptions of the statistical 
methods and approaches will be provided in the Statistical Analysis Plan (SAP) which will be 
prepared and finalized prior to breaking the blind. 
 


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11.1.2 Sample Size 
Since, the study is aimed at demonstrating safety of FX-322 the selected sample size was 
considered adequate for an initial assessment of safety and tolerability and was not based on 
formal statistical considerations such as power.  
11.1.3 Analysis Sets 
For the purpose of analysis subjects will be considered participating in the study if they are 
randomized.  Subjects signing the informed consent who are not randomized will not be 
included in any analysis. 
•
Safety Analysis Set
The safety analysis set will include all subjects exposed to study drug and will be analyzed 
according to the actual treatment received regardless of the randomized treatment.  The safety 
analysis set will be used for safety analyses. 
•
Pharmacokinetic Analysis Set
All subjects in the safety analysis set with measurable plasma concentrations will be included 
in the PK analysis set.   
•
Per Protocol Analysis Set (PPAS)
The per protocol analysis set will consist of all subjects in the SAS who meet all of the 
inclusion and none of the exclusion criteria and have no major protocol deviations impacting 
the interpretation of the study results.  Major protocol deviations potentially impacting the 
interpretation of the study results will be determined by the study team prior to breaking the 
blind.  If different from the SAS, the PPAS may be used in additional analyses.   
11.1.4 Subject Disposition 
Subject disposition will be summarized by dose cohort, randomized treatment group, and 
overall and will include the following: 
•
The number of all randomized subjects regardless of exposure to study drug or
availability of post-baseline data
•
The number and percent of subjects in each analysis set
•
The number and percent of subjects completing the study
•
The number and percent of subjects not completing the study and the reason for early
withdrawal


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11.1.5 Description of Subgroups to be Analyzed  
Give the small sample size it is not anticipated that the data will be summarized by subgroups 
other than treatment; however, analyses for some subgroups, if feasible, may be conducted. 
 
11.1.6 Subject Demographics and Baseline Characteristics 
Descriptive statistics for subject demographics and baseline disease status will be provided 
using the safety analysis set. 
 
11.1.7 Safety Analyses 
Safety will be evaluated using the safety analysis set and will include treatment emergent 
AEs (TEAEs), serious adverse events (SAE)s, vital signs, clinical laboratory measurements, 
electrocardiograms (ECGs), concomitant medications, otology and audiometry assessments.   
Summary descriptive statistics will be provided by actual treatment received and overall.  No 
safety data will be imputed except for partial dates if required to determine if an adverse 
event is treatment emergent or a medication concomitant with exposure to study drug.  
Details of partial date imputation will be provided in the SAP. 
 
11.1.7.1  Adverse Events 
Treatment emergent adverse events (TEAEs) will be summarized via the MedDRA system by 
organ class and preferred term using subject incidence rates.  Data will be tabulated by 
severity, physician assessment of relationship to study drug, serious TEAEs, and TEAEs 
leading to death or study withdrawal.  Adverse events of the ear will also be summarized by 
treated vs. non-treated ear. 
 
11.1.7.2  Clinical Laboratory Evaluations 
The analysis of laboratory parameters will include descriptive statistics for baseline and each 
post-baseline visit along with the change from baseline and/or shift tables for the proportion 
of subjects within and outside the normal ranges.  All laboratory assessments will be included 
in line listings.  Urinalysis results will be presented in a similar manner as laboratory 
assessments.  Pregnancy results will not be summarized but will be provided in data tables.   
 
11.1.7.3  Vital Signs 
Vital signs will be summarized via descriptive statistics similar to laboratory evaluations 
described above. 
 
11.1.7.4 Physical Examination and Electrocardiogram 
Baseline physical exam and electrocardiogram data, where relevant, will be summarized in 
the subject demographic and baseline characteristic table.  All other physical examination and 
electrocardiogram data results will be provided in tables.


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11.1.7.5  Prior and Concomitant Medications 
Prior and concomitant medications will be coded to identify the drug class and preferred drug 
name.  Concomitant medications will include all medications that started, or were continuing, 
during or after administration of study drug.  Prior medications will include all recorded 
medications that started and stopped prior to administration of study drug. 
The number and percent of subjects using concomitant medications will be tabulated by drug 
class and preferred drug name.  Prior medications will be presented in line listings only.  
Concomitant medications will also be provided in line listings. 
11.1.7.6 Tympanometry and Otoscopy 
Tympanometry and otoscopic results will be presented by dose cohort, treatment group and 
ear (treated vs. untreated) at baseline and each visit with changes and/or shifts from baseline.  
11.1.7.7 Comprehensive Audiometry 
Comprehensive audiometry will be summarized by frequency for each dose cohort, treatment 
group, and ear (treated vs. untreated) for baseline and each post-baseline visit including 
changes from baseline.  Separate analyses will be conducted for air and bone.   
11.2 Handling of Missing Data and Subject Withdrawals 
In general, other than partial dates to assess treatment emergent AEs and concomitant 
medications, there will be no imputation of missing data.  Subjects will be included in a 
particular analysis so long as they have no missing data for the particular endpoint and/or 
visit being evaluated.   
11.3 Protocol Deviations 
Protocol deviations will be provided in line listings.  A table of major and minor protocol 
deviations will be tabulated by dose cohort, treatment group, and overall. 
12 QUALITY ASSURANCE AND QUALITY CONTROL 
12.1 Audit and Inspection 
Study centers and study documentation may be subject to Quality Assurance audit during the 
course of the study by the sponsor or its nominated representative. In addition, inspections 
may be conducted by regulatory authorities at their discretion. 
12.2 Monitoring 
Data for each subject will be recorded on a eCRF. Data collection must be completed for 
each subject who signs an informed consent form (ICF) and is administered study drug. 
In accordance with GCP and ICH guidelines, the study monitor will carry out source 
document verification at regular intervals to ensure that the data collected in the CRF are 
accurate and reliable.  


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The investigator must permit the monitor, the IEC/IRB, the sponsor’s internal auditors, and 
representatives from regulatory authorities direct access to all study-related documents and 
pertinent hospital or medical records for confirmation of data contained within the CRFs.   
12.3 Data Management and Coding 
The sponsor or CRO will be responsible for activities associated with the data management of 
this study. This will include setting up a relevant database and data transfer mechanisms, 
along with appropriate validation of data and resolution of queries. Data generated within this 
clinical study will be handled according to the relevant standard operating procedures (SOPs) 
of the data management and biostatistics departments of sponsor or CRO. 
Study centers will enter data directly into an electronic data capture (EDC) system by 
completing the CRF via a secure internet connection. Data entered into the eCRF must be 
verifiable against source documents at the study center. Data to be recorded directly on the 
eCRF will be identified and the eCRF will be considered the source document. Any changes 
to the data entered into the EDC system will be recorded in the audit trail and will be FDA 
CFR 21 Part 11 compliant. 
Medical coding will use Medical Dictionary for Regulatory Activities (MedDRA) for 
concomitant diseases and AEs and WHO Drug for medications. 
Missing or inconsistent data will be queried in writing to the investigator for clarification. 
Subsequent modifications to the database will be documented. 
13 RECORDS AND SUPPLIES 
13.1 Drug Accountability 
On receipt of the study drug, the investigator (or deputy) will conduct an inventory of the 
supplies and verify that study drug supplies are received intact and in the correct amounts 
before completing a supplies receipt. The investigator will retain a copy of this receipt at the 
study center and return the original receipt to the study monitor. The monitor may check the 
study supplies at each study center at any time during the study. 
It is the responsibility of the study monitor to ensure that the investigator (or deputy) has 
correctly documented the amount of the study drug received, dispensed, and returned on the 
dispensing log that will be provided. A full drug accountability log will be maintained at the 
study center at all times. The study monitor will arrange collection of unused study drug 
returned by the subject. The study monitor will also perform an inventory of study drug at the 
close-out visit to the study center. All discrepancies must be accounted for and documented. 
13.2 Financing and Insurance 
Financing and insurance of this study will be outlined in a separate agreement between CRO 
and the sponsor. 


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14 ETHICS 
14.1 Independent Ethics Committee or Institutional Review Board 
Before initiation of the study at each study center, the protocol, the ICF, other written 
material given to the subjects, and any other relevant study documentation will be submitted 
to the appropriate IEC/IRB. Written approval of the study and all relevant study information 
must be obtained before the study center can be initiated or the study drug is released to the 
investigator. Any necessary extensions or renewals of IEC/IRB approval must be obtained for 
changes to the study such as amendments to the protocol, the ICF or other study 
documentation. The written approval of the IEC/IRB together with the approved ICF must be 
filed in the study files. 
The investigator will report promptly to the IEC/IRB any new information that may adversely 
affect the safety of the subjects or the conduct of the study. The investigator will submit 
written summaries of the study status to the IEC/IRB as required. On completion of the study, 
the IEC/IRB will be notified that the study has ended. 
14.2 Regulatory Authorities 
Relevant study documentation will be submitted to the regulatory authorities of the 
participating countries, according to local/national requirements, for review and approval 
before the beginning of the study. On completion of the study, the regulatory authorities will 
be notified that the study has ended. 
14.3 Ethical Conduct of the Study 
The investigator(s) and all parties involved in this study should conduct the study in 
adherence to the ethical principles based on the Declaration of Helsinki, GCP, ICH 
guidelines, and the applicable national and local laws and regulatory requirements. 
14.4 Informed Consent 
The process of obtaining informed consent must be in accordance with applicable regulatory 
requirement(s) and must adhere to GCP. 
The investigator is responsible for ensuring that no subject undergoes any study related 
examination or activity before that subject has given written informed consent to participate 
in the study. 
The investigator or designated personnel will inform the subject of the objectives, methods, 
anticipated benefits, potential risks, and inconveniences of the study. The subject should be 
given every opportunity to ask for clarification of any points s/he does not understand and, if 
necessary, ask for more information. At the end of the interview, the subject will be given 
ample time to consider the study. Subjects will be required to sign and date the ICF. After 
signatures are obtained, the ICF will be kept and archived by the investigator in the 
investigator’s study file. A signed and dated copy of the subject ICF will be provided to the 
subject or their authorized representative. 
It should be emphasized that the subject may refuse to enter the study or to withdraw from 
the study at any time, without consequences for their further care or penalty or loss of 


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CONFIDENTIAL 
benefits to which the subject is otherwise entitled. Subjects who refuse to give or who 
withdraw written informed consent should not be included or continue in the study. 
If new information becomes available that may be relevant to the subject’s willingness to 
continue participation in the study, a new ICF will be approved by the IEC(s)/IRB(s) (and 
regulatory authorities, if required). The study subjects will be informed about this new 
information and reconsent will be obtained. 
14.5 Subject Confidentiality 
Monitors, auditors, and other authorized agents of the sponsor and/or its designee, the 
IEC(s)/IRB(s) approving this research, and the United States (US) FDA, as well as that of 
any other applicable agency(ies), will be granted direct access to the study subjects’ original 
medical records for verification of clinical study procedures and/or data, without violating the 
confidentiality of the subjects to the extent permitted by the law and regulations. In any 
presentations of the results of this study or in publications, the subjects’ identity will remain 
confidential. 
All personal data collected and processed for the purposes of this study should be managed 
by the investigator and his/her staff with adequate precautions to ensure confidentiality of 
those data, and in accordance with the Health Insurance Portability and Accountability Act 
(1), applicable to national and/or local laws and regulations on personal data protection. 
Reporting and Publication, Including Archiving 
Essential documents are those documents that individually and collectively permit evaluation 
of the study and quality of the data produced. After completion of the study (end of study 
defined as the date of the last visit of the last subject), all documents and data relating to the 
study will be kept in an orderly manner by the investigator in a secure study file. This file 
will be available for inspection by the sponsor or its representatives. Essential documents 
should be retained for 2 years after the final marketing approval in an ICH region or for at 
least 2 years since the discontinuation of clinical development of the investigational product. 
It is the responsibility of the sponsor to inform the study center when these documents no 
longer need to be retained. The investigator must contact the sponsor before destroying any 
study related documentation. In addition, all subject medical records and other source 
documentation will be kept for the maximum time permitted by the hospital, institution, or 
medical practice. 
The sponsor must review and approve any results of the study or abstracts for professional 
meetings prepared by the investigator(s). Published data must not compromise the objectives 
of the study. Data from individual study centers in multicenter studies must not be published 
separately. 


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15 REFERENCES 
1.
US Department of Health and Human Services.  The Health Insurance Portability and
Accountability 
Act 
(HIPAA) 
of 
1996 
(P.L.104-191) 
[HIPAA].
http://aspe.hhs.gov/admnsimp/pl104191.htm. Effective August 21, 1996.
2.
Li, L., Chao, T., Brant, J., O'Malley Jr, B., Tsourkas, A., & Li, D. (2017). Advances in Nano-
based inner ear delivery systems for the treatment of sensorineural hearing loss. Adv Drug
Delivery. Advanced Drug Delivery Reviews.
3.
Hong , O., Kerr, M., & Poling , G. (April, 2013). Understanding and preventing noise-induced
hearing loss. ELSEVIER, 110-118.
4.
Basner, M., Brink, M., Bristow, A., de Kluezinaar, Y., Finegold , L., & Hong, J. (2015). ICBEN
review of research on the biological effects of noise 2011-2014. Noise & Health, 57-82.
5.
Lin, F., Niparko, J., & Ferrucci, L. (November, 2011). Hearing loss prevalence in the United
States. Arch Intern Med., 1851-1853.
6.
Nelson DI, Nelson RY, Concha-Barrientos M, Fingerhut M, (2005). The global burden of
occupational noise-induced hearing loss. Am J Ind Med, 48(6): 446-458.
7.
Choi, J., Betz, J., Li, L., Blake, C., Sung, Y., Contrera, K., & Lin, F. (July 2016). Association of
Using Hearing Aids or Cochlear Implants With Changes in Depressive Symptoms in Older
Adults. JAMA Orolaryngology -Head & Neck Surgery, 142(7):652–657.
8.
McGilton, K., Höbler, F., Campos, J., Dupuis, K., Labreche, T., & Guthrie, D. (2016). Hearing
and vision screening tools for long-term care residents with dementia: protocol for a scoping
review. BMJ Open.
9.
Deal, J., Betz, J., Yaffe, K., Harris, T., Purchase-Helzner, E., Satterfield, S., & Pratt, S. (2017).
Hearing impairment and incident dementia and cognitive decline in older adults: The health ABC
study. Journals of Gerontology - Series A Biological Sciences and Medical Sciences, 703-709.
10.
Thornton AR, Raffin MJM: Speech discrimination scores modified as a binomial variable. J Sp
Hear Res 1978;21:507–518.
11.
American Speech-Language-Hearing Association. (1994). Guidelines for the audiologic
management of individuals receiving cochleotoxic drug therapy. ASHA 36 (Suppl
12):11-19.
12.
World Health Organization. (2015). Hearing Loss due to recreational exposure to loud sounds:
A review.
13.
Wilson RH, Abrams, HB, Pillion AL. (2003). A word-recognition task in multitalker babble
using a descending presentation mode from 24 dB to 0 dB signal to babble. J Rehabil Res Dev.
2003 Jul-Aug;40(4):321-7.


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16 APPENDIX 1 
Word Recognition Scoring Chart 


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17 INVESTIGATOR SIGNATURE PAGE 
Protocol Title: 
A Phase 1/2 Randomized, Double-blind, Placebo-controlled Single 
Dose Study at Two Dose Levels of FX-322 Administered by 
Intratympanic Injection in Adults with Stable Sensorineural Hearing 
Loss 
Protocol Number: 
FX-322-201 
Confidentiality and GCP Compliance Statement 
I, the undersigned, have reviewed this protocol (and amendments), including appendices, and 
I will conduct the study as described in compliance with this protocol (and amendments), 
GCP, and relevant ICH guidelines. 
Once the protocol has been approved by the IEC/IRB, I will not modify this protocol without 
obtaining prior approval of Frequency Therapeutics and of the IRB. I will submit the protocol 
amendments and/or any ICF modifications to Frequency Therapeutics and IRB, and approval 
will be obtained before any amendments are implemented. 
I understand that all information obtained during the conduct of the study with regard to the 
subjects’ state of health will be regarded as confidential. No subjects’ names will be 
disclosed. All subjects will be identified by assigned numbers on all CRFs, laboratory 
samples, or source documents forwarded to the sponsor. Clinical information may be 
reviewed by the sponsor or its agents or regulatory agencies. Agreement must be obtained 
from the subject before disclosure of subject information to a third party. 
Information developed in this clinical study may be disclosed by [sponsor name], to other 
clinical investigators, regulatory agencies, or other health authority or government agencies 
as required. 
Investigator Signature 
Date 
Printed Name 
Institution 

SAP_001.pdf: 
Frequency Therapeutics 
FX-322-201 
Statistical Analysis Plan 
Version 1.0 
Issue Date 10-AUG-2018 
 
 
 
Page 1 of 26 
 
 
Confidential 
 
 
 
 
Statistical Analysis Plan 
A Phase 1/2 Randomized, Double-blind, Placebo-controlled Single Dose 
Study at Two Dose Levels of FX-322 Administered by Intratympanic 
Injection in Adults with Stable Sensorineural Hearing Loss 
Protocol Number: FX-322-201 
 
Version 1.0 
Issue Date: 10-Aug-2018 
Authors: 
  
 
 
Previous Versions 
Version 0.1, Issued: 08-Jun-2018 
Version 0.2, Issued: 22-Jun-2018 
Version 0.3, Issued: 23-Jul-2018 
 


 
Frequency Therapeutics 
 
FX-322-201 
 
Statistical Analysis Plan 
 
Version 1.0 
 
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SAP Amendments before database lock 
 
Version  Issue Date 
Section 
Revision/Addition 
Rationale 
 
Not Applicable 
 
 
 
 
 
 


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Table of Contents 
1 
INTRODUCTION ............................................................................................... 5 
2 
STUDY OBJECTIVES ....................................................................................... 5 
3 
ENDPOINTS ....................................................................................................... 5 
3.1 
Safety Endpoints ............................................................................................................. 5 
3.2 
Pharmacokinetic Endpoints ............................................................................................ 5 
4 
SAMPLE SIZE .................................................................................................... 6 
5 
RANDOMIZATION ........................................................................................... 6 
6 
PLANNED ANALYSES ..................................................................................... 6 
6.1 
Analysis Sets ................................................................................................................... 6 
6.1.1 
Randomized Set ........................................................................................................ 7 
6.1.2 
Safety Analysis Set ................................................................................................... 7 
6.1.3 
Per Protocol Analysis Set.......................................................................................... 7 
6.1.4 
PK Analysis Set ........................................................................................................ 7 
6.2 
Derived Data ................................................................................................................... 7 
6.2.1 
Race........................................................................................................................... 7 
6.2.2 
Baseline ..................................................................................................................... 7 
6.2.3 
Early Withdrawal Assessments................................................................................. 8 
6.2.4 
Duration/Study Day/Time ......................................................................................... 8 
6.2.5 
Conventions for Missing and Partial Dates .............................................................. 8 
6.2.6 
Inexact Values ........................................................................................................... 8 
6.2.7 
Unscheduled Visits ................................................................................................... 8 
6.2.8 
PK Parameters ........................................................................................................... 8 
6.2.9 
Word Recognition (Quiet) ...................................................................................... 10 
6.2.10 
Identifying the Treated Ear ..................................................................................... 10 
6.3 
Conventions .................................................................................................................. 10 
6.3.1 
Decimal Places ........................................................................................................ 11 
6.4 
Subject Disposition ....................................................................................................... 11 
6.5 
Protocol Deviations ....................................................................................................... 11 
6.6 
Baseline Comparability ................................................................................................. 12 
6.7 
Demographic data ......................................................................................................... 12 
6.8 
Medical History ............................................................................................................ 12 
6.9 
Prior and Concomitant Medications ............................................................................. 12 


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6.10 
Exposure to Study Drug ................................................................................................ 13 
6.11 
Pharmacokinetic Analyses ............................................................................................ 13 
6.12 
Biomarker Analysis ...................................................................................................... 13 
6.13 
Safety Analyses ............................................................................................................. 13 
6.13.1 
Adverse Events ....................................................................................................... 14 
6.13.2 
Laboratory Data ...................................................................................................... 15 
6.13.3 
Vital Signs ............................................................................................................... 15 
6.13.4 
Electrocardiogram Data .......................................................................................... 15 
6.13.5 
Physical Examination.............................................................................................. 16 
6.13.6 
Tympanometry and Otoscopy Assessments ........................................................... 16 
6.13.7 
Comprehensive Audiogram Assessment ................................................................ 16 
6.13.8 
Word Recognition and Words-In-Noise Assessments ............................................ 16 
6.13.9 
Interval Noise History ............................................................................................. 17 
7 
BLIND SAFETY DATA REVIEW ..................................................................17 
8 
CHANGES TO PLANNED PROTOCOL ANALYSIS ...................................17 
9 
REFERENCES ..................................................................................................18 
10 LIST OF TABLES, FIGURES AND LISTINGS .............................................19 
APPENDIX 1 ...........................................................................................................25 
 
 
 


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1 INTRODUCTION 
This document details the planned statistical analyses for the Frequency Therapeutics, protocol 
“FX-322-201” study titled “A Phase 1/2 Randomized, Double-blind, Placebo-controlled Single 
Dose Study at Two Dose Levels of FX-322 Administered by Intratympanic Injection in Adults 
with Stable Sensorineural Hearing Loss”. 
The proposed analyses are based on the amended version of the Protocol version 2.0 (dated 
29-May-2018).   
This is a Phase 1/2 randomized, double-blind, placebo-controlled, single dose study at two dose 
levels of FX-322 compared to placebo in adults with stable sensorineural hearing loss (no changes 
over six months of 10 dB or more in any frequency). The two dose levels proposed in this study, 
FX-322L and FX-322H will be dosed concurrently.  The study will have three phases: Screening, 
Treatment with Observation, and Follow-up.   
2 STUDY OBJECTIVES 
The primary objectives of the study are: 
• 
To assess the systemic safety of FX-322 
• 
To assess the plasma pharmacokinetic (PK) profile to determine the systemic exposure to 
the active pharmaceutical ingredients 
• 
To assess the effect of FX-322 on otologic and audiologic measures 
3 ENDPOINTS 
3.1 Safety Endpoints 
The following safety assessments will be evaluated at the Screening visit and throughout: 
Treatment emergent adverse events (TEAEs), clinical laboratory tests (including hematology, 
serum 
chemistry, 
coagulation, 
and 
urinalysis), 
vital 
signs, 
physical 
examination, 
electrocardiogram (ECG),  medical history, prior and concomitant medications, audiometry, and 
otoscopy.   
3.2 Pharmacokinetic Endpoints 
Serial blood samples will be collected predose and postdose through 24 hours to determine 
concentration of FX00 and FX03 in plasma.  PK parameters including Cmax, AUC0-inf, CL/F, Vz/F, 


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T1/2, and Tmax will be calculated from plasma concentrations of FX00 and FX03.  Additional PK 
parameters may be calculated if deemed appropriate.   
4 SAMPLE SIZE 
Approximately 24 subjects are planned to be randomized into the study, and 24 subjects are 
expected to complete the study. Subjects who do not comply with the protocol or who withdraw 
consent may be replaced.  Additional subjects may replace early withdrawal subjects, unless the 
reason for discontinuation was a treatment emergent adverse event (TEAE).  
Since the study is aimed at demonstrating safety of FX-322, the selected sample size was 
considered adequate for an initial assessment of safety and tolerability and was not based on formal 
statistical considerations such as power.   
5 RANDOMIZATION 
Subjects will be randomized to one of four treatment groups based on the randomization schedule 
prepared by the unblinded statistician.   
Subjects will receive a single dose of either FX-322L, placeboL, FX-322H, or placeboH. 
Randomized subjects will be allocated 1:1 to one of 2 cohorts (12 in each cohort) and 2:1 allocation 
ratio to study drug (8 FX-322:4 placebo) within each cohort.  
The otolaryngologist will be blinded to whether a patient is randomized to drug or placebo within 
a cohort, however, may not be blinded to the cohort (high dose or low dose) because low and high 
dose cohorts have different injection volumes.   
6 PLANNED ANALYSES 
No statistical analysis plan (SAP) prepared in advance of the data can be absolutely definitive and 
the final clinical study report (CSR) may contain additional tables or statistical tests if warranted 
by the data obtained.  The justification for any such additional analyses will be fully documented 
in the final CSR. 
6.1 Analysis Sets 
Subjects excluded from the analysis sets and the reason for their exclusion will be listed in 
Appendix 16.2.  For the purpose of analysis, subjects will be considered participating in the study 
if they are randomized. 


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6.1.1 
Randomized Set 
The Randomized Set includes all subjects who were randomly allocated to a treatment group.     
6.1.2 
Safety Analysis Set 
The Safety Analysis Set (SAS) will include all subjects exposed to study drug and will be analyzed 
according to the actual treatment received regardless of the randomized treatment. The SAS will 
be used for safety analyses.  
6.1.3 
Per Protocol Analysis Set  
The Per Protocol Analysis Set (PPAS) will consist of all subjects in the SAS who meet all of the 
inclusion and none of the exclusion criteria and have no major protocol deviations impacting the 
interpretation of the study results. Major protocol deviations potentially impacting the 
interpretation of the study results will be determined by the study team prior to breaking the blind. 
If different from the SAS, the PPAS may be used in additional analyses.  
6.1.4 
PK Analysis Set 
All subjects in the SAS with measurable plasma concentrations will be included in the PK analysis 
set.  
6.2 Derived Data 
This section describes the derivations required for statistical analysis. Unless otherwise stated, 
variables derived in the source data will not be re-calculated. 
6.2.1 
Race 
Where more than one race category has been selected for a subject, these race categories will be 
combined into a single category labeled “Multiple Race” in the summary tables. The listings will 
reflect the original selected categories.  
6.2.2 
Baseline 
Baseline is defined as the last non-missing value (either scheduled, unscheduled or repeat) that is 
collected before dosing. 


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6.2.3 
Early Withdrawal Assessments 
Data collected during Early Withdrawal assessments will be grouped and summarized separately 
from scheduled assessments.  All Early Withdrawal data will be displayed as “Early Withdrawal” 
in the listings.  
6.2.4 
Duration/Study Day/Time  
Study day will be calculated as the number of days from the single dose of study drug. The day of 
study drug dosing will be considered as Study Day 1. 
 Date of event – date of dose of study drug + 1, for events on or after dosing  
 Date of event – date of dose of study drug, for events before dosing.   
6.2.5 
Conventions for Missing and Partial Dates 
It is not expected that there will be any missing dates, however in the rare case that an AE start 
date or time is missing and it is unclear whether the AE is treatment emergent or not then a 
conservative approach will be taken and it will be assumed that the AE occurred after first dosing.  
All dates presented in the individual subject listings will be as recorded on the eCRF.  
6.2.6 
Inexact Values 
In the case where a safety laboratory variable is recorded as “> x”, “≥ x”, “< x” or “≤ x”, a value 
of x will be taken for analysis purposes.   Original values, as reported by the analytical laboratory, 
will be presented in the listing.    
6.2.7 
Unscheduled Visits 
Only scheduled post-baseline assessments will be tabulated. Post-baseline repeat/unscheduled 
assessments will be disregarded, although these post-baseline assessments will be listed in the 
relevant appendices to the CSR.   
6.2.8 
PK Parameters 
Concentration-time data for FX00 and FX03 will be analyzed using noncompartmental methods 
in Phoenix™ WinNonlin® (Version 6.3 or higher, Certara, L.P.)1. 
During the pharmacokinetic analysis, concentrations below the limit of quantitation (BLQ) up to 
the time of the first quantifiable concentration will be treated as zero. Embedded (values between 
2 quantifiable concentrations) and terminal BLQ concentrations will be treated as “missing”. 


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Subjects receiving active treatment will be included in the PK analysis; subjects receiving placebo 
will be listed in the concentration-time listing by subject. 
The following PK parameters will be calculated for FX00 and FX03: 
Cmax 
Maximum 
concentration 
determined 
directly 
from 
individual 
concentration-time data; reported to 3 significant figures 
Tmax 
Time to reach maximum concentration determined directly from individual 
concentration-time data; reported to 3 significant figures 
AUClast 
Area under the concentration–time curve from time zero to the time of the 
last quantifiable concentration; calculated using the linear trapezoidal rule; 
reported to 3 significant figures 
AUCinf 
Area under the concentration–time curve extrapolated to infinity, calculated 
as:  
AUCinf = AUClast + Clast/λz, 
where Clast is the last quantifiable concentration and λz is the terminal 
elimination rate constant; reported to 3 significant figures 
AUCex 
The percentage of AUCinf based on extrapolation, calculated as:  
%AUCex = (AUCinf – AUClast)/AUCinf*100; reported to 3 significant figures 
T½ 
The observed terminal elimination half-life calculated as: 
T½= ln(2)/λz; reported to 3 significant figures 
λz 
The observed elimination rate constant; estimated by linear regression in 
the terminal phase of the log concentration-time profile; see additional 
criteria below; reported to 3 significant figures 
CL/F 
Apparent total body clearance after extravascular administration, calculated 
as: 
CL/F = Dose/AUCinf; reported to 3 significant figures 
Vz/F 
Apparent volume of distribution following extravascular administration 
based on the terminal phase, calculated as: 
Vz/F = (CL/F)/λz; reported to 3 significant figures 


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No value for λz and other λz-related parameters (AUCinf, T1/2, CL/F, etc.) will be reported if λz 
cannot be estimated.   
6.2.9 
Word Recognition (Quiet) 
The percentage of words correctly identified out of a list of 50 words will be calculated for the 
Word Recognition (Quiet) test.  If a subject has completed the assessment, it will be assumed that 
all 50 words were completed.   
6.2.10 
Identifying the Treated Ear 
For assessments performed for each ear (left/right), the treated ear will be identified from the CRF 
and assessments will be presented by treated and untreated ear.   
6.3 Conventions 
All clinical data listings, summaries, figures and statistical analyses will be generated using SAS 
(Version 9.4 or higher)2. 
Summaries of the clinical data will be presented by treatment group and overall.  
PK data listings, summaries, figures, and statistical analyses will be generated using Phoenix™ 
WinNonlin® (Version 6.3 or higher) or SAS (Version 9.4 or higher).  PK concentration data will 
be summarized by analyte (FX00 and FX03), treatment (FX-322L or FX-322H), and overall at 
each nominal sample time.  
Treatment group labels will be displayed as follows:  
PlaceboL 
PlaceboH 
Pooled 
Placebo 
FX-322L 
(0.05 mL) 
FX-322H 
(0.20 mL) 
Overall 
Listings will be sorted in the following order: Treatment group [PlaceboL, PlaceboH, FX-322L 
(0.05 mL), FX-322H (0.20 mL)], subject, parameter, and visit unless otherwise stated. All data 
will be listed.   
For clinical data, continuous variables will be summarized by the number of non-missing 
observations, mean, median, standard deviation, and minimum and maximum. A 95 % confidence 
interval (CI) will also be provided where change from baseline has been calculated.    
The following summary statistics will be used to summarize the PK data (concentration-time data 
and PK parameters) by analyte (FX00 and FX03), treatment (FX-322L or FX-322H) and overall: 
the number of non-missing observations (n), mean, standard deviation (SD), median, minimum 


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(min), maximum (max), and coefficient of variation (CV%), calculated as (SD/Mean)*100.  In 
addition, the geometric mean will be reported for Cmax and AUCs. Where relevant, 95% confidence 
intervals for summary statistics will also be provided.  
Categorical variables will be summarized by presenting the frequency and percent. Percentages 
will be based on the number of non-missing observations or the subject population unless 
otherwise specified. For each variable, all categories will be shown. Zero frequencies (but not the 
percent) within a category will be presented.  
6.3.1 
Decimal Places 
For all clinical safety data, means, medians and percentiles will be displayed to one more decimal 
place than the data, dispersion statistics (e.g. standard deviation) will have two more decimal 
places, and the minimum and maximum will be displayed to the same number of decimal places 
as reported in the raw data. Percentages will be displayed with one decimal place.  
For PK data, individual concentrations and parameter values will be reported to three significant 
figures and summarized by analyte (FX00 and FX03), treatment (FX-322L or FX-322H) and 
overall. For summary statistics, n will be reported as a whole number; mean, standard deviation, 
median, minimum, maximum, and geometric mean will be reported to the same precision as for 
individual data.  CV% will be reported to two decimal places; p-values will be reported to four 
decimal places.  Percent ratios of the geometric least squares means and associated 90% confidence 
intervals will be reported to two decimal places.  
6.4 Subject Disposition 
Subject disposition will be summarized as follows:  
 The number of subjects in each analysis set will be summarized by treatment group and 
overall for the Randomized set.  
 The number of early withdrawals and the reasons for withdrawal will be tabulated by 
treatment group for the Randomized set.  
6.5 Protocol Deviations 
A summary of major and minor protocol deviations will be tabulated by treatment group and 
overall.  A detailed listing of protocol deviations will be provided within Appendix 16.2.  


 
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6.6 Baseline Comparability 
The comparability of treatment groups with respect to subject demographics and baseline 
characteristics will be assessed in a descriptive manner, but no formal statistical testing will be 
performed.  The Safety Analysis Set will be used to summarize all baseline and demographic data. 
6.7 Demographic data 
Standard continuous or categorical variable summaries will be presented by treatment group for 
the following variables based on the Safety Analysis Set. 
 Age at Informed Consent (years) 
 Gender 
 Ethnicity 
 Race, where more than one race is selected the subject will be presented under the ‘Multiple 
races’ category in the summary but each selected race will be identified in the listing. 
 Affected Ear (Left/Right/Both) 
 Worst Ear (Left/Right) 
 Treated Ear (Left/Right, Reason) 
 Weight at Screening (kg) 
 Height at Screening (cm) 
 BMI at Screening (kg/m2) 
 Fertility Status 
6.8 Medical History 
Previous and ongoing conditions at baseline will be tabulated by treatment group and overall for 
the Safety Analysis Set. Conditions will be coded using the Medical Dictionary of Regulated 
Activities (MedDRA version 20.0 March 2017) primary system organ class and preferred term.  
Any medical condition will be classed as resolved if a stop date is recorded, otherwise, the 
condition will be classed as ongoing.  Prior conditions are defined as all conditions starting and 
stopping before the date of first dose of study drug.  Ongoing conditions are defined as conditions 
present on or after the date of first dose of study drug.   
6.9 Prior and Concomitant Medications 
The number and percentage of subjects using concomitant medications will be tabulated using 
drug class and preferred drug name by treatment group and overall for the Safety Analysis Set.  


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All prior and concomitant medications will be listed. Prior medications are defined as all 
medications starting and stopping before the date of first dose of study drug. Concomitant 
medications are defined as medications taken on or after the date of first dose of study drug.   
All medications will be coded using the World Health Organization Drug Dictionary (WHO Drug 
Dictionary version March 2017) ATC Level 2 class and preferred term.  
6.10 Exposure to Study Drug 
All dosing information will be listed.  
6.11 Pharmacokinetic Analyses 
Blood samples for plasma pharmacokinetic (PK) analysis will be obtained at pre-dose, 0.5 hour 
(+/-10 min), 1 hour (+/- 15 min), 2 hour (+/- 15 min), 4 hour (+/- 15 min), 8 hour (+/- 15 min), and 
24 hours (+4 hours) post injection. 
Concentration-time data will be tabulated by analyte (FX00 and FX03), treatment and nominal 
time using descriptive statistics. For presentation of the individual data and summary statistics, 
concentrations below the limit of quantitation (BLQ) will be set to zero. 
Mean and individual plasma concentration-time data will be presented graphically on both linear 
and semi-log scales. Spaghetti plots (all subjects in one plot) will be presented by analyte (FX00 
and FX03), treatment, and overall on both linear and semi-log scales. Mean data will be plotted 
using nominal sample times, and individual data will be plotted using actual times. 
PK parameters for FX00 and FX03 will be calculated as described in Section 6.2.8.  
6.12 Biomarker Analysis 
All blood sampling information for biomarker analysis will be listed.   
6.13 Safety Analyses 
The safety analyses will be presented by the treatment received for the Safety Analysis Set. 


 
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6.13.1 
Adverse Events 
A treatment emergent adverse event (TEAE) is defined as: 
 Any AE that has an onset on or after the single dose of study drug through the final follow-
up visit (Day 90).  
 Any pre-existing AE that has worsened in severity on or after the single dose of study drug 
through the final follow-up visit (Day 90).  
Relatedness of an AE to treatment can be recorded as Unrelated, Unlikely, Possible or Probable. 
A treatment-related AE is defined as any AE classified as possibly or probably related to the study 
drug.  If an AE has missing relationship it is assumed to be related to the study drug for analysis 
purposes.  
Maximum severity will be assumed for an AE with missing severity.  
The following tables will be presented for AEs: 
 Overall incidence and the number of TEAEs, TEAEs of the Ear, Severe TEAEs, SAEs, 
TEAEs leading to withdrawal and TEAEs leading to Death 
 TEAE by system organ class and preferred term, incidence 
 Serious TEAE by system organ class and preferred term, incidence 
 TEAE by system organ class, preferred term and maximum severity,  incidence 
 TEAE by system organ class, preferred term and relationship,  incidence 
 TEAE of the Ear by system organ class, preferred term and treatment status of the ear,  
incidence 
 TEAEs leading to Early Withdrawal by system organ class and preferred term, incidence 
 Listing of Serious TEAEs (presented in the Table section of the appendices) 
 Listing of Deaths (presented in the Table section of the appendices) 
System organ class will be presented in descending order of overall frequency and then 
alphabetically.  Preferred terms will be displayed in descending order of overall frequency and 
then alphabetically.  
Subjects reporting more than one AE per system organ class and preferred term will only be 
counted once.  For analyses concerning severity of the event or relatedness to study drug, the most 
severe event or the most related event will be used.  
All AEs will be listed. 


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6.13.2 
Laboratory Data 
Descriptive statistics of the observed values and change from baseline (continuous data) will be 
presented by treatment group and visit for each hematology, serum chemistry, coagulation and   
urinalysis parameter. Each measurement (continuous data) will be classed as below, within, or 
above normal range, based on ranges supplied by the laboratory used. Shift tables in relation to the 
normal range from baseline to each follow-up visit will be presented.  Categorical data will be 
presented by treatment group and follow-up visit.   
A listing of any clinically significant laboratory measurements recorded throughout the study will 
be presented.  
All laboratory data, including serology, urine drug and alcohol screening and pregnancy data will 
be listed.   
6.13.3 
Vital Signs 
Descriptive statistics for observed values and changes from baseline in the following vital signs 
will be presented by treatment group and overall at each visit: 
 Systolic blood pressure (mmHg) 
 Diastolic blood pressure (mmHg) 
 Pulse rate (bpm)  
 Respiration rate (breath/min) 
 Body temperature (degrees Celsius) 
 Body weight (kg) 
All vital sign data will be listed. 
6.13.4 
Electrocardiogram Data 
Descriptive statistics for observed values in the following ECG variables will be tabulated by 
treatment group and overall at the baseline visit and each follow-up visit:  
 Heart rate (bpm) 
 PR interval (ms) 
 RR interval (ms)  
 QRS complex (ms)   
 QT interval (ms)  


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 QTc interval (ms)  
 QTc interval (ms) [Bazett’s formula - QTcB]  
 QTc interval (ms) [Fridericia’s formula - QTcF]  
Overall interpretation (Normal, Abnormal NCS, and Abnormal CS) will be tabulated by treatment 
group and overall for the Baseline visit and each follow-up visit.    
All ECG data, including details of any abnormalities, will be listed.  
6.13.5 
Physical Examination 
Summaries for the observed status of each of the body systems (Normal, Abnormal NCS, and 
Abnormal CS) will be tabulated by treatment group and overall for the Baseline visit and for each 
follow-up visit.  All data, including details of clinically significant findings will be listed.  
6.13.6 
Tympanometry and Otoscopy Assessments 
Summaries for each element of the tympanometry and otoscopy assessments will be tabulated by 
treatment group and overall at each visit for the treated and untreated ear.  Changes from baseline 
and shifts in status from baseline to each follow-up visit will also be tabulated. All data, including 
details of any abnormalities, will be listed.  
6.13.7 
Comprehensive Audiogram Assessment 
Summaries of data obtained for each frequency will be tabulated by treatment group and overall 
at each visit for the treated and untreated ear.  Changes from baseline will also be tabulated. All 
data will be listed including correction factors and any frequencies attempted but not obtained.  
6.13.8 
Word Recognition and Words-In-Noise Assessments 
The percentage of words correctly identified out of a list of 50 words will be summarized for the 
Word Recognition (Quiet) test.  The number of words correctly identified will be summarized for 
the Words-in-Noise Test (WIN).  Summaries will be tabulated by treatment group and overall at 
each visit.  Changes from baseline will also be tabulated.  
Categorical analysis of the change from baseline will also be performed for the Word Recognition 
(Quiet) test.  Percentage scores obtained at each follow-up visit will be classified as Improved, No 
Change or Worsening according to the table of 95% Critical Differences in Appendix 1.  Values 
within the range shown are not significantly different from the value shown in the percentage score 


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columns (p > 0.05).  Values falling below the stated range will be considered Worsening and values 
exceeding the stated range will be considered Improved.  All data will be listed.  
6.13.9 
Interval Noise History 
All interval noise history data will be listed.   
7 BLIND SAFETY DATA REVIEW 
A blind Safety Data Review will be performed when all subjects have completed the Day 15 
follow-up assessment.  Subject disposition, demographic data, medical history, treatment-
emergent adverse events, laboratory data, tympanometry, otoscopy and audiometry data will be 
summarized for all subjects. Required outputs are marked with an asterisk in the list of Tables, 
Figures and Listings.  Outputs will be produced using the same shells as for the End of Study with 
one Overall column to ensure the blind is maintained.    
8 CHANGES TO PLANNED PROTOCOL ANALYSIS 
A blind Safety Data Review will be performed that was not previously detailed in the protocol 
but was requested by the Sponsor.  The rationale for the analysis is that completion of the Day 15 
visit will be the first opportunity to review safety endpoints for all subjects.     
 
 


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9 REFERENCES 
1. Phoenix™ WinNonlin® (Version 6.3, Certara L.P.) 
2. SAS Institute Inc., Cary, NC, 27513, USA 
3. Thornton AR, Raffin MJ (1978) J Speech Hear Res., Sep; 21(3): 507-18.  
 
 


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10 LIST OF TABLES, FIGURES AND LISTINGS 
The following table includes details of the tables, figures and listings to be included within each 
section of the electronic Common Technical Document (eCTD). The eCTD section is shown in 
bold. The following validation methods maybe used  
 Independent programming of numbers and manual review of format (IP) 
 Independent programming by statistician of numbers and manual review of format (Stat 
IP) 
 Manual review (MR) 
 Code review (CR) 
Table 
Number 
Table Title 
Validation 
Method 
Shell 
Number 
(if repeat) 
Items in bold are not table titles but references to the section headings within eCTD. 
 
14.1  
Demographics Data 
 
 
14.1.1  
Disposition 
 
 
14.1.1.1* 
Subject Disposition, Early Withdrawals – Randomized 
Set 
IP 
 
14.1.2  
Demographics 
 
 
14.1.2.1* 
Demographics, Safety Analysis Set 
IP 
 
14.1.3 
Baseline Characteristics 
 
 
14.1.3.1* 
Medical History Prior to Baseline – Safety Analysis Set 
IP 
 
14.1.3.2* 
Medical History Ongoing at Baseline – Safety Analysis 
Set 
IP 
 
14.1.3.3 
Physical Examination at Screening – Safety Analysis 
Set 
IP 
 
14.1.3.4 
ECG Data at Baseline – Safety Analysis Set 
IP 
 
14.2  
Safety Data 
 
 
 
Not Applicable 
 
 
14.3  
Safety Data 
 
 
14.3.1  
Displays Of Adverse Events 
 
 
14.3.1.1* 
Summary Of Treatment-Emergent Adverse Events 
(TEAEs) – Safety Analysis Set 
IP 
 
14.3.1.2* 
TEAEs By Primary System Organ Class And Preferred 
Term – Safety Analysis Set 
IP 
 
14.3.1.3* 
Serious TEAEs By Primary System Organ Class And 
Preferred Term – Safety Analysis Set 
IP 
 


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Statistical Analysis Plan 
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Table 
Number 
Table Title 
Validation 
Method 
Shell 
Number 
(if repeat) 
14.3.1.4* 
TEAEs By Primary System Organ Class Preferred 
Term And Severity – Safety Analysis Set 
IP 
 
14.3.1.5* 
TEAEs By Primary System Organ Class Preferred 
Term And Relationship – Safety Analysis Set 
IP 
 
14.3.1.6* 
TEAEs Of The Ear By Primary System Organ Class 
Preferred Term And Treatment Status – Safety Analysis 
Set 
IP 
 
14.3.1.7* 
TEAEs Leading To Early Withdrawal By Primary 
System Organ Class And Preferred Term – Safety 
Analysis Set 
IP 
 
14.3.2  
Listings Of Deaths, Other Serious And Significant 
Adverse Events 
 
 
14.3.2.1* 
Serious TEAEs, Listing– Safety Analysis Set 
IP 
 
14.3.2.2* 
Deaths, Listing – Safety Analysis Set 
IP 
 
14.3.3  
Narratives of Deaths, Other Serious and Certain 
Other Significant Adverse Events  
(Placeholder should be maintained and tables/figures 
should not be numbered to appear in this section) 
 
 
14.3.4  
Abnormal Laboratory Values 
 
 
14.3.4.1* 
Listing of Clinically Significant Laboratory Values – 
Safety Analysis Set 
IP 
 
14.3.5  
Extent Of Exposure, Dosage Information, And 
Compliance 
 
 
 
Not Applicable 
 
 
14.3.6  
Vital Signs And Physical Examination 
 
 
14.3.6.1 
Vital Signs, Descriptive Statistics – Safety Analysis Set 
IP 
 
14.3.7  
Other Safety 
 
 
14.3.7.1* 
Hematology Data, Descriptive Statistics – Safety 
Analysis Set 
IP 
 
14.3.7.2* 
Hematology Data, Normal Range Shifts – Safety 
Analysis Set 
IP 
 
14.3.7.3* 
Serum Chemistry Data, Descriptive Statistics – Safety 
Analysis Set 
IP 
 
14.3.7.4* 
Serum Chemistry Data, Normal Range Shifts – Safety 
Analysis Set 
IP 
 


Frequency Therapeutics 
FX-322-201 
Statistical Analysis Plan 
Version 1.0 
Issue Date 10-AUG-2018 
 
 
Page 21 of 26 
 
Confidential 
Table 
Number 
Table Title 
Validation 
Method 
Shell 
Number 
(if repeat) 
14.3.7.5* 
Coagulation Data, Descriptive Statistics – Safety 
Analysis Set 
IP 
 
14.3.7.6* 
Coagulation Data, Normal Range Shifts – Safety 
Analysis Set 
IP 
 
14.3.7.7* 
Urinalysis Data, Descriptive Statistics – Safety Analysis 
Set 
IP 
 
14.3.7.8* 
Urinalysis Data, Normal Range Shifts – Safety Analysis 
Set 
IP 
 
14.3.7.9 
ECG Data, Descriptive Statistics – Safety Analysis Set 
IP 
 
14.3.7.10 
ECG Data, Overall Interpretation – Safety Analysis Set 
IP 
 
14.3.7.11* 
Tympanometry Data, Descriptive Statistics – Safety 
Analysis Set 
IP 
 
14.3.7.12* 
Otoscopy Data, Descriptive Statistics – Safety Analysis 
Set 
IP 
 
14.3.7.13* 
Comprehensive Audiometry Data, Descriptive Statistics 
– Safety Analysis Set 
IP 
 
14.3.7.14 
Word Recognition (Quiet) Data, Descriptive Statistics – 
Safety Analysis Set 
IP 
 
14.3.7.15 
Words-In-Noise Data, Descriptive Statistics – Safety 
Analysis Set 
IP 
 
14.3.8  
Concomitant Medication 
 
 
14.3.8.1 
Concomitant Medication Data – Safety Analysis Set 
IP 
 
14.4  
PK Tables 
 
 
14.4.1 
FX00 and FX03 Concentration-Time Data, Descriptive 
Statistics – PK Analysis Set 
 
 
14.4.2 
FX00 and FX03 Pharmacokinetic Parameters, Listing 
and Descriptive Statistics – PK Analysis Set 
 
 
 
 
 


 
Frequency Therapeutics 
 
FX-322-201 
 
Statistical Analysis Plan 
 
Version 1.0 
 
Issue Date 10-AUG-2018 
 
 
Page 22 of 26 
 
Confidential 
Figure 
Number 
Figure Title 
Validation 
Method 
Shell 
Number (if 
repeat) 
14.4.1 
Mean FX00 and FX03 Concentration Time Data on 
Linear and Semi-Logarithmic Scales – PK Analysis 
Set 
 
 
14.4.2 
FX00 and FX03 Concentration-Time Data, All 
Subject Profiles on Linear and Semi-Logarithmic 
Scales – PK Analysis Set 
 
 
14.4.3 
Concentration-Time Profiles for FX00 and FX03 with 
Linear Regression for Estimating the Terminal 
Elimination Rate – PK Analysis Set 
 
 
 
 


Frequency Therapeutics 
FX-322-201 
Statistical Analysis Plan 
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Issue Date 10-AUG-2018 
 
 
Page 23 of 26 
 
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Listing 
Number 
Listing Title 
Validation 
Method 
Shell 
Number 
(if repeat) 
16.2  
Subject Data Listings 
 
 
16.2.1  
Discontinued Subjects 
 
 
16.2.1.1* 
Subject Disposition, Early Withdrawals – Randomized 
Set 
IP 
 
16.2.2 
Protocol Deviations 
 
 
16.2.2.1 
Protocol Deviations – Safety Analysis Set 
IP 
 
16.2.3 
Subjects Excluded From The Efficacy Analyses 
 
 
16.2.3.1  
Analysis Sets – Randomized Set 
IP 
 
16.2.4  
Demographic Data 
 
 
16.2.4.1* 
Demographic Data  – Safety Analysis Set 
IP 
 
16.2.4.2* 
Previous and Ongoing Medical History – Safety 
Analysis Set 
IP 
 
16.2.5  
Compliance And/Or Drug Concentration Data 
 
 
16.2.5.1  
PK Sampling Data – Safety Analysis Set 
IP 
 
16.2.5.2  
Biomarker Sampling Data – Safety Analysis Set 
IP 
 
16.2.5.3 
Prior and Concomitant Medication – Safety Analysis 
Set 
IP 
 
16.2.5.4 
Dosing Information – Safety Analysis Set 
IP 
 
16.2.6  
Individual Efficacy Response Data   
 
 
16.2.6.1 
FX00 and FX03 Concentration Listing by Subject – 
Safety Analysis Set 
 
 
16.2.6.2 
Plasma Terminal Elimination Rate of FX00 and FX03 
for Individual Subjects – PK Analysis Set 
 
 
16.2.6.3 
PK Output Text 
 
 
16.2.7  
Adverse Event Listings 
 
 
16.2.7.1* 
Adverse Event Data – Safety Analysis Set 
IP 
 
16.2.8  
Individual Laboratory Measurements And Other 
Safety 
 
 
16.2.8.1* 
Hematology Data – Safety Analysis Set 
IP 
 
16.2.8.2* 
Serum Chemistry Data – Safety Analysis Set 
IP 
 
16.2.8.3* 
Coagulation Data – Safety Analysis Set 
IP 
 
16.2.8.4* 
Urinalysis Data – Safety Analysis Set 
IP 
 
16.2.8.5 
Other Clinical Laboratory Data – Safety Analysis Set 
IP 
 
16.2.8.6 
Vital Signs Data – Safety Analysis Set 
IP 
 
16.2.8.7 
Physical Examination Data – Safety Analysis Set 
IP 
 


Frequency Therapeutics 
FX-322-201 
Statistical Analysis Plan 
Version 1.0 
Issue Date 10-AUG-2018 
 
 
Page 24 of 26 
 
Confidential 
Listing 
Number 
Listing Title 
Validation 
Method 
Shell 
Number 
(if repeat) 
16.2.8.8 
ECG Data – Safety Analysis Set 
IP 
 
16.2.8.9* 
Tympanometry Data – Safety Analysis Set 
IP 
 
16.2.8.10* 
Otoscopy Data – Safety Analysis Set 
IP 
 
16.2.8.11* 
Comprehensive Audiometry Data – Safety Analysis Set 
IP 
 
16.2.8.12 
Word Recognition (Quiet) Data – Safety Analysis Set 
IP 
 
16.2.8.13 
Words-In-Noise Data – Safety Analysis Set 
IP 
 
16.2.8.14 
Interval Noise History – Safety Analysis Set   
IP 
 
 
 
 


 
Frequency Therapeutics 
 
FX-322-201 
 
Statistical Analysis Plan 
 
Version 1.0 
 
Issue Date 10-AUG-2018 
 
Page 25 of 26 
 
Confidential 
APPENDIX 1 
Lower and Upper Limits of the 95% Critical Differences for Percentage scores for Word 
Recognition (Quiet) Test3.   
Values within the range shown are not significantly different from the value shown in the 
percentage score columns (p > 0.05).   
% Score 
n = 50 
0 
0 - 4 
2 
0 - 10 
4 
0 - 14 
6 
2 - 18 
8 
2 - 22 
10 
2 - 24 
12 
4 - 26 
14 
4 - 30 
16 
6 - 32 
18 
6 - 34 
20 
8 - 36 
22 
8 - 40 
24 
10 - 42 
26 
12 - 44 
28 
14 - 46 
30 
14 - 48 
32 
16 - 50 
34 
18 - 52 
36 
20 - 54 
38 
22 - 56 
40 
22 - 58 
42 
24 - 60 
44 
26 - 62 
46 
28 - 64 
48 
30 - 66 
50 
32 - 68 
52 
34 - 70 


 
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Statistical Analysis Plan 
 
Version 1.0 
 
Issue Date 10-AUG-2018 
 
 
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54 
36 - 72 
56 
38 - 74 
58 
40 - 76 
60 
42 - 78 
62 
44 - 78 
64 
46 - 80 
66 
48 - 82 
68 
50 - 84 
70 
52 - 86 
72 
54 - 86 
74 
56 - 88 
76 
58 - 90 
78 
60 - 92 
80 
64 - 92 
82 
66 - 94 
84 
68 - 94 
86 
70 - 96 
88 
74 - 96 
90 
76 - 98 
92 
78 - 98 
94 
82 - 98 
96 
86 - 100 
98 
90 - 100 
100 
96 - 100