Evaluation of Safety and Pharmacodynamics of OP0201 Compared to Placebo
Phase: PHASE1
Conditions: Healthy Adults
Sponsor: Novus Therapeutics, Inc

 
 
 
Study Title Evaluation of Safety and Pharmacodynamics of OP0201 Compared to Placebo in 
Healthy Adults 
NCT Number NCT03828149 
Document Description Clinical Protocol, Version: V04_0 
Document Date 08 Jan 2019 
 

 
Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
 
TRIAL PROTOCOL 
EVALUATION OF SAFETY AND PHARMACODYNAMICS OF OP0201 COMPARED TO PLACEBO IN HEALTHY 
ADULTS  
 
Novus Therapeutics, Inc. 
19900 MacArthur Blvd., Suite 550 
Irvine, California 92612, U.S.A. 
(Sponsor) 
represented by  
University of Cologne 
Albertus-Magnus-Platz 
50923 Cologne Germany 
(EU Sponsor Representative) 
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Novus TherapeuticsTrial Protocol Code:  OP0201-C001 
University of Cologne Internal Trial Protocol Code: Uni-Koeln-2809 
EudraCT number: 2016-003667-19 
 
08-01-2018, Version V04_0 
The information in this trial protocol is strictly confidential. It is for the use of the Sponsor, investigator, trial 
personnel, ethics committee, the authorities, and trial subjects only. This trial protocol may not be passed on 
to third parties without the express agreement of the Sponsor or the Principal Coordinating Investigator 
(PCI, “Leiter der klinischen Prüfung (LKP)”). 
 
 
 
 
 
 
 
 

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I. Signatures 
 
 
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Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
 
II. Synopsis 
Sponsor: 
Novus Therapeutics, Inc. 
19900 MacArthur Blvd., Suite 550 
Irvine, California 92612, U.S.A. 
represented in the EU by 
University of Cologne 
Albertus-Magnus-Platz 
50923 Cologne 
Germany 
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Title of the clinical trial: 
Evaluation of Safety and Pharmacodynamics of OP0201 
Compared to Placebo in Healthy Adults  
Indication: 
Eustachian tube function  

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Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
 
Phase: 
I  
Type of trial, trial design, 
methodology: 
Single center clinical trial 
Randomized, double-blind, placebo-controlled, single dose, 
cross-over 
Number of subjects: 
16 
Primary trial objective: 
To evaluate the safety and tolerability of a single intranasal dose 
of OP0201 compared to a single intranasal dose of placebo in 
healthy adults.  
Secondary trial objectives: 
 
 
 
 
 
 
1. To assess pharmacodynamics of the Eustachian Tube 
following a single intranasal dose of OP0201 compared to a 
single intranasal dose of placebo in healthy adults. 
 
2. To explore whether a single intranasal dose of OP0201 
compared to a single intranasal dose of placebo modulates 
ear pain during the hypobaric/hyperbaric atmospheric 
pressure chamber assessment in healthy adults. 
Trial end points: 
Variables to be evaluated in support of the primary objective 
(safety and tolerability) for this study: 
Adverse events 
Otoscopy 
Tympanometry 
Nasal and Epipharynx Endoscopy 
Triplicate 12-Lead ECG 
Physical exam 
Vital signs 
Clinical laboratory tests 
Variables to be evaluated in support of the secondary objectives 
(pharmacodynamics [continuous tympanic impedance] and ear 

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Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
 
pain) for this study: 
Pharmacodynamics  
• 
Eustachian tube opening pressure (ETOP) 
• 
Eustachian tube opening duration (ETOD) 
• 
Eustachian tube closing pressure (ETCP) 
• 
Eustachian tube opening frequency (ETOF) 
Ear pain 
Diagnosis and Principal 
inclusion and exclusion 
criteria: 
Medical condition or disease to be investigated:  
None; this is a healthy volunteer trial. 
Key Inclusion criteria: 
• 
Male and female; 18 to 50 years of age at time of 
signed informed consent 
• 
Healthy subjects with no history or presence of 
significant medical condition or a clinically significant 
abnormal finding, as determined by the investigator. 
This includes study screening results from: 
• 
Medical history 
• 
Physical examination 
• 
ECG (in consultation with cardiologist) 
• 
Vital sign measurements 
• 
Clinical laboratory data 
• 
Physiologic tympanogram type A or type C1 at 
screening visit 
Key Exclusion criteria: 
• 
Upper respiratory tract infection currently or within 6 
weeks prior to screening visit 
 
1 Tympanogram Type C defined as peak pressure between -100 mm water to -200 mm water 

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Trial protocol V04_0 of 08-01-2018 
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• 
Allergy or sinus conditions (e.g., sinusitis, non-
specific nasal inflammation) currently or within 6 
weeks prior to screening visit 
• 
Clinically significant findings on ENT exam 
• 
Gastroesophageal reflux disease currently or within 6 
weeks prior to screening visit 
• 
Current diagnosis of sleep apnea 
• 
Evidence of craniofacial anomalies (eg, cleft palate, 
Down’s Syndrome) that may interfere with Eustachian 
Tube function 
• 
Disorders with decreased mucociliary clearance or 
higher viscosity of the mucous (eg, cystic fibrosis, 
primary ciliary dyskinesia, Kartagener’s syndrome) 
• 
Clinically significant ear disease currently or within 6 
weeks prior to screening visit  
• 
History of tympanostomy tubes in one or both ears 
within 48 weeks prior to screening visit 
• 
Clinically relevant blockage of one or both nasal 
passages, in the investigator’s opinion  
• 
Females who are pregnant, nursing, or planning a 
pregnancy during the trial  

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Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
 
Name of investigational 
medicinal product (IMP): 
OP0201 [dipalmitoylphosphatidylcholine (DPPC):cholesteryl 
palmitate (CP)] 
Investigational medicinal 
product – dosage and 
method of administration: 
• 
OP0201 (DPPC:CP [approximately 20:1 w/w] in HFA-134a) 
metered-dose inhaler 
• 
Intranasal administration  
• 
Dosage: 20 mg OP0201 administered as four consecutive 
sprays (2.5 mg per spray) first to the left nare, followed by 
four consecutive sprays to the right nare until a total of eight 
sprays (four to each nare) has been administered (see 
Investigational Medicinal Product (IMP) Instructions for Use). 
IMP or therapy used as a 
comparator – dosage and 
method of administration: 
• 
OP0201 Placebo (HFA-134a only) metered dose inhaler  
• 
Intranasal administration  
• 
Dosage: OP0201 Placebo administered as four consecutive 
sprays (0 mg per spray) first to the left nare, followed by four 
consecutive sprays to the right nare until a total of eight 
sprays (four to each nare) has been administered (see 
Investigational Medicinal Product (IMP) Instructions for Use). 
Duration of application: 
Trial subjects will receive a single-dose of OP0201 or OP0201 
Placebo at the Day 1 visit, followed by an approximately one-
week wash-out, and then they will receive a single-dose of the 
opposite IMP at the Day 8 cross-over visit. It is anticipated that 
the subject will be in the clinic for approximately 4 to 6 hours for 
each trial visit. 
Estimated Time plan: 
First patient first visit (FPFV): 
Q1 2019 
Last patient first visit (LPFV): 
Q1 2019 
Last patient last visit (LPLV): 
Q1 2019  
End of trial: 
Q1 2019 
Final trial report to BfArM: 
Q1 2020  
(See Chap. 3.1.1.) 
 

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Statistical methods: 
A centralised blocked 2-group randomization stratified by gender 
and “non-significant baro-challenges (yes/no)“ will be performed 
via the internet based randomization service TENALEA. 
Populations: 
The safety population will consist of all subjects who received at 
least one spray of IMP in either nare. In safety data analyses, 
subjects will be analysed according to the actual IMP received, 
regardless of randomization assignment. 
The per-protocol population (PP) will consist of all randomized  
subjects who received the two complete doses (4 sprays to each 
nare) of IMP at Day 1 and 8. Valid measures of ETOP and 
ETOD for passive phases 1 and 5 for both ears and for all three 
measurement times at visits 2 and 4 Excluded are subjects with 
major protocol deviations (determined prior to database lock and 
unblinding of the data).  
Analyses: 
The analysis will be descriptive, significance level will not be 
adjusted for multiple testing. A p-value ≤ 5% is defined as 
significant for demographic and screening characteristics and 
the assessments supporting the secondary objectives for this 
study.  
Descriptive summary tables for each treatment and period and 
for each measurement time will be performed including statistics 
depending on variable type. If variables are measured before 
and after treatment at the same visit, the difference will be 
calculated for each visit and analyses will apply to this change, if 
appropriate. 

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Novus Therapeutics, Inc. by University of Cologne 
 
Incidence of AEs will be tabulated by primary system organ class 
(SOC) and by preferred term within each primary SOC.   
Primary analysis is the evaluation of the safety variables in the 
safety population. Adequate confidence intervals will be given for 
the change from baseline, if appropriate. 
Secondary analysis consisting of pharmacodynamics and ear 
pain analyses will be performed in the safety population and 
additionally in the PP population. Non-parametric treatment 
comparisons will be done with a Grizzle’s two-stage approach for 
the cross-over design. The tests will be carried out without 
covariates. If confidence intervals (CI) are to be provided for 2-
group comparison, the 95% 2-sided Hodges-Lehmann intervals 
will be calculated. 
Missing values will not be replaced, but drop-outs may be 
replaced with additionally randomized trial subjects to ensure 
N=16 for the PP population.  
Subgroup analyses are planned for stratification factors, if 
sufficient numbers of subjects are met. 
Sample size: 
N=16 in the PP population. 
 
GCP conformance: 
This protocol is to be conducted in accordance with the 
applicable current Good Clinical Practice (GCP) regulations and 
guidelines, i.e., the International Council for Harmonization (ICH) 
Guideline on GCP. 
 
Financing: 
Novus Therapeutics, Inc. 

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Trial protocol V04_0 of 08-01-2018 
Novus Therapeutics, Inc. by University of Cologne 
III. Table of contents 
I. 
Signatures 
2 
II. 
Synopsis 
3 
III. 
Table of contents 
10 
III.a) 
List of tables 
15 
III.b) 
List of figures 
15 
IV. 
Abbreviations 
16 
1 
Introduction 
19 
1.1 
OP0201 (dipalmitoylphosphatidylcholine [DPPC] and cholesteryl palmitate [CP])19 
1.2 
Rationale for the clinical trial 
19 
1.3 
Primary objective 
20 
1.4 
Secondary and other objectives 
20 
1.5 
Benefit-Risk Analysis 
20 
2 
Organisational and administrative aspects of the trial 
22 
2.1 
Sponsor 
22 
2.2 
Principal Coordinating Investigator 
22 
2.3 
Statistics 
22 
2.4 
Data Monitoring Committee 
22 
2.5 
Trial laboratories and other technical services 
23 
2.6 
Central organisation units 
23 
2.7 
Investigators and trial sites 
23 
2.7.1 
Requirements for investigators and trial sites 
24 
2.8 
Financing 
24 
3 
Trial conduct 
25 
3.1 
General aspects of trial design 
25 
3.1.1 
Estimated Time plan 
25 
3.2 
Discussion of trial design 
25 

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3.3 
Selection of trial population 
26 
3.3.1 
Inclusion criteria 
26 
3.3.2 
Exclusion criteria 
27 
3.4 
Withdrawal of trial subjects after trial start 
28 
3.4.1 
Procedures for premature withdrawal from treatment during the trial 
29 
3.5 
Closure of trial sites/Premature termination of the clinical trial 
29 
3.5.1 
Closure of trial sites 
29 
3.5.2 
Premature termination of trial 
29 
3.5.3 
Stopping Criteria for individual subjects and for the study 
29 
3.6 
Treatment 
30 
3.6.1 
Treatments to be given 
30 
3.6.2 
Visit Schedule 
34 
3.6.2.1 
Screening 
34 
3.6.2.2 
Day 1 
35 
3.6.2.3 
Day 2 (telephone follow-up visit) 
37 
3.6.2.4 
Day 8 ± 1 (crossover visit) 
37 
3.6.2.5 
Day 9 Trial Exit (telephone follow-up visit) 
39 
3.6.3 
Description of investigational medicinal product (IMP) 
39 
3.6.3.1 
OP0201 
39 
3.6.3.2 
Placebo 
40 
3.6.3.3 
Manufacture of the IMP 
40 
3.6.3.4 
Labelling of IMP 
40 
3.6.3.5 
Storage of IMP 
40 
3.6.4 
Compliance with treatment & dispensing and return of IMP 
41 
3.6.5 
Assignment of trial subjects to treatment groups 
41 
3.6.6 
Selection of dosage of investigational medicinal product 
41 
3.6.7 
Blinding 
42 
3.6.7.1 
Unblinding 
42 

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3.6.8 
Previous and concomitant medication 
42 
3.6.9 
Pregnancy and Contraception Considerations 
43 
3.6.10 
End of Trial 
44 
3.6.11 
Continuation of treatment after the end of the clinical trial 
44 
3.7 
Efficacy and safety variables 
44 
3.7.1 
Safety variables 
44 
3.7.2 
Pharmacodynamic Assessments 
46 
3.7.3 
Exploratory Efficacy Assessment 
47 
3.7.4 
Demographics and Baseline Characteristics 
47 
3.7.5 
Rationale for assessment procedures 
47 
3.8 
Data quality assurance 
48 
3.8.1 
Monitoring 
48 
3.8.2 
Audits/Inspections 
48 
3.9 
Documentation 
49 
3.9.1 
Data management 
50 
3.9.2 
Archiving 
50 
4 
Ethical and regulatory aspects 
51 
4.1 
Independent ethics committee 
51 
4.2 
Ethical basis for the clinical trial 
51 
4.2.1 
Legislation and guidelines used for preparation 
51 
4.3 
Notification of the authorities, approval and registration 
52 
4.4 
Obtaining informed consent from trial subjects 
52 
4.5 
Insurance of trial subjects 
52 
4.6 
Data protection 
52 
5 
Statistical methods and sample size calculation 
53 
5.1 
Statistical and analytical plan 
53 
5.1.1 
Analysis populations 
53 
5.1.2 
Description of trial subject groups 
53 

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5.1.3 
Primary objectives 
54 
5.1.4 
Secondary objectives 
54 
5.1.5 
Subgroup analyses 
54 
5.1.6 
Interim analysis 
54 
5.2 
Sample size calculation 
54 
6 
Safety 
55 
6.1 
Definitions of adverse events and adverse drug reactions 
55 
6.1.1 
Adverse event 
55 
6.1.2 
Adverse drug reaction 
55 
6.1.3 
Serious adverse events or serious adverse drug reactions (SAE/SADRs) 
55 
6.1.4 
Unexpected adverse drug reaction (UADR) 
56 
6.2 
Documentation and follow-up of AEs and ADRs 
56 
6.2.1 
Severity of the AE 
57 
6.2.2 
Causal relationship between AE and IMP 
57 
6.2.3 
Documentation and reporting of SAE and pregnancy 
58 
6.2.4 
SAE reporting by the investigator to the Sponsor 
59 
6.2.5 
Assessment of Serious Adverse Event by Sponsor 
60 
6.2.6 
Notification of ethics committee and competent supreme federal authority 
60 
6.2.7 
Notification of study staff and study participants 
60 
6.2.8 
Informing the Data Monitoring Committee 
61 
6.2.9 
Known side effects caused by pressure differential during pressure chamber 
measurement protocol 
61 
7 
Use of trial findings and publication 
62 
7.1 
Reports 
62 
7.1.1 
Interim reports 
62 
7.1.2 
Final report 
62 
7.2 
Publication 
62 
7.3 
Amendments to the trial protocol 
62 

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8 
References 
63 
9 
Attachments 
64 
9.1 
Hyperbaric/hypobaric pressure chamber measurement 
64 
9.2 
Ear Pain Assessment (Example Page) 
65 
9.3 
TEED Classification 
66 
 
 
 
 
 

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III.a) List of tables 
Table 1: 
Time plan of the trial 
 
Error! Bookmark not defined. 
Table 2: 
Visit Schedule 
 32 
 
III.b) List of figures 
Figure 1: 
Trial flowchart 
 31 
Figure 2: 
Standard pressure profile during continuous tympanic impedance measurement  64 
 

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IV. Abbreviations 
Abbreviation 
meaning 
ADR 
Adverse Drug Reaction 
AOM 
Acute otitis media 
AE 
Adverse Event 
AMG 
Federal Drug Law (Arzneimittelgesetzt) 
BID 
Twice daily  
BfArM 
Federal Institute for Drugs and Medical Devices (Bundesinstitut für 
Arzneimittel und Medizinprodukte) 
CA 
Competent authority (BfArM) 
BP 
Blood pressure  
CI 
Confidence interval 
CP 
Cholesteryl Palmitate 
CRF 
Case Report Form 
CTCC 
Clinical Trial Centre Cologne 
DMC 
Data Monitoring Committee 
DPPC 
Dipalmitoylphosphatidylcholine 
EC 
Ethics Committee 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
EDC 
Electronic data capture 
ENT 
Ear, nose & throat 

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EOS 
End-of-trial 
ET 
Eustachian tube 
ETCP 
Eustachian tube closing pressure 
ETOD 
Eustachian tube opening duration 
ETOF 
Eustachian tube opening frequency 
ETOP 
Eustachian tube opening pressure 
ETD 
Eustachian tube dysfunction 
FPFV 
First patient first visit  
GCP 
Good Clinical Practice 
GLP 
Good Laboratory Practice 
GMP 
Good Manufacturing Practice 
HFA-134a 
Hydrofluoroalkane 134a (1,1,1,2-tetrafluorethane) 
IEC 
Independent Ethics Committee 
IMP 
Investigational Medicinal Product 
IMSB 
Institute for Medical Statistics and Computational Biology 
ITT 
Intent-to-treat 
ISF 
Investigator Site File 
LKP 
Leiter der klinischen Prüfung (Principal Coordinating Investigator)  
LPFV 
Last patient first visit 
LPLV 
Last patient last visit 
ME 
Middle ear 
mg 
Milligram(s) 

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mL 
Milliliters  
mmHG 
Millimeters of mercury 
OM 
Otitis media 
OME 
Otitis media with effusion 
OTC 
Over-the-counter 
PCI 
Principal Coordinating Investigator (Leiter der klinischen Prüfung) 
 
PE 
Physical Exam 
PL 
phospholipid 
PP 
Per-protocol 
PR 
Pulse rate  
QD 
Once daily 
RR 
Respiratory rate 
SADR 
Serious Adverse Drug Reaction 
SAE 
Serious Adverse Event 
SAP 
Statistical analysis plan 
SDV 
Source data verification 
SOC 
System organ class 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TM 
Tympanic membrane 
TMF 
Trial Master File 
T 
Temperature  
w/w 
Weight/weight 

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1 
Introduction 
1.1 
OP0201 (dipalmitoylphosphatidylcholine [DPPC] and cholesteryl palmitate [CP]) 
The Investigational Medicinal Product (IMP), OP0201, is comprised of two active ingredients, 
dipalmitoylphosphatidylcholine (DPPC [a phospholipid surfactant; also known as colfosceril 
palmitate]) and cholesteryl palmitate (CP [a neutral phospholipid spreading agent]), in 
approximately 20:1 (weight/weight [w/w]) ratio, and an inactive ingredient, synthetic 
chlorofluorocarbon-free propellant, hydrofluoroalkane 134a (HFA-134a). The product is delivered 
to the nasal mucosa in the anterior nares, towards the opening of the Eustachian tube (ET), as a 
dry powder in HFA-134a via an intranasal metered dose inhalant (MDI). Each 0.1 milliliter (mL) 
MDI spray of OP0201 delivers 0.1 mL of HFA-134a containing approximately 2.5 milligrams (mg) 
of the active ingredients (approximately 2.38 mg of DPPC and approximately 0.12 mg of CP).  
For further details about OP0201 see Investigator’s Brochure.   
1.2 
Rationale for the clinical trial 
The ET is a compliant liquid-lined tube between the middle ear (ME) and the nasopharynx. The ET 
serves three major physiologic functions: 1) ventilation of the ME to maintain ambient air pressure, 
2) clearance of fluid from the ME to nasopharynx, and 3) protection of the ME (1, 2). It is well 
established that Eustachian Tube Dysfunction (ETD) is a significant factor in the cause of middle 
ear diseases, such as acute and chronic otitis media (1, 3, 4).   
During the past few decades, various methods have been developed to evaluate ET function such 
as manometric, endoscopic and sonographic methods, impedance measurements and 
Tubomanometry. However, none of these methods was able to reflect physiologic conditions and 
fully assess ET function (5, 6). Another established method for evaluating ET function is by utilizing 
a pressure chamber (7, 8).   
It is established that surfactants, which are endogenous components in human ETs, are present on 
the liquid surface of the ET’s mucosal lining. Surfactants lower surface tension. Thus, when 
surfactant is administered exogenously to ET, it is hypothesized that it will spread along the 
pressure gradient from the area of high surface pressure (low surface tension) to an area of low 
surface pressure (high surface tension) and thus may act as ‘release’ agents by being directly 
absorbed onto the ET surface, causing the entire surface to become hydrophobic. As a result, ET 
function is modulated. It has been demonstrated in various animal models, including healthy 
animals, that exogenous surfactant administered through the nose to the ET reduces the ET 

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surface tension, which in turn decreases the ET passive opening pressure, facilitating opening of 
the ET (2). 
This exploratory trial is proposed as an evaluation of the safety and tolerability of intranasal 
administration of OP0201 in healthy adults. In addition, the trial will evaluate pharmacodynamics of 
the ET to guide future studies of OP0201 as a possible treatment for diseases caused by ET 
dysfunction (e.g. Otitis Media).   
1.3 
Primary objective 
The primary objective of the trial is to assess the safety and tolerability of a single intranasal dose 
of OP0201 compared to a single intranasal dose of placebo in healthy adults. 
1.4 
Secondary and other objectives 
The secondary objectives in this trial are: 
1. To assess pharmacodynamics of the ET following a single intranasal dose of OP0201 
compared to a single intranasal dose of placebo in healthy adults. 
 
2. To explore whether a single intranasal dose of OP0201 compared to a single intranasal 
dose of placebo modulates ear pain during the hypobaric/hyperbaric atmospheric pressure 
chamber assessment in healthy adults. 
1.5 
Benefit-Risk Analysis 
This trial is designed to evaluate the safety, tolerability, pharmacodynamics of the ET and potential 
effect in modulating ear pain of an investigational treatment (OP0201) in healthy volunteers. A 
benefit-risk analysis is not able to be fully established at this time. There is no expected or 
anticipated benefit to healthy subjects. The IMP is not expected to cause harm to healthy subjects. 
Both DPPC and CP are endogenous to human nasal passages and ETs. Furthermore, surfactants 
in much higher concentrations and doses are components in several pharmaceutical products that 
have received approval by Health Authorities, including US FDA and many EU countries, (e.g., 
Alveofact®2, Curosurf®, Infasurf®, Survanta®) as intratracheal treatment of premature infants with 
respiratory distress syndrome (RDS). The propellant excipient, HFA-134a, is used in approved 
metered dose oral inhalers (e.g., Proventil HFA® [albuterol], Aerospan® [flunisolide] and intranasal 
inhalers (e.g., QNASL® [beclomethasone]) containing active pharmaceutical agents for the 
 
2 Alveofact® and Curosurf® are both approved and marketed in Germany 

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management of asthma and chronic obstructive pulmonary disease (see Investigator’s Brochure, 
Table 5, Section 8.1)). The MDI device used to deliver OP0201 into the nares is similar to devices 
utilized in the delivery of other approved inhaled metered dose products.   
 
 

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2 
Organisational and administrative aspects of the trial 
2.1 
Sponsor 
Sponsor: 
Novus Therapeutics, Inc. 
 
19900 MacArthur Blvd., Ste. 550 
 
Irvine, CA  92612 
 
United States 
 
Represented by: 
University of Cologne 
Albertus-Magnus-Platz 
 
50923 Cologne 
 
Germany 
2.2 
Principal Coordinating Investigator 
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2.3 
Statistics 
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xxxxxxxxxxxxx 
xxxxxxx 
2.4 
Data Monitoring Committee 
A Data Monitoring Committee (DMC) is not planned for this trial. 

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2.5 
Trial laboratories and other technical services 
The local laboratory will perform the laboratory analyses described in this protocol. 
The local pharmacy will be responsible for storing and dispensing the OP0201 IMP during the trial. 
They will also be responsible for maintaining a detailed inventory of the OP0201 IMP including 
accounting for the number of OP0201 IMP units received, the number of OP0201 IMP units 
dispensed, and the number of unused OP0201 IMP units at the completion of the trial.   
An ECG vendor, ERT (Peterborough, United Kingdom) will be responsible for ECG data review 
and analysis.    
2.6 
Central organisation units 
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xxxxxxxxxxxxxxxxxxxxxxxxxxx 
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xxxxxxx 
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2.7 
Investigators and trial sites 
This clinical trial will be conducted at a single centre 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx  
A listing of the trial site, (principal) investigators, their deputies, subinvestigators, and other trial 
staff, will be kept and continuously updated. The final version of this list will be attached to the final 
report of the clinical trial. 

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2.7.1 Requirements for investigators and trial sites 
The clinical trial site where the trial will be conducted has to meet the following requirements as 
agreed by the Sponsor and University of Cologne: 
• 
Specialised unit for ear, nose and throat discipline 
• 
Possession of a CE-certified single-person hypobaric and hyperbaric pressure chamber 
capable of performing compression and decompression phases between 0.8 and 1.2 
bar relative to atmospheric pressure in order to assess continuous tympanic 
impedance; 
• 
Space and facilities to perform standard ENT exams including tympanogram, ear 
microscopy and nose endoscopy. 
• 
Facilities to conduct monitoring of the clinical trial.  
The investigators have to meet the following requirements for the conduct of the trial as agreed by 
the Sponsor and University of Cologne: 
• 
Specialist ENT physician with at least 3 years professional experience (principal 
coordinating investigator, PCI) 
• 
At least 2 years experience in the conduct of clinical studies according to GCP as 
member of investigator’s team or investigator (PCI) 
• 
Registered physician with at least 1 year professional ENT experience (subinvestigator) 
• 
Experienced in using the hypobaric/hyperbaric pressure chamber 
• 
Trained and/or experience in regulatory requirements of clinical trials according to the 
German Federal Medicines Law (“Arzneimittelgesetz”) and GCP 
In addition, requirements for investigator and subinvestigator will be described in detail on a 
separate qualification list.  
2.8 
Financing 
The clinical trial is funded by Novus Therapeutics, Inc. Additionally, the IMP is provided free of 
charge by Novus Therapeutics, Inc. 
 

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3 
Trial conduct 
3.1 
General aspects of trial design 
This is a randomized, double-blind, placebo-controlled, cross-over trial to evaluate the safety, 
tolerability, pharmacodynamics of the ET and potential effect in modulating ear pain of a single 
intranasal dose of OP0201 compared to a single intranasal dose of placebo in healthy adults.  
3.1.1 Estimated Time plan 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx 
First patient first visit (FPFV): 
Q1 2019 
Last patient first visit (LPFV): 
Q1 2019 
Last patient last visit (LPLV): 
Q1 2019  
End of trial 
Q1 2019 
Final trial report to BfArM: 
Q1 2020 
 
End of the clinical trial 
The end of trial is defined as the last patient last visit (LPLV).  
3.2 
Discussion of trial design 
This is a randomized, double-blind, placebo-controlled, cross-over trial designed to evaluate the 
safety, tolerability, pharmacodynamics of the ET and potential effect in modulating ear pain of a 
single intranasal dose of OP0201 compared to a single intranasal dose of placebo in healthy 
adults.   
A total of 16 subjects will be randomized to a treatment sequence – either OP0201-placebo or 
placebo-OP0201. Each subject will participate in the trial for up to five weeks and have up to three 
in-clinic visits and two telephone follow-up visits. There will be a screening period of up to 28 days, 
with a randomization visit on Day 1 where subjects will be randomized to receive a single masked 
dose of intranasal OP0201 containing a total of 20 mg OP0201 or placebo. The trial site staff will 
administer the IMP to each subject as four consecutive sprays at a time into each nare so that a 
total of eight sprays (four per nare) have been administered. On Day 8, subjects will return to the 
trial site for the cross-over visit where they will receive the opposite masked treatment 

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administered in the same way. Subjects will remain in the clinic for the duration of approximately 4 
to 6 hours at each visit.   
 
Safety will be assessed by clinical monitoring and will include a general physical examination, an 
ear, nose and throat (ENT) examination that includes otoscopy, nose and epipharynx endoscopy, 
tympanogram, clinical laboratory tests (hematology, biochemistry and urinalysis), triplicate12-lead 
ECG, vital signs (PR, RR, BP, temperature), and spontaneous adverse event reporting.  
Pharmacodynamics of OP0201 on ET function will be assessed by continuous tympanic 
impedance measures using a single-person hypobaric/hyperbaric atmospheric pressure chamber.  
An exploratory efficacy assessment of OP0201 effect to modulate ear pain during the 
hypobaric/hyperbaric atmospheric pressure chamber protocol is also planned.  
3.3 
Selection of trial population 
Prospective subjects who meet inclusion and who do not meet exclusion criteria as defined in 
Sections 3.3.1 and 3.3.2 (inclusion/exclusion, respectively) will be considered for entry into this 
trial.  
Subject eligibility should be reviewed and documented by an appropriately qualified member of the 
investigator’s trial team before subjects are included in the trial. The investigator or other medically 
qualified individual is responsible for assuring that subjects meet eligibility criteria. 
3.3.1 Inclusion criteria 
Subjects must meet all of the following inclusion criteria to be eligible for enrolment into the trial: 
• 
Male and female; 18 to 50 years of age at time of signed informed consent 
• 
Written informed consent obtained 
• 
Body Mass Index (BMI) 18 to 30 and a minimum body weight of 50 kg at the screening 
visit 
• 
Healthy subjects with no history or presence of significant medical condition or a 
clinically significant abnormal finding, as determined by the investigator. This includes 
study screening results from: 
• 
Medical history 
• 
Physical examination 
• 
ECG (in consultation with cardiologist) 
• 
Vital sign measurements 

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• 
Clinical laboratory data  
• 
Negative urine pregnancy test at screening and baseline for females of childbearing 
potential 
• 
Agree to refrain from water immersion of the ears from the time of signed informed 
consent to the end of trial 
• 
Physiologic tympanogram type A or type C3 at screening visit 
• 
Ability to follow trial instructions and complete all required visits 
3.3.2 Exclusion criteria 
If any of the following criteria are met, the subject is not eligible for participation in the trial.  
• 
Known substance abuse (e.g., alcohol, licit or illicit drugs) within 96 weeks prior to 
screening visit 
• 
Positive urine drug screen at screening visit 
• 
Upper respiratory tract infection currently or within 6 weeks prior to screening visit 
• 
Allergy or sinus conditions (e.g., sinusitis, non-specific nasal inflammation) currently 
or within 6 weeks prior to screening visit 
• 
Claustrophobia that is sufficient to prevent them tolerating assessments while in a 
hypobaric/hyperbaric atmospheric pressure chamber  
• 
Smoker (e.g., cigarettes, vapor) within the last 48 weeks prior to screening visit 
• 
Clinically significant findings on ENT exam 
• 
Gastroesophageal reflux disease currently or within 6 weeks prior to screening visit 
• 
Current diagnosis of sleep apnea 
• 
Evidence of craniofacial anomalies (eg, cleft palate, Down’s Syndrome) that may 
interfere with ET function 
• 
Disorders with decreased mucociliary clearance or higher viscosity of the mucous 
(eg, cystic fibrosis, primary ciliary dyskinesia, Kartagener’s syndrome) 
• 
Known hypersensitivity to any of the ingredients in the investigational product 
• 
Clinically significant ear disease currently or within 6 weeks prior to screening visit  
 
3 Tympanogram Type C defined as peak pressure between -100 mm water to -200 mm water 

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• 
History of tympanostomy tubes in one or both ears within 48 weeks prior to 
screening visit 
• 
Clinically relevant blockage of one or both nasal passages, in the investigator’s 
opinion  
• 
Use of any medication (either topically or systematically) for a condition related to 
ear or nose currently or within 12 weeks prior to screening visit 
• 
Use of anti-cholinergic medication (eg, anti-histamine, anti-nausea) currently or 
within 6 weeks prior to screening visit 
• 
Within seven days prior to the first study drug administration, use of any prescription 
drugs (except for oral contraceptives), and/or any over-the-counter drugs including 
herbal supplements, vitamins, dietary supplements (except for the occasional use of 
paracetamol not exceeding 3 grams per day) 
• 
Any condition or situation that would, in the investigator’s opinion, put the subject at 
significant risk, confound the trial results or interfere significantly with the subject’s 
participation  
• 
Participation in a clinical trial within 30 days prior to screening visit 
• 
Females who are pregnant, nursing, or planning a pregnancy during the trial  
• 
Females of child-bearing potential who are not willing to use an acceptable method 
of contraception during the trial (see Section 3.6.9 Pregnancy and Contraception 
Considerations). 
• 
Investigator site personnel directly affiliated with this trial and/or their immediate 
families (defined as spouse, parent, child, or sibling, whether adopted or biologic). 
• 
Persons employed by the Sponsor or investigator 
• 
Persons held in an institution by legal or official order 
3.4 
Withdrawal of trial subjects after trial start  
The subject’s participation in the trial is voluntary. The subject may refuse to participate or 
withdraw from the trial at any time, without penalty or loss of benefits to which the subject is 
otherwise entitled. Subjects may be withdrawn at the discretion of the Investigator or Sponsor for 
safety, behavioural reasons, or the inability of the subject to comply with the protocol required 
schedule of trial visits or procedures at the trial site.  
Notification of early subject withdrawal from the trial and the reason for discontinuation will be 
clearly documented in the electronic case report form (eCRF). 

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Subjects who withdraw from the trial may be replaced at the discretion of the investigator upon 
consultation with the Sponsor. 
3.4.1 Procedures for premature withdrawal from treatment during the trial 
If a subject does not return for a scheduled visit (crossover), every effort should be made to contact 
the subject. In any circumstance, every effort should be made to document subject outcome, if 
possible. The investigator should inquire about the reason for withdrawal, request the subject to 
participate in the final visit, if applicable, and follow-up with the subject regarding any unresolved 
adverse event, if any occurred. 
If the subject withdraws from the trial and also withdraws consent for disclosure of future 
information, no further evaluations should be performed, no additional data should be collected. If 
such action is taken, the date of withdrawal of consent must be documented. The Sponsor may 
retain and continue to use any data collected prior to such withdrawal of consent. 
All data collected until this point of time will be stored according to AMG §40, 2a, 3.  If a subject is 
lost to follow up, i.e. not attending the visit on day 8 (crossover), then the subject will be withdrawn 
by the investigator, and replaced. 
3.5 
Closure of trial sites/Premature termination of the clinical trial 
3.5.1 Closure of trial sites 
The trial site will be closed in case of insufficient subject recruitment in a timely manner, or by 
mutual decision of the investigator/PCI and Sponsor for any reason. 
3.5.2 Premature termination of trial 
The Sponsor has the right to terminate the trial prematurely if there are any relevant medical or 
ethical concerns, or if completing the trial is no longer practical. If such action is taken, the reasons 
for terminating the trial must be documented. All trial subjects still under treatment at the time of 
termination should undergo a final examination, which should be documented.  
3.5.3 Stopping Criteria for individual subjects and for the study 
The following stopping criteria are defined: 
1. Stopping Criteria for Individual Subjects 
Study treatment for any individual subject will be stopped if the subject experiences a serious 
adverse event (SAE) or a clinically significant possibly drug-related related AE, which in the 

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opinion of the study physician, Principal Investigator, or Sponsor’s medical representative, 
warrants discontinuation of the study for that subject’s wellbeing.   
Any female subject who becomes pregnant whilst participating in the study will be withdrawn 
from the study. 
2. Criteria for Stopping the Study 
If 1 or more subjects experience drug-related SAEs or 2 or more subjects experience any drug 
related severe AEs, the study will be stopped. 
If any of the following scenarios occur with a reasonable possibility of a causal relationship with 
the study treatment, the study will be stopped: 
• 
≥1 subject experiences SAEs considered to have a reasonable possibility of relationship 
to the study treatment. 
• 
≥2 subjects experience non-tolerable AEs that are considered to have a reasonable 
possibility of relationship to the study treatment. 
• 
≥1 subject receiving study treatment fulfills Hy’s Law defined as alanine 
aminotransferase (ALT) >3 × the upper limit of normal (ULN) and bilirubin >2 × ULN in 
the absence of significant increase in alkaline phosphatase (ALP) and in the absence of 
an alternative diagnosis that explains the increase in total bilirubin, to be assessed from 
the first administration of study treatment up to and including follow-up. 
• 
≥2 subjects who receive study treatment have >2 ULN of either ALT, bilirubin, or ALP. 
• 
≥2 subjects who receive study treatment have a QTc prolongation defined as QTcF 
>500 ms, or an increase of QTcF >60 ms above baseline on the 12-lead 
electrocardiogram (ECG), confirmed (persistent for >5 minutes) on repeated 12-lead 
ECGs. 
3.6 
Treatment 
3.6.1 Treatments to be given 
The trial site staff will administer each dose to the subject per the detailed instructions for IMP 
administration provided in the IMP Instructions for Use.   
At Day 1, subjects will receive a single masked dose of either OP0201 20 mg (2.5 mg per spray) or 
placebo (0 mg per spray) administered as four consecutive sprays first to the left nare, followed by 
four consecutive sprays to the right nare until a total of eight sprays (i.e. four to each nare) has 
been administered. At Day 8, subjects will return to the trial site where they will receive the 
opposite masked treatment administered as four consecutive sprays first to the left nare, followed 
by four consecutive sprays to the right nare until a total of eight sprays (i.e. four to each nare) has 
been administered.  

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Figure 1: Trial flowchart 
 
 
 
 
 
 
 
 
 
 
 

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Table 1: Visit Schedule 
 
Assessment Procedures 
Screening 
(-28 days 
to -1 day) 
Day 1 
(Baseline) 
-90-0 min 
Day 1 
(Trial 
Treatment) 
0 min 
Day 1 
Day 2h 
6
 
d
a
y
s
 
i
n
t
e
r
v
a
l 
 
Day 8 ± 1 
(Baseline; 
crossover) 
-90-0 min 
Day 8 ± 1 
(Trial Treatment; 
crossover) 
0 min 
Day 8 ± 1 (crossover) 
Day 9h 
(Trial Exit) 
 
0-60 
min 
 
90-150 
min 
 
150-215 
min 
(Visit 
Exit) 
 
0-60 
min 
 
90-150 
min 
 
150-215 
min 
(Visit 
Exit) 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/Exclusion 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
Medical, Surgical and Ear 
Historyi 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Physical Exama 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
Tympanogram 
X 
X 
 
 
 
X 
 
X 
 
 
 
X 
 
Continuous measure of 
tympanic impedance 
(ETOP, ETOD, ETCP, ETOF) 
in hyperbaric/hypobaric 
pressure chamber 
X 
X 
 
10 Min 
± 5g 
X 
 
 
X 
 
10 Min 
± 5g 
X 
 
 
Ear pain assessment 
(Numeric Pain Rating 
Scale 0-10 ) 
 
X 
 
X 
X 
 
 
X 
 
X 
X 
 
 
Otoscopy incl. TEED 
classificationb 
X 
X 
 
X 
X 
 
 
X 
 
X 
X 
 
 
Nose and epipharynx 
endoscopyj 
X 
X 
 
 
X 
 
 
X 
 
 
X 
 
 
12-Lead ECG 
X 
Xk 
 
 
 
Xk 
 
Xk 
 
 
 
Xk 
 
Clinical laboratory testsc 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
Vital signsd 
X 
X 
 
 
 
X 
 
X 
 
 
 
X 
 
Urine sample for drug 
screen 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Urine Pregnancy testse 
X 
X 
 
 
 
 
 
X 
 
 
 
 
 
Concomitant 
medications/procedures 
X 
X 
 
 
 
X 
X 
X 
 
 
 
X 
X 
Randomization 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
IMP Administrationf 
 
 
X 
 
 
 
 
 
X 
 
 
 
 
Adverse Events 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
ETOP = Eustachian Tube Opening Pressure; ETOD = Eustachian Tube Opening Duration; ETCP = Eustachian Tube Closing Pressure; ETOF = Eustachian Tube Opening Frequency; ECG = 
Electrocardiogram; IMP = Investigational Medicinal Product. 
 
 

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a. Includes: general appearance, overall status of the skin, head, neck, trunk, eyes, heart and lungs (eg, breathing sounds), abdomen, extremities, lymph nodes. Height and weight are recorded only on 
day of screening visit. 
b. Otoscopy involves ear endoscopy, taking a picture of the tympanic membrane and assessing TEED classification (TEED 0-4). 
c. Includes hematology, biochemistry and urinalysis   
d. Includes blood pressure (BP) [mmHg], respiration rate (RR) [breaths/min], pulse rate (PR) [beats/min] and body temperature (T) [°C]. 
e. For females of childbearing potential; urine pregnancy test must be negative. 
f. Start time of IMP administration and stop time of last spray must be recorded. 
g. Tympanic impedance measures are taken 10 ± 5 min after administration of IMP is completed. Time points as follows are recorded: (1) start time of impedance measures and (2) stop time of 
impedance measures. 
h. Telephone follow-up asking subjects about general status of well-being 24 h after IMP administration 
i. Ear history will record subject’s response (yes/no) to the following question “Have you ever had problems (eg, symptoms of aural fullness, popping or discomfort/pain) when flying or diving?”).  
j. Nose endoscopy will include a measurement of nasal cavity length (in cm) only on day of screening visit.  
k. Triplicate 12-Lead ECG 
 
 
 
 

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3.6.2 Visit Schedule 
Unless otherwise defined as in Figure 1 Flow Chart and Table 2 Visit Schedule, the following list of 
study-related procedures do not reflect a particular order. The specific order of sequence for these 
study-related procedures are referenced in an operational study procedure manual. 
3.6.2.1 Screening 
Subjects will be screened within 28 days up to 1 day prior to randomization to confirm that they 
meet the subject selection criteria for the trial. Prior to performing any study related procedure, the 
investigator (or an appropriate delegate at the trial site) will obtain informed consent from each 
subject according to the process as described in Section 5.4.  
The following procedures will be completed during the screening period: 
• 
Obtain written informed consent 
• 
Record demographics (age, race, gender)  
• 
Record height in centimeters (cm) and weight in kilograms (kg)   
• 
Assess inclusion/exclusion criteria  
• 
Record medical, surgical and ear history 
• 
Record subject’s response (yes/no) to the following question “Have you ever had problems 
(eg, symptoms of aural fullness, popping or discomfort/pain) when flying or diving?”)  
• 
Perform physical examination (general appearance, overall status of the skin, head, neck, 
trunk, eyes, heart and lungs (eg, breathing sounds), abdomen, extremities, lymph nodes)  
• 
Obtain vital signs (PR [beats/min], RR [breaths/min], BP [mmHg], T [°C]))  
• 
Perform otoscopy (separately for left and right ear, classify each ear according to TEED 
classification [Section 3.7.1] and record description of appearance of TM for each ear as 
described in Section 3.7.1)  
• 
Perform nasal endoscopy (separately for left and right nostril; assess as normal/abnormal) 
• 
Perform epipharynx endoscopy (assess as normal/abnormal)   
• 
Record measurement of nasal cavity length [cm] (separately for left and right side) 
• 
Perform tympanogram (assess as Type A, Type B or Type C) 
• 
Perform tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
(see Attachment 9.1) 
• 
Collect 12-Lead ECG  
• 
Collect clinical laboratory tests (non-fasting), (assess as normal/abnormal  – if abnormal, 
determine if clinically significant [yes/no], parameters described in Section 3.7.1) 
• 
Collect urine for drug screen  

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• 
Perform urinary pregnancy test for females of child-bearing potential performed, record as 
negative/positive 
• 
Record adverse events, if applicable  
• 
Record previous medication used within 12 weeks prior to screening and concomitant 
medications/procedures, if applicable  
• 
Confirm subjects agree that they will need to avoid immersion of ears in water for the 
course of the study. 
At the end of the screening visit, potential subjects will be instructed to; 
 
• 
Avoid immersion of ears in water between now and the time they return for the Day 1 visit 
until completion of the in-clinic visit on Day 8 
• 
Refrain from the following within 24 hours prior to Day 1 and again 24 hours prior to Day 8:  
• 
Consumption of more than six units (cups/glass) per day of coffee, tea, cola, 
energy-drinks, or other caffeinated beverages or foods containing caffeine or 
xanthine  
• 
Consumption of alcohol 
• 
Unaccustomed strenuous physical activity (e.g., weight lifting, running, bicycling, 
etc.).  
• 
Avoid the following medications throughout the duration of the trial: 
• 
Medications (either topically or systematically) for a condition related to ear or nose. 
• 
Anti-cholinergic medications (eg, anti-histamine, anti-nausea) 
• 
Any prescription drug (except oral contraceptives, see Section 3.6.9) 
• 
Any over-the-counter (OTC) drugs including herbal supplements, vitamins, dietary 
supplements (except for the occasional use of paracetamol not exceeding 3 grams 
per day)  
3.6.2.2 Day 1  
During the time subjects are in the clinic on Day 1, food or drink should not be consumed, other 
than water. 
Day 1 Baseline (-90 to 0 min pre-dose) 
• 
Confirm inclusion/exclusion criteria 
• 
Perform urinary pregnancy test for females of child-bearing potential performed, record as 
negative/positive 
• 
Obtain vital signs (PR, RR, BP, T) 
• 
Collect triplicate 12-Lead ECG  

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• 
Perform otoscopy separately for left and right ear, classify each ear according to TEED 
classification [Section 3.7.1] and record description of appearance of TM for each ear as 
described in Section 3.7.1  
• 
Perform nasal endoscopy (separately for left and right nostril; assess as normal/abnormal) 
• 
Perform epipharynx endoscopy (assess as normal/abnormal)   
• 
Perform tympanogram (assess as Type A, Type B or Type C) 
• 
Perform tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
(see Attachment 9.1) 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Obtain subject randomization 
• 
Record adverse events, if applicable  
• 
Record concomitant medications/procedures, if applicable  
Day 1 Dosing (0 min) 
• 
Trial site staff will administer IMP to subject 
• 
Record start time of IMP administration and stop time of last spray  
• 
Record adverse events, if applicable 
Day 1 Post-Dose Period 0-60 min 
• 
Perform the tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
within 10 minutes ±5 minutes of dosing (see Attachment 9.1); record start and stop times of 
impedance measures 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Perform otoscopy separately for left and right ear within 30 min after completion of the 
pressure chamber protocol, classify each ear according to TEED classification [Section 
3.7.1] and record description of appearance of TM for each ear as described in Section 
3.7.1  
• 
Record adverse events, if applicable 
Day 1 Post-Dose Period 90-150 min 
• 
Perform the tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
(see Attachment 9.1) 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Perform otoscopy separately for left and right ear within 30 min after completion of the 
pressure chamber protocol, classify each ear according to TEED classification [Section 

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3.7.1] and record description of appearance of TM for each ear as described in Section 
3.7.1  
• 
Perform nasal endoscopy (separately for left and right nostril; assess as normal/abnormal) 
• 
Perform epipharynx endoscopy (assess as normal/abnormal) 
• 
Record adverse events, if applicable 
Day 1 Exit (150-215 min) 
• 
Perform tympanogram (assess as Type A, Type B or Type C) 
• 
Obtain vital signs PR [beats/min], RR [breaths/min], BP [mmHg], T [°C]  
• 
Collect triplicate 12-Lead ECG  
• 
Record adverse events, if applicable 
• 
Record concomitant medications/procedures, if applicable 
3.6.2.3 Day 2 (telephone follow-up visit) 
• 
Record adverse events, if applicable 
• 
Record concomitant medications/procedures, if applicable 
3.6.2.4 Day 8 ± 1 (crossover visit) 
During the time subjects are in the clinic on Day 8, food or drink should not be consumed, other 
than water. 
Day 8 Baseline (-90 to 0 min pre-dose) 
• 
Perform urinary pregnancy test for females of child-bearing potential performed, record as 
negative/positive 
• 
Obtain vital signs PR [beats/min], RR [breaths/min, BP [mmHg], T [°C]  
• 
Collect triplicate 12-Lead ECG  
• 
Perform otoscopy separately for left and right ear, classify each ear according to TEED 
classification [Section 3.7.1] and record description of appearance of TM for each ear as 
described in Section 3.7.1  
• 
Perform nasal endoscopy (separately for left and right nostril; assess as normal/abnormal) 
• 
Perform epipharynx endoscopy (assess as normal/abnormal)   
• 
Perform tympanogram (assess as Type A, Type B or Type C) 
• 
Perform tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
(see Attachment 9.1) 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Record adverse events, if applicable  
• 
Record concomitant medications/procedures, if applicable 

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Day 8 Dosing (0 min) 
• 
Trial site staff will administer IMP to subject 
• 
Record start time of IMP administration and stop time of last spray 
• 
Record adverse events, if applicable 
Day 8 Post-Dose Period 0-60 min 
• 
Perform the tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
within 10 minutes ±5 minutes of dosing (see Attachment 9.1); record start and stop times of 
impedance measures 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Perform otoscopy separately for left and right ear within 30 min after completion of the 
pressure chamber protocol, classify each ear according to TEED classification [Section 
3.7.1] and record description of appearance of TM for each ear as described in Section 
3.7.1  
• 
Record adverse events, if applicable 
Day 8 Post-Dose Period 90-150 min 
• 
Perform the tympanic impedance measurements in hypobaric/hyperbaric pressure chamber 
(see Attachment 9.1) 
• 
Have subject complete ear pain assessment (0 = no pain up to 10 = worst pain imaginable) 
during pressure chamber protocol 
• 
Perform otoscopy separately for left and right ear within 30 min after completion of the 
pressure chamber protocol, classify each ear according to TEED classification [Section 
3.7.1] and record description of appearance of TM for each ear as described in Section 
3.7.1  
• 
Perform nasal endoscopy (separately for left and right nostril; assess as normal/abnormal) 
• 
Perform epipharynx endoscopy (assess as normal/abnormal) 
• 
Record adverse events, if applicable 
Day 8 Exit (150-215 min) 
• 
Perform tympanogram (assess as Type A, Type B or Type C) 
• 
Obtain vital signs PR [beats/min], RR [breaths/min], BP [mmHg], T [°C]  
• 
Collect triplicate 12-Lead ECG  
• 
Perform physical examination (general appearance, overall status of the skin, head, neck, 
trunk, eyes, heart and lungs (eg, breathing sounds), abdomen, extremities, lymph nodes)  

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• 
Collect clinical laboratory tests (non-fasting), (assess as normal/abnormal  – if abnormal, 
determine if clinically significant [yes/no], parameters described in Section 3.7.1) 
• 
Record adverse events, if applicable 
• 
Record concomitant medications/procedures, if applicable 
3.6.2.5 Day 9 Trial Exit (telephone follow-up visit) 
• 
Record adverse events, if applicable  
• 
Record concomitant medications/procedures, if applicable 
3.6.3 Description of investigational medicinal product (IMP) 
The IMP OP0201 is an intranasal aerosol, a dosage form recognized by the US Pharmacopeia, 
that contains two active ingredients, DPPC and CP, as a suspension in synthetic HFA-134a 
(1,1,1,2-tetrafluorothane [Hall 2015]), which is utilized as the propellant for the intranasal 
administration. OP0201 is supplied in mechanical packaging parts that includes a pressurized 
canister as the primary container, a metering valve that has a crimping seal for secure attachment 
to the canister, a tip to deliver the drug to the nares and a holder for the canister.   
OP0201 is manufactured using Good Manufacturing Processes with development phase 
appropriate compliance with global Health Authority guidelines (e.g., European Commission 
Volume 4, Annex 13; U.S. FDA Guidance for Industry: CGMP for Phase 1 Investigational Drugs, 
July 2008; U.S. FDA Guidance for Industry: Nasal Spray and Inhalation Solution, Suspension, and 
Spray Drug Products, July 2002). The OP0201 canisters are filled to deliver 100 actuations 
(sprays) per canister. Each metered spray of OP0201 delivers 0.1 mL of HFA-134a containing 
approximately 2.5 mg of the active ingredients.    
All the ingredients in this drug product are synthetic and do not contain any animal or human 
derivatives. 
For further details about OP0201 see Investigator’s Brochure.    
3.6.3.1 OP0201 
Trade name: N/A 
INN (International Nonproprietary Name)/active substance:  
Dipalmitoylphosphatidylcholine (DPPC) – C40H80NO8P (MW: 734.039 g/mol) 
Cholesteryl palmitate (5-cholestene 3-palmitate; CP) – C43H76O2 (MW: 625.07 g/mol) 
ATC-Code: N/A 
Presentation: Dry powder suspension in metered dose inhaler 

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Dose: 20 mg  
Manufacturer: Sciarra Laboratories, Inc., Hicksville, New Jersey, United States. 
3.6.3.2 Placebo 
Trade name:   
HFA-134a 
INN (International Nonproprietary Name)/active substance:  
1,1,1,2-Tetrafluoroethane (CH2FCF3) 
ATC-Code: N/A 
Presentation: Propellant in metered dose inhalor 
Dose: N/A 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx 
3.6.3.3 Manufacture of the IMP 
OP0201 is manufactured using Good Manufacturing Processes with development phase 
appropriate compliance with global Health Authority guidelines (e.g., European Commission 
Volume 4, Annex 13; U.S. FDA Guidance for Industry: CGMP for Phase 1 Investigational Drugs, 
July 2008; U.S. FDA Guidance for Industry: Nasal Spray and Inhalation Solution, Suspension, and 
Spray Drug Products, July 2002) by xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx in 
compliance with EU Directives 2001/20/EC and Annex 13 to the European GMPs and the 
European Clinical Trial Authorization dossier.  
See Investigator’s Brochure for further information.    
3.6.3.4 Labelling of IMP 
The IMP will be packaged and labeled in identical appearing canisters. The IMP (OP0201 and 
Placebo) will be identified as an IMP. The trial number, subject number and kit number will be 
identified on the unit label.   
The IMP will be labeled in accordance with Section 5 of the GCP regulations (GCP-V). 
3.6.3.5 Storage of IMP 
The IMP must be stored in a secured area at controlled room temperatures between 15 °C and 25 
°C (59 °F and 77 °F); excursions are permitted up to 30 °C (86 °F).  

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3.6.4 Compliance with treatment & dispensing and return of IMP 
A detailed inventory of all of the IMP units for this trial must be completed. The investigator must 
keep an accurate accounting of the number of IMP units for each IMP that are:   
• 
received from Novus Therapeutics (or Novus Therapeutics designee),  
• 
dispensed/administered to the trial subjects,  
• 
Any used IMP at the trial site will be destroyed according to regular disposal measures 
and local regulations, with the exception that any problem canister (eg, product does 
not spray out with pushing the valve, or clumps of product versus a ‘spray’ are noticed) 
may be returned to the Sponsor. 
• 
Any unused IMP at the trial site may be returned to the Sponsor or designee.  
3.6.5 Assignment of trial subjects to treatment groups 
Subjects are assigned to one of two treatment sequences (OP0201-placebo or placebo-OP0201) 
by means of the central 24/7 Internet randomization service TENALEA (FormsVisions BV, Abcoude, 
NL) on Day 1. TENALEA provides IMP kit IDs for the randomized subjects. Randomization will be 
blocked and stratified by gender and “non-significant baro-challenges (yes/no)“. Additional details 
will be documented in the randomization manual. 
3.6.6 Selection of dosage of investigational medicinal product 
In this controlled clinical trial, the proposed dose is 20 mg OP0201 . The clinical safety and 
tolerability of OP0201 for human use is guided by data from animal studies . These pre-clinical 
proof-of-concept and safety evaluations were performed in three animal species (i.e., gerbils, mice, 
and chinchillas) to assess the effects of the GLP material on ET passive opening pressure (POP), 
OM with effusion (OME), and acute OM (AOM). As compared to no treatment or propellant alone, 
the administration of the GLP surfactant material produced significant reductions in ET POP as 
compared to no treatment or propellant alone within 10 minutes of administration of a single 20 mg 
intranasal aerosolized dose (9) in healthy animals. In the ears of animals with OME, significant 
reductions in the duration and severity of middle ear effusion were observed with an average of 
10.57 days with 10 mg per day (QD group; (approximate average total exposure of 106 mg) and 
average of 8.57 days with 20 mg per day (BID group; approximate average total exposure of 171 
mg) of the GLP surfactant material (see Investigator’s Brochure, Section 4.2.1) as compared to 
baseline and other treatments (e.g., no treatment, aerosol alone, or surfactant plus phenylephrine) 
(10, 11). In the ears of animals with AOM, treatment with 40 mg per day for 10 days of the GLP 
surfactant material (total exposure of 400 mg) as compared to propellant alone, was associated 
with a greater proportion of normal tympanometry findings, a lower percentage of culture positivity, 

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a lower severity of ME disease (occurrence of labyrinthitis) and a greater proportion of animals 
without ME fluid (i.e., dry ears) (2).   
Results from these animal model studies demonstrate that single doses of 20 mg (given as 2 
sprays per nostril [5 mg per spray]) and daily repeated exposures (either 20 mg once daily or 20 
mg twice daily) resulting in total cumulative doses of up to 400 mg over 10 days (see Investigator’s 
Brochure: Section 4.2.1) applied via intranasal MDI were safe and well-tolerated and the measured 
effects on ET POP were generally consistent across animal model studies (2, 9-11) 
The clinical safety and tolerability of OP0201 manufactured by Novus Therapeutics, Inc. for human 
use is further guided by a limited amount of data from nine human cases . A total of eight cases 
were exposed to a different formulation, which was DPPC:CP 16:1 w/w see Investigator’s 
Brochure: Section 5.1.1). One case was exposed to another different formulation comprised of 
DPPC:CP 200:1 see Investigator’s Brochure: Section 5.1.1. 
3.6.7 Blinding 
Novus Therapeutics (or designee) will provide IMP in masked numbered kits that contain 
identically appearing canisters. Subjects and all site personnel will be blinded to the IMP 
assignment throughout the duration of the trial.   
3.6.7.1 Unblinding 
When necessary for the safety and proper treatment of the trial subject, the investigator can 
unblind the trial subject’s treatment assignment to determine which treatment has been assigned 
and institute appropriate follow-up care. When possible, Novus Therapeutics Medical Safety 
Physician (or designee) should be notified prior to unblinding. The investigator should inform the 
Novus Therapeutics Medical Safety Physician (or designee) of the unblinding if there is no 
notification prior to the unblinding. 
Emergency envelopes with randomization codes for specific kit IDs are provided to the site and the 
safety management by IMSB. The reason for breaking the code must be recorded in the trial 
subject’s source documents. 
3.6.8 Previous and concomitant medication 
Previous medications used within 12 weeks prior to screening will be recorded. Any concomitant 
medications used during the study will be recorded starting from time of informed consent. 
The decision to administer a prohibited medication is done with the safety of the trial subject as the 
primary consideration. When possible, Novus Therapeutics should be notified before the prohibited 
treatment or medication is administered. 

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The following medications are prohibited throughout the duration of the trial: 
• 
Medications (either topically or systematically) for a condition related to ear or nose. 
• 
Anti-cholinergic medications (eg, anti-histamine, anti-nausea) 
• 
Any prescription drug (except oral contraceptives, see Section 3.6.9) 
• 
Any over-the-counter (OTC) drugs including herbal supplements, vitamins, dietary 
supplements (except for the occasional use of paracetamol not exceeding 3 grams 
per day)  
3.6.9 Pregnancy and Contraception Considerations 
Definition of Female of Child-bearing Potential  
For purposes of this study, a female will be considered of childbearing potential unless she is 
permanently sterilized (i.e., hysterectomy) or is naturally postmenopausal.  Natural menopause is 
defined as the permanent cessation of menstrual periods, determined retrospectively after a 
woman who is at least 40 years of age has experienced 12 months of amenorrhea without any 
other obvious pathological or physiological cause. Female subjects who have had a hysterectomy 
or are naturally postmenopausal will not be required to use contraceptives during the trial. 
Pregnancy Considerations 
All female subjects who are of child-bearing potential will be advised to avoid pregnancy and are 
required to use methods of contraception considered acceptable to Novus Therapeutics during the 
trial.   
Acceptable Contraception 
The investigator will review the contraception requirements for the trial with each subject.  
The following methods of contraception with a Pearl Index lower than 1%, if properly used, are 
considered acceptable for use to avoid pregnancy during this study: oral hormonal contraception 
(‘pill’), dermal hormonal contraception, vaginal hormonal contraception (NuvaRing®), contraceptive 
plaster, long-acting injectable contraceptives, implants that release progesterone (Implanon®), 
tubal ligation (female sterilisation), intrauterine devices that release hormones (hormone spiral), 
double-barrier methods. 
This means that the following are not regarded as safe: condom plus spermicide, simple barrier 
methods (vaginal pessaries, condom, female condoms), copper spirals, the rhythm method, basal 
temperature method, and the withdrawal method (coitus interruptus). 

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3.6.10 End of Trial  
The end of trial date for each subject (i.e. completion of Visit 5/Day 9 ) will be recorded in the 
eCRF. 
3.6.11 Continuation of treatment after the end of the clinical trial 
This section does not apply for this trial.  
3.7 
Efficacy and safety variables 
3.7.1 Safety variables 
Otoscopy  
Otoscopy will be performed to assess the appearance of the TM. The TM will be rated for the 
following variables:   
1) Contour (normal, retracted, full, bulging, perforated),  
2) Color (normal, partly red, completely red),  
3) Fluid behind the TM (no/yes; if yes – yellow, translucent, red, blue or black) 
4) Translucency (translucent, semi-opaque, opaque)   
5) TEED classification (12, 13):  
• 
TEED 0 = normal otoscopy 
• 
TEED 1 = retraction and increased vascularization of manubrium and Schrapnell  
• 
TEED 2 = retraction and hyperemia of the entire eardrum  
• 
TEED 3 = fluid or blood in the middle ear  
• 
TEED 4 = perforated eardrum 
Tympanometry  
Tympanometry will be performed to assess TM mobility, ET function and middle ear function by 
measuring the amount of sound energy reflected back when a small probe is placed in the ear 
canal (14). The output of the right ear and left ear tympanometry will be a tympanogram tracing 
classified as follows (15): 
• 
Type A 
• 
Type B 
• 
Type C 

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Nasal and Epipharynx Endoscopy  
Nasal and epipharynx endoscopy will assess for normal or abnormal appearance. Abnormal 
appearance of nose and/or epipharynx will be documented and captured as applicable: swelling, 
redness, bleeding, rhinorrhoea (for the nose only) or, if applicable, other abnormal result that 
should be then specified. 
At the screeing vist, as a part of the nasal endoscopy, the length of the nasal cavity (in cm) will be 
recorded separately for the left and right side.  
12-Lead ECG 
A 12-lead ECG will be performed at the screening visit to ensure that a trial subject does not have, 
in the investigator’s clinical judgment, a clinically significant condition or a condition that may pose 
an unacceptable safety risk for study participation. Triplicate 12-lead ECGs will be performed at 
Day 1 (pre-dose, 150-215 min) and Day 8 (pre-dose, 150-215 min).  
ECGs will be assessed by a central ECG reading center (ERT). An ERT cardiologist will review 
screening, pre- and post-treatment ECGs and discuss any abnormal findings with the investigator, 
as appropriate.  
Physical Exam 
Physical exam will be performed to assess any physical abnormalities. The body systems to 
assess are: general appearance, overall status of the skin, head, neck, trunk, eyes, heart and 
lungs (eg, breathing sounds), abdomen, extremities, lymph nodes.  
Vital Signs 
Systolic and diastolic blood pressure (mm Hg), pulse rate (beats/minute), respiration rate 
(breaths/minute) and body temperature (°C) should be taken after a subject has been at rest 
(seated) for at least 2 minutes. Blood pressure will be recorded using a sphygmomanometer. Pulse 
rate and respiration rate will be measured by counting over 30 seconds and multiplying by 2. 
Respiration rate may be taken concurrently with all ECGs. 
Clinical Laboratory Tests 
Clinical laboratory tests will be analyzed at the local clinical laboratory of the University Hospital 
Cologne.  
Blood samples 
Blood samples (non-fasting) should be collected using standard laboratory instructions and 
procedures provided by the local laboratory. These blood specimens will be analyzed for the 
following: 

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• 
Hematology: hemoglobin [g/dL], % hematocrit, red blood cell count (RBC), white blood 
cell count (WBC), % neutrophils, % lymphocytes, % monocytes, % basophils, % 
eosinophils, platelet count, partial thromboplastin time [sec], and % prothrombin time. 
• 
Biochemistry: sodium, potassium, chloride, magnesium, phosphorus, calcium, 
creatinine, urea nitrogen, uric acid, total bilirubin, direct bilirubin, alkaline phosphatase 
(total), glutamic-oxaloacetic transaminase (GOT), glutamicpyruvic transaminase (GPT), 
gamma glutamyltransferase (GGT), albumin, total protein.  
General urine analysis 
pH, protein, ketones, bilirubin, blood (total Hb), nitrite, urobilinogen, specific gravity. If there are any 
abnormalities in the urinalysis that suggest the need for urine microscopy, then this should be 
performed and reported as a comment. 
Urine Drug Screen 
A urine drug screen will be performed by the local laboratory of the University of Cologne using a 
commercially available urine drug test kit obtained from möLab GmbH, Langenfeld, Germany. This 
urine drug screen will test for the following:  
Amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine, methamphetamines, 
methadone, opiates.  
No analysis is intended for this screening variable. 
3.7.2 Pharmacodynamic Assessments 
Pharmacodynamics of the ET will be assessed using continuous tympanic impedance measures 
(5). The following measurements will be recorded: 
1. ET opening pressure (ETOP) indicating the function of the ET  
Passive (phase 1 and phase 5) : only the first ETOP [mbar] measurement will be 
recorded for left and right ear, separately 
Active (phase 3): all ETOP [mbar] measurements will be recorded for left and right ear, 
separately 
 
2. ET opening duration (ETOD) indicating the duration between tube opening and closing 
Passive (phase 1 and phase 5): only the first ETOD  [s] measurement will be recorded 
for left and right ear, separately 
Active (phase 3): all ETOD [s] measurements will be recorded for left and right ear, 
separately 
 

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3. ET closing pressure (ETCP) indicating atmospheric pressure event that induces ET 
closure 
Passive (phase 1): only one ETCP [mbar] measurement will be recorded for left and 
right ear, separately 
Active (phase 3): No measurements will be recorded 
 
4. ET opening frequency (ETOF) indicating number of ET opening within a given time 
interval and depends on the rate of pressure decrease and increase measurement for 
passive and active ET function:  
Passive (phase 1 and phase 5): number of openings in phase 1 and 5 [/min] will be 
recorded for left and right ear, separately 
Active (phase 3): number of openings in phase 3 [/min] will be recorded for left and right 
ear, separately 
3.7.3 Exploratory Efficacy Assessment 
An exploratory efficacy assessment of OP0201 compared to placebo to ascertain whether the IMP 
modulates ear pain during the hypobaric/hyperbaric atmospheric pressure chamber protocol is 
planned. During the conduct of the hypobaric/hyperbaric atmospheric pressure chamber protocol 
subjects will rate the worst pain they felt in the right ear, and separately in the left ear during phase 
1, during phase 3 and during phase 5. The pain will be rated using a whole number Numeric 
Rating Scale (NRS) ranging from 0 (no pain) to 10 (worst pain imaginable).  
3.7.4 Demographics and Baseline Characteristics 
Demographic data (age, race, gender) will be collected at the screening visit. 
Medical, surgical and ear history will be collected at screening visit.  
3.7.5 Rationale for assessment procedures 
The safety of subjects participating in this phase 1 human clinical trial is the most important 
consideration as they would not normally be expected to derive any therapeutic benefit. The 
primary objective of this trial is to evaluate the safety and tolerability of a single dose of 20 mg 
OP0201 compared to placebo in healthy adult volunteers utilizing standard safety assessments 
including PE, ENT exam (i.e., otoscopy, nasal and epipharynx endoscopy, tympanogram), vital 
signs, clinical laboratory tests, triplicate 12-lead ECG  and AE monitoring. 
The two active ingredients, DPPC and CP, in OP0201 are highly endogenous in mammalian tissue 
including those of the nasopharynx and respiratory system. Therefore, it is essentially impossible 

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to measure administered concentrations of OP0201 using traditional pharmacokinetic 
assessments. Hence, pharmacodynamics assessments (continuous tympanic impedance) have 
been included to help characterize pharmacological effects on the ET. This method has been 
established as a dynamic evaluation of the ET (6). 
It has been reported that in healthy people ear pain can result from exposure to pressure 
fluctuations caused by compression and decompression manoeuvers in a pressure chamber (5). 
Thus, an ear pain assessment has been included to explore whether OP0201 compared to 
placebo modulates ear pain as a possible efficacy measure. 
3.8 
Data quality assurance 
3.8.1 Monitoring 
A representative on behalf of Novus Therapeutics, Inc., namely, CTC Cologne, will monitor the 
trial. The determination of the extent and nature of monitoring will be based on consideration such 
as the objective, purpose, design complexity, blinding, size and assessments in the trial. The 
monitor will perform 100% source data verification. Details about the extent of monitoring will be 
described in a separate monitoring manual.   
 
3.8.2 Audits/Inspections 
As part of quality assurance, the Sponsor has the right to audit the trial sites and any other 
institutions involved in the trial. The aim of an audit is to verify the validity, accuracy and 
completeness of data, to establish the credibility of the clinical trial, and to check whether the trial 
subject’s rights and trial subject safety are being maintained. The Sponsor may assign these 
activities to persons otherwise not involved in the trial (auditors). These persons are allowed 
access to all trial documentation (especially the trial protocol, case report forms, trial subjects’ 
medical records, drug accountability documentation, and trial-related correspondence). 
Authorized representatives of Novus Therapeutics, Inc. or regulatory authority representatives may 
conduct on-site visits to review, audit, and copy trial-related documents under applicable data 
protection regulations. These representatives will meet with the investigator(s) and appropriate 
staff at mutually convenient times to discuss trial-related data and questions.  
The Sponsor and the trial site involved undertake to support auditors and inspections by the 
competent authorities at all times and to allow the persons charged with these duties access to the 
necessary original documentation. 

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All persons conducting audits undertake all measures to keep all trial subject data and other trial 
data confidential. 
3.9 
Documentation 
Clinical documentation relevant to the trial includes all records in any form (including but not 
limited to written, electronic, ENT records and scans) that describe or record the methods, conduct 
and/or results of the trial, the factors affecting the trial and the actions taken. 
Data for this trial will be recorded in source documents that may include subject’s medical records, 
hospital charts, clinic charts, the investigator's trial subject trial files, as well as the results of 
diagnostic tests such as X-rays, laboratory tests, and electrocardiograms.  
The data will be entered online using an eCRF at the trial site via the Internet. Access to an eCRF 
will be provided to the trial site personnel as required. Plausibility checks are run during data entry, 
thereby detecting many discrepancies immediately. The CTC Cologne Data Management will 
conduct further checks for completeness and plausibility and will clarify any questions with the trial 
sites electronically via the trial software. These electronic queries have to be answered by the trial 
site without unreasonable delay. Further details will be specified in the data management manual. 
The following information should be entered into the trial subject’s medical record: 
• 
Trial subject’s name 
• 
Trial subject’s contact information 
• 
The date that the subject entered the trial, subject number and subject IMP kit number  
• 
Site will receive the randomization confirmation for each transaction.  All notifications 
are to be maintained with the trial source documents   
• 
The trial title and/or the protocol number of the trial and the name of Novus 
Therapeutics 
• 
A statement that informed consent was obtained including the date. A statement that 
data protection consent, or other country and local trial subject privacy required 
documentation for this trial has been obtained (including the date). 
• 
Reasons for screen failure  
• 
Dates of all trial visits 
• 
Trial procedures and assessments (e.g., ECG, labs, otoscopy, TEED, tympanogram).  
• 
All concomitant medications  
• 
All concurrent procedures and non-medication treatments  
• 
Occurrence and status of any adverse events (including onset date, assessments of 
severity, seriousness, and relationship with IMP, treatment instigated, action taken 
regarding IMP, outcome, and stop date). 

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• 
The date the subject exited the trial, and a notation as to whether the subject completed 
the trial or reason for early exit. 
• 
The results of protocol-required laboratory tests performed by the site (e.g., results of 
urine pregnancy tests). 
• 
Trial subject’s medical, surgical, and ear histories (including demographics) 
• 
For females, documentation of non-childbearing potential or results of pregnancy test 
and documentation of trial subject’s stated birth control method (if applicable) 
• 
Physical examination and vital sign results  
• 
All trial subject questionnaires (ear pain assessments)  
• 
Source notes should also include any trial subject counseling/education (when 
applicable) permissible medications/treatments, prohibited medications/treatments, and 
timing in relation to scheduled trial visits and trial compliance.   
3.9.1 Data management 
An electronic data capture (EDC) system will be used for this trial. The IT infrastructure and data 
management staff will be supplied by the CTC Cologne. The trial database will be developed and 
validated before data entry based on standard operating procedures at the CTC Cologne. The data 
management system is based on commercial trial software and stores the data in a database. All 
changes made to the data are documented in an audit trail. The trial software has a user and role 
concept that can be adjusted on a trial-specific basis. The database is integrated into a general IT 
infrastructure and safety concept with a firewall and backup system. The data are backed up daily. 
After completion and cleaning of data, the database is locked and the data exported for statistical 
analysis. 
The data will be entered online at the trial sites via the Internet. Plausibility checks are run during 
data entry, thereby detecting many discrepancies immediately. The CTC Cologne Data 
Management will conduct further checks for completeness and plausibility and will clarify any 
questions with the trial sites electronically via the trial software. These electronic queries have to 
be answered by the trial site without unreasonable delay. Further details will be specified in the 
data management manual. 
3.9.2 Archiving 
All trial related correspondence, patient records, consent forms, patient privacy documentation, 
records of the distribution and use of all investigational products, and copies of case report forms 
should be archived for at least 10 years in accordance with §13 (10) of the GCP-V (German legal 
implementation of ICH-GCP regulation). Trial site shall ensure archiving and retention of essential 

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trial documents (e.g. ISF, subject files and source data), after the termination or discontinuation of 
the trial for at least 10 years unless longer times are foreseen by local regulatory requirements.  
For countries falling within the scope of the ICH guidelines, the Sponsor-specific essential 
documents should be retained until at least 2 years after the last approval of a marketing 
application in an ICH region and until there are no pending or contemplated marketing applications 
in an ICH region or at least 2 years have elapsed since the formal discontinuation of clinical 
development of the investigational product. These documents should be retained for a longer 
period, however, if required by the applicable regulatory requirement(s) or if needed by the 
Sponsor. 
Novus Therapeutics, Inc. requires that it will be notified in writing if the investigator wishes to 
relinquish ownership of the data so that mutually agreed-upon arrangements can be made for 
transfer of ownership to a suitably qualified, responsible person. 
 
4 
Ethical and regulatory aspects 
4.1 
Independent ethics committee 
The clinical trial will not be started before approval by the competent local ethics committee. 
4.2 
Ethical basis for the clinical trial 
The present trial protocol and any amendments are prepared in accordance with the current 
version of the Declaration of Helsinki.  
4.2.1 Legislation and guidelines used for preparation 
The present clinical trial will be conducted in accordance with the published principles of the 
guidelines for Good Clinical Practice (ICH-GCP) and applicable legislation (especially the Federal 
Drug Law [AMG] and the GCP-V). These principles cover, amongst other aspects, ethics 
committee procedures, the obtaining of informed consent from trial subjects, adherence to the trial 
protocol, administrative documentation, documentation regarding the IMP, data collection, trial 
subjects’ medical records (source documents), documentation and reporting of adverse events 
(AEs), preparation for inspections and audits, and the archiving of trial documentation. All 
investigators and other staff directly concerned with the trial will be informed that domestic and 
foreign supervisory bodies, the competent federal authorities and authorised representatives of the 
Sponsor have the right to review trial documentation and the trial subjects’ medical records at any 
time. 

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4.3 
Notification of the authorities, approval and registration 
Before the start of the clinical trial, all necessary documentation will be submitted to the competent 
supreme federal authority for approval (Federal Institute for Drugs and Medical Products, 
Bundesinstitut für Arzneimittel und Medizinprodukte [BfArM]). The state authorities in the federal 
state in which the trial will be conducted will also be notified. Further, competent supreme federal 
authority, local authorities and ethics committee will be informed about the end of trial . 
4.4 
Obtaining informed consent from trial subjects 
Written informed consent is to be obtained from each trial subject prior to any trial-related activities 
or procedures in the trial. Together with the consent to take part in the trial, the subject must also 
agree to representatives of the Sponsor (e.g. monitors or auditors) or the competent supervisory or 
federal authorities having access to the data recorded within the framework of the clinical trial. The 
subject will be informed of the potential benefit and possible side effects of the IMP and placebo, 
and of the need and reasons to conduct a placebo-controlled clinical trial. It must be clear to 
subjects that he or she can withdraw his/her consent at any time without giving reasons and 
without jeopardizing his/her further course of treatment. The subjects have sufficient time for 
consideration prior to giving their consent to participate in the trial. 
The originally signed consent form is archived in the investigator site file. Trial subjects receive 
copies of the written information sheet, the confirmation of insurance with conditions, and the 
signed informed consent form.  
4.5 
Insurance of trial subjects 
All trial subjects enrolled are insured in accordance with § 40 AMG (German Medicinal Law) 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
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4.6 
Data protection 
A report of the results of this trial may be published or sent to the appropriate health authorities in 
any country in which the IMP may ultimately be marketed, but the trial subject’s name will not be 
disclosed in these documents. The trial subject's name may only be disclosed to Novus 
Therapeutics’ assigned auditor, who is obliged to confidentiality, or governing health authorities 
(e.g., US FDA, European Medicines Agency) if they inspect the trial site. Appropriate precautions 
will be taken to maintain confidentiality of medical records and personal information. 

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Data protection consent and other documentation in accordance with the relevant country and local 
privacy requirements (where applicable) is to be obtained from each trial subject prior to enrollment 
into the trial in accordance with the applicable privacy requirements (e.g., German Data Protection 
Law, European Union General Data Protection Regulation 2016/679). 
Subjects will be informed that their pseudonymised data will be passed on in accordance with 
provisions for documentation and notification pursuant to § 12 and § 13 of the GCP Regulations to 
the recipients described there.  
5 
Statistical methods and sample size calculation 
The analysis is descriptive, significance level is not adjusted for multiple testing. A p-value ≤ 0.05 is 
defined as significant for pharmacodynamic and efficacy assessments, demographic and 
screening characteristics. Confidence intervals will be calculated at the 95%-level.  
Missing values will not be replaced, but drop-outs may be replaced with additionally randomized 
trial subjects to ensure N=16 subjects for the PP population. 
5.1 
Statistical and analytical plan 
The analysis of trial data will be specified elaborately in a separate statistical analysis plan (SAP). 
5.1.1 Analysis populations 
The following populations will be used for analysis:  
The safety population will consist of all randomized trial subjects who received at least one spray of 
IMP in either nare. In safety data analyses, subjects will be analysed according to actual IMP 
received, regardless of randomization assignment.  
The per-protocol (PP) population consists of all randomized subjects who received the two 
complete doses (4 sprays to each nare) of IMP at Day 1 and Day 8 . Valid measures of ETOP and 
ETOD in passive phases 1 and 5 need to be collected for both ears and for all three measurement 
times at visits 2 and 4. Excluded are subjects with major protocol deviations (determined prior to 
database lock and unblinding of the data).  
5.1.2 Description of trial subject groups 
For all variables and analysis populations summary statistics will be calculated, grouped by 
treatment, period and measurement time, where applicable. If variables are measured before and 
after treatment at the same visit, the difference will be calculated - if appropriate - for each 
timepoint for visit 2 and separately for visit 4, and summary statistics will also be given.   

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Descriptive statistics include for quantitative variables the number of subjects, mean, standard 
deviation, median, median confidence interval, quartiles, minimum, maximum; and for qualitative 
variables: number of subjects, absolute and relative response frequencies. Confidence intervals 
may be calculated if appropriate. 
5.1.3 Primary objectives 
Primary analysis is the evaluation of the safety variables in the safety population.  
Incidence of AEs will be tabulated by primary system organ class (SOC) and by preferred term 
within each primary SOC, according to MedDRA code. The severity of the AEs will also be 
summarized. AE and SAE will be listed by subject, treatment and period.  
Analyses for single parameters will be specified in more detail in the SAP.  
5.1.4 Secondary objectives 
Pharmacodynamics of the ET and ear pain will be analysed in the safety population and 
additionally in the PP population. Treatment comparisons will be made according to Grizzle’s two-
stage approach of the cross-over design by two-sided tests, where appropriate (16). The applied 
test for comparison within the Grizzle’s two-stage concept is the Wilcoxon rank-sum test. Non-
parametric testing is planned for the main cross-over analysis because the determination of the 
distribution of variables is difficult with respect to the small sample size and the potential for 
outliers. In a sensitivity analysis, a parametric cross-over analysis will be performed. Tests will be 
carried out without covariates. Missing data will not be imputed. Exact Hodges-Lehmann 
confidence intervals will be calculated for estimates of treatment effects. Appropriate sequence-by-
period plots will be produced to present tympanic impedance variables. 
5.1.5 Subgroup analyses 
If sufficient numbers of subjects are met, subgroup analyses by stratification factors are planned. 
Details will be specified in the SAP. 
5.1.6 Interim analysis 
An interim analysis is not planned in this trial. 
5.2 
Sample size calculation 
Using a one-sample t-test with a significance level of 0.05 (2-sided) and a power of 0.8, a sample 
size of 14.3 is required in order to detect a effect size (treatment difference standardized by 

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standard deviation) of 0.8 (17). Since the ear pain and tympanic impedance variables will be 
analyzed primarily with a non-parametric method (whose asymptotic relative efficiency to the t-test 
under assumption of normal distribution is 0.955 (18)), the final sample size is adjusted to 16 
(=ceiling(14.3/0.955)) in total and 8 per sequence.  
6 
Safety 
6.1 
Definitions of adverse events and adverse drug reactions 
6.1.1 Adverse event 
An adverse event (AE) is any untoward medical occurrence in a clinical investigation subject 
administered a pharmaceutical product and that does not necessarily have a causal relationship 
with this treatment. An AE can therefore be unfavourable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease temporarily associated with the use of a 
medicinal (investigational) product, whether or not related to the medicinal (investigational) 
product.  
AEs will be assessed, documented, and recorded throughout the trial. AEs will be collected from 
the time after informed consent is signed until the end of trial for each subject (Day 9, trial exit). 
At each visit, the investigator will begin by querying for adverse events by asking each trial subject 
a general, non-directed question such as “How have you been feeling since the last visit?” Directed 
questioning and examination will then be done as appropriate. All reported adverse events will be 
documented on the appropriate eCRF. 
If, after obtaining the trial subject’s informed consent but prior to administration of trial 
product/treatment, a medical condition is identified via a trial entry protocol assessment or 
procedure (e.g., screening ECG test) which, based on the Investigator’s judgment (and in 
consultation with the trial subject), is determined to be a pre-existing condition (i.e., a condition 
existing prior to the time of the trial subject’s informed consent), it should be documented on the 
Medical History eCRF. Otherwise, the condition should be reported as a pre-treatment AE.  
6.1.2 Adverse drug reaction 
An adverse drug reaction (ADR) is any noxious and unintended response to an IMP related to any 
dose with at least a reasonably possible causal relationship with the IMP. 
6.1.3 Serious adverse events or serious adverse drug reactions (SAE/SADRs) 
A SAE/SADR is any adverse event that results in any of the following outcomes:  

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• 
Death,  
• 
A life-threatening adverse event, 
• 
Inpatient hospitalization or prolongation of existing hospitalization, 
• 
A persistent or significant disability/incapacity,  
• 
Or a congenital anomaly/birth defect. 
• 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse event, when based upon appropriate 
medical judgment may jeopardize the subject and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition.    
Any SAE must be reported immediately but no later than 24 hours after becoming aware of the 
SAE by the investigator to Sponsor or authorized representative. SAEs must be reported directly to 
the Sponsor authorized representative, namely, CTC Cologne as listed on the Trial Contacts Page 
and recorded on the SAE Form. All subjects with SAE/SADR must be followed up and their 
outcomes reported. In the case that the outcome is death, SAEs must be directly reported to the 
responsible ethics committee and competent authority according to §12 (6) GCP-V. The 
investigator must supply Novus Therapeutics, CTCC, the responsible ethics commitee and 
responsible competent authority with any additional requested information (e.g., autopsy reports, 
discharge summaries). 
6.1.4 Unexpected adverse drug reaction (UADR) 
An UADR is an ADR which, the nature or severity of which is not consistent with the applicable 
product information available for the IMP. Further information is described in the Investigator’s 
Brochure Section 6.3.1.  
All SAEs that are drug-related and unexpected (not listed as treatment-related in the current 
Investigator's Brochure) must be reported to the competent authority, responsible ethics committee 
and/ or other bodies according to local regulations.   
6.2 
Documentation and follow-up of AEs and ADRs 
The Sponsor ensures that all persons involved in the treatment of trial subjects are adequately 
informed of the responsibilities and actions required when AEs and/or ADRs occur. AEs and ADRs 
will be documented in the trial subject’s medical records and in the eCRF. 
For all AEs and ADRs the following information will be documented in the source documentation 
and the eCRF:  
• 
AE verbatim terminology and if applicable location (e.g., left ear pain) 
• 
Onset and resolution date 

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• 
Severity 
• 
Causal relationship with IMP  
• 
Seriousness 
• 
Action taken with IMP and other measures taken 
• 
Outcome 
Regardless of whether a causal relationship between the AE and the IMP is suspected, trial 
subjects who develop AEs must be monitored within the trial period until all symptoms have been 
subsided, pathological laboratory values have returned to pre-event levels, a plausible explanation 
is found for the AE/ADR, or the trial subject has died. 
 
Adverse reactions reported for other products that contain both DPPC and CP are discussed in 
Section 6.3.2 of the Investigator’s Brochure. Adverse reactions reported for intranasal HFA-134a 
(placebo) are discussed in Section 6.3.3 of the Investigator’s Brochure. 
 
For the procedure of SAE documentation and reporting see section 6.2.3 and 6.2.4, respectively. 
 
6.2.1 Severity of the AE 
A clinical determination will be made of the intensity of an AE. The severity assessment for a 
clinical AE must be completed using the following definition as guidelines: 
• 
Mild - Awareness of sign or symptom, but easily tolerated 
• 
Moderate - Discomfort enough to cause interference with usual activity 
• 
Severe - Incapacitating with inability to work or do usual activity 
6.2.2 Causal relationship between AE and IMP 
The investigator is obligated to assess causal relationship (related or not related) between study 
drug and each AE/SAE. 
Related: A causal relationship is present if a determination is made that there is a reasonable 
possibility that the AE/SAE may have been caused by the study drug. A reasonable possibility of a 
relationship conveys that there are facts, evidence, and/or arguments to suggest a causal 
relationship, rather than a relationship cannot be ruled out.  
Not related: There is not a reasonable possibility that the AE may be related to the study drugs. 

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The investigator will use clinical judgment to determine the relationship. The investigator will also 
consult the Investigator’s Brochure to aid his/her assessment. Alternative causes, such as 
underlying disease(s), concomitant therapy, and other risk factors, as well as the temporal 
relationship of the event to study treatment administration should also be considered and 
investigated. 
For each AE/SAE, the investigator must document in the source documents that he/she has 
reviewed the AE/SAE and has provided an assessment of causality. There may be situations in 
which an SAE has occurred and the investigator has minimal information to include in the initial 
report to CTCC. However, it is important that the investigator makes an assessment of causality for 
every event before the initial transmission of the SAE data to CTCC. 
The investigator may change his/her opinion of causality in light of follow-up information and send 
an SAE follow-up report with the updated causality assessment. The causality assessment is one 
of the criteria used when determining regulatory reporting requirements. 
6.2.3 Documentation and reporting of SAE and pregnancy  
Regardless of the assumed causal relationship, every SAE that occurs within the trial period must 
be documented in the source documentation and in the appropriate part of the eCRF. Additionally, 
all SAEs must be reported to CTCC by using the study specific SAE report form. SAEs after the 
reporting period must be reported if the investigator suspects a causal relationship between the 
IMP and the SAE. 
Pregnancies must also be documented in the source documentation,in the eCRF (on separate 
pregnancy reporting page and on the pregnancy outcome form) and additionally on the study 
specific pregnancy form and reported to CTC Cologne. 
Details of Safety Management are defined in a trial-specific Safety Manual, which describes 
information and communication pathways, important timeframes for reporting, interfaces and 
clarification of responsibilities and national statutory requirements. 
Pregnancy  
The investigator will inform CTC Cologne without delay about any pregnancy that occurs within the 
trial period, i.e. within 24 hours of being made aware of such. This will be documented on a 
separate “Pregnancy Report Form I”. Pregnant subjects should not receive further study treatment.  
The pregnant trial subject will be asked to give separate informed consent for pregnancy follow up 
after the completion of the trial. The investigator must document the outcome of the pregnancy and 
provide the documentation to the Sponsor. A separate “Pregnancy Report Form II” should be sent 
to CTC Cologne in a timely fashion after becoming aware of the pregnancy outcome. Both parents 

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are asked to give separate informed consent for child follow up. For live births, the health of the 
child should be monitored until the age of one month. 
If an SAE/SUSAR of mother or child occurs in the course of pregnancy or delivery, it has to be 
reported according to statutory requirements. 
6.2.4 SAE reporting by the investigator to the Sponsor 
In the event of a SAE, the investigator must: 
1. Inform the CTC Cologne, which is assigned with Safety-Management duties by the 
Sponsor, without delay (at latest 24 hours after being aware of the SAE) by sending the 
SAE report form, via Fax. 
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx 
xxxxxxxxxxxxxxxxxxxxxxxxx 
 
2. Obtain and maintain in his/her files all pertinent medical records, information, and medical 
judgments from colleagues who assisted in the treatment and follow-up of the subject. 
 
3. Provide Sponsor with a complete, written description of adverse event(s) on the SAE Form 
describing the event chronologically, including any treatment given (e.g., medications 
administered, procedures performed) for the adverse event(s). Summarize relevant clinical 
information about the event: signs, symptoms, diagnosis, was or clinical course and 
relevant clinical laboratory tests, etc. Include any additional or alternative explanation(s) for 
the causality which includes a statement as to whether the event was or was not related to 
the use of the investigational drug. 
 
Each SAE collected in the trial period must be followed up until one of the following occurs:  
 
The SAE is no longer serious. 
 
The subject dies. 
 
After the initial AE/SAE report, the Investigator is required to proactively follow each subject at 
subsequent visits/contacts. All AEs/SAEs will be followed until resolution, stabilization, the event is 
otherwise explained, or the subject is lost to follow-up.  
The recurrence, deterioration or ending of an existing SAE but also each correction of an SAE-
report will be documented as a follow-up report and will be reported to the Sponsor or its designee 
(i.e. CTC Cologne) immediately. 

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The investigator is obligated to perform or arrange for the conduct of supplemental measurements 
and/or evaluations as medically indicated or as requested to elucidate the nature and/or causality 
of the AE or SAE as fully as possible. This may include additional laboratory tests or investigations, 
histopathological examinations, or consultation with other health care professionals.  
Investigators are not obligated to actively seek AE or SAE after conclusion of the study 
participation. However, if the investigator learns of any SAE, including a death, at any time after a 
subject has been discharged from the study, and he/she considers the event to be reasonably 
related to the study treatment or study participation, the investigator must promptly notify the 
Sponsor.  
6.2.5 Assessment of Serious Adverse Event by Sponsor 
All SAEs are assessed by the Sponsor and PI with regard to seriousness and causality. The 
expectedness assessment will be done by Sponsor.  
If an AE is “serious”, “related” and “unexpected”, the criteria for an expedited report (Suspected 
Unexpected Serious Adverse Reaction, SUSAR) are fulfilled.  
When necessary for the safety and proper treatment of the subject, the investigator can unblind the 
subject’s treatment assignment to determine which treatment has been assigned and institute 
appropriate follow-up care. When possible, the Novus Therapeutics Medical Safety Physician (or 
designee, namely CTCC) should be notified prior to unblinding IMP. The investigator should inform 
the Novus Therapeutics Medical Safety Physician (or designee, CTCC) of the unblinding if there is 
no notification prior to the unblinding. 
6.2.6 Notification of ethics committee and competent supreme federal authority 
Adverse event reporting, including suspected unexpected serious adverse reactions, will be carried 
out in accordance with applicable local regulations (GCP-V §13 (1-4). CTCC will provide regulatory 
authorities with safety updates/reports (e.g. DSUR; GCP-V §13 (6)) according to local 
requirements, including suspected and unexpected serious adverse reactions, where relevant.  
6.2.7 Notification of study staff and study participants 
Safety-relevant results from ongoing studies will be communicated by the Novus Medical Safety 
Physician (or designee) to the Study Safety Physician(s) (e.g. at the CRO CTCC), the Principal 
Investigator(s) and CRO Project Managers.  Investigators and the CRO Project Manager are 
responsible for ensuring new safety information is provided to appropriate study staff in a timely 
manner. Investigators will inform study participants of any new safety findings in a timely manner. 

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The Informed Consent and the Investigator’s Brochure may be updated with new safety 
information as appropriate. 
6.2.8 Informing the Data Monitoring Committee 
A DMC is not planned in this trial. 
6.2.9 Known side effects caused by pressure differential during pressure chamber 
measurement protocol 
In very rare cases, injuries may result from pressure variations during compression or 
decompression (barotrauma) during the pressure chamber protocol including, for example, 
reddening of the tympanic membrane, bursting of a blood vessel on/near the tympanic membrane 
or accumulation of fluid behind the tympanic membrane. In rare cases, subjects can experience 
temporary ear pain, even if they do not have any prior history of ear pain when experiencing 
atmospheric pressure changes (e.g. during take-off or landing in an airplane; scuba diving).   
Serious ear injuries do not happen suddenly and, if they occur, appear slowly during the pressure 
variations and are manifested as growing ear pain prior to ear injury. Some subjects may 
experience anxiety or claustrophobia while in the chamber. 
The risk from pressure chamber exposure for the subject is minimized because the chamber 
protocol does not expose the subject to large atmospheric pressure changes for long periods of 
time. Subjects will undergo two pressure profile simulations representing (1) an airplane ascent 
and airplane descent and (2) a scuba dive and return to sea level. Subjects are exposed for a total 
of 4.5 minutes to atmospheric pressure changes per each pressure chamber assessment. At each 
of the Day 1 and Day 8 visits, the exposure will be a total of 13.5 minutes over a 4-6-hour period 
(total exposure 27 minutes across both days). Limiting the total daily time in the chamber reduces 
the risk for decompression illness, hypoxia, barotrauma or other potential hazards from excessive 
decompression.  
During the 4.5-minute chamber protocol, the investigator is constantly monitoring the pressure and 
observing the study participant visually (through the chamber window) and is able to speak to and 
hear from the study participant via an internal speaker system embedded in the chamber. If at any 
time the study participant appears to be experiencing excessive ear pain, anxiety, claustrophobia 
or for any other reason in the Investigator’s judgement, the pressure chamber protocol will be 
stopped immediately, the door of the chamber will be opened, and the study participant will be 
removed from the chamber. 
 

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7 
Use of trial findings and publication 
7.1 
Reports 
7.1.1 Interim reports 
No interim report is planned for this trial. 
7.1.2 Final report 
The competent authority and ethics committee will be informed within 90 days that the trial has 
officially ended. 
Within one year of the completion of the trial, the competent federal authority and the ethics 
committee will be supplied with a full final report on the clinical trial according to ICH E3 describing 
the results according to §13 (9) GCP-V. 
7.2 
Publication 
The Sponsor will comply with the requirements for publication of study results in accordance with 
standard editorial and ethical practice. Novus Therapeutics, Inc., as the Sponsor, has proprietary 
interest in the trial and thus will be involved in reviewing, at a minimum, any abstract or manuscript 
prior to submission in order to allow the Sponsor to protect proprietary information and to provide 
comments. For this study, authorship and abstracts or manuscripts composition will reflect joint 
cooperation between the investigator and Novus Therapeutics, Inc. personnel. Authorship will be 
1) established prior to the writing of abstracts or manuscripts, 2) determined by mutual agreement 
and 3) in line with International Committee of Medical Journal Editors authorship requirements.  
7.3 
Amendments to the trial protocol 
The investigator must not implement any deviation from or changes to the protocol without 
approval by Novus Therapeutics and prior review and documented approval/favorable opinion 
from the IEC of a protocol amendment, except where necessary to eliminate immediate hazards to 
study subjects. 
Amendments made in accordance with § 10 Secs. 1 and 4 GCP Regulations that require approval 
are submitted to the ethics committee and the competent supreme federal authority and will not be 
implemented until approved.  

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8 
References 
1. 
Seibert JW, Danner CJ. Eustachian tube function and the middle ear. Otolaryngol Clin 
North Am. 2006;39(6):1221-35. 
2. 
Chandrasekhar SS, Mautone AJ. Otitis Media: Treatment With Intranasal Aerosolized 
Surfactant. The Laryngoscope. 2004;114(3):472-85. 
3. 
Hain TC. Eustachian tube dysfunction (ETD). 2007. 
4. 
Monsell EM, Harley RE. Eustachian tube dysfunction. 1996;29:437-44. 
5. 
Meyer MF, Mikolajczak S, Korthäuer C, Jumah MD, Hahn M, Grosheva M, et al. Impact of 
Xylomethazoline on Eustachian Tube Function in Healthy Participants. Otology & Neurotology. 
2015;36(5). 
6. 
Mikolajczak S, Meyer MF, Hahn M, Korthäuer C, Jumah MD, Hüttenbrink K-B, et al. 
Characterizing the Active Opening of the Eustachian Tube in a Hypobaric/Hyperbaric Pressure 
Chamber. Otology & Neurotology. 2015;36(1). 
7. 
Ingelstedt S, Ivarsson A, Jonson B. Mechanics of the Human Middle Ear: Pressure 
Regulation in Aviation and Diving, a Nontraumatic Method: Acta oto-laryngologica; 1967. 
8. 
Münker G. Funktionsanalyse der Tuba eustachii. Albert-Ludwigs-Universität zu Freiburg i. 
Br.1972. 
9. 
Chandrasekhar SS, Connelly PE, Venkatayan N, Ammar Me-S, Tabor M, Mautone AJ. 
Intranasal Metered Dose Aerosolized Surfactant Reduces Passive Opening Pressure of the 
Eustachian Tube: Comparison Study in Two Animal Models. Otology & Neurotology. 2002;23(1). 
10. 
Venkatayan N, Connelly PE, Mautone AJ, Troublefield YL, Chandrasekhar SS. Dosage 
Regimens of Intranasal Aerosolized Surfactant on Otitis Media with Effusion in an Animal Model. 
Otolaryngology -- Head and Neck Surgery. 2001;124(4):388-93. 
11. 
Venkatayan N, Troublefield YL, Connelly PE, Mautone AJ, Chandrasekhar SS. Intranasal 
Surfactant Aerosol Therapy for Otitis Media With Effusion. The Laryngoscope. 2000;110(11):1857-
60. 
12. 
Teed RW. Factors producing obstruction of the auditory tube in submarine personnel. US 
Naval Med Bull. 1944;42:293-306. 
13. 
Edmonds C. Otological aspects of diving: Australasian Medical Publishing Company; 1973. 
14. 
Rosenfeld RM, Shin JJ, Schwartz SR, Coggins R, Gagnon L, Hackell JM, et al. Clinical 
Practice Guideline: Otitis Media with Effusion (Update). Otolaryngology -- Head and Neck Surgery. 
2016;154(1 suppl):S1-S41. 
15. 
Jerger J. Clinical experience with impedance audiometry. Archives of otolaryngology. 
1970;92(4):311-24. 
16. 
Grizzle J. E.. The Two-Period Change-Over Design and Its Use in Clinical Trials, 
Biometrics, 1965; 21(2), pp. 467-48017. 
StataCorp. Stata Statistical Software: Release 14. 
College Station, TX: StataCorp LP.2015. 
(https://www.stata.com/manuals14/psspoweronemean.pdf) 
18. 
William J Conover. Practical nonparametric statistics. 1971, p. 213. 
 
 

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9 
 Attachments 
9.1 
Hyperbaric/hypobaric pressure chamber measurement 
Pharmacodynamics is based on an exploratory analysis of ET function (by continuous tympanic 
impedance) of subjects whilst being exposed to an (semi)auto-modulated atmospheric pressure 
profile ranging from 0.8 bar to 1.2 bar in a single-person hypobaric/hyperbaric pressure chamber 
(Haux Life Support, Karsbad, Germany). The resulting pressure profile consists of five phases 
which are (Figure 2):  
• 
Phase 1 (P1): decompression atmospheric 1 bar to 0.8 bar 
• 
Phase 2 (P2): stationary at 0.8 bar (for 15 sec) 
• 
Phase 3 (P3): compression from 0.8 bar to 1.2 bar 
• 
Phase 4 (P4): stationary at 1.2 bar (for 15 sec) 
• 
Phase 5 (P5): decompression back to atmospheric 1 bar 
 
Figure 2: Standard pressure profile during continuous tympanic impedance measurement 
 
Adpated from Meyer and colleagues (5)  

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9.2 
Ear Pain Assessment (Example Page) 
 
 
 

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9.3 
TEED Classification 
TEED classification (12, 13):  
• 
TEED 0 = normal otoscopy 
• 
TEED 1 = retraction and increased vascularization of manubrium and Schrapnell  
• 
TEED 2 = retraction and hyperemia of the entire eardrum  
• 
TEED 3 = fluid or blood in the middle ear  
• 
TEED 4 = perforated eardrum 
 
Exemplary images of changes to the TM are shown for assigning classifications 1, 2, 3 and 4 
according to TEED (12) adapted by Edmonds (13)  
 
 
 
 
1
2
3
4