Clinical trial NCT05590728
Conditions: Bacterial Infection
Keywords: Apramycin Lung Pharmacokinetics Open-label Phase I Respiratory
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)
Start date: 2023-06-16; Completion date: 2023-10-22
Enrollment: 16 participants; Allocation: NON_RANDOMIZED; Intervention model: PARALLEL

Brief summary:
A Phase I, open label study of a single dose of 30 mg/kg of apramycin administered intravenously (IV) over 30 (+/- 5) minutes. Twenty subjects will be enrolled in the study to one of 5 cohorts, T1-T5, each corresponding to a timepoint after initiation of infusion at which a single fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) is performed. There will be 4 subjects per cohort. Cohort T5 will be enrolled after plasma and lung apramycin concentrations and preliminary PK data analysis are completed in cohorts T1-T4. Enrollment and dosing will be determined by bronchoscopy schedule. For each cohort, if 2 subjects are scheduled to receive study drug on the same day, the dose will be administered sequentially at least 2 hours apart. The primary objective is to assess plasma pharmacokinetic (PK) profile of apramycin and lung penetration of apramycin in epithelial lining fluid (ELF) and alveolar macrophages (AM) after single intravenous (IV) apramycin dose of 30 mg/kg in healthy subjects.

Detailed description:
A Phase I, open label study of a single dose of 30 mg/kg of apramycin administered intravenously (IV) over 30 (+/- 5) minutes. Twenty subjects will be enrolled in the study to one of 5 cohorts, T1-T5, each corresponding to a timepoint after initiation of infusion at which a single fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) is performed. There will be 4 subjects per cohort. Cohort T5 will be enrolled after plasma and lung apramycin concentrations and preliminary PK data analysis are completed in cohorts T1-T4. Enrollment and dosing will be determined by bronchoscopy schedule. For each cohort, if 2 subjects are scheduled to receive study drug on the same day, the dose will be administered sequentially at least 2 hours apart. The primary objective is to assess plasma pharmacokinetic (PK) profile of apramycin and lung penetration of apramycin in epithelial lining fluid (ELF) and alveolar macrophages (AM) after single intravenous (IV) apramycin dose of 30 mg/kg in healthy subjects. The secondary objectives are to 1) assess the safety of single IV administration of 30 mg/kg apramycin in healthy subject and 2) to assess changes in otoacoustic testing.

Arms: arm 1: Subjects 18 to 45 years of age to receive 30 mg/kg apramycin dose administered intravenously (IV) in a forearm vein over 30 min (+/- 5 min) using a syringe or infusion pump. 0.5 h (+/- 5 min) after dosing a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed to analyze concentration of apramycin in BAL. N=4 arm 2: Subjects 18 to 45 years of age to receive single 30 mg/kg apramycin dose administered intravenously (IV) in a forearm vein over 30 min (+/- 5 min) using a syringe or infusion pump. 2 h (+/- 5 min) after dosing a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed to analyze concentration of apramycin in BAL. N=4 arm 3: Subjects 18 to 45 years of age to receive single 30 mg/kg apramycin dose administered intravenously (IV) in a forearm vein over 30 min (+/- 5 min) using a syringe or infusion pump. 4 h (+/-10 min) after dosing a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed to analyze concentration of apramycin in BAL. N=4 arm 4: Subjects 18 to 45 years of age to receive single 30 mg/kg apramycin dose administered intravenously (IV) in a forearm vein over 30 min (+/- 5 min) using a syringe or infusion pump. 8 h (+/- 15 min) after dosing a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed to analyze concentration of apramycin in BAL. N=4 arm 5: Subjects 18 to 45 years of age to receive single 30 mg/kg apramycin dose administered intravenously (IV) in a forearm vein over 30 min (+/- 5 min) using a syringe or infusion pump. Cohort T5 will be enrolled after plasma and lung apramycin concentrations and preliminary PK data analysis are completed in cohorts T1-T4 24 h (+/- 1 h) after dosing a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed to analyze concentration of apramycin in BAL. N=4
Interventions: intervention 1: Apramycin (EBL-1003)
Intervention descriptions: intervention 1: A mono-substituted 2-deoxystreptamine comprising a unique bicyclic octadiose moiety. It is a crystalline free base of the amoniglycoside apramycin.

Protocol documents:
ICF_000.pdf: 
Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 1 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
INFORMED CONSENT TO TAKE PART IN A CLINICAL RESEARCH STUDY AND 
AUTHORIZATION TO USE AND DISCLOSE MEDICAL INFORMATION 
 
Sponsor / Study Title: 
 
Division of Microbiology and Infectious Diseases (DMID), 
National Institute of Allergy and Infectious Diseases (NIAID),  
National Institutes of Health (NIH) / “A PHASE I, OPEN-
LABEL STUDY TO ASSESS LUNG PHARMACOKINETICS 
AND SAFETY OF A SINGLE DOSE OF APRAMYCIN 
ADMINISTERED INTRAVENOUSLY IN HEALTHY 
ADULT SUBJECTS” 
Protocol Number: 
20-0012 
Principal Investigator: 
(Study Doctor) 
William B. Smith, M.D. 
Telephone: 
865-410-8300 
865-305-9100 (24 Hours) 
Address: 
Alliance for Multispecialty Research, LLC. 
1924 Alcoa Highway, 4 and 5 Northwest 
Knoxville, TN    37920 
 
KEY INFORMATION 
 
The following is a summary of the things you should know:  
 
• You are invited to take part in a research study of an investigational drug called apramycin. 
This means that the drug is evaluated in healthy persons like you and is not approved for use 
in people by the U.S. Food and Drug Administration (FDA). Your participation in voluntary 
and your consent is sought after you have read and understood information provided in this 
document. If you consent to participate and change your mind, you may withdraw from the 
study and notify the study doctor. If you withdraw after you received a treatment with the 
study drug, you will be asked to have standard medical physical examination and lab tests to 
ensure you are doing well and do not need more follow up. 
 
• Apramycin is a compound that is being developed for use as an antibiotic against severe 
infections that are resistant to multiple available antibiotics. The purpose of the research study 
is to evaluate how safe it is to give apramycin to people (that is, what type side-effects it 
might cause and how severe they are), to study its pharmacokinetics (that is, how the body 
handles the study drug) and if it is taken up by the lungs.  
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 2 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• If you decide to participate, you will be in the study for up to 58 days (from Screening Visit to 
Final Visit). There is a Screening Period of up to 26 days; a 5-day/4- night in-patient period, 
with 2 days of screening and baseline tests before you receive the study drug on Day 1; and 3 
days of follow up after receiving the study drug; and an out-patient follow-up period of 
approximately 27 days,  with two return visits to the Research site, about 14 days and 30 days 
after receiving the study drug.  
 
• Screening evaluations include standard physical examination and blood and urine tests, test of 
lung function, an electrocardiogram (ECG; tracing of the heart signals), ear exams and hearing 
tests, and may be completed in one or two visits. If you meet all the requirements to take part 
in the study, you will be admitted in the clinical research unit 2 days before study drug 
administration to confirm that you continue meeting the requirements to participate in the 
study. The day before you receive the study drug you will be assigned in one of 5 groups of 4 
people each who will have only one bronchoscopy and bronchoalveolar lavage at either 30 
minutes (0.5 hours), 2, 4, 8 or 24 hours after receiving the study drug. On Day 1, you will 
receive a single dose of the study drug in a dose of 30 mg/kg body weight. The study drug 
will be given in a vein in your arm over 30 min. The effects of the study drug on you will be 
assessed by vital signs (blood pressure, heart rate, breathing rate and temperature), physical 
exams and blood and urine at various timepoints after you received the study drug and until 
your last visit.   
 
• Bronchoscopy with bronchoalveolar lavage is an established procedure to examine the lung 
airways and take fluid specimens from a segment of the lung. The procedure will be done in 
the university hospital. An anesthesiologist will give you a small dose of a sedative drug or 
light anesthesia to minimized discomfort before the procedure. Then, the pulmonologist (a 
lung specialist) will pass a pneumonoscope (a thin tube with channels to look inside the lung) 
through your nose down your throat into a segment of your lung and inject and quickly 
remove for testing small amounts of sterile saline (salt solution). The procedure will take 
about 10 minutes to complete after you are sedated, and you will then be monitored in the 
recovery room before you return to the research unit.  
 
• Ear exams and hearing tests are standard procedures and will be performed in the Audiology 
Department on the hospital campus. Ear exams will consist of looking inside your external ear 
canal with an otoscope (an instrument with a light source to look into the ear). Hearing tests 
will be performed in a quiet room and evaluate your responses to sounds of different 
frequencies and volume transmitted through the air and bone.  
  
• The risks of the study can involve risks caused by the study drug and risks caused by study 
procedures. 
o In a completed research study in healthy people, apramycin was safe and well-
tolerated in doses up the dose to be used in this study. Potential side-effects of 
apramycin are an allergic reaction, and side-effects reported after multiple doses or 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 3 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
high doses of other aminoglycoside antibiotics, such as decrease in kidney function, 
hearing loss, imbalance problems and muscle weakness. 
o Bronchoscopy with bronchoalveolar lavage is a common and generally safe. Potential 
side-effects during the procedure include a drop in blood oxygen. increase in heart 
rate, and irregular heartbeats, which will be monitored continuously, and oxygen or 
medications may be used. Very rarely, injury to the airway or to the lung with 
accumulation of air or blood in chest cavity may occur and may require immediate 
intervention. Other side-effects may include nausea and dizziness from the sedative, 
and nose and throat irritation, nasal stuffiness, and cough from the pneumonoscope. 
Very rarely, a syndrome like bronchitis several hours later could occur. You will be 
followed closely in the recovery room and the research unit and appropriate treatment 
will be given if you develop any symptoms. 
o No side-effects are expected from ear and hearing testing procedures. 
o There may be side effects of drawing blood with a needle and/or IV tubes and IV 
catheter placement. Local pain, bruising, bleeding, inflammation or infection might 
occur at the site of the needle stick where blood is drawn. There is a possibility of 
dizziness or fainting while your blood is being drawn. Precautions will be taken to 
minimize the above complications. 
 
• The study has no direct benefit for you. Knowledge gained in the study could be of future 
benefit to public health and to individuals with infections, who might benefit if the study drug 
is licensed. You will be promptly notified of any abnormal clinical test results that may 
suggest previously unknown abnormalities that could be clinically significant, and you will be 
counseled to see your doctor for follow up. 
 
• There are no alternatives to the research study. You are free to consent to participate in the 
study or not to participate or to withdraw after your consented initially. You will be 
compensated for the study days and procedures you complete. You will not be penalized if 
you withdraw from the study. 
 
• Your personal health information will be protected. Your study medical record, laboratory test 
results and storage tubes containing your blood, plasma, urine or bronchoalveolar lavage 
specimens will be coded and will not identify you by name or provide any other personal 
information that can identify you.   
  
• You will be asked to consent separately to keep in long-term storage residual plasma or 
bronchoalveolar lavage samples to be used in future clinical research by the study sponsor, the 
Division of Microbiology and Infectious Diseases (DMID) of the National Institute of Allergy 
and Infectious Diseases (NIAID), part of the National Institutes of Health (NIH). DMID. 
 
INTRODUCTION 
You are being asked to be a subject in a clinical research study. You cannot be in this study if you 
are in another research study or have been in another study within the past month. Before you 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 4 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
agree to be a part of this study, it is very important that you read and understand the study plan 
and in what ways you will need to cooperate. This consent form may have words in it that you do 
not understand. You may ask the study doctor or the study staff to explain any words or 
information that you do not understand. If you sign and date this form, it means you want to be in 
this study. 
  
This form describes the reason for the study, and the way the study will be done. It also describes 
the benefits, risks, discomforts, and warnings about the study. This form will also explain how 
your medical information will be used, and who may see it. It describes your rights as a subject in 
the study. 
  
The study staff will ask you many questions. Your answers must be completely truthful. Your 
health history and any changes in the way you feel during the study are very important. You must 
tell the study staff about any changes in your health or the way you feel. If you are not truthful, 
you may harm yourself by being in this study. Your signature and date on this consent form 
serves as proof of your promise that the information you give is true and correct and that you will 
remain truthful during this research study. 
 
Alliance for Multispecialty Research, LLC (AMR, LLC.), the Research Site, is being paid by the 
Division of Microbiology and Infectious Diseases (DMID) of the National Institute of Allergy 
and Infectious Diseases (NIAID), part of the National Institutes of Health (NIH). DMID is also 
referred to as the “study sponsor” in this document. 
  
BACKGROUND AND PURPOSE:  
The study doctor and Division of Microbiology and Infectious Diseases (DMID), the sponsoring 
drug company, are conducting this research study of an investigational drug called apramycin. An 
investigational drug is a study drug that is not approved by the U.S. Food and Drug 
Administration (FDA). 
 
Apramycin, the investigational drug in this study, is a natural drug that was isolated from a fungus 
cultured in the laboratory and has been used as an antibiotic extensively in the last 40 years for the 
treatment of animal infections. It belongs to the class of aminoglycoside antibiotics that are used 
for the treatment of infections in people. It is being evaluated as an antibiotic for use in people 
because it was found to have some important properties in laboratory testing: (a) it is effective 
against infections by bacteria that are resistant to treatment by approved antibiotics; (b) it does not 
interact with genes affected by other antibiotics that cause hearing impairment or loss; and (c) it 
was safe in animal testing. Because of these findings, apramycin received approval to initiate 
testing in people by regulatory authorities, such as the European Medicines Agency (EMA) and 
the U.S. Food and Drug Administration (FDA). In a completed first clinical study in healthy 
subjects like you, apramycin was tested in doses ranging from 0.3 to 30 mg/kg body and follow 
up periods up to 30 days and was found to be safe and well tolerated. 
 
This study is being done to evaluate the effect of a single dose of the study drug, apramycin, after 
it is administered intravenously (IV; in the vein). The overall goals are to determine the safety, 
tolerability, pharmacokinetics (PK) and buildup of the study drug in the lungs. Pharmacokinetics 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 5 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
is the study of how a drug is absorbed, distributed, metabolized, and eventually eliminated by the 
body. Apramycin, the study drug, is being developed for the treatment of bacterial infections that 
are resistant to multiple current antibiotics. 
 
This study will be conducted in subjects that are healthy and between 18 to 45 years of age who 
are in good health, do not take certain medications, and, for women, if they are not pregnant or 
breastfeeding. There will be a total of approximately 20 male and female subjects in this study.  
This study will be done at one Research Site in the United States. All subjects in this study will 
receive single dose of apramycin on Day 1 of the study. 
 
The study will take up to 58 days to complete and is made up of the following periods: a 
Screening Period up to 26 days (Day -28 to Day -3), during which you will make up to 2 visits to 
the Research Site for medical testing and ear and hearing exams; a Check-in day (Day -2) to 
confirm your eligibility; a Baseline day (Day -1) to assign you to study treatment cohort (group) 
and enroll you in the study, and complete baseline assessments; an Inpatient Study Treatment 
Period lasting 3 days/2 nights, with eligibility reviewed prior to study treatment administered on 
Day 1, bronchoscopy with bronchoalveolar lavage (BAL) done once for each subject within 24 
hours after dosing, and in-patient follow up completed on Day 2 and Day 3; and an Outpatient 
Follow-up Period of 27 days (Days 4 to 30) with two site visits to the Research Site, on Day 14 
and Day 30 after study treatment. The last visit will be on Day 30. 
 
A description of this research study will be available on https://www.clinicaltrials.gov  as required 
by U.S. Law. This Web site will not include information that can identify you. At most, the Web 
site will include a summary of the study results. You can search this Web site at any time.  
 
INFORMATION ABOUT THE STUDY:  
If you are eligible to take part in this study, you will receive a dose of apramycin based on your 
body weight (30 mg for every kilogram). The study drug will be given once by IV infusion over 
30 minutes on Day 1 of the study treatment period. Each subject will take the study drug as open 
label, meaning that you will know that you are going to receive apramycin. Additional subjects 
may also be admitted to the Research Site before dosing and may serve as back-up study subjects. 
You may be selected to participate in this research study as either a study subject or a back-up 
study subject.   
 
After dosing with apramycin, you will have a lung procedure called bronchoscopy with 
bronchoalveolar lavage (BAL) to measure the amount of apramycin in the lung. Bronchoscopy is 
done under “conscious” sedation. You continue to breathe on your own but do not feel the 
discomfort of having the tube in your mouth or nose. It may feel uncomfortable, but it should not 
hurt. The entire procedure, including prep and recovery time, typically takes about four hours. 
Bronchoscopy itself usually lasts about 30 to 60 minutes.  
 
Each subject will be assigned to a different time for completing the lung procedure. The lung 
procedure will be done once in each subject at one of 5 different timepoints between 0.5 hours to 
24 hours after dosing. There will be 4 subjects at each timepoint who will have this procedure. 
This procedure will be done by specialist pneumonologists (lung physicians). Study staff will 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 6 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
accompany you to bronchoscopy and be present during the procedure as needed to collect 
required information after dosing with study drug.  
 
You will also have several ear exams and audiology tests to evaluate any effects of apramycin on 
your hearing. Appointments for hearing tests will be made by the Research Site and study staff 
will accompany you to your appointments.  
  
You will be asked to come to the Research Site 4 times: 2 times before receiving the study drug, 
and 2 times after receiving it. If it is not possible to complete all testing on some days, or if there 
is a need to do a repeat test, you will be given an appointment to complete testing on another day. 
Also, you will be admitted in the Research Site on the same day after you complete testing on the 
second visit and stay in the AMR clinical trial unit for 4 nights if you meet the requirements for 
receiving the study drug.  
 
The first visit will be a screening visit. You will be informed about the study, have your questions 
answered and, if you decide to participate in the study, as shown by signing and dating this 
consent form, you can proceed to the first part of screening to evaluate your eligibility to take part 
in the study. If you decide to participate, and sign and date this consent form, you will have 
medical tests and complete an ear and hearing tests. 
  
If you pass the screening tests, you will be asked to return to the Research Site for a second visit 
(Day -2, Check-in) to complete medical and ear exam and hearing tests to confirm your 
eligibility to be admitted. If you are eligible to continue, you will be admitted to the Research Site 
on this day (Day -2) and stay overnight. On the next day (Day -1, Baseline), your eligibility to 
enroll in the study will be evaluated before you have electrocardiograms (ECGs). If you are 
eligible, you will be enrolled and assigned to 1 of 5 study treatment cohorts (groups), T1 to T5, to 
have only one bronchoscopy with bronchoalveolar lavage (BAL) at a specified timepoint after 
you receive the study drug as follows: Cohort T1, 0.5 hours (30 min); Cohort T2, 1 hour; Cohort 
T3, 4 hours, Cohort T4, 8 hours; and Cohort T5, 24 hours. You will be told what your cohort 
number is. You will complete ECGs on the same day. An ECG is a test to monitor the electrical 
activity of your heart. If you continue being eligible to continue participating in the study, then 
you will stay in the Research Site overnight for the inpatient study treatment period.  
 
On the next day (Day 1), your eligibility to receive the study drug will be evaluated. If you are 
eligible to receive the study drug, you will begin the inpatient study treatment period. In the 
morning on Day 1, you will have three baseline ECGs taken before you receive the dose of 
apramycin. Prior to receiving a dose of Apramycin, you will be required to have no food to eat or 
drink at least 6 hours. Apramycin, dissolved in 30 mL (about 1 ounce or 2 tablespoons) of sterile 
normal saline will be slowly administered intravenously (IV) through a vein in your forearm over 
30 minutes. 
 
You will be asked for any new symptoms before dosing (taking the study drug) and monitored for 
any side effects from the time of dosing, before, during and after the lung procedure, and until you 
complete the last visit (Day 30). 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 7 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
You will have the lung procedure (bronchoscopy with bronchoalveolar lavage) at the time that 
was assigned to your cohort number after you receive apramycin. You will have no food to eat at 
least 6 hours before dosing with apramycin and 2 hours after dosing. If you have the lung 
procedure, you will need to continue the fast until the procedure is completed and 2 hours after 
the lung procedure. This means that if you are in groups T1, T2, or T3, you will continue fasting 
until at least 2 hours after the lung procedure; if you are in group T4, you will eat 2 hours after the 
dose of apramycin, but you will need to start fasting again 4 hours before the lung procedure and 
continue until 2 hours after the procedure; and if you are in group T5, you will start eating 2 hours 
after dosing, but fast overnight before the lung procedure in the morning of the next day and for 
2 hours after the procedure. You will be allowed to drink water during that time, and you may be 
given nutritional broth or a light meal, depending on what time after dosing you will have the 
bronchoscopy. 
 
You will continue to stay at the Research Site overnight for 2 days after dosing for observation 
and additional required testing. These will include medical examinations, measurements of vital 
signs (heart rate, blood pressure, respiratory rate, and oral temperature), monitoring of blood 
oxygen, ECGs, and hearing tests. Blood will be drawn from a vein for clinical laboratory tests and 
measurements of the study drug at various times up to 60 hours after dosing. Additional testing 
may be done if medically indicated. You will be discharged from the Research Site on Day 3, 
after completion of the required testing at 60 hours after dosing. You may be kept overnight at the 
Research Site and discharged in the morning of Day 4 depending on timing of the 60-hour 
collection. 
 
You will then start the Out-patient Follow Up Period. You will return for two out-patient follow 
up visits to the Research Site, on Day 14 and Day 30, to complete additional required 
observations. Additional hearing tests will be done on these days. If there is no medical reason for 
further follow-up, you will complete your participation in the study on Day 30.  
 
The schedule for screening, study treatment and all study periods are shown in the table below:   
    
Outpatient 
Screening 
Inpatient 
Inpatient 
Study Treatment 
Outpatient 
Follow Up 
Screening 
Period 
Check-in Baseline 
Dosing 
Discharge 
Follow-up 
Visit 
Final 
Visit 
Day -28 to  
Day -3 
Day -2 
Day -1 
Day 1 
Day 3 
Day 14 
Day 30 
  
MEDICAL MONITORING:  
The study staff will monitor you for side effects during your stay in the Research Site and provide 
medical care as needed if you have any medical problems. Guidelines have been created to 
evaluate the safety of the study drug and whether the side effects are acceptable. 
    
 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 8 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
WHAT WILL HAPPEN DURING THE STUDY? 
 
SCREENING PERIOD (Day -28 to Day -3): 
 
Screening Visit 
During this visit, you will be informed about the study, have your questions answered and, if you 
decide to participate in the study, as shown by signing and dating this consent form. Following 
consent, you will have your medical assessments completed for the study staff to determine if you 
are in good health and qualify to continue participation in the study. 
  
After signing and dating this consent and before you begin the study, the following screening tests 
will be done: 
• Complete medical history (including your demographic information) and medication 
review. 
• Vital signs (blood pressure, heart rate, breathing rate and oral temperature) measured. You 
will need to rest for at least 5 minutes before your vital signs are taken. 
• Height and weight measured, and Body Mass Index (BMI) calculated. 
• Physical examination. 
• 12-lead ECG (standard recording) – This will require attachment of electrodes from the 
standard ECG machine to your chest, arms, and legs. Male subjects may have to have their 
chest hair shaved for the ECG. Female subjects may not be allowed to wear a bra during 
ECG-related procedures. 
• Blood and urine collected for laboratory tests (you will be asked to not eat or drink 
anything other than water for at least 6 hours before blood collection). 
• Urine drug screen for marijuana or other illegal drugs. The test must be negative to 
qualify. 
• Alcohol test in the urine. The test must be negative to qualify. 
• Cotinine test in the urine (for use of nicotine products).  
• Blood collected for serum pregnancy test if female. The test must be negative to qualify. 
• Blood collected for measurement of follicle-stimulating hormone (FSH) if post-
menopausal female. 
• Testing for Hepatitis B, Hepatitis C, and HIV infections: Your blood will be tested for the 
HIV antibody. HIV is a virus that causes AIDS. You will also be tested for Hepatitis B 
and C Viruses. All these test results must be negative to qualify for the study. It may take 
weeks or months after being infected with HIV for the test to be positive. If either of these 
tests is positive, a confirmation test will be done by the laboratory. If the HIV or hepatitis 
test are positive, you will be notified by the Research Site and given information on how 
to follow up for further medical care. As required by law, positive test results must be 
reported by the Research Site to the State Department of Health. If you have any questions 
about what information is required to be reported, please ask the study staff. If the 
Research Site becomes aware during your participation in this study that there is any 
change in the HIV or Hepatitis Virus results, you will be withdrawn from the study. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 9 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• On this or another visit, we will also review the hearing criteria and you will have the 
following ear exam and hearing tests by an audiologist (a hearing specialist) in a nearby 
building, where you will be accompanied by study staff: 
• Ear Examination: You will have 4 tests that allow the audiologists to evaluate your 
external ear canal, the ear drum, the middle ear and the eustachian tube (the canal that 
connects the middle ear with the upper part of the throat): 
o Ear Otoscopy: inspection of your ear canal with a microscope to magnify the 
view of the canal and the ear drum. 
o Valsalva test: you will take a deep breath and then blow out fast with your 
mouth closed, as you do when you pop your ears, while the ear drum 
movement and middle ear pressure are measured. 
o Tympanometry: is a test of the condition of the middle ear, the mobility of the 
eardrum and the conduction bones by creating variations of air pressure in the 
ear canal using a small probe in your ear canal. 
o Stapedial Reflexes: a test of the lowest stimulus that activates the stapedial 
reflex at various sound pressure levels using a small probe in your ear canal. 
• Ear audiometry: The test evaluates the condition of your inner ear. It evaluates 
how sharp your hearing is by asking you to detect sound of various frequencies 
transmitted through the air or bone. The test is done in a quiet room or chamber, and 
you may wear headphones (for air conduction) or have a probe that sends sounds 
attached to your head (for bone conduction). It requires your concentration and 
attention.  
• Distortion product otoacoustic emissions (DPOAEs): This is a test of the condition 
of the inner ear. This takes a short time to complete and requires the insertion of a 
small probe in your ear canal. 
 
After you complete the ear and hearing tests, you will return to the Research Site and be told of 
the results of medical and hearing tests and whether you can continue participating in the study. 
• If you do not meet the study requirements, you will be discharged on this day. You will be 
told why you were not selected for the study. 
o If there is a medical condition or abnormal laboratory test that needs further 
medical attention, you will be counselled to follow up with your primary physician 
or contact community physicians or clinics where you can go for testing and 
follow up. You may sign and date a release form to allow the Research Site to 
release medical tests to you or send them to your own doctor or clinic. 
o If you did not pass the hearing tests, you will be told why and be counselled to see 
a specialist. You will be asked to sign and date a release form to obtain a copy of 
the hearing tests. 
• If your medical tests and hearing are good, the study staff will give you an appointment to 
return to the Research Site to complete the inpatient screening tests to confirm your 
eligibility to participate in the study. 
• You will be told to come prepared to stay for the inpatient study treatment if you pass the 
tests. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 10 of 30 
  
  
 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
• You will be counselled to avoid pregnancy, use appropriate contraception (both male and 
female), and avoid prohibited medications, tobacco and other nicotine products, alcohol, 
and marijuana and or other illegal drugs. 
• If you are male, you will be counseled to avoid sperm donation for 30 days after dosing. 
• You should avoid strenuous physical activity and exposure to loud noise until you 
complete the study if you meet the study criteria or until you are told you did not qualify. 
 
CHECK-IN VISIT (Day -2) (2 days before the study treatment period) 
On this day, the following procedures will be done: 
• Update medical history. 
• Review and update prior medications. 
• Vital signs (blood pressure, heart rate, breathing rate, oral temperature) measured. 
• Weight taken. 
• Physical examination. 
• 12-lead ECG (standard paper recording). 
• Blood and urine samples collected for lab tests (you will be asked to not eat or drink 
anything other than water for 6 hours before blood collection). 
• Urine pregnancy test (for females). The result must be negative. 
• Urine drug screen for marijuana or other illegal drugs. The result must be negative. 
• Alcohol test in urine. (The result must be negative.) 
• Cotinine test in the urine. (The result must be negative.) 
• Repeat Ear exam (ear otoscopy) and the Hearing tests (ear audiometry and distortion 
product otoacoustic emissions [DPOAEs]). 
• Review your results of the procedures to see if you still qualify. 
o If you do not meet the study requirements, you will be discharged on this day. You 
will be told why you were not selected for the study If there is a medical condition 
or abnormal laboratory test that needs further medical attention, you will be 
counselled to follow up with your primary physician or contact community 
physicians or clinics where you can go for testing and follow up. You may sign 
and date a release form to allow the Research Site to release medical tests to you or 
send them to your own doctor or clinic. 
o If you did not pass the hearing tests, you will be told why and be counselled to see 
a specialist. You will be asked to sign and date a release form to obtain a copy of 
the hearing tests. 
 
If you meet the study requirements, you will be admitted to the Research Site. After you are 
admitted, you will read the House Rules and are expected to follow them while you participate in 
the study. You will be encouraged to drink plenty of fluids to keep well hydrated. You will 
receive dinner and snacks in the Research Site. 
 
 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 11 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
INPATIENT BASELINE (Day -1): 
You will have the following assessments on this day: 
• You will be assigned to one of five study treatment cohorts (groups), T1-T5, that 
correspond to the timepoint when you will have a single bronchoscopy with BAL after 
receiving the study drug. You will be told what cohort you were assigned to.  
• Update medical history. 
• Review and update prior medications. 
• Vital signs (blood pressure, heart rate, breathing rate, oral temperature) measured. 
• Weight taken. 
• Physical examination completed. 
• 12-lead standard ECG (standard paper recording) done in triplicate at 4 timepoints during 
the day that will match the timepoints when you will have ECGs after dosing on Day 1. 
You will be resting for about 5 minutes while ECGs are recorded. 
• Review the results of the assessments to see if you still qualify. 
• You will not be allowed to eat anything for 6 hours before taking the study drug the 
following morning, but you will be allowed to drink water during that period. 
• You will be counseled to avoid strenuous exercise and loud music. 
  
INPATIENT STUDY TREATMENT PERIOD (Days 1, 2 and 3) 
You will receive the study drug on Day 1 and followed as inpatient until Day 3. The following 
procedures will be done during this period: 
 
DAY 1: Administration of study drug 
 
Before Dosing: 
• 
You will not eat any food or drink (other than water) at least 6 hours before taking the 
study drug (dosing). That means that you may have an evening snack the night before 
dosing but no food afterwards, and you will not eat breakfast before dosing. You will be 
allowed to drink water during that time before dosing. 
• 
Update medical history. 
• 
Physical exam may be done as needed depending on your symptoms. 
• 
Review and update medications taken. 
• 
Vital signs (blood pressure, heart rate, breathing rate, oral temperature) measured 
approximately 30 minutes before study treatment.  
• 
12-lead ECG (standard paper recording) within approximately 30 minutes before study 
treatment. 
• 
Review the results of the assessments to see if you still qualify to receive the study drug. 
• 
An IV line (a short intravenous catheter) will be inserted in a vein in your hand or forearm 
to study drug infusion. Another IV line will be inserted in your other arm to draw blood.  
• 
Blood and urine samples collected for lab tests (you will be asked to not eat or drink 
anything other than water for 6 hours before blood collection).  
• 
Blood drawn for analysis of study drug. 
• 
You will be kept well hydrated by drinking water starting before the study drug infusion 
and continue until 24 hour later. You will drink about 2 liters during this period (about 64 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 12 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
fluid ounces or 8 8-ounce glasses). You will be asked to document the time and volume of 
water consumed. 
 
Dosing: 
• 
Apramycin, 30 mg per kg of body weight, will be infused IV (in a vein) in a volume of 
30 mL (1 oz, 2 tablespoons) over 30 minutes using a syringe or infusion pump. 
• 
Report any side effects that you may experience. 
• 
Document your use of any medications. 
• 
Physical exam may be done as needed depending on your symptoms. 
 
After Dosing, the following procedures and assessments will be performed: 
 
Safety procedures: 
• 
Report any side effects that you may experience. 
• 
Document your use of any medications. 
• 
Physical exam may be done as needed depending on your symptoms. 
• 
12-lead ECG (standard) approximately 1, 4, and 16 hours after dosing. 
• 
Vitals signs (blood pressure, heart rate, breathing rate, oral temperature) measured 
approximately 1, 4 and 16 hours after dosing and as needed. 
• 
Blood drawn to measure the concentration of study drug approximately at 0.5, 1, 2, 4, 8 
and 16 hours after dosing. 
• 
The infusion site will be checked at 1 hour and 16 hours after the infusion and as needed 
because of symptoms that you might have. 
• 
You will not have food to eat until approximately 2 hours after dosing unless you are 
scheduled to have a bronchoscopy on this day (Day 1). In that case, you will not have 
solid food at least 4 hours before the procedure and start eating after you recover and are 
able to swallow. You will receive liquids before the procedure and as soon as you can 
swallow after the procedure.  
• 
You will continue drinking water until 24 hours after he infusion. Document time and 
volume of water consumed. 
• 
You should avoid strenuous physical exercise and exposure to loud noises after dosing and 
until the study end. 
 
Bronchoscopy with Bronchoalveolar Lavage (BAL): 
• 
Bronchoscopy with Bronchoalveolar lavage will be done only once after dosing on Day 1 
if you were assigned to study treatment groups T1–T4. 
• 
You will be accompanied by study staff to the bronchoscopy procedure. 
• 
The procedure will be explained to you. The entire procedure, including prep and recovery 
time, typically takes about four hours. Bronchoscopy itself usually lasts about 30 to 60 
minutes. 
• 
Safety monitoring will be initiated by bronchoscopy staff. Your vital signs (blood 
pressure, heart rate, respiratory rate) and ECG will be monitored continuously from before 
the procedure to the time you have fully recovered. 
• 
Your blood oxygen will be monitored continuously with a pulse oximeter, a small probe 
attached like a clamp to one of your fingers. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 13 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• 
An intravenous catheter, if not already in place, will be inserted in one of your hand or 
forearm veins to provide IV fluids, and medication as needed. 
• 
You may receive oxygen through a nasal canula as needed. 
• 
You will be given a sedative or light anesthesia by vein. You will be told what drug will 
be used. 
• 
The bronchoscopist will perform the bronchoscopy and collect bronchoalveolar lavage 
(BAL) samples. 
o A flexible bronchoscope (a thin instrument, having the width of a pencil, with 
several channels that allow to see inside the lung and inject and/or remove fluid) 
will be inserted through your nose or mouth, and then past your vocal cords into 
the bronchus (large airpipe) of only a lobe (a segment) of your lung. A local 
anesthetic will be given as the bronchoscope passes through your airway. 
o Lavage will be done with sterile sodium chloride (salty water for injection). While 
the bronchoscopist ensures that the tip of the bronchoscope is in the selected lung 
lobe, an assistant will inject into the lung lobe a measured volume, about 50 mL 
(about 1.7 ounces or 3 tablespoons), of sterile sodium chloride with a syringe 
through one channel of the bronchoscope and then aspirate it (remove it) with 
gentle suction with the same syringe. The same lavage will be repeated 3 more 
times. 
o The procedure ends when the bronchoscopist removes the bronchoscope from your 
lung and airways. 
• 
You will then be transferred to the recovery room. 
o Your vital signs, ECG and oxygen will be monitored continuously. 
o You may receive oxygen through a nasal cannula as needed 
o You will receive medication as needed. 
o Study staff may perform additional assessments according to the study plan, such 
as take blood for analysis of the study drug or record an ECG, while you are in the 
bronchoscopy suite or recovery room. 
o You will be given fluids to drink as you tolerate. 
o You will be discharged from the recovery room when you are stable and return to 
the Research Site accompanied by study staff. 
• 
Your monitoring will continue in the Research Site according to instructions by the 
pulmonologist (lung doctor) who did the bronchoscopy. 
o Your vital signs and oxygen will continue to be monitored frequently. 
o You will receive oxygen and medication as needed. 
o You will continue drinking water (about 2 liters) for 24 hours after dosing.  
Document time and volume of water consumed. 
o You will eat solid food when you are able to swallow well approximately 4 hours 
after the procedure. 
 
DAY 2: Inpatient Follow-up 
If you were assigned to study treatment group T5 (24 hours): 
• You will have bronchoscopy with BAL at 24 after dosing and be monitored as described 
above for subjects in Groups 1-4 on Day 1. 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 14 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
For all subjects: 
• 12-lead standard ECG with 10-second rhythm strip taken at 24 hours and 36 hours after 
dosing. 
• Vital signs measured at 24 hours and 36 hours. 
• Blood drawn to measure the concentration of study drug at 24 hours and 36 hours.  
• Blood and urine samples collected for clinical laboratory tests. 
• Infusion site examined at 24 hours and 36 hours. 
• Report any side effects that you may experience. 
• Physical exam may be done as needed depending on your symptoms. 
• Document medications taken. 
• Drink at least 2 L water daily – Document time and volume of water consumed. 
• Counsel on avoidance of vigorous exercise and exposure to loud noises. 
 
DAY 3: Inpatient Follow-up 
• 12-lead standard ECG with 10-second rhythm strip at 48 hours and 60 hours after dosing. 
• Vital signs measured at 48 hours and 60 hours. 
• Blood drawn to measure the concentration of study drug at 48 hours and 60 hours. 
• Blood and urine samples collected for clinical laboratory tests. 
• Infusion site examined at 48 hours and 60 hours after dosing.  
• Report any side effects that you may experience.  
• Document medications taken. 
• Have a physical examination. 
• Repeat ear exam and hearing tests.  
• Ear otoscopy. 
• Audiometry. 
• Distortion product otoacoustic emissions (DPOAE). 
• You will be discharged from the Research Site after review of clinical laboratory tests, 
ECGs, and other assessments by PI or authorized clinician and audiologist.  
• Note: If the time of discharge is late in the evening, due to delays in starting procedures on 
Day 1 or personal reasons (such as distance from residence), you may stay in the Research 
Site overnight and be discharged the following morning. 
• Counsel on the avoidance of pregnancy, use of contraception, avoidance of prohibited 
medication, alcohol, coffee, or caffeinated drinks (not more than 3 cups daily), and 
recreational drugs. 
• If you are male, you will be counseled to avoid sperm donation for 30 days after dosing. 
• Counsel on avoidance of vigorous exercise and exposure to loud noises. 
• Instruct on the next scheduled visit. 
 
OUTPATIENT FOLLOW-UP PERIOD (Days 4 to 30) 
• 
You will be provided contact information and instructions to report to the Research Site 
any side effects or new symptoms that you develop during these periods and any 
medication that you take. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 15 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• 
You will be given appointments to return to the Research Site to have follow-up tests on 
Day 14 and Day 30 after dosing: 
 
DAY 14 Follow-up  
• Obtain 12-lead standard ECG with 10-second rhythm strip. 
• Vital Signs measured. 
• Weight taken. 
• Blood and urine samples collected for clinical laboratory tests. 
• Repeat ear exam and hearing tests. 
• Ear microscopy. 
• Audiometry. 
o Distortion product otoacoustic emissions (DPOAE). 
• Infusion site checked.  
• Report any side effects that you may experience.  
• Document medications taken. 
• Have a physical examination. 
• Counsel on the avoidance of pregnancy, use of contraception, avoidance of prohibited 
medication, alcohol, coffee, or caffeinated drinks (not more than 3 cups daily), and 
recreational drugs. 
• If you are male, you will be counseled to avoid sperm donation for 30 days after dosing.  
• Counsel on avoidance of vigorous exercise and exposure to loud noises. 
• Instruct on the next scheduled visit.  
 
DAY 30– FINAL VISIT (Discharge Visit) 
• Report any side effects that you may experience. 
• Physical exam may be done as needed depending on your symptoms. 
• Document medications taken. 
• Blood and urine samples collected for clinical laboratory tests. 
• Repeat ear exam and hearing tests. 
o Ear microscopy. 
o Audiometry. 
o Distortion product otoacoustic emissions (DPOAE). 
• You will be told if your study participation has completed. 
• If you have any side effects that need further follow up, you will be told when to return to 
the Research Site. 
 
EARLY TERMINATION 
If you withdraw or are terminated by the study doctor before you complete the study, you will 
have the following tests and procedures:  
• Vital signs measured. 
• Have a physical examination. 
• Check infusion site. 
• Weight taken. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
• Report any side effects that you may experience.  
• Document medications taken. 
• Obtain 12-lead standard ECG with 10-second rhythm strip. 
• Blood and urine samples collected for clinical laboratory tests. 
• Obtain blood for measuring the concentration of the study drug (if your early termination 
occurs within 24 hour of dosing). 
• Have ear exam and hearing tests. 
• You will be counseled to avoid pregnancy and use contraception for 30 days after dosing. 
• If you are male, you will be counseled to avoid sperm donation for 30 days after dosing.  
 
UNSCHEDULED VISIT (if needed) 
You may return to the Research Site for an unscheduled visit at any time after your discharge on 
Day 3 if you have any symptoms that you want to discuss or because the study staff asked you to 
come in for an evaluation. The following activities at a minimum will be performed:  
• Collect information on the side-effects you may experience. 
• Document medication taken. 
• Vital signs measured. 
• Have a physical examination, as needed. 
• Have a 12-lead ECG as needed.  
• Obtain blood and/or urine for clinical laboratory tests, if applicable. 
• Have ear exam and hearing tests, as needed. 
• The study staff will give you information and tell you if you need to return for follow up 
before you next scheduled visit. 
 
BLOOD SAMPLES 
Blood samples will be drawn by single needle-sticks or through an IV catheter, as determined by 
the study staff. An IV catheter is a small plastic tube inserted into your arm vein by a needle. The 
needle is removed, but the tube temporarily remains in your vein. This tube allows the removal of 
blood without having to stick you each time. The total amount of blood drawn will be up to about 
188.5 mL (about -6.5 oz) during the in-patient study treatment period (Days 1 to Day 3) and 262.1 
mL (about 8.7 oz or less than a cup of water) during the entire study (from screening to the final 
visit on Day 30). For comparison, a standard blood donation at a blood collection center, once in a 
56-day period, is about 500 mL (2 cups) of blood. 
 
Additional blood samples may be drawn during the study, if the study staff considers it necessary 
for monitoring your health. 
 
Blood samples and collected data will only be used for this study. Blood samples left after all 
routine clinical laboratory testing is done will be discarded by the clinical safety laboratory. 
 
YOUR ROLE IN THIS STUDY: 
Taking part in a research study can be an inconvenience to your daily life. Please consider the 
study time commitments and responsibilities as a research subject when you are deciding to 
participate. Your responsibilities as a subject include the following: 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 17 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• You cannot participate in this study if you have participated in a prior study within the 
past 30 days. You must agree that under no circumstance will you take another 
investigational drug during this study, because it may cause serious physical harm or death 
and result in permanent restriction for any future studies. 
• You must provide accurate responses to questions about your past medical and surgical 
history (illnesses and surgeries you had including abnormal lab tests, X-rays, and other 
procedures), medications you are taking, medications you are allergic to and your family 
history. Failure to respond accurately may result in side-effects that may be harmful after 
you receive the study drug or have during and after bronchoscopy with alveolar lavage 
(BAL). 
• You must read, and agree to follow, the House Rules at the Research Site if you want to 
take part in this study. House Rules discuss proper behavior at the Research Site and are 
important to ensure your safety and make sure the study results are accurate. Failure to 
follow these rules may result in reduced compensation or involuntary discharge from this 
study. 
• You must report any side effects and medical problems to the study staff. 
• You must give true and complete answers to any questions. 
• You must be able to comply with the study procedures and visit schedule. 
• You must follow all instructions from the study staff. 
• You must inform the study staff if you decide to discontinue your participation in the 
study. If you decide to discontinue your participation in the study, you may be asked to 
consent to stay in the study and complete any remaining procedures for the most recent 
study treatment period. If you do not consent to complete any remaining procedures, you 
will be asked to complete the early termination visit procedures as described in this 
consent form. 
• You must agree to use an effective method of contraception, as defined later in this 
consent form, throughout the study. If you are male, you must agree not to donate sperm 
for 30 days after dosing. 
• You must not do any vigorous activity for 2 days before the dosing until discharge from 
the Research Site on Day 3, and for 24 hours before each out-patient follow up visit on 
Day 14 and Day 30. Or at any time during the study. 
• You must avoid exposure to loud noises (for example, construction sites, visiting concerts 
or dance events, fireworks, listening to music with headphones) from screening until Day 
30 (final visit). 
• You must not consume alcohol for 2 days before check-in on Day -2 until your discharge 
from the Research Site on Day 3, and it is recommended to avoid alcohol during the out-
patient follow-up until Day 30 (final visit). Tell the study doctor if you consumed any 
alcoholic beverages during this time. 
• You must not use marijuana or other illegal drugs during the study.  
• You must not drink liquids or eat foods that contain caffeine within 2 days before dosing 
and during the inpatient study treatment period, and no more than 3 cups of coffee or 
caffeine-containing drinks during the out-patient follow-up period to the end of the study. 
Tell the study doctor if you consumed any coffee or caffeine-containing beverages during 
this time. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Page 18 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• You will be advised to avoid smoking after screening. Smoking is prohibited during the 
in-patient period of the study (Day -2 to Day 3) and recommended to avoid smoking 
during the follow-up out-patient period to the end of the study (Day 4 to Day 30). Tell the 
study doctor if you consumed any coffee or caffeine-containing beverages during this time. 
• You must not participate in another research study at any time during this study or 
continue participating in another study and have blood draws. This includes studies of a 
drug, biologic (such as vaccine or proteins), device, or blood product. 
  
PROHIBITED MEDICATIONS, VACCINATIONS, and BLOOD DONATIONS:  
Additional restrictions to ensure the health and safety of study subjects are listed below:  
• 
You must not take any prescription medications within 30 days before dosing or during 
the study. Certain drugs should be avoided for a longer period before taking the study 
drug. (See Risks of Apramycin below)  
• 
You will continue taking birth control medications that you were already using. Certain 
other medications may be permitted if approved by the study staff. Please report all 
prescription medications you are taking. 
• 
You must not take any non-prescription medications, herbs, vitamins, or nutritional 
supplements from the time of screening (about 28 days before dosing) to 14 days after 
dosing, with the exception of single doses of acetaminophen up to 1,000 mg. Certain non-
prescription medications are prohibited if taken 15 days before dosing, and others may be 
permitted if approved by the study staff. Please report all non-prescription medications, 
herbs, vitamins, or nutritional supplements you are taking. 
• 
You must not donate any blood or blood products (red cells, white cells, platelets, plasma) 
during the course of this study. Tell the study doctor if you donated any blood or blood 
products within 3 months before dosing. 
• 
You must not receive any blood or blood products during this study. 
  
RISKS of APRAMYCIN: 
Observed risks: 
There were no identified risks associated with apramycin in the completed first in human study in 
30 healthy subjects who were given IV single apramycin doses ranging from 0.3 to 30 mg/kg 
body weight. One subject had decreased hearing at one test after dosing but not on a repeat and 
the effect was not considered related to the study drug. Apramycin was safe and well-tolerated 
and there were no clinically important changes in laboratory tests and ECGs during that study. 
 
Potential Risks:  
Until more clinical experience with apramycin is available, similar undesirable effects as 
observed for other aminoglycosides should be anticipated. Special attention will be paid to signs 
and symptoms of kidney injury, hearing loss and balance disorders. 
 
Based on the knowledge on other aminoglycosides, the following contraindications to the use of 
apramycin will be assumed until further testing, because of potentially higher risk if you have a 
history of the following: 
• 
Hypersensitivity or an allergic reaction to aminoglycosides. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Page 19 of 30 
  
  
 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
• 
Myasthenia gravis (a chronic autoimmune, neuromuscular disease that causes weakness in 
the skeletal muscles that worsens after periods of activity and improves after periods of 
rest). 
• 
Kidney failure. 
• 
Hearing loss. 
• 
Balance problems, especially those caused by a disorder of the inner ear (for example, 
Meniere’s disease), vertigo (a sensation of dizziness and loss of balance, with a feeling 
like you are spinning or that the world around you is spinning), ataxia (impaired 
coordination of movement). 
 
Potential Drug Interactions: 
There are no clinical data on drug interactions with apramycin. However, based on the knowledge 
about other aminoglycosides, concomitant administration of the following drugs should be 
avoided for the indicated period before dosing to the end of the study: 
• 
Drugs affecting kidney function for 2 weeks before dosing with apramycin: blood pressure 
and medications for heat failure (such as enalapril, lisinopril, ramipril captopril, 
benazepril), spironolactone, and epleronone); and anti-inflammatory drugs (such as 
aspirin, ibuprofen, naproxen, indomethacin, bextra, celebrex, vioxx). 
• 
Drugs with potential ear toxicity and permanent hearing loss for 3 months before dosing 
with apramycin: vancomycin, loop diuretics (bumetanide [bumex], ethacrynic acid 
(edecrin), furosemide [Lasix]), quinine and quinidine derivatives including mefloquine. 
• 
Neuromuscular blocking agents (drugs that block the transmission of nerve signals to the 
muscles and cause muscle paralysis) within a week before dosing: tracrium, norcuron, 
pavulon, and others. 
• 
Other aminoglycoside antibiotics within 3 months before dosing with study drug: 
gentamicin, tobramycin, kanamycin, streptomycin, neomycin. 
 
Allergic Reaction Risks 
As with taking any drug, there is a risk of allergic reaction.  If you have a very serious allergic 
reaction, you may be at risk of death.  Some symptoms of allergic reactions are: 
• Rash 
• Wheezing and difficulty breathing 
• Dizziness and fainting 
• Swelling around the mouth, throat or eyes 
• A fast pulse 
• Sweating 
 
Please seek treatment immediately and tell the study doctor and study staff if you have any of 
these symptoms.  
 
Use During Pregnancy and Lactation: 
Aminoglycosides pass the placenta barrier and may cause fetal harm. Therefore, you should not 
participate in the study if you are pregnant or plan to become pregnant during the study. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 20 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
Aminoglycosides have been reported to be detected in maternal milk. Therefore, you should not 
participate in the study if you are breastfeeding. 
 
Overdose 
There is no specific antidote in case of apramycin overdose. The same measures as recommended 
for other aminoglycosides (treatment cessation, adequate hydration, dialysis) will be considered 
until clinical data are available for apramycin. 
 
Drug Abuse and Dependency 
No clinical experience available. 
 
RISKS OF BRONCHOSCOPY WITH BRONCHOALVEOLAR LAVAGE: 
Bronchoscopy with bronchoalveolar lavage has a proven safety record in both clinical 
applications as well as human research during more than 40 years of clinical use of this procedure 
in hundreds of thousands of subjects, without any serious lung troubles and side effects. However, 
like all procedures, there are some risks associate with it. 
 
The most common side effect related to the procedure is transient hypoxemia (drop in oxygen 
content in the blood), which may require administration of oxygen. Transient tachycardia 
(increased heart rate) and premature contractions, which may require no treatment or appropriate 
intervention. 
 
Other side effects that could occur are nausea and dizziness, due to the drugs used for sedation or 
anesthesia, and nasal stuffiness and irritation, throat irritation, hoarseness and cough, due to the 
passage of the bronchoscope and the bronchoalveolar lavage (BAL; instilling saline into a lung 
segment (lobe). 
 
A post-bronchoscopy fever (PBF), usually less than 40°C / 100°F may occur. The incidence varies 
from 0% in large series to up to 10% reports in smaller series. It may occur about 8 hours after 
BAL and lasts about 14 hours. It is not accompanied by bacterial infection or pneumonia with 
abnormal chest X-ray, and it could be a rare natural reaction to the procedure. Antibiotics are not 
used for prevention and treatment targets the symptoms (drugs to lower fever, good hydration). 
Additional tests may be needed to guide treatment. 
 
Rare complications, with incidence 0.1 to 0.3%, are perforation of the bronchus with 
pneumothorax (collection of air in the chest cavity), and atelectasis (collapse of the segment of 
the lung where the bronchoscope is inserted), and infection. Bleeding has been described for 
bronchoscopy with biopsy of the lung but is very rare (0.1% or less) if only bronchoalveolar 
lavage is done. These are related to mechanical trauma, especially in subjects who have a disorder 
of blood coagulation or take anticoagulants (heparin, warfarin, new anticoagulants) and non-
prescription non-steroidal anti-inflammatory drug (such as aspirin, and ibuprofen). 
 
These risks are mitigated by adherence to the study plan. As mentioned above, you are expected 
to provide accurate and truthful information about past and current illnesses and use of 
medications. You are going to have a thorough physical examination and clinical lab testing to 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
exclude any abnormalities that are contraindications to receiving the study drug and having the 
procedure. Finally, the procedure is performed by experienced pulmonary physicians (lung 
specialists) that are certified to perform this procedure, assisted by experienced and certified 
personnel in specially equipped hospital rooms equipped to handle emergencies. 
 
RISKS OF AUDIOLOGY TESTS: 
There are no risks associated with the planned procedures to examine your ears and hearing. The 
procedure will be performed by experienced and certified audiologists. 
 
BLOOD SAMPLE RISKS:  
There may be side effects of drawing blood with a needle and/or IV tubes and IV catheter 
placement. Local pain, bruising, bleeding, inflammation or infection might occur at the site of the 
needle stick where blood is drawn. There is a possibility of dizziness or fainting while your blood 
is being drawn. Precautions will be taken to minimize the above complications. There is a small 
risk your blood counts will decrease slightly during the course of the study, but not expected to 
cause anemia (low blood counts that cause symptoms and need treatment). Any symptoms that 
may occur are usually reversible with good nutrition, drinking fluids, vitamins, and iron. 
 
ECG RISKS: 
ECG patched may case a skin reaction such as redness or itching. You may also experience 
localized skin discomforts and/or hair loss associated with the placement of ECG patches. 
 
UNKNOWN RISKS:  
You may have side effects or discomforts that are not listed in this form. Some side effects may 
not be known yet. New ones could happen to you. Tell the study staff right away if you have any 
problems. 
  
INFORMATION ON BIRTH CONTROL:  
  
Females: 
The effect of apramycin on the unborn child is unknown at this time. Therefore, it is important 
that women who are pregnant, may become pregnant or who are breastfeeding NOT participate in 
this study. If you are a woman who can get pregnant, you must use an acceptable form of birth 
control that is very effective in preventing pregnancy. You must agree to use birth control for at 
least 30 days after dosing. You agree to avoid becoming pregnant during this study.  
  
The following are acceptable forms of birth control for this study: 
• 
Progestin implants. 
• 
Intrauterine devices (IUD). 
• 
Surgery to prevent pregnancy in females (such as bilateral tubal ligation/occlusion, 
salpingectomy [removal of the tubes], oophorectomy [removal of the ovaries], or 
hysterectomy [removal of the uterus]). 
• 
Abstinence. 
 
Tell the study staff if you use any other forms of birth control. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Page 22 of 30 
  
  
 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
  
If at any time during this study you think you may be pregnant, you are required to immediately 
contact the study doctor at the telephone number on the first page of this form. 
 
Males:  
The effect of apramycin on the unborn child is unknown at this time. The effect of the study drug 
on the male reproductive system is unknown. Therefore, you must agree to use an acceptable 
form of birth control from dosing with study drug through at least 30 days after dosing. 
Acceptable birth control means not having sex (abstinence) or having a vasectomy. If you are 
sexually active, you should agree to use a condom throughout this study and for at least 30 days 
after dosing. Tell the study staff if you use any other form of birth control. 
  
You should also not donate sperm for 30 days after dosing. 
 
It is also recommended that you make certain that your female partner is also using an acceptable 
form of birth control if you have not had a vasectomy and are using a condom. If you think that 
your partner may be pregnant at any time during this study to 30 days after dosing, you are 
required to immediately contact the study doctor at the telephone number on the first page of this 
form. 
 
REPORTING PREGNANCY:  
If you are a woman, you should report to the study staff if you miss your period and suspect you 
are pregnant or if you become pregnant during the 30 days after dosing. With your permission, 
blood samples will be collected, and you will continue to be followed for safety during this study. 
You should provide pregnancy outcome to the study staff upon delivery or pregnancy 
termination. 
  
If you are a male and your female partner becomes pregnant during the 30 days after dosing, you 
should inform study staff of the pregnancy and provide pregnancy outcome upon delivery or 
pregnancy termination. 
  
INFORMATION ON USE OF ALCOHOL, MARIJUANA OR OTHER ILLEGAL 
DRUGS: 
You are not allowed to use alcohol, marijuana or other illegal drugs (such as cocaine, etc.), 
because they may cause you serious harm and are strictly forbidden while you are in this study. 
You will be tested for drugs and alcohol at Screening, and on Day -2. If you are found to be in 
possession of or using marijuana or other illegal drugs at the Research Site, then AMR-Knoxville 
is obligated to report the use of marijuana or other illegal drugs to the appropriate legal 
authorities. 
 
INFORMATION ON ACTIVITIES TO AVOID: 
You should avoid vigorous exercise from at least 2 days before your admission to the Research 
Site to after the discharge on Day 3 after dosing and 24 hours before each follow-up visit (Day 14 
and Day 30). 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Page 23 of 30 
  
  
 
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Advarra IRB Approved Version 6 May 2022 
 
 
 
You should avoid exposure to loud sounds (listening to music or playing videogames with 
headphones on, attending indoor concerts and other entertainment venues, etc.) from signing and 
dating informed consent until Final Visit (Day 30). 
 
NEW INFORMATION ABOUT APRAMYCIN: 
The study doctor or study staff will discuss with you any important new information, findings or 
changes to the way the research will be performed that may affect your decision to remain in this 
study. If this happens, you will be asked to decide if you want to keep taking part in this study. 
You may be given an additional informed consent and will be asked to agree in writing that you 
were told about these new findings. 
  
BENEFITS TO THE STUDY SUBJECT: 
The goal of this research study is to provide scientific information. You will not receive any 
medical benefits. This study may help doctors and scientists learn things about the study drug that 
will help others. 
  
ALTERNATE THERAPY:  
This is a clinical research study. It is not treatment or therapy. The other choice is to not 
participate in this study. 
  
PAYMENT FOR PARTICIPATION:  
If, after screening, you are enrolled in the study and complete all of the study visits, you will 
receive a total of up to $4,050.00 at your completion of the study to help cover the costs of your 
participation and compensate you for your time. If, after screening, it is determined that you 
qualify to be enrolled in the study, but you choose not to enroll, you will not be compensated for 
completing the screening process.  Compensation will be made at the following intervals: 
- 
Completion of Day 3 at:       
 
$2,000.00           
- 
Completion of Day 30/End of Study at:         $2,050.00 
 
If after enrollment, your participation is ended early, or if you withdraw for any reason, the 
amount you will be paid will be prorated and the payment made when you complete your 
participation in the study. This means that you will receive payment only for the portion of the 
study that you completed and only after your participation in the study has been completed. 
Payment will be prorated as follows: 
 
Visit 1 
Screening  
$400.00  
Visit 2 
Check-in, Day -2 
$425.00  
 
Baseline, Day-1 Baseline 
$425.00  
 
Study Treatment Period, Day 1, Dosing 
$1,500.00  
 
Study Treatment Period, Day 2, In-patient Follow up 
$425.00  
 
Study Treatment Period, Day 3, In-patient Follow-up / 
Discharge 
$425.00  
Visit 3  
Outpatient Follow Up Period, Day 14 
$150.00  


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
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Page 24 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
Visit 4 
Outpatient Follow Up Period, Day 30 (Final Visit) 
$300.00  
  
Total 
$4,050.00  
 
Compensation for participation in the study will be made using a reusable, pre-loaded card 
(similar to a debit card). You must not lose the card during your participation in the study. If the 
card is lost, stolen, or destroyed, you will be charged a $5.00 fee to issue a replacement card. 
 
If you test positive for illegal drugs at any time during the study, you will be withdrawn from the 
study and not be given any amount of compensation. 
 
If you have any questions regarding your compensation for participation, please contact the study 
doctor at the telephone number listed on page one of this consent document. 
 
If, after enrollment, your participation is terminated or you decide to withdraw for any reason, 
you will be compensated $150.00 for completing the early termination/withdrawal visit 
procedures. If you have an unscheduled visit, you will be compensated $100.00 for completing 
the unscheduled visit procedures. 
 
In addition, you will be advised of, and asked to sign and date a document entitled “Research Site 
Guidelines”. These rules outline penalties for certain violations of our rules for when you are 
staying at the clinic. Violations, should they occur, may result in reduction of the compensation 
described above. 
 
If the amount of compensation from your participation is over a certain amount, the law requires 
Alliance for Multispecialty Research, LLC to report the compensation you receive to the Internal 
Revenue Service. No deductions from your compensation for taxes will be made by Alliance for 
Multispecialty Research, LLC. It is your responsibility to report this compensation on your 
federal tax returns and to pay any taxes that are due. 
 
COMPENSATION FOR INJURY:  
• 
In the event that you experience an adverse reaction (side-effect), illness or injury during 
this study, you should immediately seek treatment. It is important that you tell your study 
doctor if you have experienced one of these events. You must contact the study staff at the 
telephone number on the first page of this form as soon as you are able. You may obtain 
medical care in the same way as you would ordinarily receive any other medical 
treatment. Immediate necessary care, emergency treatment, and professional services will 
be available to research subjects just as they are to the community generally. In general, 
no long-term medical care or financial compensation for research-related injuries will be 
provided by the NIH or the Federal Government. 
• 
Medical care will be given to you, at no cost, if you are injured because of being in this 
study. 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 25 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• 
While you are in this study, you may be injured because of your personal conduct or 
during activities, which are not part of this study. There are no plans to pay you for these 
types of injuries. 
 
COSTS TO YOU:  
There is no cost to you to participate in this study. You do not have to pay for any study-related 
drugs, procedures, or treatment at the Research Site during this study. 
  
SOURCE OF FUNDING: 
This research study is being funded by NIAID, which has a contract with a company named DVC 
to do studies like this one. AMR-Knoxville has signed and dated a contract with IQVIA, Inc., a 
subcontractor of DVC, to conduct this research study under the direction of the study doctor. 
  
 
 
VOLUNTARY PARTICIPATION IN THIS STUDY:  
Your participation in this study is voluntary. You will be starting this study of your own free will 
and without any kind of pressure. You may quit this study any time you wish. If you withdraw 
from this study, you will not be penalized. You will not lose any benefits to which you were 
otherwise entitled.  
 
If you have health problems, you will get medical attention. You may be withdrawn from this 
study by the study staff and/or the sponsor of this study, without your consent, if it is in your best 
medical interest. It is possible that the study staff and/or sponsor may think you can stay in this 
study. You must decide what you want to do.  
   
The study staff or the sponsor may stop your participation in this study at any time without your 
consent for any of the following reasons:  
• 
If you don’t follow the study staff’s instructions;  
• 
Something serious happens to you which may require treatment;  
• 
The study staff decides it is in the best interest of your health, and welfare to discontinue 
further participation;  
• 
The sponsor stops this study for other reasons not known now;  
• 
You do not later consent to any future changes that may be made in the study plan; or  
• 
For any other reason.  
  
If you received any amount of the study drug but withdraw from the trial within 24 hours after 
dosing, you will be encouraged to continue follow-up (with your consent) for safety assessments 
and PK sample collection. 
 
The FDA, the Safety Monitoring Committee (SMC), and Advarra IRB may also terminate this 
study if there are safety concerns.  
  
If you do not complete this study for any reason, you will be asked to undergo a physical exam, 
vital signs measurement, 12-lead ECG, blood and urine laboratory tests, a pregnancy test (females 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 26 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
only), ear and hearing tests and tests for the measurement of apramycin in your blood if early 
termination occurs within 24 hours of dosing. This is needed to identify any changes that may 
have occurred after you began to take the study drug and to protect your safety, health, and 
welfare. 
 
CERTIFICATE OF CONFIDENTIALITY: 
To help us protect your privacy, we have a Certificate of Confidentiality from NIH. The study 
staff can use this Certificate to legally refuse to give information that may identify you in any 
federal, state, or local civil, criminal, administrative, legislative, or other proceedings, for 
example, if there is a court subpoena. The study staff will use the Certificate to resist any 
demands for information that would identify you, except for reporting of communicable diseases, 
such as Hepatitis Virus and HIV infections, to state and local health departments, or for reporting 
possession or use of marijuana or other illegal drugs while at the Research Site to the appropriate 
legal authorities. 
  
A Certificate of Confidentiality does not prevent disclosure of your information to the NIH, FDA, 
or federal funding agency. Information from this study will be given to DMID and Juvabis AG, 
the pharmaceutical organization developing apramycin and other drugs for the treatment of 
infectious diseases. Any persons or companies, which are contracted by the sponsor for 
conducting this study, measuring drug level in blood samples, monitoring the execution of this 
study, and analyzing the data, will have access to the research information during and after this 
study. 
  
The information will also be given to the FDA. It may be given to governmental agencies in other 
countries where the study drug may be considered for approval. 
  
Authorization to Use and Disclose Protected Health Information 
 
If you decide to be in this study, the study doctor and study staff will use and share health data 
about you to conduct the study. Health data may include: 
• Your name. 
• Address. 
• Phone number. 
• Date of birth. 
• Medical history. 
• Information from your study visits, including all test results.  
 
Health data may come from your study records or from existing records kept by your doctor or 
other health care workers. 
 
Medical records which identify you and the consent form signed and dated by you will be looked 
at and/or copied for research or regulatory purposes by: 
• 
The sponsor; 
• 
An agent for the sponsor; 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 27 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
• 
An agent for the study doctor; 
• 
The FDA; 
• 
Department of Health and Human Services (DHHS) agencies; and 
• 
The Institutional Review Board, Advarra IRB. 
  
Your health data will be used to conduct and oversee the research, including for instance: 
• To see if the study drug works and is safe. 
• For other research activities related to the study drug 
 
Absolute confidentiality cannot be guaranteed because of the need to give information to these 
parties. The results of this research study may be presented at meetings or in publications. Your 
identity will not be disclosed in those presentations. 
• 
AMR-Knoxville will handle the medical information obtained in this study with the 
strictest confidence. It will be protected as required by laws and/or regulations. It will not 
be made publicly available. 
• 
If the results of this study are published, you will not be identified by name. 
• 
Your identity will not be disclosed to anyone else, unless required by law. 
  
By signing and dating this consent, you authorize AMR-Knoxville to verify your study 
participation history with other businesses that conduct clinical research studies. 
  
A Certificate of Confidentiality does not prevent you or a member of your family from voluntarily 
releasing information about yourself or your involvement in this research. If you give your 
consent to release information to a medical care provider, an insurer, or other person to receive 
research information, then the researchers will not withhold that information. 
 
Your permission to use and share health data about you will end in 50 years unless you revoke it 
(take it back) sooner.  
 
You may revoke (take back) your permission to use and share health data about you at any time 
by writing to the study doctor at the address listed on the first page of this form. If you do this, 
you will not be able to stay in this study. No new health data that identifies you will be gathered 
after your written request is received. However, health data about you that has already been 
gathered may still be used and given to others as described in this form.  
 
Your right to access your health data in the study records will be suspended during the study to 
keep from changing the study results. When the study is over, you can access your study health 
data. 
 
If you decide not to sign and date this form, you will not be able to take part in the study. 
 
PRIMARY CARE PROVIDER NOTIFICATION  
We would like your permission to contact the primary care provider you see regularly to let them 
know that you are taking part in this study. It is important for all your healthcare providers to 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 28 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
know that you may be taking an experimental drug. Your healthcare provider will want to know 
and think about all the drugs you are taking before giving you any new ones. While you are in the 
study, the study doctor will ask about your symptoms. If you have symptoms after the study ends, 
your primary care provider may want to contact the study personnel. 
  
If you agree, your primary care provider (regular provider) will be informed of your participation 
in the study.  Indicate your choice below by initialing only one (1) section below. 
 
 
Initials 
I do not have a primary care provider. 
 
Initials 
I do not want my primary care provider notified. 
 
Initials 
I would like my primary care provider notified and below is my physician’s 
contact information. 
 
 
  
 
 
FUTURE USE OF RESIDUAL PLASMA AND BRONCHOALVEOLAR LAVAGE (BAL) 
SPECIMENS 
 
We would like to ask your permission to store any residual (left-over) blood (plasma) and 
bronchoalveolar lavage fluid collected for measuring the concentration of apramycin for use in 
future studies. You can choose whether to allow residual collections of blood (plasma) and BAL 
fluid to be stored for future use or destroyed at the end of your participation in the study. 
 
These samples will be stored in test tubes labeled with a code number without information which 
could identify you. It does not identify you by name or provide any other personal information 
that can lead to your identification by someone who does not have access to the code (see Section 
on Confidentiality of Records). The code number on the test tube label is used to track the day 
and time of plasma or BAL fluid collection and your assigned study number. All stored samples 
will be used only for related research purposes. The samples will not be sold or used directly for 
production of any commercial product.  Human genetic (DNA) tests will not be performed on 
these blood samples. Blood and BAL fluid samples obtained in this study may result in the 
development of a product that could be patented or licensed. There are no plans to provide 
financial compensation to you should this occur.  
 
Your decision may be changed at any time by notifying the study doctor at the telephone number 
on the first page of this form. You will be asked to sign and date a new form to demonstrate your 
new decision. Your decision regarding these specimens will not affect your participation in this 
study or other studies. 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 29 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
If you agree/disagree to your blood and BAL fluid specimens being stored for future studies, 
please initial and date your choice below. 
 
 
 
(initial)                (date) 
YES, you may store my unused blood (plasma) and BAL fluid 
samples in coded tubes samples for an indefinite period of time for 
future research without removing the link to my personal 
identifying information. 
 
 
(initial)                (date) 
YES, you may store my unused blood (plasma) and BAL fluid 
samples in coded tubes samples for an indefinite period of time 
only after removing the link to my personal identifying 
information. 
 
 
(initial)                (date) 
NO, you may not store my unused blood (plasma) or BAL future 
samples for future research.  Destroy all unused samples at the end 
of the study.  
 
 
WHOM TO CONTACT ABOUT THIS STUDY 
During the study, if you experience any medical problems, suffer a research-related injury, or 
have questions, concerns or complaints about the study, please contact the study doctor at the 
telephone number listed on the first page of this consent document. If you seek emergency care, 
or hospitalization is required, alert the treating physician that you are participating in this research 
study.  
 
An institutional review board (IRB) is an independent committee established to help protect the 
rights of research subjects. If you have any questions about your rights as a research subject, 
and/or concerns or complaints regarding this research study, contact: 
 
• By mail: 
Study Subject Adviser 
Advarra IRB 
6100 Merriweather Dr., Suite 600 
Columbia, MD 21044 
• or call toll free:    877-992-4724 
• or by email:          adviser@advarra.com 
 
Please reference the following number when contacting the Study Subject Adviser: Pro00059918. 
 
 


Division of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID), National Institutes of Health (NIH)) / Protocol Number 20-0012 
ICF version 3.0  – 02 May 2022 
Page 30 of 30 
  
  
 
William Smith, M.D. 
Advarra IRB Approved Version 6 May 2022 
 
 
 
CONSENT and AUTHORIZATION 
 
I have read all information in this document, and all my questions have been answered to my 
satisfaction. I voluntarily consent to participate in this study. I will be given a signed and dated 
copy of this consent form and have been told the description of the study drug, and the name, 
address and phone number of the study doctor. My signature and date give my consent to 
participate. I authorize the release of my medical records and health information related to this 
study, including my signed and dated consent form and any addendum, to the sponsor and its 
representatives, the FDA, Advarra IRB and other regulatory agencies as described above. By 
signing and dating this form, I have not given up any of my legal rights as a research subject. 
 
 
 
Printed Volunteer’s Name 
 
Date 
Volunteer’s Signature 
 
 
Person Obtaining Informed Consent Printed Name 
 
 
Person Obtaining Informed Consent Signature  
 
Date 
Witness Printed Name 
 
 
Witness Signature 
 
Date 
Investigator’s Signature* 
 
Date 
*The Investigator’s signature evidences only that the Investigator has reviewed the consent form 
and the signatures and dates of the subject, person obtaining consent, and witness, for compliance.  
 

SAP_002.pdf: 
CLINICAL RESEARCH IN INFECTIOUS DISEASES 
 
 
 
STATISTICAL ANALYSIS PLAN 
for 
DMID Protocol: 20-0012 
 
Study Title: 
 
A Phase I, Open-Label Study to Assess Lung 
Pharmacokinetics and Safety of a Single Dose of 
Apramycin Administered Intravenously in Healthy 
Adult Subjects 
 
NCT05590728 
 
Version 1.0 
 
7 February 2023 
RESTRICTED 
 


Statistical Analysis Plan - DMID Protocol: 20-0012  
Version 1.0 
7 February 2023 
- 2 - 
RESTRICTED 
STUDY TITLE 
Protocol Number Code: 
DMID Protocol: 20-0012 
Development Phase: 
Phase 1 
Products: 
Apramycin 
Form/Route: 
Intravenous (IV) infusion 
Indication Studied: 
Multi-drug resistant bacterial infections 
Sponsor: 
Division of Microbiology and Infectious Diseases (DMID) 
National Institute of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
Clinical Trial Initiation Date: 
TBD 
Clinical Trial Completion Date: 
TBD 
Date of the Analysis Plan: 
7 February 2023 
Version Number: 
v1.0 
This study was performed in compliance with Good Clinical Practice. 
Information contained in this publication is the property of Division of Microbiology and Infectious Diseases and is confidential. This 
information may not be disclosed to third parties without written authorization from Division of Microbiology and Infectious Diseases. This 
report may not be reproduced, stored in a retrieval system or transmitted in any form or by any means - electronic, mechanical, recording or 
otherwise - without the prior authorization from Division of Microbiology and Infectious Diseases. This document must be returned to Division 
of Microbiology and Infectious Diseases upon request. 
 


Statistical Analysis Plan - DMID Protocol: 20-0012  
Version 1.0 
7 February 2023 
- 3 - 
RESTRICTED 
TABLE OF CONTENTS 
STUDY TITLE ................................................................................................................................2 
TABLE OF CONTENTS .................................................................................................................3 
LIST OF ABBREVIATIONS ..........................................................................................................6 
1. 
PREFACE ...................................................................................................................11 
2. 
INTRODUCTION ......................................................................................................12 
2.1. 
Purpose of the Analyses ..............................................................................................12 
3. 
STUDY OBJECTIVES AND ENDPOINTS ..............................................................13 
3.1. 
Study Objectives .........................................................................................................13 
3.2. 
Endpoints ....................................................................................................................13 
3.3. 
Study Definitions and Derived Variables ...................................................................14 
3.3.1. 
Timepoints for Safety and PK Endpoints ...................................................................14 
3.3.2. 
BAL Pharmacokinetics Parameters ............................................................................15 
3.3.2.1. 
Apramycin Concentrations in ELF .............................................................................15 
3.3.2.2. 
Apramycin Concentrations in AM ..............................................................................15 
3.3.3. 
Baseline .......................................................................................................................16 
3.3.4. 
Study Day ...................................................................................................................16 
4. 
INVESTIGATIONAL PLAN .....................................................................................17 
4.1. 
Overall Study Design and Plan ...................................................................................17 
4.2. 
Discussion of Study Design, Including the Choice of Control Groups ......................18 
4.3. 
Selection of Study Population ....................................................................................18 
4.4. 
Treatments ..................................................................................................................21 
4.4.1. 
Treatments Administered ............................................................................................21 
4.4.2. 
Identity of Investigational Product(s) .........................................................................21 
4.4.3. 
Method of Assigning Participants to Treatment Groups ............................................22 
4.4.4. 
Selection of Doses in the Study ..................................................................................22 
4.4.5. 
Selection and Timing of Dose for Each Participant ...................................................22 
4.4.6. 
Blinding ......................................................................................................................23 
4.4.7. 
Prior and Concomitant Therapy ..................................................................................23 
4.4.8. 
Treatment Compliance ................................................................................................24 
4.5. 
Safety and Pharmacokinetic Variables .......................................................................24 
4.5.1. 
Safety Variables ..........................................................................................................24 


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4.5.2. 
Pharmacokinetic Variables .........................................................................................26 
5. 
SAMPLE SIZE CONSIDERATIONS .......................................................................27 
6. 
GENERAL STATISTICAL CONSIDERATIONS ....................................................28 
6.1. 
General Principles .......................................................................................................28 
6.2. 
Timing of Analyses .....................................................................................................28 
6.3. 
Analysis Populations ..................................................................................................28 
6.3.1. 
Safety Population ........................................................................................................29 
6.3.1.1. 
Safety Subset Population ............................................................................................29 
6.3.2. 
PK Populations ...........................................................................................................29 
6.3.2.1. 
Lung PK Population ...................................................................................................29 
6.3.2.2. 
Plasma PK Population ................................................................................................29 
6.3.2.3. 
PK Subset Population .................................................................................................29 
6.4. 
Covariates and Subgroups ..........................................................................................29 
6.5. 
Missing Data ...............................................................................................................29 
6.6. 
Interim Analyses and Data Monitoring ......................................................................29 
6.7. 
Multicenter Studies .....................................................................................................30 
6.8. 
Multiple Comparisons/Multiplicity ............................................................................30 
7. 
STUDY PARTICIPANTS ..........................................................................................31 
7.1. 
Disposition of Participants ..........................................................................................31 
7.2. 
Protocol Deviations ....................................................................................................31 
8. 
EFFICACY EVALUATION ......................................................................................32 
9. 
SAFETY EVALUATION ..........................................................................................33 
9.1. 
Demographic and Other Baseline Characteristics ......................................................33 
9.1.1. 
Prior and Concurrent Medical Conditions ..................................................................33 
9.1.2. 
Prior and Concomitant Medications ...........................................................................33 
9.2. 
Measurements of Treatment Compliance ...................................................................33 
9.3. 
Adverse Events ...........................................................................................................33 
9.3.1. 
Treatment Emergent Adverse Events .........................................................................34 
9.4. 
Deaths, Serious Adverse Events and other Significant Adverse Events ....................34 
9.5. 
Pregnancies .................................................................................................................34 
9.6. 
Clinical Laboratory Evaluations .................................................................................34 


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9.6.1. 
Exploratory Biomarkers for Renal Injury ...................................................................36 
9.7. 
Vital Signs and Physical Evaluations .........................................................................36 
9.8. 
12-Lead Standard Electrocardiogram .........................................................................37 
9.9. 
Otoacoustic Testing ....................................................................................................38 
10. 
PHARMACOKINETICS ...........................................................................................40 
10.1. 
Graphical and Tabular Summaries of Pharmacokinetic Profiles ................................40 
10.2. 
Noncompartmental Analysis ......................................................................................42 
10.2.1. 
Ratio of Lung PK to Plasma PK .................................................................................44 
11. 
IMMUNOGENICITY ................................................................................................45 
12. 
OTHER ANALYSES .................................................................................................46 
13. 
REPORTING CONVENTIONS ................................................................................47 
14. 
TECHNICAL DETAILS ............................................................................................48 
15. 
SUMMARY OF CHANGES IN THE CONDUCT OF THE STUDY OR 
PLANNED ANALYSES ............................................................................................49 
16. 
REFERENCES ...........................................................................................................50 
17. 
LISTING OF TABLES, FIGURES, AND LISTINGS ..............................................51 
APPENDICES ...............................................................................................................................52 
APPENDIX 1. TABLE MOCK-UPS..........................................................................................53 
APPENDIX 2. FIGURE MOCK-UPS ......................................................................................119 
APPENDIX 3. LISTINGS MOCK-UPS ...................................................................................155 
 


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LIST OF ABBREVIATIONS 
AC 
Alveolar Cell 
AE 
Adverse Event 
ALT 
Alanine Aminotransferase 
AM 
Alveolar Macrophage 
AMR 
Alliance for Multispecialty Research 
AP 
Alkaline Phosphatase 
aPTT 
Activated Partial Thromboplastin Time 
AST 
Aspartate Aminotransferase 
ATC 
Anatomical Therapeutic Classification 
AUC 
Area Under the Concentration-Time Curve 
AUC(0-24) 
Area Under the Concentration-Time Curve from Time of Dosing to 24 h 
AUC(0-last) 
Area Under the Concentration-Time Curve from Time of Dosing to Time of 
Last Measurable Concentration 
AUC(0-t) 
Area Under the Concentration-Time Curve from Time of Dosing to Time t 
AUC(0-∞) 
Area Under the Concentration-Time Curve Extrapolated to Infinity 
β-HCG 
Beta Human Chorionic Gonadotropin 
BAL 
Bronchoalveolar Lavage 
BAL AC 
Number of total alveolar cells per mL of BAL volume 
BAL AM 
Bronchoalveolar Lavage Alveolar Macrophage 
BAL ELF 
Bronchoalveolar Lavage Epithelial Lining Fluid 
BAL SUP 
Bronchoalveolar Lavage Supernatant 
BMI 
Body Mass Index 
BP 
Blood Pressure  
bpm 
Beats per Minute 
BQL 
Below the Quantification Limit 
BUN 
Blood Urea Nitrogen 
C 
Celsius 
CHEM 
Chemistry 
CI 
Confidence Interval 
CKD-EPI 
Chronic Kidney Disease Epidemiology collaboration equation for estimating 
GFR 


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CLT 
Total Clearance 
Cmax 
Maximum Concentration 
COAG 
Coagulation 
ConMed 
Concomitant Medication 
CRF 
Case Report Form 
CS 
Clinically Significant 
CSR 
Clinical Study Report 
CTU 
Clinical Trial Unit 
CV 
Coefficient of Variation 
ΔQTcF 
Pre-dose, baseline difference in QTcF interval 
ΔΔQTcF 
Time-matched, baseline-adjusted difference in QTcF interval 
dB 
Decibel 
DBP 
Diastolic Blood Pressure 
dL 
Deciliter 
DMID 
Division of Microbiology and Infectious Diseases 
DPOAE 
Distortion Product Otoacoustic Emission 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
EDC 
Electronic Data Capture 
eGFR 
Estimated Glomerular Filtration Rate 
ELF 
Epithelial Lining Fluid 
F 
Fahrenheit 
FDA 
Food and Drug Administration 
FEV1 
Forced Expiratory Volume in the first second 
FSH 
Follicle Stimulating Hormone 
FVC 
Forced Vital Capacity 
g 
Gram(s) 
GM 
Geometric Mean 
h 
Hour(s) 
HBsAg 
Hepatitis B-virus Surface Antigen 
HCV 
Hepatitis C-virus 
HEENT 
Head, Eyes, Ears, Nose, and Throat 


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HEM 
Hematology 
HIV 
Human Immunodeficiency Virus 
HLGT 
High Level Group Term 
HPF 
High-Powered Field 
HR 
Heart Rate 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IMP 
Investigational Medical Product 
INR 
International Normalized Ratio 
IUD 
Intrauterine Device 
IV 
Intravenous 
Ke 
Elimination Rate Constant 
kg 
Kilogram(s) 
kHz 
Kilohertz 
KIM-1 
Kidney Injury Molecule 1 
L 
Liter(s) 
LC-MS/MS 
Liquid Chromatography Tandem-Mass Spectrometry 
LLOQ 
Lower Limit of Quantification 
Max 
Maximum Value 
MedDRA 
Medical Dictionary for Regulatory Activities  
µg 
Microgram(s) 
mg 
Milligram(s) 
MH 
Medical History 
Min 
Minimum Value 
mL 
Milliliter(s) 
mmHg 
Millimeters of Mercury 
mmol 
Millimole(s) 
MOP 
Manual or Procedures 
msec 
Millisecond(s) 
N 
Number (typically refers to participants) 
NCA 
Noncompartmental Analysis 
NCS 
Not Clinically Significant 


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NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
ONR 
Outside Normal Range 
OTC 
Over the Counter 
PE 
Physical Examination 
PI 
Principal Investigator 
PK 
Pharmacokinetics 
Protime 
Prothrombin Time 
PT 
Preferred Term 
QNS 
Quantity Not Sufficient 
QTcF 
Corrected QT Interval of the ECG using Fridericia’s Formula 
RBC 
Red Blood Cell 
RR 
Respiratory Rate 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SBP 
Systolic Blood Pressure 
SD 
Standard Deviation 
SDCC 
Statistical and Data Coordinating Center 
SDTM 
Study Data Tabulation Model 
sec 
Second(s) 
SMC 
Safety Monitoring Committee 
SNR 
Signal-to-Noise Ratio 
SOC  
System Organ Class 
t½ 
Terminal Elimination Half-Life 
T3 
Triiodothyronine 
T4 
Thyroxine 
TEAE 
Treatment-Emergent Adverse Event 
Tmax 
Time of Maximum Concentration 
TSH 
Thyroid Stimulating Hormone 
UA 
Urinalysis 
Vd 
Volume of Distribution 
VS 
Vital Signs 


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WBC 
White Blood Cell 
WHO 
World Health Organization 
 


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1. 
PREFACE 
The Statistical Analysis Plan (SAP) for “A Phase I, Open-Label Study to Assess Lung Pharmacokinetics and 
Safety of a Single Dose of Apramycin Administered Intravenously in Healthy Adult Subjects” (DMID 
Protocol 20-0012) describes and expands upon the statistical information presented in the protocol. 
The document describes all planned analyses and provides reasons and justifications for these analyses. It also 
includes sample tables, listings, and figures planned for the final analyses. Regarding the final analyses and 
Clinical Study Report (CSR), this SAP follows the International Conference on Harmonisation of Technical 
Requirements for Registration of Pharmaceuticals for Human Use (ICH) Guidelines, as indicated in Topic E3 
(Structure and Content of Clinical Study Reports) [1], and more generally is consistent with Topic E8 
(General Considerations for Clinical Trials) [2], and Topic E9 (Statistical Principles for Clinical Trials) [3]. 
The structure and content of the SAP provides sufficient detail to meet the requirements identified by the 
Food and Drug Administration (FDA) and ICH, while all work planned and reported for this SAP will follow 
internationally accepted guidelines published by the American Statistical Association [4] and the Royal 
Statistical Society of statistical practice [5].  
This document contains four sections: (1) a review of the study design, (2) general statistical considerations, 
(3) comprehensive statistical analysis methods for the safety and pharmacokinetic (PK) outcomes, and (4) a 
list of proposed tables and figures. Within the table, figure, and listing mock-ups (Appendix 1, Appendix 2, 
and Appendix 3), references to CSR sections are included. Any deviation from this SAP will be described and 
justified in protocol amendments and/or in the CSR, as appropriate. The reader of this SAP is encouraged to 
also review the study protocol for details on conduct of the study and the operational aspects of clinical 
assessments. 


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2. 
INTRODUCTION 
Aminoglycosides are a class of antibiotics established with the introduction of streptomycin in the 1940s for 
the treatment of tuberculosis and have defended a strong clinical presence ever since and until today. Mainly 
due to the common recognition of the rapid bactericidal activity of aminoglycosides – also at very high 
bacterial burden – the drug class has established itself as an important therapeutic component in the treatment 
of serious hospital infections worldwide. Despite the introduction of several new chemical classes of 
antibacterial agents over the past few decades, aminoglycosides still remain an extremely important 
therapeutic option.  
The therapeutic efficacy of standard-of-care aminoglycosides currently in clinical use is compromised by 
significant rates of antimicrobial resistance acquired by horizonal gene transfer. Infections with antibiotic-
resistant bacteria caused an estimated 33,000 deaths in Europe in 2015 [6]. According to the CDC’s Antibiotic 
Resistance Threats in the United States, 2019, more than 2.8 million antibiotic-resistant infections occur in the 
United States each year, and more than 35,000 people die as a result [7]. Bacterial pathogens highly resistant 
to various classes of antibiotics are estimated to account for a total of about 700,000 deaths annual worldwide. 
Without successful intervention, the burden of continued emergence and global spread of antimicrobial 
resistance is expected to multiply by the year 2050 [7,8]. New antibiotics are needed to adequately respond to 
the increasing prevalence of drug-resistant infections. 
Apramycin is a mono-substituted 2-deoxystreptamine compound that is significantly different from the di-
substituted 2-deoxystreptamines which include all the standard-of-care aminoglycoside antibiotics 
(gentamicin, amikacin, tobramycin, arbekacin, plazomicin and other) and, therefore, represents a subclass of 
aminoglycosides [9]. Apramycin is active against a wide variety of pathogen Gram-positive and Gram-
negative bacteria including pathogens with resistance to gentamicin and all other clinical aminoglycosides of 
relevance. 
This is a Phase 1, open-label study of a single dose of apramycin conducted at a single center. A total of 20 
participants will be enrolled to one of 5 cohorts, each corresponding to a timepoint after initiation of infusion 
at which a single fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) is performed. There will be 4 
participants per cohort. 
2.1. 
Purpose of the Analyses 
These analyses will assess the safety and plasma and intrapulmonary PK of a single dose of 30 mg/kg 
apramycin administered as an IV infusion over a period of 30 (± 5) minutes.  


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3. 
STUDY OBJECTIVES AND ENDPOINTS 
3.1. 
Study Objectives 
Primary: 
• To assess plasma pharmacokinetic profile of apramycin and lung penetration of apramycin in 
epithelial lining fluid (ELF) and alveolar macrophages (AM) after single IV apramycin dose of 30 
mg/kg in healthy participants. 
Secondary: 
• To assess the safety of single IV administration of 30 mg/kg apramycin in healthy participants. 
• To assess changes in otoacoustic testing.  
Exploratory: 
• To assess changes in kidney function biomarkers. 
3.2. 
Endpoints 
Primary: 
• Lung concentration and PK parameters of total apramycin: 
o ELF: Area under the concentration-time curve (AUC) from time of dosing to 24 h (AUC(0-24)), 
AUC from time of dosing to time t (AUC(0-t)), AUC extrapolated to infinity (AUC(0-∞)), maximum 
concentration (Cmax), time to maximum concentration (Tmax), terminal elimination half-life (t½) 
o AM: AUC(0-24), AUC(0-t), AUC(0-∞), Cmax, Tmax, t½ 
• Plasma concentration and PK parameters of total apramycin:  
o AUC(0-∞), AUC(0-t), AUC(0-24), Cmax, Tmax, t½, total clearance (CLT), volume of distribution (Vd) 
central 
• Ratio of exposure parameters of Lung PK to Plasma PK: 
o ratio of Cmax in ELF over Cmax in plasma  
o ratio of Cmax in AM over Cmax in plasma  
o ratio of AUC (0-24), AUC(0-t), and AUC(0-∞) in ELF over AUC (0-24), AUC(0-t), and AUC(0-∞) in plasma  
o ratio of AUC (0-24), AUC(0-t), and AUC(0-∞) in AM over AUC (0-24), AUC(0-t), and AUC(0-∞) in plasma  
Secondary: 
• Type and incidence of treatment-emergent adverse events (TEAEs) and serious AEs (SAEs) to Day 30 
(± 4 days) after dosing. 
• Frequency of clinically significant TEAEs in physical examination (PE) findings and of changes from 
baseline in vital signs and clinical laboratory parameters until Day 14 (± 3 days) after dosing. 
• Frequency of changes in electrocardiogram (ECG) intervals (QTcF, PR, QRS, heart rate [HR]), and of 
morphological changes from baseline until 24 ± 1 h after dosing. 


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• Type and incidence of TEAEs related to auditory (cochlear) function tests (pure-tone audiometry and 
distortion product otoacoustic emissions [DPOAEs]) to Day 30 (± 4 days) after dosing.  
Exploratory: 
• Changes from baseline in urinary kidney injury molecule 1 (KIM-1) and serum cystatin C 
concentrations until Day 30 (± 4 days) after dosing. 
3.3. 
Study Definitions and Derived Variables  
3.3.1. 
Timepoints for Safety and PK Endpoints 
Table 1 shows the analysis timepoints in the order that will appear in tables and figures along with the safety 
and PK endpoints measured at each timepoint for all participants. Endpoints analyzed by timepoint include 
chemistry clinical laboratory results (CHEM), hematology clinical laboratory results (HEM), coagulation 
clinical laboratory results (COAG), and urinalysis laboratory results (UA); urinary KIM-1 and serum cystatin 
C; vital signs (VS); ECG; ENT examination and otoacoustic (pure-tone audiometry and DPOAE) tests (listed 
in the table below as Audiology); and blood (plasma) PK and lung BAL PK results (see Section 3.3.2 for 
derivations of BAL PK parameters). Serology, toxicology, and pregnancy testing at Screening and/or Check-
In will not be included in tables or figures but will be presented in listings. 
Table 1: 
Analysis Timepoints for Safety and PK Endpoints 
Analysis Timepoint 
Abbreviated Label 
Endpoints 
Screening 
N/A 
CHEM, HEM, COAG, UA, VS, ECG, Audiology 
Check-In 
D-2 
CHEM, HEM, COAG, UA, KIM-1, Cystatin C, VS, ECG, Audiology 
Day -1 
D-1 
VS, ECG 
Day 1, Pre-Dose 
0 h 
CHEM, HEM, COAG, UA, KIM-1, Cystatin C, VS, ECG, Plasma PK 
Maximum Severity Post Baseline 
N/A 
CHEM, HEM, COAG, UA, VS, ECG, Audiology 
Day 1, 0.5 h Post-Dose 
0.5 h 
VS, Plasma PK, BAL PK 
Day 1, 1 h Post-Dose 
1 h 
VS, ECG, Plasma PK 
Day 1, 2 h Post-Dose 
2 h 
Plasma PK, BAL PK 
Day 1, 4 h Post-Dose 
4 h 
VS, ECG, Plasma PK, BAL PK 
Day 1, 8 h Post-Dose 
8 h 
Plasma PK, BAL PK 
Day 1, 16 h Post-Dose 
16 h 
VS, ECG, Plasma PK 
Day 2, 24 h Post-Dose 
24 h 
CHEM, HEM, COAG, UA, VS, ECG, Plasma PK, BAL PK 
Day 2, 36 h Post-Dose 
36 h 
VS, ECG, Plasma PK 
Day 3, 48 h Post-Dose 
48 h 
CHEM, HEM, COAG, UA, KIM-1, Cystatin C, VS, ECG, Plasma PK 
Day 3, 60 h Post-Dose 
60 h 
VS, ECG, Plasma PK 
Day 3a 
D3 
Audiology 
Day 14 
D14 
CHEM, HEM, COAG, UA, KIM-1, Cystatin C, VS, ECG, Audiology 
Day 30 
D30 
KIM-1, Cystatin C, Audiology 
a Day 3 refers to any time on Day 3. 


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3.3.2. 
BAL Pharmacokinetics Parameters 
There are two BAL components collected in this study, ELF (Section 3.3.2.1) and AM (Section 3.3.2.2), and 
apramycin concentrations and PK parameters will be measured and estimated separately in each component. 
3.3.2.1. 
Apramycin Concentrations in ELF 
The estimated concentration of apramycin in the ELF component of the BAL supernatant (Apramycin ELF) 
will be calculated as follows using the urea method [10, 11, 12]: 
- 
Volume ELF = Volume BAL SUP x (Urea BAL SUP / Urea Plasma) 
 
 
 
(equation 1) 
- 
Apramycin ELF = Apramycin BAL SUP x (Volume BAL SUP / Volume ELF)  
 
(equation 2) 
- 
Apramycin ELF = Apramycin BAL SUP x (Urea Plasma / Urea BAL SUP)  
 
 
(equation 3) 
Where, 
 
 Volume ELF is the estimated volume of ELF in the BAL supernatant (BAL SUP), which is calculated 
from the following parameters:  
o Volume BAL SUP is the measured total volume of BAL SUP after centrifugation of BAL#2 
(combined volume of aspirated BAL in tubes 2, 3 and 4)1; and 
o Urea Plasma and Urea BAL SUP are the measured concentrations of urea in plasma and the 
BAL supernatant, respectively2. 
 Apramycin BAL SUP is the measured apramycin concentration in the BAL supernatant fluid2.  
 Note: Equation 3 is the result of merge of equations 1 and 2 
3.3.2.2. 
Apramycin Concentrations in AM 
The estimated concentration of apramycin in the AM component of the BAL cell pellet (Apramycin AM) will 
be calculated as follows:  
- 
Apramycin BAL AC = Concentration BAL AC x total volume of the BAL Pellet in suspension  (equation 4) 
- 
Volume AM = BAL AC x BAL Volume x % AM  x 2.42 µL / 106  
 
 
 
(equation 5) 
- 
Apramycin AM = Apramycin BAL AC / Volume AM  
 
 
 
 
 
(equation 6) 
Where,  
 Apramycin BAL AC is the measured amount of apramycin in the total volume of alveolar cell (AC) 
suspension, which is calculated from the following parameters: 
o Concentration BAL AC is the apramycin concentration in BAL Pellet2; and  
o Total volume of BAL Pellet in suspension (the total volume in which the cell pellet is 
suspended in after centrifugation of BAL#2)1.  
 Volume AM is the total volume of alveolar macrophages in the BAL pellet, which is calculated from 
the following parameters: 
 
 
 
 
 
1 Reported by the study site (Alliance for Multispecialty Research [AMR]) 
2 Reported by the bioanalytical laboratory (KCAS) 


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o BAL AC is the counted number of total alveolar cells per mL of BAL volume3; 
o BAL volume is the measured total volume of BAL#2 (combined volume of aspirated BAL in 
tubes 2, 3 and 4) before centrifugation3;  
o % AM is the percentage of alveolar macrophages in total alveolar cells (BAL cell pellet)3; and 
o 2.42 µL is the mean cell volume of 106 AM cells4.  
3.3.3. 
Baseline 
Baseline for clinical laboratory, VS, and audiology results will be defined as the last value recorded before 
administration of study drug.  
For ECG measurements, pre-dose baseline is defined as the last value recorded before administration of study 
drug, and time-matched baseline is defined as the value recorded at the same timepoint on the day before 
administration of study drug (Day -1) as on Day 1. If triplicate ECG measurements were performed, the mean 
value of the ECG measurements will be used as the pre-dose and time-matched baseline values.  
Baseline height, weight, and body mass index (BMI) will be the measurements obtained at Screening. Age 
will be based on age at the time of enrollment. 
3.3.4. 
Study Day 
Study day will primarily be used in listings to refer to the timing of assessments and events relative to study 
drug administration. The day that the administration of study drug is received is considered Study Day 1 for 
all participants. The day prior to the administration of study drug is considered Study Day -1; there is no 
Study Day 0. 
 
 
 
 
 
3 Reported by the study site (Alliance for Multispecialty Research [AMR]) 
4 Reported in the literature [11,12] 


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4. 
INVESTIGATIONAL PLAN 
4.1. 
Overall Study Design and Plan 
This is a Phase 1, single center, open-label study to evaluate the plasma and lung PK, safety and tolerability of 
a single IV dose of apramycin in 20 healthy male and female participants 18 to 45 years of age (inclusive). 
Each participant will be enrolled into one of five cohorts (T1-T5) before dosing on Day 1. Four participants 
will be allocated to each cohort. Each cohort corresponds to a timepoint after dosing when a single 
bronchoscopy with BAL will be performed: 0.5 h (± 5 min) (Cohort T1), 2 h (± 5 min) (Cohort T2), 4 h (± 10 
min) (Cohort T3), 8 h (± 15 min) (Cohort T4), and 24 (± 1 h) (Cohort T5). The timing of bronchoscopy and 
BAL in Cohort T5 is nominal. The final timepoint will be determined after analysis of apramycin 
concentrations in BAL in Cohorts T1-T4. If apramycin is detectable in BAL from Cohorts T1-T4, then T5 
will be enrolled. If it is not detectable, then Cohort T5 will not be enrolled. 
A diagram of the overall study design is shown in Figure 1. Participants will participate in the study for 
approximately 58 days, including a 28-day screening period, consisting of an out-patient period of 26 days 
(Day -28 to Day -3) with 1 to 2 site visits and 2 in-patient days; Day -2 and Day -1, to confirm eligibility for 
admission (Day -2) and eligibility for enrollment (Day -1), complete baseline assessments for ENT and 
otoacoustic tests (Day -2), and complete time-matched ECGs (Day -1); a 3-day in-patient treatment period, to 
confirm eligibility for dosing (Day 1), complete baseline VS, clinical labs, and exploratory labs before dosing 
(Day 1), administer treatment (Day 1), perform bronchoscopy with BAL within 24 hours after dosing 
(timepoints T1-T4 on Day 1 and T5 on Day 2), and complete in-patient follow up on Days 2 and 3; and a 27-
day out-patient follow-up period to complete site visits on Day 14 (± 3 days) and Day 30 (± 4 days), the last 
follow-up visit. It is anticipated that the duration of the study will be about 6 months to enroll Cohort T1-T4 
and 10 months to complete Cohort T5, if enrolled, following an interim data analysis to review lung PK data. 
Enrollment will be rolling and will be coordinated with the bronchoscopy service. Tentative reservations will 
be made and confirmed once participant eligibility is determined. Enrollment and treatment of participants 
will be staggered and no more than two participants will be dosed on a single day not less than 2 hours apart. 
Additional participants may be admitted to the Clinical Trial Unit (CTU) before dosing and may serve as 
back-up study participants. A participant assigned to one of the T1 to T4 cohorts may be reassigned to another 
cohort depending on the availability of a bronchoscopy time slot (Section 4.4.3). 
A schedule of all study events can be found in Table 2. Safety monitoring during the inpatient period will 
include: (a) Scheduled assessments per protocol: a PE, VS; 12-lead standard ECG (in triplicate at scheduled 
timepoints on Day-1 matching planned timepoints on Day 1); clinical safety laboratory tests (HEM, COAG, 
CHEM, including estimated Glomerular Filtration rate [eGFR], and UA); and audiology testing (middle ear 
examination and cochlear testing [hearing testing, consisting of pure-tone audiometry and DPOAEs). 
Exploratory plasma and urinary nephrotoxicity biomarkers (plasma cystatin C and urinary KIM-1) will also 
be measured. In addition, (b) Unscheduled assessments after bronchoscopy will be performed according to 
orders by the attending pulmonologist and as needed. Participants with on-going TEAEs will be followed 
until resolution or stability of the TEAE as assessed by the principal investigator (PI). 
A Safety Monitoring Committee (SMC) will be appointed to oversee the safe conduct of the trial, review 
safety data and provide recommendations on safety monitoring in the current and future clinical trials with 
apramycin. A scheduled interim SMC meeting will be held after all participants in Cohorts T1-T4 complete 
all assessments. If the last cohort (T5) will be enrolled for dosing, a final SMC will be held to review 
cumulative data on all participants. If Cohort T5 is not dosed, the interim meeting will be considered final. If 


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criteria for halting the trial are met, enrollment and dosing of new participants will be suspended, and an ad 
hoc SMC meeting will be held to review all available safety data. 
4.2. 
Discussion of Study Design, Including the Choice of Control Groups 
As the main focus of this protocol is to evaluate the PK, safety, and tolerability of a single dose of apramycin, 
and there is no control group, all participants will receive the same dose of apramycin with the same PK blood 
collection timepoints. Participants will be assigned to one of five BAL sampling timepoints with a total of 4 
participants assigned to each BAL sampling time. The timing of bronchoscopy and BAL in Cohort T5 will be 
determined after analysis of apramycin concentrations in BAL in Cohorts T1-T4. If apramycin is detectable in 
BAL from Cohorts T1-T4, then T5 will be enrolled. If it is not detectable, then Cohort T5 will not be enrolled. 
4.3. 
Selection of Study Population 
The study population for this trial is 20 healthy male or female participants 18 to 45 years of age (inclusive) at 
the time of dosing. Only participants who consent to participate and meet all of the inclusion and none of the 
exclusion criteria will be eligible for enrollment into this study. No exemptions are granted on Inclusion/ 
Exclusion Criteria in DMID-sponsored studies. 
Eligibility criteria from v3.0 of the protocol are listed below: 
Inclusion Criteria: 
All must be answered YES for the participant to be eligible for study participation: 
 
1. Participant reads and signs the Informed Consent Form (ICF) and agree to have bronchoscopy with 
bronchoalveolar lavage under sedation or light anesthesia and comply with study procedures. 
2. Healthy male or non-pregnant, non-lactating female participants 18 to 45 years of age (both inclusive) 
at the time of dosing.  
• Note 1: Determined by medical history (MH), medication use, physical examination (PE), and 
vital signs, clinical laboratory tests and 12-lead ECG within reference ranges at Screening and 
Day-2. (See protocol Sections 8.1 and 8.2, and Appendix B, Table 2, Table 3, and Table 4). 
Exceptions to blood pressure (BP), HR and laboratory test values being within normal ranges are: 
o Abnormal HR and BP on first measurement may be repeated twice more with the participant 
resting between measurements for at least 5 min according to protocol Section 8.1.6. 
o Participants with baseline HR ≥ 45 to 50 bpm may be accepted if otherwise healthy adults with 
known history of asymptomatic bradycardia.  
o Participants with baseline systolic BP up to 140 mmHg and diastolic BP up to 90 mmHg may 
be accepted if otherwise healthy.  
o A laboratory value that is Grade 1 will be allowed if not considered to be clinically significant 
by the investigator, with the exception of alanine aminotransferase (ALT), aspartate 
aminotransferase (AST), alkaline phosphatase (AP), blood urea nitrogen (BUN), urine protein, 
serum creatinine or estimated glomerular filtration rate (eGFR) <70 mL/min /1.73 m2 by the 
Chronic Kidney Disease Epidemiology collaboration (CKD-EPI) equation. 
3. Female participants of childbearing potential should use highly effective methods of contraception 
from the time of screening to 30 days after dosing. 


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• Note 1: A female is considered of childbearing potential unless post-menopausal (defined as 
history of ≥1 year of spontaneous amenorrhea and a follicle stimulating hormone [FSH] level >40 
IU/L), or permanently surgically sterilized. 
• Note 2: Highly effective contraceptive methods include: (a) surgical sterilization methods, such as 
tubal ligation, bilateral oophorectomy, salpingectomy, hysterectomy, or successful tubal 
obliteration (e.g., Essure®) with documented radiological confirmation test at least 90 days after 
the procedure, or (b) long-acting reversible contraception, such as progestin-releasing subdermal 
implants, copper intrauterine devices (IUDs), levonorgestrel-releasing IUDs. 
• Note 3: A participant who is not sexually active and abstains from sexual intercourse can be 
enrolled and abstinence documented.   
4. Males, including vasectomized men, having sexual intercourse with women of childbearing potential 
must agree to consistent use of condoms from investigational medical product (IMP) administration 
through at least 30 days after dosing, and must also agree to not donate sperm during this same time 
period. 
• Note: A participant who is not sexually active and abstains from sexual intercourse can be enrolled 
and abstinence documented. 
5. BMI 18.0 to 32.0 kg/m2 (inclusive) and body weight not less than 50 kg.  
6. Participants with normal hearing, i.e., symmetric hearing with air conduction thresholds no worse than 
20 dB hearing loss for the frequencies 0.5-1-2-4-6-8 kHz bilaterally. 
7. Normal (reproducibility 70% or better) of distortion product otoacoustic emissions (DPOAEs). 
8. Normal otoscopic findings in the ears, normal tympanic membrane mobility and stapedial reflex 
present. 
9. From the signing of the informed consent until the last follow-up visit, participants must be willing to 
avoid exposure to loud music or noise.  
• Note: Noise avoidance to include continuous usage of earpieces at high volume, attending loud 
concerts or dance events, or using firearms or attending fireworks. 
10. Normal lung function with Forced Expiratory Volume in the first second (FEV1) predicted ≥ 80% and 
FEV1/Forced Vital Capacity (FVC) > 70%. 
11. Participants must be willing to avoid excessive physical exercise within 48 h prior to dosing until 
discharge from the CTU on Day 3, and 24 h before each follow-up visit  
(Day 14 ± 3 days and Day 30 ± 4 days). 
12. No history of acute febrile or infectious illness for at least 7 days prior to the administration of the 
IMP.  
13. No history of lower respiratory tract infection within 4 weeks prior to screening. 
14. Have adequate venous access for infusion and blood draws. 
Exclusion Criteria: 
All must be answered NO for the participant to be eligible for study participation: 
1. Lactating females. 


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Medical and surgical history: 
2. Any history of hypersensitivity to aminoglycosides. 
3. Any history of drug hypersensitivity, asthma, urticaria or other severe allergic diathesis.  
4. Any history of seasonal allergies with ongoing symptoms for more than a week prior to dosing 
requiring glucocorticoids and/or frequent use of antihistamines for treatment. 
5. Any history of a chronic condition that may increase risk to participant or interfere with endpoint 
assessment, or any unstable chronic disease.  
• Note 1: Unstable chronic disease is defined by need for frequent medical interventions that lead to 
a change in medications and/or required hospitalization, surgery or an invasive procedure or 
emergency department/urgent care visit. 
• Note 2: Any chronic disease, that has been diagnosed within 90 days of screening is excluded. 
6. History of any psychiatric medical condition that has required hospitalization in the last 5 years or 
participant is considered psychologically unstable by the investigator. 
7. History of acute or chronic problems with hearing and/or balance in the last 24 months. 
• Note: These include but not limited to use of hearing aid, head injury leading to otologic damage, 
tumor of the head or neck, autoimmune disease of the inner ear, tinnitus, vestibular disease, 
auditory neurinoma, endolymphatic hydrops and/or Meniere’s disease, perilymphatic fistula, otitis 
media, labyrinthitis, sudden hearing loss, known retrocochlear hearing impairment, conductive 
hearing loss exceeding 10 dB at any frequency, ear canal and/or middle ear disease including 
inflammation or effusion, pathological tympanometry. 
8. Past injury or surgery to the middle or inner ears.  
• Note: Myringotomy or tympanic tube insertion in childhood with complete healing and normal 
hearing test are excluded. 
9. Family history of hearing loss before the age of 60. 
10. Participants who have had previous intolerance or contraindications to medications applied for 
sedation or anesthesia during bronchoscopy. 
• Note: These include benzodiazepines or topical anesthetic agents (lidocaine or xylocaine) 
including reversal agents such as flumazenil.  
Laboratory examinations: 
11. Positive serum pregnancy test for women or urine pregnancy test at check-in. 
12. Positive test for human immunodeficiency virus (HIV) antibodies, hepatitis B-virus surface antigen 
(HBsAg), or anti-hepatitis C-virus antibodies (anti-HCV). 
Prior medication: 
13.  Use of any prescription or non-prescription medication prior to the dose of IMP as described in 
protocol Section 6.6. 
• Note: Exceptions are hormonal contraceptives, which are permitted throughout the study, and 
solitary doses of up to 1,000 mg paracetamol.  


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14. Use of any investigational drug product within 30 days or 5 half-lives (whichever is longer) before 
dosing.  
15. Planned participation in a clinical research study that requires treatment with a study drug or blood 
draws or other invasive assessments during the study period (screening until final visit). 
Lifestyle restrictions: 
16. More than low-risk alcohol consumption (men: ≥ 24 g of pure alcohol regularly per day; women: ≥12 
g of pure alcohol regularly per day) for the previous 3 months. 
17. Any history of alcohol or drug abuse or positive urine alcohol screening test. 
18. Suspicion of illicit drug use / abuse or positive urine drug screen test (See protocol Section 6.7.4 and 
Section 8.2.4 for prohibited substances). 
19. History of ≥10 pack-years smoking, or history of any nicotine use in the 6 months before check-in 
(Day -2) or positive urine cotinine screen at check-in. 
• Note 1: Nicotine products include cigarettes, e-cigarettes, pipe, cigar, chewing tobacco, nicotine 
patch. 
• Note 2: A positive urine cotinine at screening is allowed if negative at check-in (Day -2).  
20. Caffeinated beverages/foods are prohibited within 48 hours before dosing to Day 3 of the trial. During 
the follow up period, consumption is restricted to not more than 3 cups or equivalent per day. 
21. Judged by the investigator to have occupational noise exposure of high risk during the trial (e.g., 
construction site workers, military workers, etc.). 
22. Blood or plasma donation of 500 mL within 3 months or more than 100 mL within 30 days before 
signing informed consent or planned donation prior to completion of this trial.  
4.4. 
Treatments 
4.4.1. 
Treatments Administered  
Participants will receive a single 30 mg/kg dose of apramycin administered as an IV infusion of 30 mL in a 
forearm vein over 30 min (± 5 min) using a syringe or infusion pump. 
4.4.2. 
Identity of Investigational Product(s) 
Apramycin for infusion, the IMP, is formulated as 150 mg free base /mL in sterile aqueous solution, pH 5.5 to 
6.0 adjusted with sulfuric acid. The study product is supplied in 20 mL glass vials. A volume of ≥18 mL can 
be withdrawn from each vial. To protect the solution from light, the vials are stored in folding boxes. 
The clinical research site will provide the diluent, 0.9% sodium chloride injection, USP. The diluent will be 
commercial source, USP grade 0.9% Sodium Chloride Injection, or normal saline, a sterile, nonpyrogenic, 
clear in color isotonic solution; each mL contains sodium chloride 9 mg. It contains no bacteriostatic agent, 
antimicrobial agent, preservatives, or added buffer and is supplied only in single-dose vials. The solution may 
contain hydrochloric acid and/or sodium hydroxide for pH adjustment (pH 5.3, range 4.5 to 7.0). This product 
should be used to dilute apramycin to the desired concentration.  


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4.4.3. 
Method of Assigning Participants to Treatment Groups 
Twenty healthy participants who consent to participate in the trial and meet the eligibility criteria will be 
enrolled following admittance to the CTU and confirmation of eligibility. Participants will be registered using 
Advantage eClinical®, developed and maintained by the Statistical and Data Coordinating Center (SDCC).  
Since this is an open-label study of a single dose of IMP, participants will be enrolled on a rolling basis. Four 
participants will be assigned to each one of five cohorts, T1-T5, corresponding to a single Lung PK sampling 
timepoint (Section 4.1) with equal allocation (1:1:1:1:1). Each cohort corresponds to a single Lung PK 
sampling timepoint. It is planned to enroll participants into cohorts T1 to T4 according to availability of the 
bronchoscopy suite. However, on the day of bronchoscopy, a participant may be reassigned to another cohort 
than planned on the same day, depending on emergencies at the bronchoscopy suite at the time originally 
planned, or cancelled. 
Appointments for the bronchoscopy suite will be made in advance and every effort will be made to admit, 
dose, and perform bronchoscopy with BAL according to the schedule. However, if a scheduled bronchoscopy 
cannot be performed according to schedule on a dosing day, the following steps may be taken according to the 
time of cancellation before or after dosing: 
• If the procedure is cancelled before administration of study drug, an effort will be made to reschedule 
the procedure. 
• If the procedure is cancelled after administration of the study drug, every effort will be made to 
reschedule the bronchoscopy to a later time (up to 8 hours after dosing) to accommodate an available 
Lung PK timepoint or the procedure will be cancelled. 
o If bronchoscopy is rescheduled, the registration in the Advantage eClinical® will be updated to 
indicate the cohort that the participant completed and a future participant would be assigned in the 
registry to complete the missing Lung PK timepoint. 
o If it is not feasible to reschedule bronchoscopy, the participant will be terminated from 
participation in the study and will be replaced. The participant will be asked to remain in the study 
to complete safety and otoacoustic testing until Final Visit (Day 30 ± 4 days). If the participant 
declines to remain in the study for safety assessments, appropriate early termination assessment 
will be completed prior to discharge from the CTU (see protocol v3.0 Section 5.3.3 and Section 
7.7).  
4.4.4. 
Selection of Doses in the Study 
A single dose of 30 mg/kg had been selected for this study based on the favorable safety and tolerability 
profile observed in the first in human clinical trial in healthy participants [13]. In addition, based on the 
translational PK-PD modeling study to assess efficacy, 30 mg/kg of EBL-1003 (apramycin) is the anticipated 
therapeutic dose for future clinical investigations in patients [14]. 
4.4.5. 
Selection and Timing of Dose for Each Participant 
This trial will evaluate the plasma and lung PK and safety of a single 30 mg/kg dose of apramycin 
administered as a 30-minute IV infusion on Day 1. A single bronchoscopy with BAL will be performed after 
initiation of dosing at one of the following timepoints in the respective cohorts: 30 min (± 5 min) (T1), 2 h (± 
5 min) (T2), 4 h (± 10 min) (T3), 8 h (± 15 min) (T4), and 24 h (± 1 h) (T5). Refer to Figure 1 for additional 
details regarding the timing schedule for the BAL timepoints following dosing. 


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4.4.6. 
Blinding 
This is an open-label, unblinded study. Blinding (masking) is not needed. 
4.4.7. 
Prior and Concomitant Therapy  
Medications include the following: prescription drugs, birth control hormonal preparations, non-prescription 
medication, herbs, vitamins, nutritional supplements, and illicit and recreational substances.  
Medications taken before or after dosing will be reported as Prior Medications or Concomitant Medications 
(ConMeds), respectively.  
Prior prescription medications will be recorded at Screening Visit. All prior medications are not allowed 
during the study period with the exception of oral contraceptives, which are permitted throughout the study, 
and solitary doses of up to 1,000 mg acetaminophen (paracetamol). 
The following medications are prohibited for the indicated periods prior to dosing: 
• Use of any investigational drug product within 30 days or 5 half-lives (whichever is longer) before 
dosing or planned use during the study period (screening until Final Visit). 
• Use of aminoglycosides within 3 months prior to dosing. 
• Use of neuromuscular blocking agents within 1 week or 5 half-lives (whichever is longer) prior to 
dosing. 
• Use of potentially nephrotoxic medication 2 weeks prior to dosing: Renin– angiotensin-aldosterone 
system inhibitors: angiotensin-converting enzyme-inhibitors, angiotensin II-receptor antagonists 
(enalapril, lisinopril, ramipril captopril, benazepril), spironolactone, eplerenone; non-steroidal anti-
inflammatory drugs (aspirin, ibuprofen, naproxen, indomethacin, etc.) including cyclooxygenase 2 
selective inhibitors (Bextra, Celebrex, Vioxx).  
• Use of potentially ototoxic medication 3 months prior to dosing: vancomycin, loop diuretics 
(bumetanide, ethacrynic acid, furosemide), quinine and quinidine derivatives including mefloquine 
with the exception of occasional intake of quinine containing beverages. 
Non-prescription medications, herbs, vitamins, and nutritional supplements will not be taken within 15 
days before dosing and during the trial. Exceptions: vitamins and over the counter (OTC) medications taken 
for < 48 h for the treatment of common symptoms (e.g., headache, indigestion, muscle pain) may be allowed 
if approved by the designated study clinician. 
Blood/blood products (red blood cells [RBCs], white blood cells [WBCs], platelets, and plasma) donation of 
500 mL within 3 months or more than 100 mL within 30 days before signing informed consent to this trial is 
not allowed, and it is prohibited during the course of this trial. 
Following dosing, each new ConMed and changes to existing medications will be recorded. Participants will 
be required not to utilize non-study medications during the trial except those deemed necessary by the Site PI 
or sub-investigator. 
Any drug (e.g., non-prescription medications, herbal supplements, vitamins, or prescription medications) or 
vaccines or blood/blood products used by the participant during the trial will be recorded in the participant’s 
source documents and on the appropriate electronic case report form (eCRF), and the PI or authorized study 
clinician (listed on FDA Form 1572) will note whether the use was medically indicated and immediately 
necessary. Any use of medications not authorized by the study PI or authorized clinician will be recorded as a 
deviation. 


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4.4.8. 
Treatment Compliance 
Since each dose of apramycin will be administered by site personnel, participant compliance is not anticipated 
to be an issue. Complete information regarding any partial or interrupted dosing will be documented. 
Participants unable to receive the full volume of apramycin infusion will be withdrawn from data analysis and 
followed for safety as described in protocol v3.0 section 5.3.3. 
Infusion interruptions, not exceeding 10 minutes in total, are permitted per protocol: 
• For non-drug safety related issues, infusion interruption(s) causing a delay of 9 minutes or less is 
permitted. 
o The allowable window for the infusion will be increased to 30 minutes (-5/+10) to account for the 
interruption 
o Participant will remain in the PK analysis population. 
• For infusion interruption delays of 10 minutes or greater in total, causing the participant not to 
receive the entire infusion volume: 
o The participant will be withdrawn from the bronchoscopy and pharmacokinetics procedures but 
encouraged to remain in the study to be followed for safety. 
o Participant removed from the PK analysis population. 
o All participants withdrawn due to infusion interruptions will be replaced. 
4.5. 
Safety and Pharmacokinetic Variables 
The following section describes the safety and PK endpoints of the study. As this study is a Phase 1 clinical 
trial in healthy adult participants, there will be no assessment of drug efficacy. For a detailed schedule of 
study procedures, refer to Table 2. Refer to Section 3 for a list of primary, secondary, and exploratory 
objectives and endpoints.  
4.5.1. 
Safety Variables 
Incidence, relatedness, and severity of TEAEs and SAEs will be recorded from the time of dosing through the 
Final Visit on Day 30 (± 4 days), or early termination. All AEs will be graded for severity and the relationship 
to the study product, as described in protocol v3.0 Section 9.2. Clinical AEs will be graded using the toxicity 
grading scales in Table 3.  
Abnormal clinical laboratory, VS, and ECG results will be assessed using the toxicity grading scales in 
Table 4, Table 5, and Table 6, respectively. Audiology results will be graded according to the toxicity grading 
scales in Table 3. 
In addition to AEs, the following safety endpoints will be assessed at baseline and each post-dose timepoint 
collected. For continuous parameters, change from baseline will also be summarized for all post-dose 
timepoints. For definitions of baseline for safety endpoints, refer to Section 3.3.3. 
• Clinical safety laboratory evaluations (CHEM, HEM, COAG, UA) 
o Clinical laboratory tests will be performed at Screening, Check-In (Day -2), pre-dose on Day 1, 
Day 2 (24 h post-dose), Day 3 (48 h post-dose), and Day 14. 
o The following parameters will be measured: 


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 CHEM: sodium, potassium, glucose (fasting), BUN, creatinine, eGFR, calcium, chloride, 
magnesium, total carbon dioxide, albumin, total protein, AP, ALT, AST, total bilirubin, direct 
bilirubin, thyroid stimulating hormone (TSH), free thyroxine (T4), and free triiodothyronine 
(T3) 
 HEM: hemoglobin, hematocrit, platelets, WBC count, and differential WBC count 
(neutrophils, lymphocytes, eosinophils, basophils, and monocytes) 
 COAG: activated partial thromboplastin time (aPTT), prothrombin time (Protime), and 
international normalized ratio (INR) 
 UA: urine dipstick, including protein, glucose, occult blood, and urine microscopy, including 
WBC count, RBC count, and bacteria, if urine dipstick is abnormal  
o Additionally, tests for exploratory biomarkers for renal injury, serum cystatin C, urinary KIM-1, 
and KIM-1 normalized per mg urinary creatinine, will be performed at Check-In (Day -2), pre-
dose on Day 1, Day 3 (48 hours post-dose), Day 14, and Day 30. 
• VS 
o VS will be measured at Screening, Check-In (Day -2), Day -1, Day 1 (pre-dose and 0.5 h, 1 h, 4 h, 
and 16 h post-dose), Day 2 (24 h and 36 h post-dose), Day 3 (48 h and 60 h post-dose), and Day 
14. 
o The following parameters will be measured: systolic BP, diastolic BP, HR, respiratory rate, and 
oral temperature. 
• 12-lead standard ECG 
o ECG will be performed in either triplicate or single at Screening, Check-In (Day -2), Day -1 (at 
timepoints matching those on Day 1), Day 1 (pre-dose and 1 h, 4 h, and 16 h post-dose), Day 2 (24 
h and 36 h post-dose), Day 3 (48 h and 60 h post-dose), and Day 14.  
 For ECG measurements performed in triplicate, the mean of the replicates will be used for 
analysis, including analyses of change from baseline. 
o At each timepoint, the overall interpretation of the ECG will be reported as Normal; Abnormal, 
Not Clinically Significant (NCS); or Abnormal, Clinically Significant (CS). For post-dose 
timepoints, the change from baseline will be also reported as No change from baseline; NCS, 
change from baseline; or CS, change from baseline. 
o The following parameters will be measured: PR interval, QRS interval, QT interval, corrected QT 
interval using Fridericia’s Formula (QTcF correction), RR internal, and ventricular rate. 
 For replicate ECGs recorded on Day -1 and Day 1, the mean time-matched change-from-
baseline QTcF values (ΔΔQTcF) will be evaluated to quantify the QTcF prolongation. For 
single ECG recoded after dosing, the mean pre-dose change-from-baseline (ΔQTcF) will 
also be analyzed. 
• Audiology 
o The following audiology tests will be conducted:  
 Valsalva test, Tympanometry and Stapedial Reflexes at Screening only. 
 Ear otoscopy at Screening, Check-In (Day -2), Day 3, Day 14, and Day 30. 


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 Pure tone audiometry at standard frequencies (0.5 to 8.0 kHz) at Screening and at both 
standard and higher frequencies (9.0 to 20.0 kHz) at Check-In (Day -2), Day 3, Day 14, and 
Day 30. 
 DPOAEs (1.5 to 6.0 kHz) at Screening, Check-In (Day -2), Day 3, Day 14, and Day 30. 
• Physical Examination 
o A complete PE to assess general appearance; head, eyes, ears, nose, and throat (HEENT); heart, 
lungs, abdomen, skin, musculoskeletal system, and lymph nodes; and a neurological exam will be 
performed at Screening, Check-In (Day -2), Day 3 (60 h post-dose), and Day 14. 
o An abbreviated PE will be performed at Day -1. An abbreviated PE differs from a complete PE in 
that the abdomen and neurological system are not evaluated. 
o A symptom-directed (focused) PE may be performed on Day 1 and at any time after dosing for 
evaluation of TEAEs. 
o Height and weight will be measured, and BMI calculated, at Screening only. Weight only will be 
measured on Day -2 and Day 14. 
4.5.2. 
Pharmacokinetic Variables 
A valid liquid chromatography tandem-mass spectrometry (LC-MS/MS) method will be used to determine 
concentrations of apramycin in plasma and BAL samples as well as urea in plasma and BAL (ELF). 
• Blood (Plasma) for apramycin concentrations 
o Blood (plasma) samples for assay of apramycin PK will be collected at the following study days 
and timepoints: On Day 1 within 30 min before dosing, and 0.5 h (±5 min, immediately at end of 
infusion), 1 h (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h (±15 min), and 16 h (±15 min) after 
dosing; on Day 2 at 24 h (± 1 h) and 36 h (± 1 h) after dosing; and on Day 3 at 48 h (± 1 h) and 60 
h (± 1 h) after dosing, or ET if occurs within 24 h of dosing. 
o If BAL is delayed, blood (plasma) for apramycin will be collected within the prespecified window 
of the nominal timepoint and a second sample will be collected within 5 min of the last BAL 
sample. 
• Bronchoscopy with BAL 
o During bronchoscopy with BAL, samples will be obtained at the following timepoints during the 
study: 0.5 h ± 5 min, 2 h ± 5 min, 4 h ± 10 min, 8 h ±15 min and 24 h ± 1 h after the start of the 
infusion. Each participant will have a bronchoscopy with BAL at one timepoint only, for a total of 
4 samples at each timepoint. 
• Urea 
o Urea will be assayed in plasma samples collected at baseline (as part of the total apramycin plasma 
PK) and at 0.5 h (± 5 min) 2 h (± 5 min), 4 h (± 10 min), 8 h (± 15 min) and 24 h (± 1 h) after the 
start of the infusion), and in BAL SUP samples corresponding to plasma PK timepoints. If BAL is 
delayed and a second plasma sample is collected within 5 min of the last BAL sample, plasma urea 
will be measured only in the second sample. 
 


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5. 
SAMPLE SIZE CONSIDERATIONS 
This is Phase 1 study to assess the safety and PK of apramycin. Since the study is primarily aimed at safety 
and PK, no formal power calculations based on testing a statistical hypothesis were constructed. Enrollment 
of 20 participants assigned into 5 cohorts, T1-T5, with 4 participants per cohort is based on clinical 
experience and judgment and should provide adequate clinical information to meet the objectives of the study.  


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6. 
GENERAL STATISTICAL CONSIDERATIONS 
6.1. 
General Principles 
Summary statistics for continuous data will include the number of participants included in the analysis (N), 
mean, standard deviation (SD), median, minimum (min), and maximum (max). Summary statistics for 
categorical data will include frequencies and proportions and may include confidence intervals (CIs) for the 
proportion. When 95% CIs are given for a proportion, exact (Clopper-Pearson) CIs will be used, unless 
otherwise specified.  
All safety and PK analyses will be performed for all participants combined across cohorts, within the 
respective analysis populations. No analyses will be performed by cohort. The Safety Population and Safety 
Subset Population will both be used for summaries of safety endpoints, and the PK Analysis Subset 
Population will be used for summaries of PK endpoints. For the safety endpoints, the population used will be 
noted in the table, figure, or listing title. 
Denominators for safety endpoints will be the number of participants in the Safety Population or Safety 
Subset Population. Denominators for clinical laboratory, VS, ECG, and audiology results at planned study 
timepoints will be the number of participants with available results at the specified timepoint for that 
parameter. Denominators for the conceptual “Maximum Severity Post Baseline” timepoint for clinical 
laboratory, VS, ECG, and audiology results will be the number of participants with an observed result for that 
parameter obtained post-dose. 
The sort order for listings is indicated in the implementation note for each listing shell (Appendix 3). The sort 
order of clinical laboratory tests, VS, ECG, and audiology parameters is described in Section 9.  
6.2. 
Timing of Analyses 
The final analysis will be performed after database lock. 
One interim PK review is planned following Cohort T4 to analyze the plasma and BAL (ELF and AM) 
apramycin concentrations and PK data from cohorts T1 to T4.  
The SMC will review safety data at the following times: 
• Following Cohort T4 to review cumulative, interim safety data of all participants in cohorts T1 to T4  
• After completion of dosing and final visit for all participants, including Cohort T5 
• Ah hoc meeting(s): as required in response to halting criteria (see Protocol v3.0 Section 9.5) or at the 
request of DMID to review a potential safety concern identified by either the Site PI or DMID medical 
monitor 
6.3. 
Analysis Populations 
All analysis populations to be used in the final analysis are described in this section. A tabular listing of all 
enrolled participants excluded from an analysis population (a Safety Population or a PK Analysis Population) 
will be included in the CSR (Listing 4). Although there may be multiple reasons for exclusion from an 
analysis population, only one reason will be counted when summarizing reasons for exclusion from analysis 
populations in Table 11. The order that reasons will be considered is the same as the order shown in Table 11. 


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6.3.1. 
Safety Population 
The Safety Population will include all participants that received any amount of study product (apramycin).  
6.3.1.1. 
Safety Subset Population 
The per protocol Safety Subset Population will be based on the Safety Population and will include all 
participants who received a complete dose of apramycin. Participants who had an infusion interruption lasting 
10 minutes or longer will be withdrawn and excluded from the Safety Subset Population if they did not 
receive the full dose of apramycin.  
6.3.2. 
PK Populations 
6.3.2.1. 
Lung PK Population 
The Lung PK Analysis Population will consist of all participants who received a complete dose of apramycin, 
underwent BAL at the assigned sampling timepoint with BAL return volume adequate for analysis, and have 
a quantifiable post-dose drug concentration separately for BAL SUP (for calculation of ELF) and BAL AC 
(for calculation of AM).  
6.3.2.2. 
Plasma PK Population 
The Plasma PK Analysis Population will consist of all participants who received a complete dose of 
apramycin and have at least one quantifiable post-dose plasma drug concentration measured.  
6.3.2.3. 
PK Subset Population 
The PK Analysis Subset Population will be based on the Lung PK and Plasma PK analysis populations, which 
include all participants who completed the lung PK and plasma PK parts of the trial without any protocol 
deviations that would likely affect the PK results and who have evaluable plasma PK and BAL PK 
concentration data for apramycin from which at least a subset of the designated PK parameters can be 
determined. Potential protocol deviations include infusion interruptions or samples collected out of window. 
Participants who had an infusion interruption lasting 10 minutes or longer will be withdrawn and excluded 
from the PK Analysis Subset Population. If any participants are found to be noncompliant with respect to 
dosing or have incomplete data, a decision to include them in the analysis will be made on a case-by-case 
basis.   
6.4. 
Covariates and Subgroups 
The protocol does not define any formal subgroup analyses. 
6.5. 
Missing Data 
All attempts will be made to collect all data per protocol. No imputation will be performed for missing values.  
6.6. 
Interim Analyses and Data Monitoring 
An interim analysis of PK data is planned to follow the completion of Cohort T4 (see Section 6.2). 
Concentration-time data will be presented tabularly and graphically for participants in the PK Analysis 
Population for both plasma and BAL (ELF and AM) apramycin concentrations. Non-compartmental analysis 
(NCA) PK parameters will be summarized for the plasma and BAL PK data. Tables and figures for analyses 
described in Section 10 will be used for the interim analysis. 


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An SMC meeting to review cumulative safety data is planned after Cohort T4. Safety data, including TEAEs, 
SAEs, clinical laboratory results (CHEM, HEM, COAG, and UA), VS, ECGs, and audiology testing results 
will be presented. The SMC may also review safety data during an ad hoc meeting if a halting rule (see 
Protocol v3.0 Section 9.5) is met.  
6.7. 
Multicenter Studies 
This is a single-site study. 
6.8. 
Multiple Comparisons/Multiplicity 
This is a Phase 1 study with multiple primary endpoints. Because analyses of primary endpoints are 
descriptive rather than hypothesis tests, no adjustments for multiple testing are planned. 


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7. 
STUDY PARTICIPANTS 
7.1. 
Disposition of Participants 
Screened participants who were ineligible for enrollment in the study (screen failures) or eligible but not 
enrolled will be summarized by inclusion and exclusion criteria and reason not enrolled (Table 10). Enrolled 
participants who were ineligible for inclusion in analysis populations will be summarized by reason for 
exclusion and Cohort (Table 11). Individual reasons for participants who were excluded from a Safety or a PK 
Analysis Population will be listed (Listing 4). 
Participant disposition will be summarized (Table 8), showing the number of participants who were screened, 
enrolled, started infusion, completed infusion, completed bronchoscopy with BAL, completed all PK blood 
draws, completed all plasma urea blood draws, completed follow-up on Day 14, and completed the final study 
visit on Day 30. Enrolled participant who discontinued treatment or terminated early will be summarized by 
reason and Cohort in Table 9. 
Participants who discontinued treatment or terminated early from the study will be listed (Listing 1). 
A flowchart displaying the disposition of study participants will be included (Figure 2). This figure will 
present the number of participants screened, enrolled, lost to follow-up, and analyzed by Cohort. 
7.2. 
Protocol Deviations 
A summary of participant-specific protocol deviations will be presented by deviation category, deviation type, 
and Cohort (Table 7). This table will provide both the number of participants and the number of deviations for 
each deviation category and deviation type. All participant-specific protocol deviations and non-participant 
specific protocol deviations will be listed in Listing 2 and Listing 3, respectively. 


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8. 
EFFICACY EVALUATION 
There are no efficacy endpoints for this trial. 


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9. 
SAFETY EVALUATION 
All safety analyses will be performed using the Safety Population and Safety Subset Population. 
Any medical condition that is present at the time the participant is screened will be considered baseline and 
not reported as an AE, unless it worsens in severity or increases in frequency during the study. The 
denominators for proportion values will be indicated within the table or table header. AEs will be summarized 
for the number of participants who experienced an AE and the number of events by Medical Dictionary for 
Regulatory Activities (MedDRA®) category. Toxicity grading scales for clinical adverse events are provided 
in Table 3. All AEs reported will be included in the summaries and analyses. 
9.1. 
Demographic and Other Baseline Characteristics 
Sex, ethnicity, and race will be summarized by Cohort (Table 12). Ethnicity is categorized by “Hispanic or 
Latino”, “Not Hispanic or Latino”, “Not Reported”, or “Unknown”. In accordance with NIH reporting policy, 
participants may self-designate as belonging to more than one race or may refuse to identify a race, the latter 
reflected in the case report form (CRF) as “No” to each race option. Age at enrollment, height, weight, and 
BMI at Screening will be summarized by Cohort (Table 13). Individual participant listings will be presented 
for all demographic and baseline characteristics (Listing 5).  
9.1.1. 
Prior and Concurrent Medical Conditions 
All current illnesses and past pre-existing medical conditions will be MedDRA coded using MedDRA 
dictionary version 23.1 or higher. Summaries of participants’ pre-existing medical conditions by MedDRA 
system organ class (SOC) and Cohort will be presented in Table 14. Individual participant listings will be 
presented for all medical conditions (Listing 6). 
9.1.2. 
Prior and Concomitant Medications  
All medications will be coded to the Anatomical Therapeutic Classification (ATC) using the current version 
of the WHO Drug Dictionary. The use of prior and concomitant medications taken during the study will be 
summarized by ATC 1 and ATC 2 (separately, in Table 81 for prior medications and Table 82 for ConMeds). 
Individual participant listings will be presented for all prior and concomitant medications (separately in 
Listing 27 for prior medications and Listing 28 for ConMeds).  
9.2. 
Measurements of Treatment Compliance 
Infusion start and end dates and times, including the amount of study product received and infusion 
interruptions, will be listed (Listing 7). 
9.3. 
Adverse Events 
Refer to Section of 9.2 of the protocol for definitions of TEAEs, severity, and relatedness. When calculating 
the proportions of participants with AEs within a given MedDRA category, each participant will be counted 
once and any repetitions of AEs within a participant will be ignored, and the event will be reported according 
to the highest severity recorded (separately for related and unrelated AEs, when both severity and relatedness 
are tabulated). The denominators for percent values will be indicated within the table or table header. 


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9.3.1. 
Treatment Emergent Adverse Events 
An overall summary of TEAEs will be presented in Table 15 for the Safety Population and Table 16 for the 
Safety Subset Population, including the number of participants with at least one TEAE, the number of 
participants with at least one related TEAE, and the number of participants with at least one SAE. 
All AEs will be presented in Listing 8. A listing of non-serious TEAEs of moderate or greater severity will 
also be reported (Table 22). 
The following summaries for TEAEs will be presented by SOC, high level group term (HLGT), and preferred 
term (PT): 
• The total number of TEAEs and the number and proportion of participants reporting at least one 
TEAE, regardless of severity or relationship to study product. Ninety-five percent (95%) CIs will be 
presented for proportions (Table 17 for the Safety Population and Table 18 for the Safety Subset 
Population). 
• The number and proportion of participants reporting at least one TEAE by maximum severity and 
relationship to study product (Table 19 for the Safety Population and Table 20 for the Safety Subset 
Population). 
• Bar chart displaying the proportion of participants with related serious or non-serious TEAEs by 
maximum severity and SOC (Figure 3 for the Safety Population and Figure 4 for the Safety Subset 
Population). This figure describes the number of participants with an event (each participant is 
counted once per SOC). 
• Bar chart displaying the number of related serious and non-serious TEAEs by severity and SOC 
(Figure 5 for the Safety Population and Figure 6 for the Safety Subset Population). This figure 
described the total number of occurrences of each event, including multiple occurrences per 
participant. 
9.4. 
Deaths, Serious Adverse Events and other Significant Adverse Events 
Individual data listings of deaths and other SAEs will be provided (Table 21). The listing will include 
participant ID, Cohort, AE Number, AE description, SOC, HLGT, PT, duration of AE, reason reported as an 
SAE, severity, relationship to treatment, alternate etiology if not related, action taken with study treatment, 
whether the participant discontinued due to the AE, and AE outcome. 
9.5. 
Pregnancies 
An individual data listings of pregnancy reports will be presented if any pregnancies occur (Listing 29, 
Listing 30, Listing 31, Listing 32, and Listing 33). 
Birth control method(s) will be listed for each participant with start and end dates (Listing 34). 
9.6. 
Clinical Laboratory Evaluations 
Toxicity grading criteria for clinical laboratory results can be found in Table 4. Unscheduled clinical 
laboratory evaluations will be included in listings but excluded from tabular and graphical summaries, except 
when calculating the maximum severity post-baseline. Any pre-existing abnormal lab results at Screening and 
Baseline will be graded and presented in listings but will not be reported as an AE unless it is treatment-
emergent (i.e., severity worsens [increases to Grade 2 or higher] after dosing).  


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The following parameters will be presented (in order): 
• CHEM: sodium, potassium, glucose (fasting), BUN, creatinine, eGFR, calcium, chloride, magnesium, 
total carbon dioxide, albumin, total protein, AP, ALT, AST, total bilirubin, direct bilirubin, TSH, free 
T4, and free T3 
• HEM: hemoglobin, hematocrit, platelets, WBC count, neutrophils, lymphocytes, eosinophils, 
basophils, and monocytes 
• COAG: aPTT, Protime, and INR 
• UA: protein by dipstick, glucose by dipstick, occult blood by dipstick, WBC by microscopy, RBC by 
microscopy, and bacteria by microscopy (microscopy only completed if urine dipstick is abnormal) 
All safety laboratory results (CHEM, HEM, COAG, and UA) will be listed in Listing 9, Listing 10, 
Listing 11, and Listing 12, respectively. Abnormal laboratory results, including results outside of the normal 
range (ONR), will be listed in Table 23, Table 24, Table 25, and Table 26 for chemistry, hematology, 
coagulation, and urinalysis, respectively. Abnormal laboratory results that do not have toxicity grading ranges 
defined in the protocol may not be reported as AEs by the site and will not have severity indicated in tables 
and listings of abnormal laboratory results, except as ONR.  
Laboratory parameters that have grading criteria for both decreases (results lower than normal range) and 
increases (result higher than normal range) will be summarized separately by direction of change. For 
example, sodium will be summarized separately as “Sodium, Decrease” and “Sodium, Increase.” 
Laboratory results will be summarized in tables and figures: 
• The proportion of participants with abnormal laboratory results by parameter and timepoint will be 
summarized for CHEM (Table 27 for the Safety Population and Table 28 for the Safety Subset 
Population), HEM (Table 33 for the Safety Population and Table 34 for the Safety Subset Population), 
COAG (Table 39 for the Safety Population and Table 40 for the Safety Subset Population), and UA 
(Table 45 for the Safety Population and Table 46 for the Safety Subset Population). 
• The proportion of participants with mild, moderate, or severe laboratory results by parameter and 
timepoint will be summarized for CHEM (Table 29 for the Safety Population and Table 30 for the 
Safety Subset Population), HEM (Table 35 for the Safety Population and Table 36 for the Safety 
Subset Population), COAG (Table 41 for the Safety Population and Table 42 for the Safety Subset 
Population), and UA (Table 47 for the Safety Population and Table 48 for the Safety Subset 
Population). 
• Summary statistics of measurements and change from baseline by parameter and timepoint for CHEM 
(Table 31 for the Safety Subset Population and Table 32 for the Safety Population), HEM (Table 37 
for the Safety Population and Table 38 for the Safety Subset Population), and COAG (Table 43 for the 
Safety Population and Table 44 for the Safety Subset Population). UA character results will be 
excluded from change from baseline summaries. 
• Change from baseline at each scheduled timepoint will be visualized using box plots for each 
parameter. The safety population used for each figure will be noted in the title. UA character results 
will be excluded from graphical summaries.  
o CHEM: Beginning at Figure 7 and continuing through Figure 46 
o HEM: Beginning at Figure 47 and continuing through Figure 64 


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o COAG: Figure 65, Figure 66, Figure 67, Figure 68, Figure 69, and Figure 70 
Screening laboratory results will be listed. Serology result will be shown in Listing 14. Urine drug and 
cotinine and alcohol breathalyzer test results will be shown in Listing 15. Pregnancy testing and FSH test 
results will be shown in Listing 16. 
9.6.1. 
Exploratory Biomarkers for Renal Injury 
Kidney function biomarkers, serum cystatin C, urinary KIM-1, and KIM-1 normalized per mg urinary 
creatinine, will be summarized for measurements and change from baseline by parameter and timepoint in 
Table 49 for the Safety Population and in Table 50 for the Safety Subset Population. Additionally, results will 
be visualized using box plots showing the change from baseline at each scheduled timepoint for serum 
cystatin C in Figure 71 and Figure 72, KIM-1 in Figure 73 and Figure 74, and KIM-1 normalized per mg 
urinary creatinine in Figure 75 and Figure 76 for the Safety Population and Safety Subset Population, 
respectively. 
Results for all kidney function biomarker tests will be listed (Listing 13). 
9.7. 
Vital Signs and Physical Evaluations 
Toxicity grading criteria for VS results can be found in Table 5. Unscheduled VS measurements will be listed 
but excluded from tabular and graphical summaries by timepoint, except when calculating the maximum 
severity post-baseline. VS parameters that have grading criteria for both decreases (results lower than normal 
range) and increases (result higher than normal range) will be summarized separately by direction of change.  
The following VS parameters will be summarized (in order): systolic BP, diastolic BP, HR, respiratory rate, 
and oral temperature. 
VS parameters will be summarized in tables and figures: 
• The proportion of participants with mild, moderate, or severe VS results by parameter and timepoint 
will be summarized (Table 51 for the Safety Population and Table 52 for the Safety Subset 
Population). 
• Summary statistics of measurements and change from baseline by parameter and timepoint will be 
summarized (Table 53 for the Safety Population and Table 54 for the Safety Subset Population). 
• Change from baseline at each scheduled timepoint will be visualized using box plots for each 
parameter (Beginning at Figure 77 and continuing through Figure 86). 
All VS measurements, including height, weight, and BMI, will be presented in Listing 17. 
In the case of an abnormal VS measurement, measurements may be repeated up to 2 more times, for a 
maximum of 3 measurements including the initial measurement at each timepoint. The following rules will be 
used to decide which measurement to include in the analysis if more than one replicate was entered into the 
clinical database. VS replicates not used in the analysis will still be included in listings. 
1. If the first replicate is normal, then it will be used for analysis. 
2. If the first and second replicates are both abnormal, then the first replicate will be used if it has a 
severity greater or equal to the second replicate, or the second replicate will be used if the second 
replicate has a higher severity than the first replicate. 
3. If the first replicate is abnormal, the second replicate is normal, and the third replicate was not 
performed, then the first replicate will be used in the analysis. 


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4. If the first replicate is abnormal, the second replicate is normal, and the third replicate is normal, then 
the second replicate will be used in the analysis. 
5. If the first replicate is abnormal, the second replicate is normal, and the third replicate is abnormal, 
then the first replicate will be used if it has a severity greater or equal to the third replicate, or the third 
replicate will be used if the third replicate has a higher severity than the first replicate. 
Abnormal PE findings will be presented in Listing 18.  
9.8. 
12-Lead Standard Electrocardiogram 
Toxicity grading criteria for 12-lead standard ECG parameters (QTcF and PR Intervals) can be found in 
Table 6. Unscheduled 12-lead ECG measurements will be listed but excluded from tabular and graphical 
summaries, except when calculating the maximum severity post-baseline. When ECG measurements are 
conducted in triplicate, the mean of the triplicate values will be used for analysis. 
The following ECG parameters will be presented (in order): PR interval, QRS duration, QT interval, QTcF 
correction (refer to protocol Section 11.4.6.1 for formula), RR interval, and ventricular rate.  
12-lead standard ECG parameters will be summarized in tables and figures: 
• Summary of 12-lead ECG categorical change in overall interpretation from baseline will be presented 
by timepoint (Table 55 for the Safety Population and Table 56 for the Safety Subset Population). 
• Summary of 12-lead ECG categorical change from pre-dose baseline will be presented by timepoint 
(Table 57 for the Safety Population and Table 58 for the Safety Subset Population). 
• The proportion of participants with mild, moderate, or severe 12-lead ECG results will be presented 
for QTcF Interval and PR Interval by parameter and timepoint (Table 59 for the Safety Population and 
Table 60 for the Safety Subset Population). 
• Summary statistics of measurements and change from pre-dose baseline, including the 90% CI (see 
equation below), by parameter and timepoint will be summarized (Table 61 for the Safety Population 
and Table 62 for the Safety Subset Population). 
• For ECGs collected on Day 1, summary statistics of change from time-matched baseline, including the 
90% CI (see equation below), by parameter and timepoint will be summarized (Table 63 for the Safety 
Population and Table 64 for the Safety Subset Population). 
• A categorical summary of QTcF Interval (Table 65 for the Safety Population and Table 66 for the 
Safety Subset Population) and PR interval, QRS duration, and RR interval (Table 67 for the Safety 
Population and Table 68 for the Safety Subset Population) will be presented by timepoint. 
• Change from pre-dose baseline at each scheduled timepoint will be visualized using box plots for each 
parameter (Beginning at Figure 87 and continuing through Figure 98). 
Due to the small sample size (N < 30) and unknown population variance, the 90% CI for the change from 
baseline will be calculated as follows: 
ቌ𝑥𝑥̅ −𝑡𝑡𝑛𝑛−1,1−𝛼𝛼2
⁄ × ඨ𝜎𝜎2
𝑛𝑛,    𝑥𝑥̅ + 𝑡𝑡𝑛𝑛−1,1−𝛼𝛼2
⁄ × ඨ𝜎𝜎2
𝑛𝑛 ቍ 


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Where, 
 
𝑥𝑥̅ is the mean change from baseline 
 
𝑡𝑡𝑛𝑛−1,1−𝛼𝛼2
⁄  is the t-value with n-1 degrees of freedom at a confidence level of α (for 90% CI, α = 0.1) 
 
𝜎𝜎 is the standard deviation 
 
𝑛𝑛 is the number of observations 
All individual ECG measurements will be presented in Listing 19. Overall interpretations and changes from 
baseline will be presented in Listing 20. 
9.9. 
Otoacoustic Testing 
Toxicity grading criteria for pure-tone audiometry testing can be found in Table 3. Unscheduled otoacoustic 
testing will be listed but excluded from tabular and graphical summaries, except when calculating the 
maximum severity post-baseline. 
Otoacoustic test results, including ear otoscopy, pure-tone audiometry, and DPOAEs, will be summarized in 
tables and figures: 
• For ear otoscopy, summary of change in results from baseline will be presented by ear and timepoint 
(Table 69 for the Safety Population and Table 70 for the Safety Subset Population). 
• For pure-tone audiometry: 
o The proportion of participants with mild, moderate, or severe pure-tone audiology test results will 
be presented by timepoint (Table 71 for the Safety Population and Table 72 for the Safety Subset 
Population). 
o Summary statistics of measurements and change from baseline by timepoint, frequency, and ear 
will be summarized (Table 73 for the Safety Population and Table 74 for the Safety Subset 
Population). 
o Change from baseline at each timepoint and ear will be visualized using line plots with standard 
error bars for each frequency (Figure 99 and Figure 100 for frequencies 0.5, 1.0, 2.0, 4.0, 6.0 and 
8.0 kHz; Figure 101 and Figure 102 for frequencies 9.0, 10.0, 11.0, 12.5, 14.5, 16.0,18.0, and 20.0 
kHz for the Safety Population and Safety Subset Population, respectively).  
• For DPOAEs, including DPOAE, signal-to-noise ratio (SNR), and overall result: 
o Summary statistics of change from baseline by timepoint, frequency, and ear will be summarized 
(Table 75 and Table 76 for DPOAEs and Table 79 and Table 80 for SNR for the Safety Population 
and Safety Subset Population, respectively). 
o Summary of categorical DPAOE test results (present/absent) by timepoint, frequency, and ear will 
be presented (Table 77 for the Safety Population and Table 78 for the Safety Subset Population). 
o Change from baseline at each frequency and ear will be visualized using line plots with standard 
error bars for each scheduled timepoint and result (Figure 103 and Figure 104 for DPOAE results 
and Figure 105 and Figure 106 for SNR results for the Safety Population and Safety Subset 
Population, Respectively).  
All otoacoustic testing results will be listed: ear otoscopy test results will be presented in Listing 21; pure-tone 
audiometry test results will be presented in Listing 22; and DPOAE test results will be presented in 


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Listing 23. Additionally, screening Valsalva, tympanometry, and stapedial reflex test results will be presented 
in Listing 24, Listing 25, and Listing 26, respectively. 


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10. 
PHARMACOKINETICS 
10.1. 
Graphical and Tabular Summaries of Pharmacokinetic Profiles 
The PK Subset Population will be used when summarizing plasma and lung PK concentrations. Participants 
enrolled who did not complete dosing will not be included in the PK analysis, but the participant will have 
concentrations and PK parameters included in listings.  
For plasma concentrations, results below the limit of quantification (BQL) collected before the first 
measurable plasma concentration above the lower limit of quantification (LLOQ) will be treated as zero (0) 
for plotting, all calculations (including noncompartmental analysis [NCA]) and summary statistics. All other 
plasma BQL values observed after the first measurable concentration will be treated as missing. Similarly, all 
BQL lung PK concentrations will be treated as missing. There will be no imputation of missing 
concentrations. The geometric mean (GM) of concentrations will be treated as missing for sets of data points 
containing a BQL value. 
Collection of plasma or lung samples outside of the protocol defined time window for the timepoint will not 
result in exclusion of the sample results from NCA. Plasma or lung samples collected out of window will be 
evaluated on a case-by-case basis. Results from PK samples that were collected substantially outside of the 
protocol defined time window will be excluded from concentration summary statistics by nominal timepoints 
and plots of mean concentration by nominal timepoint. Substantially out-of-window samples are defined to be 
samples that were collected outside twice the size of the protocol required windows and are as follows: ±10 
min of the nominal timepoint for the 0.5 h, 1, h, and 2 h post-dose samples; ±20 min for the 4 h post-dose 
sample; ±30 min for the 8 h and 16 h post-dose samples; and ±2 h for the 24 h and 36 post-dose samples on 
Day 2 and 48 h and 60 h post-dose samples on Day 3.  
If the exact time of PK sample collection is not recorded then the collection time will be imputed as the 
planned time for analysis, as long as it is not known that the sample was collected outside of the protocol 
defined window. If the exact collection time is not known, but it is known that the sample was collected 
outside of the protocol defined time window, then the timepoint may be excluded from analysis at the 
discretion of the PK analyst. Rationale for excluding results from analysis will be described in the CSR. 
Results from samples with imputed collection times will be indicated in listings of PK sample concentrations.  
In the case the bronchoscopy with BAL procedure is delayed, an additional plasma PK sample will be 
collected within 5 minutes of the last BAL sample. These samples will only be included in the lung PK 
analysis when calculating the lung PK/plasma PK concentration ratios and PK parameters, and not included in 
the NCA. 
The bioanalytical lab will report concentrations of total and free apramycin in plasma, total apramycin in BAL 
SUP and BAL AC, and concentrations of urea in plasma in units of µg/mL. All concentrations will be 
reported in the same units in the CSR. Apramycin concentrations in BAL ELF and BAL AM will be 
calculated as described in Section 3.3.2. The ratio of free plasma concentration in apramycin to total plasma 
concentration in apramycin will be calculated to determine the free portion (fu, % unbound) of plasma 
apramycin. Drug concentrations will be summarized and listed by participant: 
• Participant level concentrations of total apramycin and free apramycin in plasma (Listing 35 and 
Listing 36, respectively). 
• Participant level free portion (fu, % unbound) of plasma apramycin (Listing 37). 
• Participant level parameters used to derive apramycin concentrations in ELF (see Section 3.3.2.1) and 
calculated apramycin concentrations in ELF (Listing 39). 


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• Participant level parameters used to derive apramycin concentrations in AM (see Section 3.3.2.2) and 
calculated apramycin concentrations in AM (Listing 40). 
• Participant level ELF and AM to total apramycin in plasma concentration ratios (Listing 41). 
• Participant level BAL cell counts (Listing 42). 
Participant level PK parameter concentration listings will include separate columns for concentrations 
reported by the lab and concentrations used for analysis. The lab reported concentrations may also include 
codes such as “BQL” and “QNS” (Quantity Not Sufficient), while the analysis concentrations will contain 
numeric data only, including imputed values, such as 0 for BQL samples collected prior to the first 
quantifiable sample. Listings will also indicate the nominal time (i.e., the planned sampling timepoint) and the 
actual post-dose collection time in hours and will note which samples were collected out of window, 
substantially out of window, or imputed. 
PK plasma concentrations of apramycin will be presented in tables and figures by nominal timepoint: 
• Individual concentrations in plasma and summary statistics will be presented tabularly for total 
apramycin in plasma (Table 83) and free apramycin in plasma (Table 84). 
• Summary statistics for the free portion of plasma apramycin (fu, % unbound), calculated as the ratio of 
free to total apramycin concentrations in plasma (Table 85). 
• Linear plots of individual participant concentration-time profiles for total apramycin in plasma 
(Figure 107). 
• Semi-log plots of individual participant concentration-time profiles for total apramycin in plasma 
(Figure 108). 
• Linear plots of mean concentration-time profiles for total apramycin in plasma, with error bars 
representing ± 1 SD (Figure 109). 
• Semi-log plots of mean concentration-time profiles for total apramycin in plasma (Figure 110). 
Additionally, lung PK (ELF and AM) apramycin concentrations in plasma will be presented in tables and 
figures by nominal timepoint. ELF and AM concentrations will be calculated as described in Section 3.3.2. 
For tables and figures of lung PK concentrations that include plasma concentrations, only the plasma samples 
collected at the same time as the BAL procedure will be used.  
• Summary statistics by BAL sampling timepoint for apramycin in ELF concentrations, total apramycin 
in plasma concentrations, and the concentration ratio of ELF to plasma concentrations will be 
presented (Table 87). 
• Summary statistics by BAL sampling timepoint for apramycin in AM concentrations, total apramycin 
in plasma concentrations, and the concentration ratio of AM to plasma concentration will be presented 
(Table 88). 
• Plot of individual total apramycin in plasma, ELF, and AM concentrations at each BAL sampling 
timepoint (Figure 111).  
• Linear plots of GM apramycin concentrations by compartment (total apramycin in plasma, apramycin 
in ELF, and apramycin in AM) at each BAL sampling timepoint (Figure 112). 
• Semi-log plots of GM apramycin concentrations by compartment (total apramycin in plasma, 
apramycin in ELF, and apramycin in AM) at each BAL sampling timepoint (Figure 113).  


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• Plot of individual ELF and AM to total apramycin in plasma concentration ratios at each BAL 
sampling timepoint (Figure 114). 
• Plot of GM ELF and AM to total apramycin in plasma concentration ratios at each BAL sampling 
timepoint (Figure 115).  
10.2. 
Noncompartmental Analysis 
PK parameters for plasma and lung PK data will be estimated through NCA using version 8.2 or higher of 
Phoenix WinNonlin®. Actual post-dose times will be used for the estimation of plasma PK parameters, and 
nominal time will be used for the estimation lung (ELF and AM) PK parameters. In the case of unknown 
sample collection times, the imputed time will be included in NCA. Any outliers identified in the PK analysis 
will be discussed in the analysis report. Outliers will not be excluded from the PK analysis.  
Individual plasma PK parameter estimates for total apramycin will be listed (Listing 38). Plasma PK 
parameters will be summarized in Table 86 for total apramycin. Summary statistics will include mean, SD, 
min, max, GM, and coefficient of variation as a percent (CV%).  
The CV% will be calculated using the method for log-normally distributed data: 
𝐶𝐶𝐶𝐶% = ඥexp(𝑠𝑠2) −1 × 100% 
Where s2 is the variance of the natural log-transformed data. 
Phoenix WinNonlin NCA will use the following settings to compute parameters from plasma and lung PK 
data: 
• Linear Up Log Down calculation method 
• Uniform weighting 
• IV infusion 
• Lambda Z Acceptance Criteria 
o Rsq _adjusted ≥ 0.90 
o Span ≥ 3 half-lives 
o Includes at least 3 timepoint after Tmax 
If an insufficient number of participants meet the Lambda Z Acceptance Criteria for computing PK 
parameters, then relaxed Lambda Z Acceptance Criteria may be used and will be described in the CSR. 
Cmax  
Cmax is defined as the maximum drug or concentration observed over all PK sample concentrations. It will be 
obtained from the Cmax parameter calculated by WinNonlin. If there is no measurable concentration in the 
participant’s PK profile, then Cmax will be missing for that participant. Cmax will be reported in units of µg/mL. 
Tmax  
Time of maximum concentration (Tmax) is defined as the time at which the Cmax occurs. It will be obtained 
from the Tmax parameter calculated by WinNonlin. If there is no measurable Cmax in the participant’s PK 
profile, then Tmax will be missing for that participant. Tmax will be reported in units of h. 


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Ke 
The terminal phase elimination rate constant (Ke) is defined as the first-order rate constant describing the rate 
of decrease of drug or metabolite concentration in the terminal phase (defined as the terminal region of the PK 
curve where drug or metabolite concentration follows first-order elimination kinetics). Ke will be computed as 
the slope of a terminal region consisting of > 3 successive points in the plot of log-transformed concentration 
data versus time. Ke will be estimated using uniform weighting.  
Timepoints used in the estimation of Ke will be initially selected using the WinNonlin automatic algorithm. 
Manually chosen timepoints may be used at the discretion of the PK analyst after examination of the 
automatically chosen points in the context of the semi-log profile to improve estimation of Ke on a case-by-
case basis. The set of points chosen must satisfy the Lambda Z Acceptance Criteria described above. 
Otherwise, the elimination rate constant and all derived parameters (apparent terminal elimination half-life 
[t½], AUC Extrapolated to Infinity [AUC(0-∞)], clearance [CL], and volume of distribution [Vd]) will be treated 
as missing.  
Drug concentration used to calculate Ke will be indicated in Listing 35. This parameter will be obtained from 
the Lambda_z parameter calculated by WinNonlin. Ke will be reported in units of 1/h.  
t½ 
The t½ is defined as the time required for the drug or metabolite concentration to decrease by a factor of one-
half in the terminal phase. The t½ can be estimated as ln(2)/Ke. It will be obtained from the HL_Lambda_z 
parameter calculated by WinNonlin. Half-life will be reported in units of h. If the Lambda Z Acceptance 
Criteria is not met, t½ will be treated as missing. 
AUC 
AUC(0-8) and AUC(0-24) are defined as the areas under the concentration-time curve from dosing (time 0) to 8 h 
and 24 h post infusion start, respectively. AUC(0-last) is defined as the area under the concentration-time curve 
from dosing (time 0) to the time of the last measured concentration. AUC(0-8), AUC(0-24), and AUC(0-last) will 
be estimated using the Linear Up Log Down calculation method and obtained from the AUClast parameter 
calculated by WinNonlin (for AUC(0-8) and AUC(0-24), data will be subset to only include results collected up 
to and including the 8 h and 24 h post-dose timepoint, respectively, and a separate NCA will be run). 
AUC(0-∞) is defined as the total area under the concentration-time curve from dosing (time 0) taken to the limit 
as the end time becomes arbitrarily large. AUC(0-∞) can be calculated by adding AUC(0-last) to an extrapolated 
value equal to the last measured concentration greater than the LLOQ divided by Ke: 
AUC(0-∞)  = AUC(0-last) + Clast
 Ke
 
Where Clast is the last measured concentration > LLOQ. AUC(0-∞) will be obtained from the AUCINF_obs 
parameter calculated by WinNonlin®. If the Lambda Z Acceptance Criteria are not met, AUC(0-∞) will be 
treated as missing. 
%AUCex is defined as percentage of AUC(0-∞) obtained by extrapolation from time of the last measured 
concentration to infinity. %AUCex can be calculated by dividing AUC from time of the last measured 
concentration to infinity by AUC(0-∞):  
%AUC𝑒𝑒𝑒𝑒=
AUC(0−∞) −AUC(0−last)
AUC(0−∞)
, 
 


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If %AUCex is >20% or the Lambda Z Acceptance Criteria is not met, the estimated AUC(0-∞) will be excluded 
from statistical summaries of PK parameter estimates and downstream calculations. %AUCex will be obtained 
from the AUC_%Extrap_obs parameter calculated by WinNonlin.  
All AUCs will be reported in units of µg*h/mL. 
CL 
Clearance (CL) is defined as the volume of plasma completely cleared of drug per unit time and is estimated 
in trials of an IV-administered drug as the dose divided by the AUC(0-∞). It will be obtained from the CL_obs 
parameter calculated by WinNonlin. If %AUCex is >20% or the Lambda Z Acceptance Criteria are not met, 
the estimated CL value will be flagged when listed in the report and will be excluded from statistical 
summaries of parameter estimates and downstream calculations. CL will be reported in units of L/h.  
Vd 
Volume of distribution (Vd) central is estimated in trials of an IV-administered drug of CL divided by Ke.  It 
will be obtained from the Vz_obs parameter calculated in WinNonlin. If %AUCex is >20% or the Lambda Z 
Acceptance Criteria is not met, the estimated Vd value will be flagged when listed in the report and will be 
excluded from statistical summaries of parameter estimates and downstream calculations. Vd will be reported 
in units of L/h.  
10.2.1. 
Ratio of Lung PK to Plasma PK  
Only plasma samples collected at the same time as the BAL procedure will be used in the calculations of the 
Lung PK to Plasma PK ratios. The GM concentrations of apramycin in ELF, AM, and plasma from the BAL 
sampling timepoints will be used to estimate the population average Cmax, AUC(0-∞), AUC(0-24), and AUC(0-last). 
AUC(0-∞), AUC(0-24), and AUC(0-last) will be estimated using the Linear Up Log Down calculation method. 
Since the calculated concentrations used at each timepoint will be based on measurements from multiple 
participants, nominal timepoints will be used when estimating these parameters. 
The population estimates of AUC(0-24), AUC(0-t), AUC(0-∞), Cmax, Tmax, and t½ for apramycin in ELF and AM 
will be presented in Table 89. The population estimates of Cmax, AUC(0-∞), AUC(0-24), and AUC(0-last) in 
plasma, ELF, and AM, as well as the ratios of ELF-to-plasma and AM-to-plasma exposure parameters will be 
presented in Table 90 for total apramycin.  
 


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11. 
IMMUNOGENICITY 
There are no immunogenicity endpoints in this trial. 


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12. 
OTHER ANALYSES 
No applicable. 


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13. 
REPORTING CONVENTIONS 
The mean, median, SD, and other statistics will be reported to 1 decimal place greater than the original data. 
The min and max will use the same number of decimal places as the original data. Proportions will be 
presented to 2 decimal places; values greater than zero but < 0.01 will be presented as “<1”. Percentages will 
be reported to the nearest whole number; values greater than zero by < 1% will be presented as “<1”; values 
greater than 99% but less than 100% will be reported as “>99%”.  
For PK parameters, AUCs will be reported using 3 significant digits. t½, Tmax, CL, and Vd values will be 
reported using 2 significant digits. Ke values will be reported to 3 significant digits. Cmax will be reported with 
the same number of significant digits as the measurement. 
Listings of individual participant data will include a Participant ID column. The participant identifiers 
assigned by site staff are replaced throughout this report with the Study Data Tabulation Model (SDTM) 
variable USUBJID to protect the confidentiality of those who volunteered to participate in this protocol. 
USUBJID has been created as a composite of the 3-letter EDC platform code followed by a numeric identifier 
assigned chronologically to enrolled participants as well as Screening failures across all sites and protocols in 
the EDC platforms. Any data sharing activities will include the USUBJID and not the participant identifiers 
assigned at the study site. 


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14. 
TECHNICAL DETAILS 
SAS version 9.4 or above or R version 4.1 or above will be used to generate all tables, figures, and listings. 
PK parameters will be estimated through NCA using Phoenix® WinNonlin version 8.2 or later. 


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15. 
SUMMARY OF CHANGES IN THE CONDUCT OF THE STUDY OR 
PLANNED ANALYSES 
If there are any changes to the planned analysis prior to final data lock and after finalization of the SAP, they 
may be added to the SAP as an addendum. The SAP will not be amended after final data lock. 


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16. 
REFERENCES 
1. ICH Harmonised Tripartite Guideline. Structure and Content of Clinical Study Reports E3. International 
Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human 
Use. 1995. Retrieved from: https://database.ich.org/sites/default/files/E3_Guideline.pdf. 
2. ICH Harmonised Tripartite Guideline. General Considerations for Clinical Trials E8. International 
Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human 
Use. 1997. Retrieved from: https://database.ich.org/sites/default/files/E8_Guideline.pdf. 
3. ICH Harmonised Tripartite Guideline. Statistical Principles for Clinical Trials E9. International 
Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human 
Use. 1998. Retrieved from: https://database.ich.org/sites/default/files/E9_Guideline.pdf. 
4. American Statistical Association Guideline. Ethical Guidelines for Statistical Practice. American 
Statistical Association. 2022. Retrieved from: https://www.amstat.org/docs/default-source/amstat-
documents/ethicalguidelines.pdf?sfvrsn=bdeeafdd_3. 
5. Royal Statistical Association Guideline. Code of Conduct. The Royal Statistical Association. 2014. 
Retrieved from: https://rss.org.uk/RSS/media/File-library/About/2019/RSS-Code-of-Conduct-2014.pdf.  
6. Cassini A, Högberg LD, Plachouras D, Quattrocchi A, Hoxha A, Simonsen GS, et al.; Burden of AMR 
Collaborative Group. Attributable deaths and disability-adjusted life-years caused by infections with 
antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: a population-level 
modelling analysis. Lancet Infect Dis. Lancet Infect Dis. 2019;19(1):56-66. 
7. CDC. Antibiotic Resistance Threats in the United States, 2019. Atlanta, GA: U.S. Department of Health 
and Human Services, CDC (CDC 2019); 2019.  Accessed in: 
https://www.cdc.gov/drugresistance/pdf/threats-report/2019-ar-threats-report-508.pdf 
8. O’Neill, J. (2014). Antimicrobial Resistance: Tackling a Crisis for the Future Health and Wealth of 
Nations. HM Government. Accessed in: https://wellcomecollection.org/works/rdpck35v 
9. O’Connor S, Lam LKT, Jones ND, Chaney MO. Apramycin, a unique aminocyclitol antibiotic. L Org 
Chem 1976; 41 (12): 2087-2092.  https://doi.org/10.1021/jo00874a003 
10. Tenero D, Bowers G, Rodvold KA, Patel A, Kurtinecz K, Dumont E, Tomayko J, Patel P. Intrapulmonary 
Pharmacokinetics of GSK2251052 in Healthy Volunteers. Antimicrob Agents Chemother 2013; 57(7): 
3334-3339 | DOI: 10.1128/AAC.02483-12. 
11. Gotfried MH, Horn K, Garrity-Ryan L, Villano S, Tzanis E, Chitra S, Manley A, Tanaka SK, Rodvold 
KA. Comparison of omadacycline and tigecycline pharmacokinetics in the plasma, epithelial lining fluid, 
and alveolar cells of healthy adult subjects. Antimicrob Agents Chemother 2017; 61:e01135-17. 
https://doi.org/10.1128/AAC.01135-17. 
12. Rizk ML, Rhee EG, Jumes PA, Gotfried MH, Zhao T, Mangin E, Bi S, Chavez-Eng CM, Zhang Z, 
Butterton JR. Intrapulmonary pharmacokinetics of relebactam, a novel β-lactamase inhibitor, dosed in 
combination with imipenem-cilastatin in healthy subjects. Antimicrob Agents Chemother 2018; 
62:e01411-17. https://doi.org/10.1128/AAC.01411-17. 
13. Investigator’s Brochure, EBL-1003 (apramycin). V8.0, Juvabis AG, 28 January 2021. 
14. Sou T, Hansen J, Liepinsh E, et al. Model-Informed Drug Development for Antimicrobials: Translational 
PK and PK/PD Modeling to Predict an Efficacious Human Dose for Apramycin. Clinical Pharmacology 
and Therapeutics 2021; 109(4); 1063-1073. doi:10.1002/cpt.2104. 


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17. 
LISTING OF TABLES, FIGURES, AND LISTINGS  
Table, figure, and listing shells are presented in Appendix 1, Appendix 2 and Appendix 3. 
 


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APPENDICES 


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APPENDIX 1. TABLE MOCK-UPS 
LIST OF TABLES 
Table 1: 
Analysis Timepoints for Safety and PK Endpoints ....................................................14 
Table 2: 
Schedule of Study Procedures ....................................................................................58 
Table 3: 
Clinical Adverse Event Toxicity Grading Scales .......................................................61 
Table 4: 
Clinical Laboratory Reference Ranges and Toxicity Grading Scales ........................64 
Table 5: 
Vital Signs Toxicity Grading Scales ...........................................................................66 
Table 6: 
 ECG Toxicity Grading Scale .....................................................................................67 
Table 7: 
Distribution of Protocol Deviations by Category, Type, and Cohort – All 
Enrolled Participants ...................................................................................................68 
Table 8: 
Participant Disposition by Cohort – All Enrolled Participants ...................................70 
Table 9: 
Treatment Discontinuations or Early Terminations by Reason and Cohort – 
All Enrolled Participants .............................................................................................71 
Table 10: Ineligibility Summary of Screen Failures ...................................................................72 
Table 11: Analysis Population Exclusions by Cohort – All Enrolled Participants .....................73 
Table 12: Summary of Categorical Demographic and Baseline Characteristics by 
Cohort – All Enrolled Participants .............................................................................74 
Table 13: Summary of Continuous Demographic and Baseline Characteristics by 
Cohort – All Enrolled Participants .............................................................................75 
Table 14: Summary of Participants with Pre-Existing Medical Conditions by MedDRA 
System Organ Class and Cohort – All Enrolled Participants .....................................76 
Table 15: Overall Summary of Adverse Events – Safety Population .........................................77 
Table 16: Overall Summary of Adverse Events – Safety Subset Population .............................77 
Table 17: Summary of Treatment Emergent Adverse Events by MedDRA System 
Organ Class, High Level Group Term, and Preferred Term – Safety 
Population ...................................................................................................................78 
Table 18: Summary of Treatment Emergent Adverse Events by MedDRA System 
Organ Class, High Level Group Term, and Preferred Term – Safety Subset 
Population ...................................................................................................................78 
Table 19: Treatment Emergent Adverse Events by MedDRA System Organ Class, 
High Level Group Term, and Preferred Term, Maximum Severity and 
Relationship – Safety Population................................................................................79 
Table 20: Treatment Emergent Adverse Events by MedDRA System Organ Class, 
High Level Group Term, and Preferred Term, Maximum Severity and 
Relationship – Safety Subset Population ....................................................................80 
Table 21: Listing of Serious Adverse Events .............................................................................81 


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Table 22: Listing of Non-Serious, Moderate or Severe Treatment Emergent Adverse 
Events .........................................................................................................................82 
Table 23: Listing of Abnormal Laboratory Results – Chemistry ...............................................84 
Table 24: Listing of Abnormal Laboratory Results – Hematology ............................................85 
Table 25: Listing of Abnormal Laboratory Results – Coagulation ............................................86 
Table 26: Listing of Abnormal Laboratory Results - Urinalysis ................................................87 
Table 27: Chemistry Abnormal Results by Parameter and Timepoint – Safety 
Population ...................................................................................................................88 
Table 28: Chemistry Abnormal Results by Parameter and Timepoint – Safety Subset 
Population ...................................................................................................................88 
Table 29: Chemistry Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Population ......................................................................................89 
Table 30: Chemistry Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Subset Population ...........................................................................89 
Table 31: Chemistry Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Population ...........................................................90 
Table 32: Chemistry Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Subset Population ...............................................90 
Table 33: Hematology Abnormal Results by Parameter and Timepoint – Safety 
Population ...................................................................................................................91 
Table 34: Hematology Abnormal Results by Parameter and Timepoint – Safety Subset 
Population ...................................................................................................................91 
Table 35: Hematology Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Population ......................................................................................91 
Table 36: Hematology Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Subset Population ...........................................................................91 
Table 37: Hematology Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Population ...........................................................91 
Table 38: Hematology Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Subset Population ...............................................91 
Table 39: Coagulation Abnormal Results by Parameter and Timepoint – Safety 
Population ...................................................................................................................92 
Table 40: Coagulation Abnormal Results by Parameter and Timepoint – Safety Subset 
Population ...................................................................................................................92 
Table 41: Coagulation Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Population ......................................................................................92 


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Table 42: Coagulation Laboratory Toxicity Grade by Parameter, Timepoint, and 
Severity – Safety Subset Population ...........................................................................92 
Table 43: Coagulation Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Population ...........................................................92 
Table 44: Coagulation Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Subset Population ...............................................92 
Table 45: Urinalysis Abnormal Results by Parameter and Timepoint – Safety 
Population ...................................................................................................................93 
Table 46: Urinalysis Abnormal Results by Parameter and Timepoint – Safety Subset 
Population ...................................................................................................................93 
Table 47: Urinalysis Laboratory Toxicity Grade by Parameter, Timepoint, and Severity 
– Safety Population .....................................................................................................93 
Table 48: Urinalysis Laboratory Toxicity Grade by Parameter, Timepoint, and Severity 
– Safety Subset Population .........................................................................................93 
Table 49: Kidney Function Biomarker Summary Statistics Measurement and Change 
from Baseline by Parameter and Timepoint – Safety Population ...............................94 
Table 50: Kidney Function Biomarker Summary Statistics Measurement and Change 
from Baseline by Parameter and Timepoint – Safety Subset Population ...................94 
Table 51: Vital Sign Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population ...................................................................................................................95 
Table 52: Vital Sign Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Subset Population .......................................................................................................95 
Table 53: Vital Sign Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Population ...........................................................96 
Table 54: Vital Sign Summary Statistics Measurement and Change from Baseline by 
Parameter and Timepoint – Safety Subset Population ...............................................96 
Table 55: ECG Overall Interpretations, Post-Dose Compared to Pre-Dose Baseline by 
Timepoint – Safety Population ...................................................................................97 
Table 56: ECG Overall Interpretations, Post-Dose Compared to Pre-Dose Baseline by 
Timepoint – Safety Subset Population .......................................................................97 
Table 57: ECG Summary in Change from Pre-Dose Baseline by Timepoint – Safety 
Population ...................................................................................................................98 
Table 58: ECG Summary in Change from Pre-Dose Baseline by Timepoint – Safety 
Subset Population .......................................................................................................98 
Table 59: ECG Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population ...................................................................................................................99 
Table 60: ECG Toxicity Grade by Parameter, Timepoint, and Severity – Safety Subset 
Population ...................................................................................................................99 


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Table 61: ECG Measurement and Change from Pre-Dose Baseline by Parameter and 
Timepoint – Safety Population .................................................................................100 
Table 62: ECG Measurement and Change from Pre-Dose Baseline by Parameter and 
Timepoint – Safety Subset Population .....................................................................100 
Table 63: ECG Change from Time-Matched Baseline by Parameter and Timepoint – 
Safety Population ......................................................................................................101 
Table 64: ECG Change from Time-Matched Baseline by Parameter and Timepoint – 
Safety Subset Population ..........................................................................................101 
Table 65: Categorical Summary of QTcF Interval by Timepoint – Safety Population ............102 
Table 66: Categorical Summary of QTcF Interval by Timepoint – Safety Subset 
Population .................................................................................................................102 
Table 67: Categorical Summary of PR Interval, QRS Duration, and RR Interval by 
Timepoint – Safety Population .................................................................................103 
Table 68: Categorical Summary of PR Interval, QRS Duration, and RR Interval by 
Timepoint – Safety Subset Population .....................................................................103 
Table 69: Ear Otoscopy Result, Post-Dose Compared to Baseline by Timepoint and 
Ear – Safety Population ............................................................................................104 
Table 70: Ear Otoscopy Result, Post-Dose Compared to Baseline by Timepoint and 
Ear – Safety Subset Population.................................................................................104 
Table 71: Pure-Tone Audiometry Toxicity Grade by Timepoint and Severity – Safety 
Population .................................................................................................................105 
Table 72: Pure-Tone Audiometry Toxicity Grade by Timepoint and Severity – Safety 
Subset Population .....................................................................................................105 
Table 73: Pure-Tone Audiometry Threshold Measurement and Change from Baseline 
by Timepoint, Frequency, and Ear – Safety Population ...........................................106 
Table 74: Pure-Tone Audiometry Threshold Measurement and Change from Baseline 
by Timepoint, Frequency, and Ear – Safety Subset Population ...............................107 
Table 75: DPOAE Measurement and Change from Baseline by Timepoint, Frequency, 
and Ear – Safety Population .....................................................................................108 
Table 76: DPOAE Measurement and Change from Baseline by Timepoint, Frequency, 
and Ear – Safety Subset Population ..........................................................................108 
Table 77: Summary of DPOAE Results by Timepoint, Frequency, and Ear – Safety 
Population .................................................................................................................109 
Table 78: Summary of DPOAE Results by Timepoint, Frequency, and Ear – Safety 
Subset Population .....................................................................................................109 
Table 79: SNR Change from Baseline by Timepoint, Frequency, and Ear – Safety 
Population .................................................................................................................110 


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Table 80: SNR Change from Baseline by Timepoint, Frequency, and Ear – Safety 
Subset Population .....................................................................................................110 
Table 81: Number and Percentage of Participants with Prior Medications by WHO 
Drug Classification and Cohort – Safety Population ................................................111 
Table 82: Number and Percentage of Participants with Concomitant Medications by 
WHO Drug Classification and Cohort – Safety Population .....................................112 
Table 83: Total Apramycin Concentrations in Plasma .............................................................113 
Table 84: Free Apramycin Concentrations in Plasma ..............................................................113 
Table 85: Summary Statistics for the Free Portion of Plasma Apramycin ...............................114 
Table 86: Summary Statistics for Total Apramycin in Plasma PK Parameters ........................115 
Table 87: Summary Statistics for Apramycin in ELF Concentrations and Apramycin in 
ELF to Total Apramycin in Plasma Concentration Ratios .......................................116 
Table 88: Summary Statistics for Apramycin in AM Concentrations and Apramycin in 
AM to Total Apramycin in Plasma Concentration Ratios ........................................116 
Table 89: Apramycin in ELF and Apramycin in AM Population PK Parameter 
Estimates ...................................................................................................................117 
Table 90: Apramycin in ELF to Total Apramycin in Plasma and Apramycin in AM to 
Total Apramycin in Plasma PK Parameter Ratios by PK Parameter .......................118 
 


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9.5.1 Pharmacokinetic and Safety Measurements Assessed and Flow Chart 
Table 2: 
Schedule of Study Procedures 
Procedures and Evaluations 
Out- 
patient 
In- 
patient 
In- 
patient 
Treatment Period (In-patient) 
Follow-up  
(out-patient) 
Early 
Termina-
tion 
Unsche-
duled 
Day -28  
to -3 
Day -2 
Day -1 
Day 1 
Day 2 
Day 3 
Day 14 
 (± 3 
days) 
Day 
30 
(±4 
days) 
Screening 1 Check-in Baseline Pre-
dose 
0 h 
(dose) 
0.5 h 
(±5 
min) 
1 h  
(±5 
min) 
2 h  
(±5 
min) 
4 h  
(±10 
min) 
8 h 
(±15 
min) 
16 h 
(±15 
min) 
24 h 
(±1 h) 
36 h  
(±1  h) 
48 h 
(±1 h) 
60 h (±1 
h) 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion and exclusion criteria 
X 2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Review of Inclusion and exclusion criteria 3 
 
X 3 
X 3 
X 3 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history (including family history of 
auditory loss, renal disorders and sudden death) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical History update 4 
 
X 4 
X 4 
X 4 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Prior medication 
X 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Concomitant medication 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Complete Physical examination (PE) 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
 
X 
 
Abbreviated PE 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Symptom-directed (Focused) PE 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Vital signs (BP, HR, RR, T) 5 
X 
X 
X 
X 
 
X 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
Height, Weight, BMI calculation 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Weight 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
Spirometry (FEV1 and FEV1/FVC) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12-lead ECG (single recording) 6 
X 
X 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
X 
 
X 
X 
12-lead ECG (triplicate recording)7 
 
 
X 7 
X 7 
 
 
X 7 
 
X 7 
 
X 7 
 
 
 
 
 
 
 
 
Viral Serology 8 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Alcohol breathalyzer test 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine drug toxicity and  cotinine 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical laboratory tests 9 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
Calculation of glomerular filtration rate (CKD-
EPI) 10 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
 
Urinalysis 11 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
Serum β-HCG pregnancy test (females only) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serum FSH (post-menopausal females only) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Table 2: 
Schedule of Study Procedures (Continued) 
 
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Procedures and Evaluations 
Out- 
patient 
In- 
patient 
In- 
patient 
Treatment Period (In-patient) 
Follow-up  
(out-patient) 
Early 
Termina-
tion 
Unsche-
duled 
Day -28  
to -3 
Day -2 
Day -1 
Day 1 
Day 2 
Day 3 
Day 14 
 (± 3 
days) 
Day 
30 
(±4 
days) 
Screening 1 Check-in Baseline Pre-
dose 
0 h 
(dose) 
0.5 h 
(±5 
min) 
1 h  
(±5 
min) 
2 h  
(±5 
min) 
4 h  
(±10 
min) 
8 h 
(±15 
min) 
16 h 
(±15 
min) 
24 h 
(±1 h) 
36 h  
(±1  h) 
48 h 
(±1 h) 
60 h (±1 
h) 
Urine pregnancy test (females only) 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood sample for exploratory biomarkers 12 
 
X 
 
X 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
X 
 
Urine sample for exploratory biomarkers 13 
 
  X 
 
X 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
X 
 
Ear otoscopy, Valsalva test, tympanometry, 
stapedial reflexes 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Ear otoscopy 
 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
 
Ear audiometry and otoacoustic tests 14 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
 
Admission 15 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Enrollment and Assignment to T cohorts 16 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Discharge 17 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
IV administration of apramycin 18 
 
 
 
 
X-------
21 
 
 
 
 
 
 
 
 
 
 
 
 
 
Check of infusion site 
 
 
 
 
X 
X 
X 
 
 
 
X 
X 
X 
X 
X 
X 
 
X 
 
Treatment- emergent adverse events (TEAE) 
and serious AEs (SAEs) 19 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Blood (plasma) sampling for apramycin PK 20 
 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
X 
 
Protein Binding Test 21 
 
 
 
 
 
X 
 
 
 
 
 
 
X 
 
 
 
 
 
 
Administration of sedative or anesthetic 
medication 22 
 
 
 
 
 
X 
 
X 
X 
X 
 
X 
 
 
 
 
 
 
 
Bronchoscopy /BAL for ELF and AM PK) 23 
 
 
 
 
 
X 
 
X 
X 
X 
 
X 
 
 
 
 
 
 
 
Oxygenation support (TBC) 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
 
 
 
 
Drinking schedule 24 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
X 
 
Counseling 25 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
1. Screening including Ear -Nose-Throat (ENT)/Audiology assessments are planned to be completed in a single Screening Visit. If schedule cannot be accommodated, ENT/Audiology assessments will be 
completed on a second visit within the screening period. 
2. Inclusion and exclusion criteria regarding ENT/Audiology assessments to be reviewed by audiologist. 
3. Review of inclusion and exclusion criteria on Day -2 to determine eligibility for admission; on Day -1, before 12-lead ECG, to determine eligibility for enrollment; and on Day 1 pre-dose to confirm the 
participant remains eligible for dosing. 
4 Medical history update to include pre-dose events.  
5. Vital signs include systolic and diastolic BP, heart rate, respiratory rate and oral temperature. 
6. 12-lead ECG single recording with 10 sec rhythm strip.   


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Table 2: 
Schedule of Study Procedures (Continued) 
 
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Procedures and Evaluations 
Out- 
patient 
In- 
patient 
In- 
patient 
Treatment Period (In-patient) 
Follow-up  
(out-patient) 
Early 
Termina-
tion 
Unsche-
duled 
Day -28  
to -3 
Day -2 
Day -1 
Day 1 
Day 2 
Day 3 
Day 14 
 (± 3 
days) 
Day 
30 
(±4 
days) 
Screening 1 Check-in Baseline Pre-
dose 
0 h 
(dose) 
0.5 h 
(±5 
min) 
1 h  
(±5 
min) 
2 h  
(±5 
min) 
4 h  
(±10 
min) 
8 h 
(±15 
min) 
16 h 
(±15 
min) 
24 h 
(±1 h) 
36 h  
(±1  h) 
48 h 
(±1 h) 
60 h (±1 
h) 
7. 12-lead ECG: triplicate safety ECGs with 10 sec rhythm strips on Day -1 at the same timepoints as the projected timepoints for ECGs to be recorded on Day 1 within 30 min of dosing, and at 1h (± 5 
min), 4h (± 10 min) and 16 h (± 15 min) after starting the apramycin dose).  Record ECG before VS and PK draws and, at 4 h post-dosing, before administration of sedation or anesthetic for BAL in cohort 
T3. 
8. Hepatitis B surface antigen (HBsAg), hepatitis virus C antibody (HCV) and human immunodeficiency virus (HIV) antibody. 
9. Clinical laboratory tests under fasting conditions: hematology, coagulation, and clinical chemistry panels and thyroid stimulating hormone (TSH) & free T3/free T4.  
10.The CKD-EPI equation, expressed as a single equation, is GFR = 141 × min(Scr/κ, 1)α × max(Scr/κ, 1)-1.209 × 0.993Age × 1.018 [if female] _ 1.159 [if black], where Scr is serum creatinine, κ is 0.7 
for females and 0.9 for males, α is -0.329 for females and -0.411 for males, min indicates the minimum of Scr/κor 1, and max indicates the maximum of Scr/κ or 1 [Levey 2009].(Formula will be used and 
result will be included in the lab report) 
11. Urinalysis (UA) by dipstick. If abnormal for blood, protein and glucose, perform microscopic UA. The results of microscopic UA supersede those of UA by dipstick. UA may be postponed in a 
menstruating female. Abnormal UA in a menstruating female is not graded as a TEAE; the UA should be completed after the end of menstruation. 
12. Cystatin C in serum. 
13 Kidney-injury molecule 1 (KIM-1) in urine. 
14 Pure-tone audiometry with standard frequencies and distortion product otoacoustic emissions (DPOAEs) AE assessments at screening. Pure-tone audiometry including high frequencies and DPOAEs at 
check-in (Day -2) and at Days 3, 14 (± 3 days), and 30 (± 4 days), or ET.  
15 Participants will be admitted to the ward on Day -2 after confirmation of eligibility. 
16 Assignment to Lung PK timepoint cohorts (T1 – T5) and Enrollment to be performed before 12-lead ECG on Day -1 - Participants with ECGs on Day-1 and pre-dose on Day 1 with clinically significant 
abnormalities according to the PI should not be eligible for receiving study drug and should be withdrawn and replaced. 
17 Participant may be discharged from the CTU after Day 3 collection of the 60-h plasma PK sample. 
18.Duration of apramycin infusion is 30 ± 5 min. 
19. Adverse events from the start of dosing to the end of the trial are considered treatment-emergent adverse events (TEAE) or serious AEs (SAE). 
20. Blood samples for plasma PK are taken pre-dose, and at 30 (± 5) minutes (end of infusion) and 1 h (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h (±15 min), 16 h (±15 min), 24 h (±1 h), 36 h (±1 h), 48 h 
(±1 h) and 60 h (±1 h) after start of the infusion - Blood samples for plasma PK are taken at the nominal timepoints. If collection of BAL is delayed, plasma PK will be collected at the nominal timepoint 
and again 5 min after collection of the last BAL sample. In that case, total apramycin concentration will be measured in both samples, but only the drug concentration in the delayed plasma PK sample will 
be used to calculate the Lung PK/Plasma PK ratio. 
21. A protein binding test for the measurement of free apramycin will be performed using aliquots of plasma PK samples collected at 0.5 h (± 5 min) and 36 h (± 1 h) after 
start of infusion. 
22. Type of sedation or light anesthesia medication for bronchoscopy/BAL procedure and local anesthetic TBD by the anesthesia and bronchoscopy protocols. 
23. Each participant is assigned to a single bronchoscopy/BAL procedure. Lung PK samples (for measurement of total apramycin concentration) after initiation of infusion and corresponding cohorts (T) 
are: (Cohort T1), 30 min (±5 min); (Cohort T2), 2 h (±5 min); (Cohort T3), 4 h (±10 min); (Cohort T4), 8h (± 15 min); and (Cohort T5), 24 h (± 1 h).  Lung PK timepoints are aligned with plasma PK 
samples for measurement of total and free apramycin concentration at 30 min (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h (±15 min) and 24 h (±1 h) after start of apramycin infusion. If BAL is delayed, 
plasma sample will be collected at the nominal timepoint and again within 5 minutes after the last BAL sample collection. Urea will be measured at the same timepoints in plasma and BAL (ELF) samples. 
24 Participants to be kept hydrated water starting 30 min before until 24 h after start of infusion (document time and volume of water intake). 
25. Provide counseling at the end of each encounter with the participant. Counsel on use of contraception (males and females), avoidance of pregnancy (women of childbearing potential), avoidance of 
sperm donation, and avoidance of prohibited medications, illicit drugs, alcohol, nicotine products, vigorous exercise, and exposure to loud noises. 


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12.2.2 Displays of Adverse Events 
Table 3: 
Clinical Adverse Event Toxicity Grading Scales 
Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
CARDIOVASCULAR 
Arrhythmia 
 
Asymptomatic or transient signs; 
no medical intervention required. 
Recurrent and/or persistent signs; 
symptomatic medical intervention 
required. 
Hemorrhage 
Estimated blood loss <100 mL. 
Estimated blood loss >100 mL; no 
transfusion required. 
Blood transfusion required. 
RESPIRATORY 
Cough  
Transient cough; no treatment 
required. 
Persistent cough; treatment 
required. 
Interferes with daily activities. 
Bronchospasm, Acute 
Transient bronchospasm; no 
treatment required; FEV1 71-80% 
of predicted peak flow. 
Requires treatment; normalizes 
with bronchodilator; FEV1 60-70% 
of predicted peak flow. 
No normalization with bronchodilator; 
FEV1 <60% of predicted peak flow. 
Dyspnea   
Does not interfere with usual and 
social activities. 
Interferes with usual and social 
activities; no treatment. 
Prevents usual and social activities 
OR requires treatment. 
EAR AND LABYRINTH DISORDERS  
Hearing Impairment 
 
(Adult enrolled on a 
Monitoring Program (on 
a 1, 2, 4, 3, 6, and 8 kHz 
audiogram) 
Threshold shift of 15 - 25 dB 
averaged at 2 contiguous test 
frequencies in at least one ear. 
Threshold shift of >25 dB averaged 
at 2 contiguous test frequencies in 
at least one ear. 
Threshold shift of >25 dB averaged at 
3 contiguous test frequencies in at 
least one ear; therapeutic intervention 
indicated. 
Tinnitus 
Mild symptoms; intervention not 
indicated. 
Moderate symptoms; limiting 
instrumental ADL. 
Severe symptoms; limiting self-care 
ADL. 
Vertigo  
Mild symptoms. 
Moderate symptoms; limiting 
instrumental ADL. 
Severe symptoms; limiting self-care 
ADL. 
GASTROINTESTINAL   
Nausea  
No interference with normal 
activity. 
Some interference with normal 
activity. 
Prevents daily activities. 
Vomiting  
No interference with activity OR 
1-2 episodes in a 24-h period. 
Some interference with activity OR 
>2 episodes in a 24-h period. 
Prevents daily activity OR requires 
medical intervention. 
Diarrhea  
2-3 loose or watery stools in a 24-
h period. 
4-5 loose OR watery stools in a 24-
h period. 
6 or more loose or watery stools in a 
24-h period OR requires IV hydration 
OR requires medical intervention. 
Oral Discomfort / 
Dysphagia 
Mild discomfort; no difficulty 
swallowing. 
Some limits on eating /drinking. 
Eating/talking very limited; unable to 
swallow solid foods. 


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12.2.2 Displays of Adverse Events 
Table 3: 
Clinical Adverse Event Toxicity Grading Scales (Continued) 
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Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
LOCAL IV CATHETER REACTION 
IV site reaction 
Not applicable. 
Erythema with associated 
symptoms (e.g., edema, pain, 
induration, phlebitis). 
Ulceration or necrosis; severe tissue 
damage; operative intervention 
indicated. 
SYSTEMIC REACTIONS 
Anaphylaxis ** 
-- 
-- 
Symptomatic bronchospasm, with or 
without urticaria; parenteral 
intervention indicated; allergy-related 
edema or angioedema; 
hypotension. 
**Definition: A disorder characterized by an acute inflammatory reaction resulting from the release of histamine and histamine-like substances 
from mast cells, causing a hypersensitivity immune response. Clinically, it presents with breathing difficulty, dizziness, hypotension, cyanosis, 
and loss of consciousness, and may lead to death. 
Allergic Reaction  
Pruritus without rash. 
Localized urticaria OR requires 
oral therapy. 
Generalized urticaria OR angioedema 
OR anaphylaxis OR requires 
epinephrine 
Hypersensitivity 
(including drug fever) 
Transient flushing or rash; 
temperature 38.0-38.4°C (100.4-
101.1°F). 
Rash; flushing; urticaria; dyspnea; 
temperature 38.5 - 38.9°C (101.2 – 
102.0°F). 
Symptomatic bronchospasm with or 
without urticaria; parenteral 
medication indicated; allergy-related 
edema or angioedema; hypotension; 
temperature >38.9°C (>102.0°F).  
Headache  
No interference with activity. 
Repeated use of non-narcotic pain 
reliever for more than 24 h OR 
some interference with activity. 
Significant; any use of narcotic pain 
reliever OR prevents daily activity 
OR requires triptans. 
Fatigue  
No interference with activity. 
Some interference with activity. 
Significant; prevents daily activity. 
Myalgia 
No interference with activity. 
Some interference with activity. 
Significant; prevents daily activity. 
SKIN 
Mucocutaneous 
Erythema, pruritus. 
Diffuse, maculo-papular rash, dry 
desquamation. 
Vesiculation OR moist desquamation 
OR ulceration. 
Pruritus 
No or minimal interference with 
usual social and functional 
activities.  
Greater than minimal interference 
with usual social and functional 
activities.  
Inability to perform usual social and 
functional daily activities. 


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12.2.2 Displays of Adverse Events 
Table 3: 
Clinical Adverse Event Toxicity Grading Scales (Continued) 
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Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
ALL OTHER CONDITIONS 
Illness or clinical AE (as 
defined according to 
applicable regulations) 
Require minimal or no treatment; 
does not interfere with the 
participant’s daily activities. 
Result in a low level of 
inconvenience or concern with 
therapeutic measures; may cause 
some interference with functioning. 
Interrupt a participant’s usual daily 
activity and may require systemic 
drug therapy or other treatment; are 
usually incapacitating. 
 


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12.4.1 Individual Laboratory Measurements and Abnormal Laboratory Values 
Table 4: 
Clinical Laboratory Reference Ranges and Toxicity Grading Scales 
Laboratory AEs 
Reference 
Ranges 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Blood, serum, or plasma 
HEMATOLOGY 
 
 
 
 
Hemoglobin decrease, female – g/dL 
11.1 – 15.9 
9.6 – 11.0 
8.0 – 9.5 
<8.0 
Hemoglobin decrease, male – g/dL 
13.0 – 17.7 
11.0 – 12.9 
8.5 – 10.9 
<8.5 
WBC increase – x103/ µL 
3.4 – 10.8 
>10.8 – 15.0 
>15.0 – 20.0 
>20.0 
WBC decrease – x103/ µL 
3.4 – 10.8 
2.4 – <3.4 
1.4 – <2.4 
<1.4 
Neutrophils decrease – x103/ µL 
1.4 – 7.0 
1.0 – <1.4 
0.75 – <1.0 
<0.75  
Lymphocytes decrease – x103/ µL 
0.7 – 3.1 
<0.7 –0.4 
<0.4 – 0.3 
<0.30 
Monocytes increase – x103/ µL 
0.1 - 0.9 
>0.9 – 2.0 
>2.0 – 3.0 
>3.0 
Eosinophils increase – x103/ µL 
0.0 – 0.4 
>0.4 – 0.75 
>0.75 – 1.0 
>1.0 
Basophils increase – x103/ µL 
0.0 – 0.2 
>0.2 – 0.5 
>0.5 – 0.8 
>0.8 
Platelets decrease – x103/ µL 
150 - 450 
90 – <150 
55 – <90 
<55 
COAGULATION 
PT INR 
0.9 – 1.2 
>1.2 – 1.8 
>1.8 – 2.1 
>2.1 
Prothrombin Time 
9.1 – 12.0 
>12.0 – 15.0 
>15.0 – 18.6 
>18.6 
Activated Partial Thromboplastin Time (APTT)  
24 - 33 
>33.0 – 54.0 
>54.0 – 75.0 
>75.0 
CHEMISTRY 
Sodium decrease – mmol/L 
134 – 144 
130 - 133 
124 –129 
<124 
Sodium increase – mmol/L 
134 – 144 
145 - 150 
 151 – 156 
>156 
Potassium increase – mmol/L 
3.5 – 5.2 
5.3 – 6.0 
6.1 – 6.5 
>6.5 
Potassium decrease – mmol/L 
3.5 – 5.2 
3.4– 3.2  
2.5 – 2.9 
<2.5 
Total Carbon Dioxide (CO2) increase – mmol/L 
20 – 29 
30 - 35 
36 - 37 
>37 
Total Carbon Dioxide (CO2) decrease – mmol/L 
20 – 29 
17 - 19 
14 - 16 
<14 
Calcium decrease – mg/dL 
8.7 – 10.2 
7.8 – 8.6 
7.0 – 7.7 
<7.0 
Calcium increase – mg/dL 
8.7 – 10.2 
10.3 – 11.4 
11.5 – 12.5 
>12.5 
Magnesium decrease – mg/dL 
1.6 – 2.3 
1.2 – 1.5 
1.1 – 0.9 
<0.9 
Blood urea nitrogen (BUN) increase, 
18-39Y - mg/dL 
6 – 20 
21 - 58 
59 – 120 
>120 
Blood urea nitrogen (BUN) increase, 
40-59Y – mg/dL  
6 – 24 
25 - 58 
59 – 120 
>120 
Glucose decrease, fasting – mg/dL 
65 – 99 
47 - 64 
42 –46 
<42 
Glucose increase, fasting – mg/dL 
65 – 99 
100 – 160 
161 – 250 
>250 
Creatinine increase, male – mg/dL 
0.76 – 1.27 
1.28 – 1.90 
1.91 – 3.80 
>3.80 
Creatinine increase, female – mg/dL 
0.57 – 1.00 
0.58 – 1.50 
1.51 – 3.00 
>3.00 
Direct bilirubin 
0.0 – 0.40 
0.41 – 0.70 
0.71 – 1.20 
>1.20 
Total bilirubin (serum) increase – mg/dL 
(with other LFTs in the normal range) 
0.0 – 1.2 
1.3 – 2.0 
2.1 – 2.5 
>2.5 
Total bilirubin (serum) increase – mg/dL 
(accompanied by a >3 x ULN increase in ALT or AST)  
0.0 – 1.2 
1.3 – 1.7 
1.8 – 2.4 
>2.4 
Total protein decrease – g/dL 
6.0 – 8.5 
5.1 – 5.9 
4.6 – 5.0 
<4.6 
Albumin, decrease, Male, 18-30Y - g/dL 
4.1 – 5.2 
4.0 – 3.5 
2.0– 2.9 
<2.0 


Statistical Analysis Plan - DMID Protocol: 20-0012  
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12.4.1 Individual Laboratory Measurements and Abnormal Laboratory Values 
Table 4: 
Clinical Laboratory Reference Ranges and Toxicity Grading Scales (Continued) 
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Laboratory AEs 
Reference 
Ranges 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Albumin decrease, Female, 18-30Y - g/dL 
3.9 – 5.0 
3.8 – 3.5 
2.0– 2.9 
<2.0 
Albumin decrease, Male, 31-50Y – g/dL 
4.0 – 5.0 
3.9 – 3.5 
2.0– 2.9 
<2.0 
Albumin decrease, Female, 31-50Y – g/dL 
3.8 – 4.8 
3.7 – 3.5 
2.0– 2.9 
<2.0 
AST increase – U/L 
0 – 40 
41 – 80 
81 – 120 
>120 
ALT increase, male – U/L 
0 – 44 
45– 88 
89 - 132 
>132 
ALT increase, female – U/L 
0 – 32 
33 – 64 
65 – 96 
>96 
Alkaline phosphatase (AP) increase, males 18-20Y – U/L 
51 – 125 
126 – 250 
251 – 375 
>375 
Alkaline phosphatase (AP) increase, females, 18-20Y – U/L 
42 – 106 
107 – 250 
251 – 375 
>375 
Alkaline phosphatase (AP) increase, 21-150Y – U/L 
44 – 121 
122 – 250 
251 – 375 
>375 
Urine  
URINALYSIS by Dipstick 
Protein 
negative / 
trace 
1+ 
2+ 
>2+ 
Blood (occult) 
negative 
1+ 
2+ 
>2+ 
Glucose 
negative 
1+ 
2+ 
>2+ 
URINE MICROSCOPY 
Red blood cells (RBC) per HPF 
0 - 2 
6-10 
11-50 
>50 and/or gross blood 
WBC (microscopic) – WBC per HPF 
0 - 5 
6-10 
11-50 
>50 
Bacteria (microscopic) 
none / few 
few 
moderate 
many 
Note 1: With the exception of AST, ALT, AP, bilirubin, electrolytes, urine protein, BUN and creatinine, which should be within reference 
range, lab values of other analytes in the grade 1 range are acceptable for enrollment if (a) they are not considered to be clinically significant by 
the investigator and (b) there is no cluster of abnormal labs that combined are suggestive of an underlying disorder. 
Note 2: Other Exceptions to screening laboratory tests’ normal reference ranges are: 
a. Racially based low total WBC or neutrophil counts up to toxicity Grade 1 are allowed, but toxicity Grades 2 or 3 are exclusionary. 
b. Labs performed as part of a panel but not listed above are to be recorded in the database. If abnormal, they are not exclusionary and are not to 
be graded per Toxicity table, however, the investigator would make a clinical decision about their clinical significance and, if clinically 
significant, they will be graded according to the criteria in Protocol Section 9.2.1. (Examples include mean corpuscular volume (MCV), mean 
corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), mean platelet 
volume (MPV), and nucleated red blood cell count (NRBC CT), which are included in a CBC with differential.)  
Note 3: If a participant was accepted into the trial with a laboratory value of an analyte that overlaps with values used for grading Grade 1 
laboratory abnormalities, a TEAE will be reported if the on-study value of the same analyte increases to Grade 2 or higher. 
Note 4: If the dipstick UA is abnormal, a microscopic UA will be performed, and the results will supersede the results of the dipstick UA. 
Note 5: If the laboratory reports RBC/HPF as a range and not absolute values, then the range 3-10 will be graded as Grade 1. 
Note 6:  Menstruating females with a positive dipstick UA or microscopic UA may be retested following cessation of menses. 
Note 7: Isolated laboratory values that are outside the range of eligibility but are thought to be due to an acute condition or due to collection or 
laboratory error may be repeated once. 
 
 


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Table 5: 
Vital Signs Toxicity Grading Scales 
Vital Sign AEs 
Reference Ranges 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Fever - °C 
Fever - °F 
36.1 - 37.2 1 
97.0 - 99.0 1 
37.3 - 38.4 
 99.1 - 101.1 
38.5 - 38.9 
101.2 - 102.0 
>38.9 
>102.0 
Tachycardia - bpm 
50 - 100 2,3,4,5 
101 - 115 
116 - 130 
>130 or ventricular 
dysrhythmias 
Bradycardia – bpm 
45 - 49 
40 - 44 
<40 
Hypertension (systolic) - 
mmHg 
130/89 2,3,4,5 
131 - 150 
151 - 160 
>160 
Hypertension (diastolic) - 
mmHg 
90 - 95 
96 - 100 
>100 
Hypotension (systolic) - 
mmHg 
85 - 88 
80 - 84 
 <80 
Tachypnea – breaths per 
min 
10 - 20 2,3,4,5 
21 - 25 
26 - 30 
>30 
Note 1:   No recent hot or cold beverages or smoking. A protocol should select either 0C or 0F for inclusion 
Note 2:   Assume awake and in supine position for 5 min at rest. For TEAE, measurements at least 3 times with 2 concordant results (See 
Protocol Section 8.1.6) 
Note 3:   Abnormal HR and BP on first measurement may be repeated twice more with the participant resting between measurements for at 
least 5 min (See Protocol Section 8.1.6). 
Note 4:   Exceptions to screening BP and HR reference range are: 
(a) Participants with baseline HR ≥ 45 to 50 bpm may be accepted if they do not have symptomatic bradycardia, including syncope, heart 
disease, ECG abnormalities, or history of syncope or use of medication  
(b) Participants with baseline SBP up to 140 mmHg and DBP up to 90 mmHg may be accepted if they do not have MH of symptomatic 
hypertension, including paroxysmal hypertension, heart disease, renal disease, CNS disease, visual abnormalities or evidence of 
peripheral vascular disease, ECG abnormalities, use of medication, use of illicit drugs or FH of hypertension  
Note 5:   Isolated/individual abnormalities of VS would not be considered toward halting criteria. Abnormalities of VS will be described as 
“increased X” or “decreased X” (X = HR, BP, RR, temperature) if asymptomatic, transient and not associated with a systemic or organ-specific 
disorder and coded by MedDRA within the System Organ Class (SOC) “Investigations.” These abnormalities will be graded per criteria in this 
table, but not considered in determining whether study stopping criteria have been met. On the other hand, abnormalities of VS that are either 
secondary to systemic or organ-specific clinical syndrome or primary disorders will be coded in the appropriate SOC (e.g., "cardiac disorders", 
respiratory disorders", "immunological disorders", etc.). These abnormalities will be considered in determining whether stopping criteria have 
been met. 


Statistical Analysis Plan - DMID Protocol: 20-0012  
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12.4.1 Individual Laboratory Measurements and Abnormal Laboratory Values 
Table 4: 
Clinical Laboratory Reference Ranges and Toxicity Grading Scales (Continued) 
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Table 6: 
 ECG Toxicity Grading Scale 
ECG interval 
abnormality 
Reference 
range 
Grade 1 
Grade 2 
Grade 3 
QTcF interval 
prolonged (msec): 
• 
Male 
• 
Female 
 
 
• 
≤450 msec 
• 
≤470 msec 
Asymptomatic, 
QTcF 
• 
451 - 479 msec 
• 
471 - 479 msec 
Asymptomatic, 
QTcF 480-500 msec 
OR 
increase in interval 30-59 msec 
above baseline 
Asymptomatic, 
QTcF >500 msec 
OR 
increase in interval ≥60 msec 
above baseline 
PR interval 
prolonged (msec) 
≤210 msec 
211-250 msec 
>250 msec 
Type II 2nd degree AV block  
OR  
ventricular pause >3.0 sec 
Note 1: Events will be coded as treatment-emergent SAEs if there are life-threatening associated symptoms or signs (arrhythmia, CHF, 
hypotension, syncope, TdP, etc.). 
Note 2: If a male participant was accepted into the trial with a QTcF value that overlaps with values used for grading Grade 1 QTcF 
prolongation, a TEAE will be reported if the QTcF value is higher than the baseline value. 
 


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10.2 
Protocol Deviations 
Table 7: 
Distribution of Protocol Deviations by Category, Type, and Cohort – All Enrolled Participants 
[Implementation Note: Only deviations reported during the course of the study will be included.] 
Category 
Deviation Type 
Cohort 1  
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All 
Participants 
(N=X) 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
Eligibility/enrollment 
Any type 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
 
Did not meet inclusion criterion 
 
 
 
 
 
 
 
 
 
 
 
 
 
Met exclusion criterion 
 
 
 
 
 
 
 
 
 
 
 
 
 
ICF not signed prior to study 
procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment administration 
schedule 
Any type 
 
 
 
 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
 
 
 
 
Incomplete treatment 
administration 
 
 
 
 
 
 
 
 
 
 
 
 
 
Delayed treatment administration 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 
 
Follow-up visit schedule 
Any type 
 
 
 
 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 
 
Protocol procedure/assessment 
Any type 
 
 
 
 
 
 
 
 
 
 
 
 
 
Incorrect version of ICF signed 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood not collected 
 
 
 
 
 
 
 
 
 
 
 
 


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Protocol Deviations 
Table 7: 
Distribution of Protocol Deviations by Category, Type, and Cohort – All Enrolled Participants (Continued) 
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Category 
Deviation Type 
Cohort 1  
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All 
Participants 
(N=X) 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
 
Urine not collected 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other specimen not collected 
 
 
 
 
 
 
 
 
 
 
 
 
 
Too few aliquots obtained 
 
 
 
 
 
 
 
 
 
 
 
 
 
Specimen result not obtained 
 
 
 
 
 
 
 
 
 
 
 
 
 
Required procedure not conducted 
 
 
 
 
 
 
 
 
 
 
 
 
 
Required procedure done 
incorrectly 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study product temperature 
excursion 
 
 
 
 
 
 
 
 
 
 
 
 
 
Specimen temperature excursion 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment administration 
Any type 
 
 
 
 
 
 
 
 
 
 
 
 
 
Required procedure done 
incorrectly 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study product temperature 
excursion 
 
 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of participants enrolled. 
 
 


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14.1 
Description of Study Participants 
14.1.1 Disposition of Participants 
Table 8: 
Participant Disposition by Cohort – All Enrolled Participants 
Participant Disposition  
Cohort 1  
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Screened 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
x 
-- 
Enrolled 
x 
100 
x 
100 
x 
100 
x 
100 
x 
100 
x 
100 
Started Infusion 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed Infusiona 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed Bronchoscopy with BAL 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed All PK Blood Draws 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed All Plasma Urea Blood Draws 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed Follow-up (Study Day 14) a 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Completed Final Study Visit (Study Day 30)a 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Early Terminationa 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Note: N = Number of participants enrolled. 
a Refer to Listing 16.2.1 for reasons participants discontinued or terminated early. 
 
 


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Table 9: 
Treatment Discontinuations or Early Terminations by Reason and Cohort – All Enrolled Participants 
[Implementation Note: Only reasons for treatment discontinuation or early termination reported during the study will be included.] 
Reason for Treatment Discontinuation or Early Termination  
Cohort 1  
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Treatment Discontinuation 
Serious Adverse Event (other than death) 
x 
100 
x 
100 
x 
100 
x 
100 
x 
100 
x 
100 
Adverse Event, Other than Serious Adverse Event 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Protocol Deviation 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Withdrawal by Participant 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Withdrawal by Investigator 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
COVID-19 Pandemic 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Termination of Site by Sponsor 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Termination of Study by Sponsor 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Death 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Technical Problems 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Not Eligible at Enrollment 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Became Ineligible After Enrollment 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Failure to Receive the Entire Volume of Study Drug Infusion 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Other 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Early Termination 
Serious Adverse Event (other than death) 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Adverse Event, Other than Serious Adverse Event 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Protocol Deviation 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Note: N = Number of participants enrolled. 
 


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Table 10: 
Ineligibility Summary of Screen Failures 
Inclusion/Exclusion Category 
Inclusion/Exclusion Criterion 
Participants Excluded 
na 
%b 
Inclusion and Exclusion 
Number of participants failing any eligibility criterion or eligible but not enrolled 
x 
100 
Inclusion 
Any inclusion criterion 
x 
xx 
 
[inclusion criterion 1] 
x 
xx 
 
[inclusion criterion 2] 
x 
xx 
 
[inclusion criterion 3] 
x 
xx 
Exclusion 
Any exclusion criterion 
x 
xx 
 
[exclusion criterion 1] 
x 
xx 
 
[exclusion criterion 2] 
x 
xx 
 
[exclusion criterion 3] 
x 
xx 
Eligible but Not Enrolled 
Any reason eligible but not enrolled 
x 
xx 
 
[reason 1] 
x 
xx 
 
[reason 2] 
x 
xx 
 
[reason 3] 
x 
xx 
a More than one criterion may be marked per participant. 
b Denominator for percentages is the total number of screen failures. 
 


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Table 11: 
Analysis Population Exclusions by Cohort – All Enrolled Participants 
Analysis Populations 
Reason Participants Excluded 
Cohort 1  
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Safety Population 
Did not receive any amount of study product 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Safety Subset Population 
Did not receive full amount of study product 
 
 
 
 
 
 
 
 
 
 
 
 
Lung PK Analysis Population 
Any Reason 
 
 
 
 
 
 
 
 
 
 
 
 
 
Did not receive full amount of study product 
 
 
 
 
 
 
 
 
 
 
 
 
 
Did not undergo BAL at assigned sampling timepoint 
 
 
 
 
 
 
 
 
 
 
 
 
 
BAL return volume not adequate for analysis 
 
 
 
 
 
 
 
 
 
 
 
 
 
Has no measurable drug concentration for ELF or 
AM 
 
 
 
 
 
 
 
 
 
 
 
 
Plasma PK Analysis Population 
Any Reason 
 
 
 
 
 
 
 
 
 
 
 
 
 
Did not receive full amount of study product 
 
 
 
 
 
 
 
 
 
 
 
 
 
Has no measurable drug concentration in plasma 
 
 
 
 
 
 
 
 
 
 
 
 
PK Analysis Subset Population 
Any Reason 
 
 
 
 
 
 
 
 
 
 
 
 
 
Excluded from Lung PK Analysis Population  
 
 
 
 
 
 
 
 
 
 
 
 
 
Excluded from Plasma PK Analysis Population 
 
 
 
 
 
 
 
 
 
 
 
 
 
Has protocol deviations that potentially impact PK 
 
 
 
 
 
 
 
 
 
 
 
 
 
Plasma PK and BAL PK data insufficient to estimate 
any PK parameters 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of participants enrolled. 
 


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14.1.2 Demographic Data by Study Group 
Table 12: 
Summary of Categorical Demographic and Baseline Characteristics by Cohort – All Enrolled Participants 
Variable 
Characteristic 
Cohort 1 
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Sex  
Male 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Female 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Ethnicity  
Not Hispanic or Latino 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Hispanic or Latino 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Not Reported 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Unknown 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Race  
American Indian or Alaska Native 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Asian 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Native Hawaiian or Other Pacific Islander 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Black or African American 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
White 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Multi-Racial 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Unknown 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Note: N = Number of participants enrolled. 
 
 


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Table 13: 
Summary of Continuous Demographic and Baseline Characteristics by Cohort – All Enrolled Participants 
Variable 
Statistic 
Cohort 1 
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
Age (years) 
Mean 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
SD 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
Median 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
Min 
x 
x 
x 
x 
x 
x 
 
Max 
x 
x 
x 
x 
x 
x 
Height (cm) 
Mean 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
SD 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Median 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Min 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
Max 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
Weight (kg) 
Mean 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
SD 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Median 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Min 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
Max 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
BMI (kg/m2) 
Mean 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
SD 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Median 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
x.xx 
 
Min 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
 
Max 
x.x 
x.x 
x.x 
x.x 
x.x 
x.x 
Note: N = Number of participants enrolled. 
 
 


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14.1.3 Prior and Concurrent Medical Conditions 
Table 14: 
Summary of Participants with Pre-Existing Medical Conditions by MedDRA System Organ Class and Cohort – All Enrolled 
Participants 
MedDRA System Organ Class 
Cohort 1 
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
[SOC 1] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Notes: N = Number of participants enrolled. n = Number of participants reporting medical history within the specified SOC. A participant is only counted once per SOC. 
 
 


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14.3 
Safety Data 
14.3.1 Displays of Adverse Events 
Table 15: 
Overall Summary of Adverse Events – Safety Population 
Participantsa with 
All Participants 
(N=X) 
n 
% 
At least one TEAE 
x 
xx 
At least one related TEAE 
x 
xx 
At least one Mild (Grade 1) related TEAE 
x 
xx 
At least one Moderate (Grade 2) related TEAE 
x 
xx 
At least one Severe (Grade 3) related TEAE 
x 
xx 
At least one SAEb 
x 
xx 
At least one related, SAE 
x 
xx 
At least one TEAE leading to early terminationc  
x 
xx 
Note: N = Number of participants in the Safety Population. 
a Participants are counted once for each category regardless of the number of events. 
b A listing of Serious Adverse Events is included in Section 14.3.2. 
c As reported on the Adverse Event eCRF. 
Table with similar format: 
Table 16: 
Overall Summary of Adverse Events – Safety Subset Population 
 


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14.3.1.2 
Unsolicited Adverse Events 
Table 17: 
Summary of Treatment Emergent Adverse Events by MedDRA System Organ Class, High Level Group Term, and Preferred 
Term – Safety Population 
[Implementation Note: If there is only 1 PT for an SOC, HLGT, there will be no “Any PT” row. Data will be presented in Alphabetical order by SOC, 
HLGT, and PT.] 
MedDRA System Organ Class 
MedDRA High Level Group Term 
MedDRA Preferred Term 
All Participants 
(N=X) 
n 
% 
95% CIa 
Number of Events 
Any SOC 
Any HLGT 
Any PT 
x 
xx 
xx, xx 
x 
[SOC 1] 
Any HLGT 
Any PT 
 
 
 
 
 
[HLGT 1] 
Any PT 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
[SOC 2] 
Any HLGT 
Any PT 
 
 
 
 
 
[HLGT 1] 
Any PT 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
Notes: N = Number of participants in the Safety Population. This table presents number and percentage of participants and the number of events. For the number of participants (n) and corresponding 
percent (%), a participant is only counted once per Preferred Term. 
a Exact (Clopper-Pearson) confidence interval was used. 
Table with similar format: 
Table 18: 
Summary of Treatment Emergent Adverse Events by MedDRA System Organ Class, High Level Group Term, and Preferred 
Term – Safety Subset Population 
 


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Table 19: 
Treatment Emergent Adverse Events by MedDRA System Organ Class, High Level Group Term, and Preferred Term, 
Maximum Severity and Relationship – Safety Population 
[Implementation Note: If there is only 1 HLGT for a SOC, there will be no “Any HLGT” row. Similarly, if there is only 1 PT for an HLGT, there will be 
no “Any PT” row. Data will be presented in alphabetical order by SOC, HLGT, and PT.] 
MedDRA System Organ 
Class 
MedDRA High Level 
Group Term 
MedDRA Preferred Term 
Severity 
All Participants 
(N = X) 
Related 
Not Related 
Total 
n 
% 
n 
% 
n 
% 
Any SOC 
Any HLGT 
Any PT 
Any Severity 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Mild 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Moderate 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Severe 
x  
xx 
x  
xx 
x  
xx 
SOC 1 
Any HLGT 
Any PT 
Any Severity 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Mild 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Moderate 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Severe 
x  
xx 
x  
xx 
x  
xx 
 
[HLGT 1] 
Any PT 
Any Severity 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Mild 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Moderate 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Severe 
x  
xx 
x  
xx 
x  
xx 
 
 
[PT 1] 
Any Severity 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Mild 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Moderate 
x  
xx 
x  
xx 
x  
xx 
 
 
 
Severe 
x  
xx 
x  
xx 
x  
xx 
Note: N = Number of participants in the Safety Population. This table presents number and percentage of participants by relationship. For the number of participants (n) and corresponding percent (%), 
a participant is only counted once per relatedness and Preferred Term. 
 


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Table with similar format: 
Table 20: 
Treatment Emergent Adverse Events by MedDRA System Organ Class, High Level Group Term, and Preferred Term, 
Maximum Severity and Relationship – Safety Subset Population 
 


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14.3.2 Listing of Deaths, Other Serious Adverse Events 
Table 21: 
Listing of Serious Adverse Events  
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. If the event is ongoing (no stop 
date), indicate “Ongoing” for the “Duration (Days)”. If more than one reason is selected for the reason reported as an SAE, list all reasons in the column, 
separated by a comma. In the “If Not Related, Alternate Etiology” column, merge the 2 data fields for collecting alternate etiology, separated by a colon. 
Listing should be sorted by Cohort, Participant ID, and AE Number.] 
Adverse 
Event 
MedDRA 
System 
Organ 
Class 
MedDRA 
High 
Level 
Group 
Term 
MedDRA 
Preferred 
Term 
Study 
Day of 
AE Onset 
No. of 
Days 
Post 
Dose the 
Event 
Became 
Serious 
Duration 
(Days) 
Reason 
Reported 
as an SAE 
Severity 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action 
Taken 
with 
Study 
Treatment 
Participant 
Discontinued 
Due to AE 
Outcome 
Cohort: , Participant ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
Cohort: , Participant ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 


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Table 22: 
Listing of Non-Serious, Moderate or Severe Treatment Emergent Adverse Events 
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. If the event is ongoing (no stop 
date), indicate “Ongoing” for the “Duration (Days)”. In the “If Not Related, Alternate Etiology” column, merge the 2 data fields for collecting alternate 
etiology, separated by a colon. Listing should be sorted by Cohort, Participant ID, and AE Number.] 
Adverse 
Event 
MedDRA 
System 
Organ Class 
MedDRA 
High Level 
Group Term 
MedDRA 
Preferred 
Term 
Study Day of 
AE Onset 
Duration 
(Days) 
Severity 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Participant 
Discontinued 
Due to AE 
Outcome 
Cohort: , Participant ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
Cohort: , Participant ID: ,  AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 


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14.3.3 Narratives of Deaths, Other Serious and Significant Adverse Events 
(not included in SAP, but this is a placeholder for the CSR) 
 


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14.3.4 Abnormal Laboratory Value Listings (by Participant) 
Table 23: 
Listing of Abnormal Laboratory Results – Chemistry 
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. This listing includes all abnormal 
Chemistry results (laboratory results outside of the normal range defined in the protocol). Normal chemistry results for other parameters that occurred on 
the same visit as the abnormal result will not be listed. Results that are outside the normal range but not mild, moderate, or severe will have “(ONR)” 
included after the result. Similarly, results that meet grading criteria of mild, moderate, or severe but which are not treatment emergent (not worse than 
baseline) will be shown as ONR. Listing should be sorted by Cohort, Participant ID, Timepoint, and Parameter.] 
Cohort 
Participant 
ID 
Sex 
Age 
(years) 
Planned Study 
Day 
Actual Study 
Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Clinical 
Significance 
Relationship 
to Treatment 
If Not 
Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Participant 
Discontinued 
Due to 
Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Table 24: 
Listing of Abnormal Laboratory Results – Hematology 
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. This listing includes all abnormal 
Hematology results (laboratory results outside of the normal range defined in the protocol). Normal hematology results for other parameters that 
occurred on the same visit as the abnormal result will not be listed. Results that are outside the normal range but not mild, moderate, or severe will have 
“(ONR)” included after the result. Similarly, results that meet grading criteria of mild, moderate, or severe but which are not treatment emergent (not 
worse than baseline) will be shown as ONR. Listing should be sorted by Cohort, Participant ID, Timepoint, and Parameter.] 
Cohort 
Participant 
ID 
Sex 
Age 
(years) 
Planned 
Study Day 
Actual Study 
Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Clinical 
Significance 
Relationship 
to Treatment 
If Not Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Participant 
Discontinued 
Due to 
Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Table 25: 
Listing of Abnormal Laboratory Results – Coagulation 
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. This listing includes all abnormal 
Coagulation results (laboratory results outside of the normal range defined in the protocol). Normal coagulation results for other parameters that occurred 
on the same visit as the abnormal result will not be listed. Results that are outside the normal range but not mild, moderate, or severe will have “(ONR)” 
included after the result. Similarly, results that meet grading criteria of mild, moderate, or severe but which are not treatment emergent (not worse than 
baseline) will be shown as ONR. Listing should be sorted by Cohort, Participant ID, Timepoint, and Parameter.] 
Cohort 
Participant 
ID 
Sex 
Age 
(years) 
Planned 
Study Day 
Actual Study 
Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Clinical 
Significance 
Relationship 
to Treatment 
If Not Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Participant 
Discontinued 
Due to 
Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Table 26: 
Listing of Abnormal Laboratory Results - Urinalysis 
[Implementation Note: This listing is included in the table shells document, as it is included in the body of the CSR. This listing includes all abnormal 
Urinalysis results (laboratory results outside of the normal range defined in the protocol). Normal urinalysis results for other parameters that occurred on 
the same visit as the abnormal result will not be listed. Results that are outside the normal range but not mild, moderate, or severe will have “(ONR)” 
included after the result. Similarly, results that meet grading criteria of mild, moderate, or severe but which are not treatment emergent (not worse than 
baseline) will be shown as ONR. Listing should be sorted by Cohort, Participant ID, Timepoint, and Parameter.] 
Cohort 
Participant 
ID 
Sex 
Age 
(years) 
Planned 
Study Day 
Actual Study 
Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Clinical 
Significance 
Relationship 
to Treatment 
If Not Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Participant 
Discontinued 
Due to 
Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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14.3.5 Displays of Clinical Laboratory Results 
14.3.5.1 
Chemistry Results 
Table 27: 
Chemistry Abnormal Results by Parameter and Timepoint – Safety Population 
Parameter 
Timepoint 
N 
Abnormal, Low 
Abnormal, High 
n 
% 
n 
% 
Chemistry – Any Parameter 
Baseline 
x 
x 
xx 
x 
xx 
 
Day 2, 24 h Post-Dose 
x 
x 
xx 
x 
xx 
 
Day 3, 48 h Post-Dose 
x 
x 
xx 
x 
xx 
 
Day 14 
x 
x 
xx 
x 
xx 
Sodium 
Baseline 
x 
x 
xx 
x 
xx 
 
Day 2, 24 h Post-Dose 
x 
x 
xx 
x 
xx 
 
Day 3, 48 h Post-Dose 
x 
x 
xx 
x 
xx 
 
Day 14 
x 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with the laboratory result assessed at the respective timepoint. Baseline = The most 
recent measurement prior to start of infusion. 
Table with similar format: 
Table 28: 
Chemistry Abnormal Results by Parameter and Timepoint – Safety Subset Population 
 


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Table 29: 
Chemistry Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Chemistry parameters without 
grading criteria are not included in this table. Toxicities representing an increase in the laboratory result will 
be summarized separately from decreases.] 
Parameter 
Time Point 
N 
Mild / 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
n 
% 
n 
% 
n 
% 
Any Chemistry Parameter 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 2, 24 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 3, 48 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 14 
x 
x 
xx 
x 
xx 
x 
xx 
Sodium, Decrease 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with the laboratory result assessed at the respective timepoint. Baseline = The most 
recent measurement prior to start of infusion. The “Maximum Severity Post Baseline” rows indicate the maximum severity experienced by each 
participant at any time point post baseline, including unscheduled assessments.  
Table with similar format: 
Table 30: 
Chemistry Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Subset Population 
 


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Table 31: 
Chemistry Summary Statistics Measurement and Change from Baseline by Parameter and Timepoint – Safety Population 
[Implementation Note: BUN should by summarized overall and separately by age (where specified in the toxicity table, for example, “BUN (mg/dL), 
Overall”; “BUN (mg/dL), 18-39 years”; “BUN (mg/dL), 40-59 years”). Creatinine and ALT should be summarized overall and separately by sex (for 
example, “Creatinine (mg/dL), Overall”; “Creatinine (mg/dL), Male”; “Creatinine (mg/dL), Female”). Albumin and AP should be summarized overall 
and separately by age and sex. Other parameters are not age or sex dependent and will be summarized overall.]  
Parameter 
Timepoint 
N 
Measurement 
Change from Baseline 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
Sodium (mmol/L) 
Baseline 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
- 
- 
- 
- 
 
Day 2, 24 h Post-Dose 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
Day 3, 48 h Post-Dose 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
Day 14 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
Potassium (mmol/L) 
Baseline 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
- 
- 
- 
- 
 
Day 2, 24 h Post-Dose 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
Notes: N = Number of participants in the Safety Population with the laboratory result assessed at the respective timepoint. Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 32: 
Chemistry Summary Statistics Measurement and Change from Baseline by Parameter and Timepoint – Safety Subset Population


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14.3.5.2 
Hematology Results 
Table 33: 
Hematology Abnormal Results by Parameter and Timepoint – Safety Population 
This table will repeat Table 27 for Hematology Parameters. 
Table 34: 
Hematology Abnormal Results by Parameter and Timepoint – Safety Subset Population 
This table will repeat Table 28 for Hematology Parameters. 
Table 35: 
Hematology Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Toxicities representing an 
increase in the laboratory result will be summarized separately from decreases.] 
This table will repeat Table 29 for Hematology Parameters. 
Table 36: 
Hematology Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Subset Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Toxicities representing an 
increase in the laboratory result will be summarized separately from decreases.] 
This table will repeat Table 30 for Hematology Parameters. 
Table 37: 
Hematology Summary Statistics Measurement and Change from Baseline by Parameter 
and Timepoint – Safety Population 
[Implementation Note: Hemoglobin should be summarized overall and separately by sex (where specified in 
the toxicity table, for example, “Hemoglobin (g/dL), Overall”; “Hemoglobin (g/dL), Male”; “Hemoglobin 
(g/dL), Female”). Other parameters are not sex dependent and will be summarized overall.] 
This table will repeat Table 31 for Hematology Parameters. 
Table 38: 
Hematology Summary Statistics Measurement and Change from Baseline by Parameter 
and Timepoint – Safety Subset Population 
[Implementation Note: Hemoglobin should be summarized overall and separately by sex (where specified in 
the toxicity table, for example, “Hemoglobin (g/dL), Overall”; “Hemoglobin (g/dL), Male”; “Hemoglobin 
(g/dL), Female”). Other parameters are not sex dependent and will be summarized overall.] 
This table will repeat Table 32 for Hematology Parameters. 
 
 


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14.3.5.3 
Coagulation Results 
Table 39: 
Coagulation Abnormal Results by Parameter and Timepoint – Safety Population 
This table will repeat Table 27 for Coagulation Parameters. 
Table 40: 
Coagulation Abnormal Results by Parameter and Timepoint – Safety Subset Population 
This table will repeat Table 28 for Coagulation Parameters. 
Table 41: 
Coagulation Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Toxicities representing an 
increase in the laboratory result will be summarized separately from decreases.] 
This table will repeat Table 29 for Coagulation Parameters. 
Table 42: 
Coagulation Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Subset Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Toxicities representing an 
increase in the laboratory result will be summarized separately from decreases.] 
This table will repeat Table 30 for Coagulation Parameters. 
Table 43: 
Coagulation Summary Statistics Measurement and Change from Baseline by Parameter 
and Timepoint – Safety Population 
This table will repeat Table 31 for Coagulation Parameters. 
Table 44: 
Coagulation Summary Statistics Measurement and Change from Baseline by Parameter 
and Timepoint – Safety Subset Population 
This table will repeat Table 32 for Coagulation Parameters. 
 
 


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14.3.5.4 
Urinalysis Results 
Table 45: 
Urinalysis Abnormal Results by Parameter and Timepoint – Safety Population 
This table will repeat Table 27 for Urinalysis Parameters. 
Table 46: 
Urinalysis Abnormal Results by Parameter and Timepoint – Safety Subset Population 
This table will repeat Table 28 for Urinalysis Parameters. 
Table 47: 
Urinalysis Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Urinalysis parameters without 
grading criteria are not included in this table. Toxicities representing an increase in the laboratory result will 
be summarized separately from decreases.] 
This table will repeat Table 29 for Urinalysis Parameters. 
Table 48: 
Urinalysis Laboratory Toxicity Grade by Parameter, Timepoint, and Severity – Safety 
Subset Population 
[Implementation Note: If there is not at least one mild, moderate, or severe event, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter. Urinalysis parameters without 
grading criteria are not included in this table. Toxicities representing an increase in the laboratory result will 
be summarized separately from decreases.] 
This table will repeat Table 30 for Urinalysis Parameters. 
 
 


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14.3.5.5 
Kidney Function Biomarker Results 
Table 49: 
Kidney Function Biomarker Summary Statistics Measurement and Change from 
Baseline by Parameter and Timepoint – Safety Population 
Parameter 
Timepoint 
N 
Measurement 
Change from Baseline 
Mean 
SD 
Median Min, Max 
Mean 
SD 
Median 
Min, Max 
Serum Cystatin C 
(mg/L) 
Baseline 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
- 
- 
- 
- 
 
Day 3, 48 h Post-Dose 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
Day 14 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
Day 30 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
Urinary KIM-1 
(ng/mL) 
Baseline 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
- 
- 
- 
- 
 
Day 3, 48 h Post-Dose 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
 
 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
Urinary KIM-1 
Normalized (ng/mg 
creatinine) 
Baseline 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
- 
- 
- 
- 
 
 
x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
xx.xx 
xx.xx 
xx.xx 
xx.x, xx.x 
Notes: N = Number of participants in the Safety Population with the laboratory result assessed at the respective timepoint. Baseline = The most 
recent measurement prior to start of infusion. 
Table with similar format: 
Table 50: 
Kidney Function Biomarker Summary Statistics Measurement and Change from 
Baseline by Parameter and Timepoint – Safety Subset Population 
 


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14.3.6 Displays of Vital Signs 
Table 51: 
Vital Sign Toxicity Grade by Parameter, Timepoint, and Severity – Safety Population 
[Implementation Note: If there is not at least one mild, moderate, or severe result with a parameter, then only 
the “Baseline” and “Maximum Severity Post Baseline” rows will be shown for that parameter. Toxicities 
representing an increase in the laboratory result will be summarized separately from decreases.] 
Parameter 
Time Point 
N 
Mild / 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
n 
% 
n 
% 
n 
% 
Any Parameter 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 0.5 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 1 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 4 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 16 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 2, 24 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 2, 36 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 3, 48 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 3, 60 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 14 
x 
x 
xx 
x 
xx 
x 
xx 
Systolic Blood Pressure, Decrease 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with the vital sign assessed at the respective timepoint. Baseline = The most recent 
measurement prior to start of infusion. The “Maximum Severity Post Baseline” rows indicate the maximum severity experienced by each 
participant at any time point post baseline, including unscheduled assessments.  
Table with similar format: 
Table 52: 
Vital Sign Toxicity Grade by Parameter, Timepoint, and Severity – Safety Subset 
Population 
 


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Table 53: 
Vital Sign Summary Statistics Measurement and Change from Baseline by Parameter and Timepoint – Safety Population 
Parameter 
Timepoint 
N 
Measurement 
Change from Baseline 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
Systolic Blood Pressure (mmHg) 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
Day 1, 0.5 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 1, 1 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 1, 4 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 1, 16 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 2, 24 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 2, 36 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 3, 48 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 3, 60 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 14 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
Diastolic Blood Pressure (mmHg) 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
Day 1, 0.5 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
Notes: N = Number of participants in the Safety Population with the vital sign assessed at the respective timepoint. Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 54: 
Vital Sign Summary Statistics Measurement and Change from Baseline by Parameter and Timepoint – Safety Subset Population 


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14.3.7 
 
Displays of ECG Measurements 
Table 55: 
ECG Overall Interpretations, Post-Dose Compared to Pre-Dose Baseline by Timepoint – Safety Population 
Timepoint 
N 
Change from Pre-Dose Baseline in ECG Interpretation 
Normal at Both Times  
n (%) 
Normal to Abnormal, 
NCS 
n (%) 
Normal to Abnormal, 
CS 
n (%) 
Abnormal, NCS at 
Both Times 
n (%) 
Abnormal, NCS to 
Abnormal, CS 
n (%) 
Abnormal, NCS to 
Normal 
n (%) 
Day 1, 1 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 4 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 16 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 24 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 36 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 48 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 60 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 14 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. Pre-Dose Baseline = The most recent measurement prior to start of infusion. 
CS = Clinically Significant. NCS = Not Clinically Significant. 
Table with similar format: 
Table 56: 
ECG Overall Interpretations, Post-Dose Compared to Pre-Dose Baseline by Timepoint – Safety Subset Population 
 


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Table 57: 
ECG Summary in Change from Pre-Dose Baseline by Timepoint – Safety Population 
Timepoint 
N 
Change from Pre-Dose Baseline 
No Change from Baseline 
n (%) 
NCS, Change from Baseline 
n (%) 
CS, Change from Baseline 
n (%) 
Day 1, 1 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 1, 4 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 1, 16 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 2, 24 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 2, 36 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 3, 48 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 3, 60 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
Day 14 
x 
x (xx) 
x (xx) 
x (xx) 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. Pre-Dose Baseline 
= The most recent measurement prior to start of infusion. CS = Clinically Significant. NCS = Not Clinically Significant. 
Table with similar format: 
Table 58: 
ECG Summary in Change from Pre-Dose Baseline by Timepoint – Safety Subset 
Population 
 


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Table 59: 
ECG Toxicity Grade by Parameter, Timepoint, and Severity – Safety Population 
[Implementation Note: If there is not at least one mild, moderate, or severe result with a parameter, then only 
the “Baseline” and “Maximum Severity Post Baseline” rows will be shown for that parameter. ECG 
parameters without grading criteria are not included in this table.] 
Parameter 
Time Point 
N 
Mild / 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
n 
% 
n 
% 
n 
% 
Any Parameter 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 1 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 4 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 1, 16 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 2, 24 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 2, 36 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 3, 48 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 3, 60 h Post-Dose 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 14 
x 
x 
xx 
x 
xx 
x 
xx 
QTcF Interval 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. Pre-Dose Baseline 
= The most recent measurement prior to start of infusion. The “Maximum Severity Post Baseline” rows indicate the maximum severity 
experienced by each participant at any time point post baseline, including unscheduled assessments.  
Table with similar format: 
Table 60: 
ECG Toxicity Grade by Parameter, Timepoint, and Severity – Safety Subset Population 


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Table 61: 
ECG Measurement and Change from Pre-Dose Baseline by Parameter and Timepoint – Safety Population 
Parameter 
Timepoint 
N 
Measurement 
Change from Pre-Dose Baseline 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
90% CI 
PR Interval (msec) 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
- 
 
Day 1, 1 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 1, 4 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 1, 16 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 2, 24 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 2, 36 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 3, 48 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 3, 60 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
Day 14 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
QRS Duration (msec) 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
- 
 
Day 1, 1 h Post-Dose 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. Pre-Dose Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 62: 
ECG Measurement and Change from Pre-Dose Baseline by Parameter and Timepoint – Safety Subset Population 
 


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Table 63: 
ECG Change from Time-Matched Baseline by Parameter and Timepoint – Safety Population 
Parameter 
Timepoint 
Study 
Day 
N 
Measurement 
Change from Time-Matched Baseline 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
90% CI 
PR Interval (msec) 
1 h Post-Dose 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
- 
 
 
Day 1 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
4 h Post-Dose 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
- 
 
 
Day 1 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
16 h Post-Dose 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
- 
 
 
Day 1 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
QRS Duration (msec) 1 h Post-Dose 
Baseline 
x 
xx.x 
xx.x 
xx.x 
xx, xx  
- 
- 
- 
- 
- 
 
 
Day 1 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
(xx.x, xx.x) 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. Time-Matched Baseline = The measurements recorded on Day -1. 
Table with similar format: 
Table 64: 
ECG Change from Time-Matched Baseline by Parameter and Timepoint – Safety Subset Population 
 


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Table 65: 
Categorical Summary of QTcF Interval by Timepoint – Safety Population 
Timepoint 
N 
QTcF Category 
QTcF >450 - 480 
msec 
n (%) 
QTcF >480 - 500 
msec 
n (%) 
QTcF >500 
msec 
n (%) 
QTcF increase ≥30 – 59 
msec from baseline 
n (%) 
QTcF increase ≥60 
msec from baseline 
n (%) 
Day 1, 1 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 4 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 16 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 24 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 36 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 48 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 60 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 14 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Notes: N = Number of participants in the Safety Population with the ECG measurement assessed at the respective timepoint. n = Number of 
participants who experienced the event. A participant may be counted in more than one category. Pre-Dose Baseline = The most recent 
measurement prior to start of infusion. 
Table with similar format: 
Table 66: 
Categorical Summary of QTcF Interval by Timepoint – Safety Subset Population 
 


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Table 67: 
Categorical Summary of PR Interval, QRS Duration, and RR Interval by Timepoint – 
Safety Population 
Timepoint 
N 
PR Interval, QRS Duration, and RR Interval Category 
PR change from 
baseline >25% 
increase resulting in 
PR >200 msec 
n (%) 
QRS change from 
baseline >25% 
increase resulting in 
QRS >120 msec 
n (%) 
HR change from 
baseline >25% 
decrease resulting in 
HR <50 bpm 
n (%) 
HR change from 
baseline >25% 
increase resulting in 
HR >100 bpm 
n (%) 
Day 1, 1 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 4 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 1, 16 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 24 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 2, 36 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 48 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 3, 60 h Post-Dose 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Day 14 
x 
x (xx) 
x (xx) 
x (xx) 
x (xx) 
Notes: N = Number of participants in the Safety Population with ECG measurements assessed at the respective timepoint. n = Number of 
participants who experienced the event. Participants may be counted in more than one category. Pre-Dose Baseline = The most recent 
measurement prior to start of infusion.  
Table with similar format: 
Table 68: 
Categorical Summary of PR Interval, QRS Duration, and RR Interval by Timepoint – 
Safety Subset Population 
 


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14.3.8 
 
Displays of Audiology Results 
Table 69: 
Ear Otoscopy Result, Post-Dose Compared to Baseline by Timepoint and Ear – Safety Population 
[Implementation Note: If there are no problems at baseline (blockage of wax or foreign body or any problem other than wax or foreign body), only keep 
the “No Apparent Problem” rows.] 
Ear 
Time Point 
Baseline Result 
N 
Post-Baseline Result 
No Apparent Problem 
Blockage of Wax or 
Foreign Body 
Any Problem Other than 
Wax or Foreign Body 
n 
% 
n 
% 
n 
% 
Right 
Day 3 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
Blockage of Wax or Foreign Body 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
Any Problem Other than Wax or Foreign Body 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 14 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
Blockage of Wax or Foreign Body 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
Any Problem Other than Wax or Foreign Body 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 30 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
Left 
Day 3 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 14 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
 
Day 30 
No Apparent Problem 
x 
x 
xx 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with ear otoscopy results at the respective timepoint. Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 70: 
Ear Otoscopy Result, Post-Dose Compared to Baseline by Timepoint and Ear – Safety Subset Population 


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Table 71: 
Pure-Tone Audiometry Toxicity Grade by Timepoint and Severity – Safety Population 
[Implementation Note: If there is not at least one mild, moderate, or severe result, then only the “Baseline” 
and “Maximum Severity Post Baseline” rows will be shown for that parameter.] 
Time Point 
N 
Any Graded Event 
Mild / 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Maximum Severity Post Baseline 
x 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Day 3 
x 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Day 14 
x 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Day 30 
x 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Population with pure-tone audiometry assessed at the respective timepoint. Baseline = The most 
recent measurement prior to start of infusion. The “Maximum Severity Post Baseline” rows indicate the maximum severity experienced by each 
participant at any time point post baseline, including unscheduled assessments.  
Table with similar format: 
Table 72: 
Pure-Tone Audiometry Toxicity Grade by Timepoint and Severity – Safety Subset 
Population 
 


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Table 73: 
Pure-Tone Audiometry Threshold Measurement and Change from Baseline by Timepoint, Frequency, and Ear – Safety 
Population 
Ear 
Timepoint 
Frequency (kHz) 
N 
Measurement (dB) 
Change from Baseline (dB) 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
Right 
Baseline 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
1.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
2.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
4.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
6.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
8.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
9.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
10.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
11.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
12.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
14.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
16.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
18.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
20.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
Day 3 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
1.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 14 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 30 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
Left 
Baseline 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
Notes: N = Number of participants in the Safety Population with the pure-tone audiometry assessed at the respective timepoint and frequency. Baseline = The most recent measurement prior to start of 
infusion. 
 


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Table with similar format: 
Table 74: 
Pure-Tone Audiometry Threshold Measurement and Change from Baseline by Timepoint, Frequency, and Ear – Safety Subset 
Population 
 


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Table 75: 
DPOAE Measurement and Change from Baseline by Timepoint, Frequency, and Ear – Safety Population 
Ear 
Timepoint 
Frequency (kHz) 
N 
Measurement (dB) 
Change from Baseline (dB) 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
Right 
Baseline 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
2.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
3.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
4.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
6.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
Day 3 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 14 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 30 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
Left 
Baseline 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
Notes: N = Number of participants in the Safety Population with the DPAOEs assessed at the respective timepoint and frequency. Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 76: 
DPOAE Measurement and Change from Baseline by Timepoint, Frequency, and Ear – Safety Subset Population


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Table 77: 
Summary of DPOAE Results by Timepoint, Frequency, and Ear – Safety Population 
Ear 
Timepoint 
Frequency 
(kHz) 
N 
DPOAE Result 
Present 
Absent 
n 
% 
n 
% 
Right 
Baseline 
1.5 
x 
x 
xx 
x 
xx 
 
 
2.0 
x 
x 
xx 
x 
xx 
 
 
3.0 
x 
x 
xx 
x 
xx 
 
 
4.0 
x 
x 
xx 
x 
xx 
 
 
6.0 
x 
x 
xx 
x 
xx 
 
Day 3 
1.5 
x 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
 
Day 14 
1.5 
x 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
 
Day 30 
1.5 
x 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
Left 
Baseline 
1,5 
x 
x 
xx 
x 
xx 
 
 
 
x 
x 
xx 
x 
xx 
Notes: N = Number of participants in the Safety Subset Population with the DPAOEs assessed at the respective timepoint and frequency. 
Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 78: 
Summary of DPOAE Results by Timepoint, Frequency, and Ear – Safety Subset 
Population 
 


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Table 79: 
SNR Change from Baseline by Timepoint, Frequency, and Ear – Safety Population 
Ear 
Timepoint 
Frequency (kHz) 
N 
Measurement 
Change from Baseline (dB) 
Mean 
SD 
Median 
Min, Max 
Mean 
SD 
Median 
Min, Max 
Right 
Baseline 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
2.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
3.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
4.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
6.0 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
Day 3 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 14 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
Day 30 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
xx.x 
xx.x 
xx.x 
xx, xx 
Left 
Baseline 
1.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
 
 
 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
- 
- 
- 
- 
Notes: N = Number of participants in the Safety Population with the DPAOEs assessed at the respective timepoint and frequency. Baseline = The most recent measurement prior to start of infusion. 
Table with similar format: 
Table 80: 
SNR Change from Baseline by Timepoint, Frequency, and Ear – Safety Subset Population 
 


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14.4 
Summary of Concomitant Medications 
Table 81: 
Number and Percentage of Participants with Prior Medications by WHO Drug Classification and Cohort – Safety Population 
[Implementation Note: Include prior medications (medications with an end date prior to date of infusion) only.] 
WHO Drug Code 
Level 1, Anatomic Group 
WHO Drug Code 
Level 2, Therapeutic Subgroup 
Cohort 1 
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any Level 1 Codes 
Any Level 2 Codes 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
[ATC Level 1 - 1] 
Any [ATC 1 – 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC Level 1 – 2] 
[ATC 2 - 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
 
 
 
 
 
 
Notes: N = Number of participants in the Safety Population. n = Number of participants reporting taking at least one medication in the specific WHO Drug Class. 
 
 


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Table 82: 
Number and Percentage of Participants with Concomitant Medications by WHO Drug Classification and Cohort – Safety 
Population 
[Implementation Note: Include concomitant medications (medications that are ongoing or have an end date after date of infusion) only.] 
WHO Drug Code 
Level 1, Anatomic Group 
WHO Drug Code 
Level 2, Therapeutic Subgroup 
Cohort 1 
(N=X) 
Cohort 2 
(N=X) 
Cohort 3 
(N=X) 
Cohort 4 
(N=X) 
Cohort 5 
(N=X) 
All Participants 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any Level 1 Codes 
Any Level 2 Codes 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
[ATC Level 1 - 1] 
Any [ATC 1 – 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC Level 1 – 2] 
[ATC 2 - 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
 
 
 
 
 
 
Notes: N = Number of participants in the Safety Population. n = Number of participants reporting taking at least one medication in the specific WHO Drug Class. 
 


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14.2 
Pharmacokinetics Data 
Table 83: 
Total Apramycin Concentrations in Plasma 
[Implementation Note: Mark concentrations collected out of window with an asterisk (*) next to the concentration and include a footnote: “Samples 
collected out of window are noted by an asterisk (*)”. Mark concentrations collected substantially out of window with two asterisks (**) next to the 
concentration and include a footnote: “Samples collected substantially out of window are noted by two asterisks (**)”. Mark imputed concentration 
times with three asterisks (***) next to the concentration and include a footnote: “Samples with imputed collection times are noted by three asterisks 
(***)”.] 
  
Nominal Timea (h) 
Participant ID 
0 
0.5 
1 
2 
4 
8 
16 
24 
36 
48 
60 
PH2.00123 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
PH2.00124 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
PH2.00125 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
 
  
  
  
  
  
  
  
  
  
  
  
Statistics 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Nb 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Mean 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
SD 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
GM 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
CV% 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Min 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Max 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Notes: Concentrations are reported in units of µg/ml. 
[Out of window footnotes here, if applicable] 
a Times are relative to time of dosing. 
b Number of data points used to compute the summary statistics. For calculation of summary statistics, BQL values were imputed as 0 if the sample was taken before the first measurable PK sample 
with a concentration above the LLOQ.  BQL values were treated as missing otherwise.  
Table with Similar Format: 
Table 84: 
Free Apramycin Concentrations in Plasma 


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Table 85: 
Summary Statistics for the Free Portion of Plasma Apramycin 
Nominal Timea (h) 
N 
Summary Statistics 
Mean 
SD 
Median 
Min, Max 
0.5 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
36 
x 
xx.x 
xx.x 
xx.x 
xx, xx 
Note: N = Number of data points used to calculate the summary statistics. Concentrations are reported in units of µg/mL. 
a Times are relative to time of dosing. 
 


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Table 86: 
Summary Statistics for Total Apramycin in Plasma PK Parameters 
Statistics 
Cmax 
(µg/mL) 
Tmax 
(h) 
AUC(0-24) 
(µg *h/mL) 
AUC(0-last) 
(µg*h/mL) 
AUC(0-∞) 
(µg*h/mL) 
Ke 
(1/h) 
t1/2 
(h) 
CL 
(L/h) 
Vd 
(L) 
N 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Mean 
x 
x 
x 
x 
x 
x 
x 
x 
x 
SD 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Min 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Max 
x 
x 
x 
x 
x 
x 
x 
x 
x 
CV% 
x 
x 
x 
x 
x 
x 
x 
x 
x 
GM 
x 
x 
x 
x 
x 
x 
x 
x 
X 
Note: N = Number of data points used to compute the summary statistics. 
 


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Table 87: 
Summary Statistics for Apramycin in ELF Concentrations and Apramycin in ELF to Total Apramycin in Plasma Concentration 
Ratios 
Statistics 
Total Apramycin in Plasma Concentration (µg/mL) 
Apramycin in ELF Concentration (µg/mL) 
Concentration Ratio Apramycin in ELF to Total 
Apramycin in Plasma 
0.5 h 
2 h 
4 h 
8 h 
24 h 
0.5 h 
2 h 
4 h 
8 h 
24 h 
0.5 h 
2 h 
4 h 
8 h 
24 h 
N 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Mean 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
SD 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Min 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Max 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
CV% 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
GM 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Note: N = Number of data points used to compute the summary statistics. 
Tables with Similar Format: 
Table 88: 
Summary Statistics for Apramycin in AM Concentrations and Apramycin in AM to Total Apramycin in Plasma Concentration 
Ratios 
 


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Table 89: 
Apramycin in ELF and Apramycin in AM Population PK Parameter Estimates 
PK Parameter 
Assay 
N 
Population Estimate 
Cmax (µg/mL) 
ELF 
x 
x 
AM 
x 
x 
AUC(0-24) (µg*h/mL) 
ELF 
x 
x 
AM 
x 
x 
AUC(0-last) (µg*h/mL) 
ELF 
x 
x 
 
AM 
x 
x 
AUC(0-∞) (µg*h/mL) 
ELF 
x 
x 
 
AM 
x 
x 
Tmax (h) 
ELF 
x 
x 
 
AM 
x 
x 
t1/2 (1/h) 
ELF 
x 
x 
 
AM 
x 
x 
Note: N = Number of participants used to compute the PK parameter.  
 


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Table 90: 
Apramycin in ELF to Total Apramycin in Plasma and Apramycin in AM to Total 
Apramycin in Plasma PK Parameter Ratios by PK Parameter 
PK Parameter 
Assay 
N 
Population Estimate 
Lung PK to Plasma PK Ratio 
Cmax (µg/mL) 
Total Plasma 
x 
x 
NA 
ELF 
x 
x 
x 
AM 
x 
x 
x 
AUC(0-24) (µg*h/mL) 
Total Plasma 
x 
x 
NA 
ELF 
x 
x 
x 
AM 
x 
x 
x 
AUC(0-last) (µg*h/mL) 
Total Plasma 
x 
x 
NA 
 
ELF 
x 
x 
x 
 
AM 
x 
x 
x 
AUC(0-∞) (µg*h/mL) 
Total Plasma 
x 
x 
NA 
 
ELF 
x 
x 
x 
 
AM 
x 
x 
x 
Note: N = Number of participants used to compute the PK parameter.  
 


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APPENDIX 2.  FIGURE MOCK-UPS 
LIST OF FIGURES 
Figure 1: 
Schematic of Study Design .......................................................................................126 
Figure 2: 
CONSORT Flow Diagram .......................................................................................127 
Figure 3: 
Incidence of Related Adverse Events by MedDRA System Organ Class and 
Maximum Severity – Safety Population ...................................................................128 
Figure 4: 
Incidence of Related Adverse Events by MedDRA System Organ Class and 
Maximum Severity – Safety Subset Population .......................................................128 
Figure 5: 
Frequency of Related Adverse Events by MedDRA System Organ Class and 
Severity – Safety Population ....................................................................................129 
Figure 6: 
Frequency of Related Adverse Events by MedDRA System Organ Class and 
Severity – Safety Subset Population .........................................................................129 
Figure 7: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Sodium - Safety Population ....................................130 
Figure 8: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Sodium – Safety Subset Population ........................131 
Figure 9: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Potassium – Safety Population ................................131 
Figure 10: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Potassium – Safety Subset Population ....................131 
Figure 11: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Glucose – Safety Population ...................................131 
Figure 12: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Glucose – Safety Subset Population .......................131 
Figure 13: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Blood Urea Nitrogen – Safety Population ..............131 
Figure 14: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Blood Urea Nitrogen – Safety Subset 
Population .................................................................................................................131 
Figure 15: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Creatinine – Safety Population ...............................131 
Figure 16: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Creatinine – Safety Subset Population ....................131 
Figure 17: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – eGFR – Safety Population.......................................131 


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Figure 18: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – eGFR – Safety Subset Population ...........................131 
Figure 19: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Calcium – Safety Population ...................................131 
Figure 20: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Calcium – Safety Subset Population .......................131 
Figure 21: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Chloride – Safety Population ..................................131 
Figure 22: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Chloride – Safety Subset Population ......................131 
Figure 23: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Magnesium – Safety Population .............................131 
Figure 24: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Magnesium – Safety Subset Population ..................131 
Figure 25: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint –Total Carbon Dioxide – Safety Population ..............131 
Figure 26: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint –Total Carbon Dioxide – Safety Subset 
Population .................................................................................................................131 
Figure 27: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Albumin – Safety Population ..................................132 
Figure 28: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Albumin – Safety Subset Population ......................132 
Figure 29: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Total Protein – Safety Population ...........................132 
Figure 30: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Total Protein – Safety Subset Population ...............132 
Figure 31: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Alkaline Phosphatase – Safety Population..............132 
Figure 32: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Alkaline Phosphatase – Safety Subset 
Population .................................................................................................................132 
Figure 33: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Alanine Aminotransferase – Safety 
Population .................................................................................................................132 
Figure 34: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Alanine Aminotransferase – Safety Subset 
Population .................................................................................................................132 


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Figure 35: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Aspartate Aminotransferase – Safety 
Population .................................................................................................................132 
Figure 36: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Aspartate Aminotransferase – Safety Subset 
Population .................................................................................................................132 
Figure 37: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Total Bilirubin – Safety Population ........................132 
Figure 38: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Total Bilirubin – Safety Subset Population .............132 
Figure 39: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Direct Bilirubin – Safety Population .......................132 
Figure 40: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Direct Bilirubin – Safety Subset Population ...........132 
Figure 41: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – TSH – Safety Population.........................................132 
Figure 42: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – TSH – Safety Subset Population .............................132 
Figure 43: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Free T4 – Safety Population ....................................132 
Figure 44: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Free T4 – Safety Subset Population ........................132 
Figure 45: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Free T3 – Safety Population ....................................132 
Figure 46: Chemistry Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Free T3 – Safety Subset Population ........................133 
Figure 47: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Hemoglobin – Safety Population ............................134 
Figure 48: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Hemoglobin – Safety Subset Population ................135 
Figure 49: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Hematocrit – Safety Population ..............................135 
Figure 50: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Hematocrit – Safety Subset Population ..................135 
Figure 51: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Platelets – Safety Population ...................................135 
Figure 52: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Platelets – Safety Subset Population .......................135 


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Figure 53: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – White Blood Cell Count – Safety Population .........135 
Figure 54: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – White Blood Cell Count – Safety Subset 
Population .................................................................................................................135 
Figure 55: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Neutrophils – Safety Population .............................135 
Figure 56: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Neutrophils – Safety Subset Population ..................135 
Figure 57: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Lymphocytes – Safety Population ..........................135 
Figure 58: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Lymphocytes – Safety Subset Population ...............135 
Figure 59: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Eosinophils – Safety Population .............................135 
Figure 60: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Eosinophils – Safety Subset Population..................135 
Figure 61: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Basophils – Safety Population ................................135 
Figure 62: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Basophils – Safety Subset Population .....................135 
Figure 63: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Monocytes – Safety Population ..............................135 
Figure 64: Hematology Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Monocytes – Safety Subset Population ...................135 
Figure 65: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Activated Partial Thromboplastin Time – 
Safety Population ......................................................................................................136 
Figure 66: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Activated Partial Thromboplastin Time – 
Safety Subset Population ..........................................................................................137 
Figure 67: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Prothrombin Time – Safety Population ..................137 
Figure 68: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – Prothrombin Time – Safety Subset Population .......137 
Figure 69: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – International Normalized Ratio – Safety 
Population .................................................................................................................137 


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Figure 70: Coagulation Laboratory Results by Scheduled Visits: Change from Baseline 
by Parameter and Timepoint – International Normalized Ratio – Safety 
Subset Population .....................................................................................................137 
Figure 71: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Serum Cystatin C – Safety 
Population .................................................................................................................138 
Figure 72: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Serum Cystatin C – Safety Subset 
Population .................................................................................................................139 
Figure 73: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Urinary KIM-1 – Safety Population .........139 
Figure 74: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Urinary KIM-1 – Safety Subset 
Population .................................................................................................................139 
Figure 75: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Urinary KIM-1 Normalized – Safety 
Population .................................................................................................................139 
Figure 76: Kidney Function Biomarker Results by Scheduled Visits: Change from 
Baseline by Parameter and Timepoint – Urinary KIM-1 Normalized – Safety 
Subset Population .....................................................................................................139 
Figure 77: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Systolic Blood Pressure – Safety Population .......................................140 
Figure 78: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Systolic Blood Pressure – Safety Subset Population ...........................141 
Figure 79: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Diastolic Blood Pressure – Safety Population .....................................141 
Figure 80: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Diastolic Blood Pressure – Safety Subset Population ..........................141 
Figure 81: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Heart Rate – Safety Population ............................................................141 
Figure 82: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Heart Rate – Safety Subset Population ................................................141 
Figure 83: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Respiratory Rate – Safety Population ..................................................141 
Figure 84: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Respiratory Rate – Safety Subset Population ......................................141 
Figure 85: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Temperature – Safety Population .........................................................141 


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Figure 86: Vital Signs by Scheduled Visits: Change from Baseline by Parameter and 
Timepoint – Temperature – Safety Subset Population .............................................141 
Figure 87: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – PR Interval – Safety Population ....................................................142 
Figure 88: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – PR Interval – Safety Subset Population ........................................143 
Figure 89: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QRS Interval – Safety Population .................................................143 
Figure 90: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QRS Interval – Safety Subset Population .....................................143 
Figure 91: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QT Interval – Safety Population ....................................................143 
Figure 92: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QT Interval – Safety Subset Population ........................................143 
Figure 93: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QTcF Interval – Safety Population................................................143 
Figure 94: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – QTcF Interval – Safety Subset Population ....................................143 
Figure 95: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – RR Interval – Safety Population ....................................................143 
Figure 96: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – RR Interval – Safety Subset Population ........................................143 
Figure 97: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – Ventricular Rate – Safety Population ............................................143 
Figure 98: ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter 
and Timepoint – Ventricular Rate – Safety Subset Population ................................143 
Figure 99: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 kHz – Pure-Tone 
Audiometry – Safety Population ..............................................................................144 
Figure 100: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 kHz – Pure-Tone 
Audiometry – Safety Subset Population ...................................................................145 
Figure 101: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – 9.0, 10.0, 11.0, 12.5, 14.5, 16.0, 18.0 and 20 kHz – 
Pure-Tone Audiometry – Safety Population .............................................................145 
Figure 102: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – 9.0 10.0, 11.0, 12.5, 14.5, 16.0, 18.0 and 20 kHz – 
Pure-Tone Audiometry – Safety Subset Population .................................................145 


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Figure 103: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – DPOAE – Safety Population .................................................145 
Figure 104: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – DPOAE – Safety Subset Population ......................................145 
Figure 105: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – SNR – Safety Population .......................................................145 
Figure 106: Audiology by Scheduled Visits: Change from Baseline by Timepoint, 
Frequency, and Ear – SNR – Safety Subset Population ...........................................145 
Figure 107: Individual Total Apramycin Concentration - Time  Profiles in Plasma ...................146 
Figure 108: Semi-Log Individual Total Apramycin Concentration-Time Profiles in 
Plasma .......................................................................................................................147 
Figure 109: Mean Total Apramycin Concentration-Time Profiles in Plasma .............................148 
Figure 110: Mean Semi-Log Total Apramycin Concentration-Time Profiles in Plasma ............149 
Figure 111: Total Apramycin Concentration in Plasma, ELF, and AM ......................................150 
Figure 112: Population Geometric Mean Total Apramycin Concentrations in Plasma, 
ELF, and AM ............................................................................................................151 
Figure 113: Population Semi-Log Geometric Mean Total Apramycin Concentrations in 
Plasma, ELF, and AM ..............................................................................................152 
Figure 114: Individual Apramycin in ELF to Total Apramycin in Plasma and Apramycin 
in AM to Total Apramycin in Plasma Concentration Ratios ....................................153 
Figure 115: Geometric Mean Apramycin in ELF to Total Apramycin in Plasma and 
Apramycin in AM to Total Apramycin in Plasma Concentration Ratios .................154 
 


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9.1 
Overall Study Design and Plan Description 
Figure 1: 
Schematic of Study Design 
 


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10.1 
Disposition of Participants 
Figure 2: 
CONSORT Flow Diagram 
 
 
 
Assessed for Eligibility (n=x) 
Enrollment 
Excluded* (n=x) 
¨   [Reason 1] (n=x) 
¨   Concern of potential risk (n=x) 
Enrolled (n=x) 
Allocation 
Analysis 
Follow-Up 
Allocated to Cohort 1 (n=x) 
♦Received infusion (n=x) 
♦Completed BAL (n=x) 
Allocated to Cohort 3 (n=x) 
♦Received infusion (n=x) 
♦Completed BAL (n=x) 
Allocated to Cohort 5 (n=x) 
♦Received infusion (n=x) 
♦Completed BAL (n=x) 
♦
Allocated to Cohort 4 (n=x) 
♦Received infusion (n=x) 
♦Completed BAL (n=x) 
Allocated to Cohort 2 (n=x)  
♦Received infusion (n=x) 
♦Completed BAL (n=x) 
Discontinued Intervention 
(n=x) 
[Reason 1] (x) 
  
Early Termination (n=x) 
[Reason 1] (x) 
Discontinued Intervention 
(n=x) 
[Reason 1] (x) 
  
Early Termination (n=x) 
[Reason 1] (x) 
Discontinued Intervention 
(n=x) 
[Reason 1] (x) 
  
Early Termination (n=x) 
[Reason 1] (x) 
Discontinued Intervention 
(n=x) 
[Reason 1] (x) 
  
Early Termination (n=x) 
[Reason 1] (x) 
Discontinued Intervention 
(n=x) 
[Reason 1] (x) 
  
Early Termination (n=x) 
[Reason 1] (x) 
Analyzed for safety (n=x) 
♦Excluded (n=x) 
[Reason 1] (x)  
Analyzed for PK (n=x) 
♦Excluded (n=x) 
[Reason 1] (x) 
(Include for all analysis 
populations) 
Analyzed for safety (n=x) 
♦Excluded (n=x) 
[Reason 1] (x)  
Analyzed for PK (n=x) 
♦Excluded (n=x) 
[Reason 1] (x) 
(Include for all analysis 
populations)  
Analyzed for safety (n=x) 
♦Excluded (n=x) 
[Reason 1] (x)  
Analyzed for PK (n=x) 
♦Excluded (n=x) 
[Reason 1] (x) 
(Include for all analysis 
populations) 
Analyzed for safety (n=x) 
♦Excluded (n=x) 
[Reason 1] (x)  
Analyzed for PK (n=x) 
♦Excluded (n=x) 
[Reason 1] (x) 
(Include for all analysis 
populations)  
Analyzed for safety (n=x) 
♦Excluded (n=x) 
[Reason 1] (x)  
Analyzed for PK (n=x) 
♦Excluded (n=x) 
[Reason 1] (x) 
(Include for all analysis 
populations)  


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14.3.1.2 
Unsolicited Adverse Events 
Figure 3: 
Incidence of Related Adverse Events by MedDRA System Organ Class and Maximum 
Severity – Safety Population  
[Implementation Note: Replace “Group A” with “All Participants (N=X)”, where X is the number of 
participants in the Safety Population. The font and size of text in the figure should be the same as in the 
report.] 
 
Figure with similar format: 
Figure 4: 
Incidence of Related Adverse Events by MedDRA System Organ Class and Maximum 
Severity – Safety Subset Population  


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Figure 5: 
Frequency of Related Adverse Events by MedDRA System Organ Class and Severity – 
Safety Population 
[Implementation Note: Replace “Group A” with “All Participants (N=X)”, where X is the number of 
participants in the Safety Subset Population. The font and size of text in the figure should be the same as in 
the report.] 
 
Figure with similar format: 
Figure 6: 
Frequency of Related Adverse Events by MedDRA System Organ Class and Severity – 
Safety Subset Population 


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14.3.5 
Displays of Laboratory Results 
14.3.5.1  
Chemistry Results 
Figure 7: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Sodium - Safety Population 
 
 
 


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Figures with Similar Format: 
Figure 8: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Sodium – Safety Subset Population 
Figure 9: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Potassium – Safety Population 
Figure 10: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Potassium – Safety Subset Population 
Figure 11: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Glucose – Safety Population 
Figure 12: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Glucose – Safety Subset Population 
Figure 13: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Blood Urea Nitrogen – Safety Population 
Figure 14: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Blood Urea Nitrogen – Safety Subset Population 
Figure 15: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Creatinine – Safety Population 
Figure 16: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Creatinine – Safety Subset Population 
Figure 17: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – eGFR – Safety Population 
Figure 18: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – eGFR – Safety Subset Population 
Figure 19: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Calcium – Safety Population 
Figure 20: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Calcium – Safety Subset Population 
Figure 21: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Chloride – Safety Population 
Figure 22: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Chloride – Safety Subset Population 
Figure 23: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Magnesium – Safety Population 
Figure 24: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Magnesium – Safety Subset Population 
Figure 25: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint –Total Carbon Dioxide – Safety Population 
Figure 26: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint –Total Carbon Dioxide – Safety Subset Population 


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Figure 27: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Albumin – Safety Population 
Figure 28: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Albumin – Safety Subset Population 
Figure 29: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Total Protein – Safety Population 
Figure 30: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Total Protein – Safety Subset Population 
Figure 31: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Alkaline Phosphatase – Safety Population 
Figure 32: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Alkaline Phosphatase – Safety Subset Population 
Figure 33: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Alanine Aminotransferase – Safety Population 
Figure 34: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Alanine Aminotransferase – Safety Subset Population 
Figure 35: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Aspartate Aminotransferase – Safety Population 
Figure 36: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Aspartate Aminotransferase – Safety Subset Population 
Figure 37: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Total Bilirubin – Safety Population 
Figure 38: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Total Bilirubin – Safety Subset Population 
Figure 39: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Direct Bilirubin – Safety Population 
Figure 40: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Direct Bilirubin – Safety Subset Population 
Figure 41: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – TSH – Safety Population 
Figure 42: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – TSH – Safety Subset Population 
Figure 43: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Free T4 – Safety Population 
Figure 44: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Free T4 – Safety Subset Population 
Figure 45: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Free T3 – Safety Population 


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Figure 46: 
Chemistry Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Free T3 – Safety Subset Population 


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14.3.5.2  
Hematology Results 
Figure 47: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Hemoglobin – Safety Population 
 
 


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Figures with Similar Format: 
Figure 48: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Hemoglobin – Safety Subset Population 
Figure 49: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Hematocrit – Safety Population 
Figure 50: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Hematocrit – Safety Subset Population 
Figure 51: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Platelets – Safety Population 
Figure 52: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Platelets – Safety Subset Population 
Figure 53: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – White Blood Cell Count – Safety Population 
Figure 54: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – White Blood Cell Count – Safety Subset Population 
Figure 55: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Neutrophils – Safety Population 
Figure 56: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Neutrophils – Safety Subset Population 
Figure 57: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Lymphocytes – Safety Population 
Figure 58: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Lymphocytes – Safety Subset Population 
Figure 59: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Eosinophils – Safety Population 
Figure 60: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Eosinophils – Safety Subset Population 
Figure 61: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Basophils – Safety Population 
Figure 62: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Basophils – Safety Subset Population 
Figure 63: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Monocytes – Safety Population 
Figure 64: 
Hematology Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Monocytes – Safety Subset Population 
 


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14.3.5.3  
Coagulation Results 
Figure 65: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Activated Partial Thromboplastin Time – Safety Population 
 
 


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Figures with Similar Format: 
Figure 66: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Activated Partial Thromboplastin Time – Safety Subset 
Population  
Figure 67: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Prothrombin Time – Safety Population  
Figure 68: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Prothrombin Time – Safety Subset Population 
Figure 69: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – International Normalized Ratio – Safety Population 
Figure 70: 
Coagulation Laboratory Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – International Normalized Ratio – Safety Subset Population 
 


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14.3.5.5  
Kidney Function Biomarker Results 
Figure 71: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Serum Cystatin C – Safety Population 
 
 
 


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Figures with Similar Format: 
Figure 72: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Serum Cystatin C – Safety Subset Population  
Figure 73: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Urinary KIM-1 – Safety Population  
Figure 74: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Urinary KIM-1 – Safety Subset Population 
Figure 75: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Urinary KIM-1 Normalized – Safety Population 
Figure 76: 
Kidney Function Biomarker Results by Scheduled Visits: Change from Baseline by 
Parameter and Timepoint – Urinary KIM-1 Normalized – Safety Subset Population 
 


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14.3.6 
 
Displays of Vital Signs 
Figure 77: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Systolic Blood Pressure – Safety Population 
 


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Figures with Similar Format: 
Figure 78: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Systolic Blood Pressure – Safety Subset Population 
Figure 79: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Diastolic Blood Pressure – Safety Population 
Figure 80: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Diastolic Blood Pressure – Safety Subset Population  
Figure 81: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Heart Rate – Safety Population 
Figure 82: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Heart Rate – Safety Subset Population 
Figure 83: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Respiratory Rate – Safety Population 
Figure 84: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Respiratory Rate – Safety Subset Population 
Figure 85: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Temperature – Safety Population 
Figure 86: 
Vital Signs by Scheduled Visits: Change from Baseline by Parameter and Timepoint – 
Temperature – Safety Subset Population 
 


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14.3.7 
 
Displays of ECG Measurements 
Figure 87: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– PR Interval – Safety Population 
 


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Figures with Similar Format: 
Figure 88: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– PR Interval – Safety Subset Population 
Figure 89: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QRS Interval – Safety Population 
Figure 90: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QRS Interval – Safety Subset Population 
Figure 91: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QT Interval – Safety Population 
Figure 92: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QT Interval – Safety Subset Population 
Figure 93: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QTcF Interval – Safety Population 
Figure 94: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– QTcF Interval – Safety Subset Population 
Figure 95: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– RR Interval – Safety Population 
Figure 96: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– RR Interval – Safety Subset Population 
Figure 97: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– Ventricular Rate – Safety Population 
Figure 98: 
ECG by Scheduled Visits: Change from Pre-Dose Baseline by Parameter and Timepoint 
– Ventricular Rate – Safety Subset Population 
 


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14.3.8 
 
Displays of Audiology Results 
Figure 99: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 kHz – Pure-Tone Audiometry – Safety Population 
 


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Figures with Similar Format: 
Figure 100: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 kHz – Pure-Tone Audiometry – Safety Subset Population 
Figure 101: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– 9.0, 10.0, 11.0, 12.5, 14.5, 16.0, 18.0 and 20 kHz – Pure-Tone Audiometry – Safety 
Population 
Figure 102: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– 9.0 10.0, 11.0, 12.5, 14.5, 16.0, 18.0 and 20 kHz – Pure-Tone Audiometry – Safety Subset 
Population  
Figure 103: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– DPOAE – Safety Population 
Figure 104: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– DPOAE – Safety Subset Population 
Figure 105: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– SNR – Safety Population 
Figure 106: 
Audiology by Scheduled Visits: Change from Baseline by Timepoint, Frequency, and Ear 
– SNR – Safety Subset Population 
 


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14.3.8 
 
Displays for Pharmacokinetics Data 
Figure 107: 
Individual Total Apramycin Concentration-Time Profiles in Plasma 
[Implementation Note: Different lines (types and colors) will be used for each cohort.] 
  
 


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Figure 108: 
Semi-Log Individual Total Apramycin Concentration-Time Profiles in Plasma 
[Implementation Note: Different line types and colors will be used for each cohort.] 
 
 


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Figure 109: 
Mean Total Apramycin Concentration-Time Profiles in Plasma 
[Implementation Note: Different symbols and line types will be used for each cohort. Error bars represent ± 1 
SD.] 
 
 


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Figure 110: 
Mean Semi-Log Total Apramycin Concentration-Time Profiles in Plasma 
[Implementation Note: Different symbols and line types will be used for each cohort.] 
 
 


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Figure 111: 
Total Apramycin Concentration in Plasma, ELF, and AM 
 
 


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Figure 112: 
Population Geometric Mean Total Apramycin Concentrations in Plasma, ELF, and AM 
[Implementation Note: Different symbols and line types will be used for each parameter.] 
 
 


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Figure 113: 
Population Semi-Log Geometric Mean Total Apramycin Concentrations in Plasma, ELF, 
and AM 
[Implementation Note: Different symbols and line types will be used for each parameter.] 
 
 


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Figure 114: 
Individual Apramycin in ELF to Total Apramycin in Plasma and Apramycin in AM to 
Total Apramycin in Plasma Concentration Ratios 
 
 


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Figure 115: 
Geometric Mean Apramycin in ELF to Total Apramycin in Plasma and Apramycin in 
AM to Total Apramycin in Plasma Concentration Ratios 
[Implementation Note: Different symbols and line types will be used for each parameter.] 
 
 


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APPENDIX 3. LISTINGS MOCK-UPS 
LISTINGS 
Listing 1: Early Terminations or Discontinued Participants .....................................................158 
Listing 2: Participant-Specific Protocol Deviations ..................................................................159 
Listing 3: Non-Participant-Specific Protocol Deviations .........................................................160 
Listing 4: Participants Excluded from Analysis Populations ....................................................161 
Listing 5: Demographic Data ....................................................................................................162 
Listing 6: Pre-Existing and Concurrent Medical Conditions ....................................................163 
Listing 7: Treatment Administration .........................................................................................164 
Listing 8: Treatment Emergent Adverse Events .......................................................................165 
Listing 9: Clinical Laboratory Results – Chemistry .................................................................166 
Listing 10: Clinical Laboratory Results – Hematology ..............................................................167 
Listing 11: Clinical Laboratory Results – Coagulation ...............................................................168 
Listing 12: Clinical Laboratory Results – Urinalysis ..................................................................169 
Listing 13: Kidney Function Biomarker Results – Cystatin C and KIM-1 .................................170 
Listing 14: Screening Laboratory Results – Serology ................................................................171 
Listing 15: Screening Laboratory Results – Urine Drug and Cotinine and Alcohol 
Breathalyzer ..............................................................................................................172 
Listing 16: Screening Laboratory Results – Pregnancy and FSH ...............................................173 
Listing 17: Vital Signs ................................................................................................................174 
Listing 18: Abnormal Physical Exam Findings ..........................................................................175 
Listing 19: Listing of ECG Interval Measurements ....................................................................176 
Listing 20: Listing of ECG Overall Interpretation and Change from Baseline ..........................177 
Listing 21: Otoacoustic Test Results – Ear Otoscopy .................................................................178 
Listing 22: Otoacoustic Test Results – Pure-Tone Audiometry .................................................179 
Listing 23: Otoacoustic Test Results – DPOAE .........................................................................180 
Listing 24: Screening Otoacoustic Test Results – Valsalva ........................................................181 
Listing 25: Screening Otoacoustic Test Results – Tympanometry .............................................182 
Listing 26: Screening Otoacoustic Test Results – Stapedial Reflexes ........................................183 
Listing 27: Prior Medications ......................................................................................................184 
Listing 28: Concurrent Medications ............................................................................................185 


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Listing 29: Pregnancy Reports – Maternal Information .............................................................186 
Listing 30: Pregnancy Reports – Gravida and Para ....................................................................186 
Listing 31: Pregnancy Reports – Live Birth Outcomes ..............................................................187 
Listing 32: Pregnancy Reports – Still Birth Outcomes ...............................................................187 
Listing 33: Pregnancy Reports – Spontaneous, Elective, or Therapeutic Abortion 
Outcomes ..................................................................................................................187 
Listing 34: Birth Control Listing.................................................................................................188 
Listing 35: Participant Level Total Apramycin Concentrations in Plasma .................................189 
Listing 36: Participant Level Free Apramycin Concentrations in Plasma ..................................190 
Listing 37: Participant Level Free Portion of Plasma Apramycin ..............................................191 
Listing 38: Participant Specific Total Apramycin in Plasma PK Parameters .............................192 
Listing 39: Participant Level Apramycin Concentrations in ELF ..............................................193 
Listing 40: Participant Level Apramycin Concentrations in AM ...............................................194 
Listing 41: Apramycin in ELF and AM to Total Apramycin in Plasma Concentration 
Ratios ........................................................................................................................195 
Listing 42: Bronchoalveolar Lavage Cell Counts .......................................................................195 
 


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16.1.6 Listing of Participants Receiving Investigational Product  
(Not included in SAP, but this is a placeholder for the CSR) 
 


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16.2 
Database Listings by Participant 
16.2.1 Discontinued Participants 
Listing 1: 
Early Terminations or Discontinued Participants 
[Implementation Note: Category will be either “Early Termination” or “Treatment Discontinuation.” In the “Reason” column, concatenate any “specify” 
fields, including AE number and DV number. Sort order: Cohort, Participant ID, Category (in the case a participant both terminates early and 
discontinues treatment).] 
Cohort 
 Participant ID 
Category 
Reason for Early Termination or Treatment Discontinuation 
Study Day 
 
 
 
 
 
 
 
 


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16.2.2 Protocol Deviations 
Listing 2: 
Participant-Specific Protocol Deviations 
[Implementation Note: In the “Deviation” column, concatenate any and all “specify” fields (including visit number, etc.). If “Reason for Deviation” is 
“Other,” concatenate “specify” field, separated by a colon, e.g., “Other: Participant refusal.” Sort order: Cohort, Participant ID, DV Number.] 
Cohort 
Participant 
ID 
DV Number 
Deviation 
Deviation 
Category 
Study Day 
Reason for 
Deviation 
Deviation 
Resulted 
in AE? 
Deviation Resulted 
in Participant 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 3: 
Non-Participant-Specific Protocol Deviations 
[Implementation Note: In the “Deviation” column, concatenate any and all “specify” fields (including visit number, etc.). If the “Reason for Deviation” 
is “Other,” concatenate “specify” field, separated by a colon, e.g., “Other: Participant refusal.” In “Comments” column, replace any occurrences of 
PATID with USUBJID. Sort order: Start Date, Deviation.]  
Start Date  
Deviation 
End Date 
Reason for 
Deviation 
Deviation Resulted in 
Participant 
Termination? 
Deviation Affected 
Product Stability? 
Deviation 
Category 
Deviation 
Resolution 
Comments 
ddMMMyyyy 
 
ddMMMyyyy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.3 Participants Excluded from the Efficacy Analysis 
Listing 4: 
Participants Excluded from Analysis Populations 
[Implementation Note: The data in this listing should be congruent with the “Analysis Population by Cohort” table. If a participant was not excluded 
from any analysis population, the participant will not appear in the listing. If the participant is excluded from multiple analysis population, they will have 
one row per analysis population excluded from in the listing. If no specifications are required for a reason for exclusion, then exclude the last column 
“Reason Participant Excluded Specification”. Sort Order: Cohort, Participant ID, Analysis from which participant is excluded (order: Safety, Safety 
Subset, Lung PK, Plasma PK, PK Analysis Subset).] 
Cohort 
Participant ID 
Analyses in which 
Participant is Excluded 
Results Available? 
Reason Participant Excluded 
Reason Participant Excluded 
Specification 
Cohort 1 
PH2.00001 
PK Analysis Subset 
Yes 
Has protocol deviations that potentially 
impact PK 
Participant consumed food 1 hour prior to 
dosing. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: “Yes” in the “Results available” column indicates that available data were removed from the analysis. “No” indicates that no data were available for inclusion in the analysis. 
 
 


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16.2.4 Demographic Data 
Listing 5: 
Demographic Data 
[Implementation Note: If a participant is multi-racial, in “Race” column, note “Multiple: (list races, separated by a comma).” Sort order: Cohort, 
Participant ID.] 
Cohort 
 Participant ID 
Sex 
Age at Enrollment (years) 
Ethnicity 
Race 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 6: 
Pre-Existing and Concurrent Medical Conditions 
[Implementation Note: “Condition Start Day” and “Condition End Day” are relative to date of treatment administration (which is Day 1, day before 
treatment administration is Day -1). Rather than use exact study days, categorize as follows: 
• 5 years prior to enrollment 
• 1-5 years prior to enrollment 
• 1-12 months prior to enrollment 
• Within 1 month of enrollment 
• During study 
• If ongoing, display “Ongoing” in the “Condition End Day” column 
Sort order: Cohort, Participant ID, MH Number.] 
Cohort 
 Participant 
ID 
MH Number 
Medical History Term 
MedDRA System Organ Class 
MedDRA Preferred Term 
Condition Start Day  
Condition End Day 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.5 Compliance and/or Drug Concentration Data (if available) 
Listing 7: 
Treatment Administration 
[Implementation Note: Sort order: Cohort, Participant ID.] 
Cohort  
Participant ID  
Planned 
Dose (mg)  
Volume 
Administered 
(mL) 
Actual Dose 
Administered 
(mg) 
Infusion 
Date 
Infusion Start 
Time 
(hh:mm) 
Infusion End 
Time 
(hh:mm) 
Interruptions in 
study product 
administration? 
Interruption 
Start Time 
(hh:mm) 
Interruption 
End Time 
(hh:mm) 
 
 
 
 
 
ddMMMyyyy 
hh:mm 
hh:mm 
Yes/No 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.7 Adverse Events 
Listing 8: 
Treatment Emergent Adverse Events 
[Implementation Note: If the event is ongoing (no stop date), indicate “ongoing” in the duration column. This listing includes all treatment emergent 
adverse events. Sort order: Cohort, Participant ID, AE Number.] 
Adverse 
Event 
MedDRA 
System 
Organ 
Class 
MedDRA 
High Level 
Group 
Term 
MedDRA 
Preferred 
Term 
Study Day 
of AE 
Onset 
Duration 
(Days) 
Severity 
SAE? 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Participant 
Discontinued 
Due to AE 
Outcome 
Cohort: , Participant ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
 
Cohort: , Participant ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 


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16.2.8 Individual Laboratory Measurements 
Listing 9: 
Clinical Laboratory Results – Chemistry 
[Implementation Note: These listings (for hematology, chemistry, coagulation, and urinalysis) include all laboratory results, scheduled and unscheduled. 
These listings are not color-coded, but the severity should be included in parentheses after the result for abnormal results, e.g., “16.2 (Mild)”. Results 
outside the reference range but not graded as Mild, Moderate, or Severe, should have ONR shown as the Severity Grade. “Change from Baseline” 
column will be blank for parameters that are not numeric. Sort Order: Parameter (same order described in Section 9.6), Cohort, Participant ID, and 
Timepoint.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study 
Day  
Sex  
Age (years) 
Laboratory Parameter 
(Units) 
Result  
(Severity Grade) 
Change from 
Baseline 
Reference Range 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 10: 
Clinical Laboratory Results – Hematology 
[Implementation Note: Sort Order: Parameter, Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study 
Day  
Sex  
Age (years) 
Laboratory Parameter 
(Units) 
Result  
(Severity Grade) 
Change from 
Baseline 
Reference Range 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 11: 
Clinical Laboratory Results – Coagulation 
[Implementation Note: Sort Order: Parameter, Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study 
Day  
Sex  
Age (years) 
Laboratory Parameter 
(Units) 
Result  
(Severity Grade) 
Change from 
Baseline 
Reference Range 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 12: 
Clinical Laboratory Results – Urinalysis 
[Implementation Note: Sort Order: Parameter, Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day  
Sex  
Age (years) 
Laboratory Parameter 
(Units) 
Result 
(Severity Grade) 
Reference Range 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 13: 
Kidney Function Biomarker Results – Cystatin C and KIM-1 
[Implementation Note: Sort Order: Parameter, Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day  
Sex  
Age (years) 
Laboratory Parameter 
(Units) 
Result  
Change from Baseline 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 14: 
Screening Laboratory Results – Serology 
[Implementation Note: Sort Order: Cohort, Participant ID.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day  
HBsAg 
HCV Antibodies 
HIV Antibodies 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 15: 
Screening Laboratory Results – Urine Drug and Cotinine and Alcohol Breathalyzer 
[Implementation Note: Sort Order: Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day  
Drug 
Result 
 
 
 
 
Amphetamines 
Positive/Negative 
 
 
 
 
Barbiturates  
 
 
 
 
 
Benzodiazepines 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 16: 
Screening Laboratory Results – Pregnancy and FSH 
[Implementation Note: Sort Order: Cohort, Participant ID, and Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day  
Urine Pregnancy Result 
Serum HCG Result 
FSH Test Result 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.9 Vital Signs and Physical Exam Findings 
Listing 17: 
Vital Signs 
[Implementation Note: This listing includes all vital signs assessments, scheduled and unscheduled. All height, weight, and BMI measurements will be 
included in this listing as well. These listings are not color-coded, but the severity should be included in parentheses after the result for abnormal 
assessments, e.g., “100.7 (Mild)”. Sort order: Cohort, Participant ID, Parameter (same order described in Section 9.6), Date of Assessment, Time of 
Assessment.] 
Cohort 
Participant ID 
Timepoint 
Actual Study Day 
Date of Assessment 
Time of Assessment 
Result (Severity) 
Change from Baseline 
 
 
 
 
ddMMMyyyy 
hh:mm 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 18: 
Abnormal Physical Exam Findings 
[Implementation Note: This listing includes all physical exam findings, scheduled and unscheduled. If a participant does not have any findings upon 
examination, they will not be included in this listing. If reported as an AE, display “Yes” with the AE Number in parenthesis, e.g., “Yes (7)”. Sort order: 
Cohort, Participant ID, Date of Assessment, Time of Assessment, Body System, Abnormal Finding.] 
Cohort 
Participant ID 
Timepoint 
Actual Study Day 
Date of Assessment 
Time of 
Assessment 
Body System 
Abnormal Finding 
Reported as an AE? 
(AE Description; 
Number) 
 
 
 
 
ddMMMyyyy 
hh:mm 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.10 ECG Results and Findings 
Listing 19: 
Listing of ECG Interval Measurements 
[Implementation Note: This listing includes all ECG assessments, scheduled or unscheduled. These listings are not color-coded, but the severity should 
be included in parentheses for abnormal assessment, e.g., “500 (Mild)”. For the mean of the triplicate readings, no assessment time should be presented, 
and the replicate number should specify “Mean”. Sort order: Cohort, Participant ID, Parameter, Date of Assessment, Time of Assessment.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study 
Day 
Date of 
Assessment 
Time of 
Assessment 
Sex 
Parameter (Units) 
Replicate 
Number 
Result 
(Severity) 
Pre-Dose 
Change from 
Baseline 
Time-Matched 
Change from 
Baseline 
 
 
 
 
ddMMMyyyy 
hh:mm 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Notes: Pre-Dose Baseline = The most recent measurement prior to start of infusion. Time-Matched Baseline = The measurement recorded on the matching timepoint on Day -1. 
 


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Listing 20: 
Listing of ECG Overall Interpretation and Change from Baseline 
[Implementation Note: This listing includes all ECG assessments, scheduled or unscheduled. Sort order: Cohort, Participant ID, Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day 
Date of Assessment 
Sex 
Interpretation 
Change from Baseline 
Comments 
 
 
 
 
ddMMMyyyy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.11 Otoacoustic Test Results 
Listing 21: 
Otoacoustic Test Results – Ear Otoscopy 
[Implementation Note: This listing includes all ear otoscopy assessment, scheduled and unscheduled. Sort order: Cohort, Participant ID, Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day 
Date of Assessment 
Right Ear Result 
Left Ear Result 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 22: 
Otoacoustic Test Results – Pure-Tone Audiometry 
[Implementation Note: This listing includes all pure-tone audiometry results, scheduled and unscheduled. Sort order: Cohort, Participant ID, Timepoint, 
and Frequency.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study Day 
Date of Assessment 
Frequency (kHz) 
Threshold (dB) 
Change from Baseline (dB) 
Right Ear  
Left Ear 
Right Ear  
Left Ear 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 23: 
Otoacoustic Test Results – DPOAE 
[Implementation Note: This listing includes all pure-tone audiometry results, scheduled and unscheduled. “Change from Baseline” will be missing for 
categorical results. Sort order: Cohort, Participant ID, Timepoint, Parameter, and Frequency.] 
Cohort 
Participant 
ID 
Timepoint  
Actual Study Day 
Date of Assessment 
Parameter 
Frequency 
(kHz) 
Result 
Change from Baseline 
Right Ear  
Left Ear 
Right Ear  
Left Ear 
 
 
 
 
 
DPOAE, SNR, 
Result 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 24: 
Screening Otoacoustic Test Results – Valsalva 
[Implementation Note: Sort Order: Cohort, Participant ID, Timepoint.] 
Cohort 
Participant ID 
Timepoint  
Actual Study Day 
Date of Assessment 
Result 
 
 
 
 
 
Normal/Abnormal  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 25: 
Screening Otoacoustic Test Results – Tympanometry 
[Implementation Note: Sort Order: Cohort, Participant ID, Timepoint.] 
Cohort 
Participant 
ID 
Timepoint 
Actual Study Day 
Date of Assessment 
Peak Pressure 
(Positive/Negative) 
Peak Pressure (daPa) 
Result 
Right Ear 
Left Ear 
Right Ear 
Left Ear 
Right Ear 
Left Ear 
 
 
 
 
 
 
 
 
 
Normal/Abnormal Normal/Abnormal 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 26: 
Screening Otoacoustic Test Results – Stapedial Reflexes 
[Implementation Note: Sort Order: Cohort, Participant ID, Timepoint, Frequency.] 
Cohort 
Participant ID 
Timepoint 
Actual Study Day 
Date of Assessment 
Frequency (kHz) 
Ipsilateral Reflex 
Contralateral Reflex 
Right Ear 
Left Ear 
Right Ear 
Left Ear 
 
 
 
 
 
0.5, 1.0, 2.0, 4.0 
Present/Absent 
Present/Absent 
Present/Absent 
Present/Absent 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.12 Concomitant Medications 
Listing 27: 
Prior Medications 
[Implementation Note: Include prior medications (medications with an end date prior to dosing) only. If start date is more than 30 days before 
enrollment, then categorize as follows:  
• 5 years prior to enrollment 
• 1-5 years prior to enrollment 
• 1-12 months prior to enrollment 
If taken for MH, display “Yes” with the MH Number in parentheses, e.g., “Yes (7)”. Sort order: Cohort, Participant ID, and CM Number.] 
Cohort 
Participant 
ID 
CM Number 
Medication 
Medication Start Day 
Medication End Day 
Indication 
Taken for a condition on 
Medical History? 
(MH Description; Number) 
ATC Level 1 
(ATC Level 2) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 28: 
Concurrent Medications 
[Implementation Note: Include concomitant medications (medications that are ongoing or have an end date after date of infusion) only. “Medication 
Start Day” and “Medication End Day” are relative to date of infusion (which is Day 1, day before date of treatment administration is Day -1). For 
medication start dates that are > 30 days prior to enrollment, rather than use the exact study days, categorize as follows: 
• 5 years prior to enrollment 
• 1-5 years prior to enrollment 
• 1-12 months prior to enrollment 
If ongoing, display “Ongoing” in the “Medication End Day” column. If taken for an AE or MH, display “Yes” with the AE or MH Number in 
parentheses, e.g., “Yes (7)”. Sort order: Cohort, Participant ID, CM Number.] 
Cohort 
Participant 
ID 
CM 
Number 
Medication 
Medication Start 
Day 
Medication End 
Day 
Indication 
Taken for an 
AE? 
(AE 
Description; 
Number) 
Taken for a condition 
on Medical History? 
(MH Description; 
Number) 
Authorized as 
medically 
indicated/ 
Immediately 
necessary by study 
clinician? 
ATC Level 1 
(ATC Level 2) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.13 Pregnancy Reports 
Listing 29: 
Pregnancy Reports – Maternal Information 
Cohort 
Participant 
ID 
Pregnancy 
Number 
Study Day 
Corresponding 
to Estimated 
Date of 
Conception 
Source of 
Maternal 
Information 
Pregnancy 
Status 
Mother’s 
Pre-
Pregnancy 
BMI 
Mother’s 
Weight 
Gain 
During 
Pregnancy 
Tobacco, 
Alcohol, or 
Drug Use 
During 
Pregnancy? 
Medications 
During 
Pregnancy? 
Maternal 
Complications 
During 
Pregnancy? 
Maternal 
Complications 
During Labor, 
Delivery, or 
Post-Partum? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Maternal Complications are included in the Adverse Event listing. Medications taken during pregnancy are included in the Concomitant Medications Listing. 
Listing 30: 
Pregnancy Reports – Gravida and Para 
 
 
 
 
Live Births 
 
 
 
 
 
Cohort 
Participant 
ID 
Pregnancy 
Number 
Gravida 
Extremely 
PBa 
Very 
Early 
PBa 
Early 
PBa 
Late 
PBa 
Early 
TBb 
Full 
TBb 
Late 
TBb 
Post 
TBb 
Still 
Births 
Spontaneous 
Abortion/ 
Miscarriage 
Elective 
Abortions 
Therapeutic 
Abortions 
Major 
Congenital 
Anomaly 
with 
Previous 
Pregnancy? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Gravida includes the current pregnancy, para events do not. 
a Preterm Birth 
b Term Birth 
 
 


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Listing 31: 
Pregnancy Reports – Live Birth Outcomes 
Cohort 
Participant 
ID 
Pregnancy 
Number 
Fetus 
Number 
Pregnancy 
Outcome 
(for this 
Fetus) 
Fetal 
Distress 
During 
Labor and 
Delivery? 
Delivery 
Method 
Gestational 
Age at Live 
Birth  
Size for 
Gestational 
Age 
Apgar 
Score, 
1 
minute 
Apgar 
Score, 5 
minutes 
Cord 
pH 
Congenital 
Anomalies? 
Illnesses/ 
Hospitalizations 
within 1 Month 
of Birth? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Congenital Anomalies are included in the Adverse Event listing. 
Listing 32: 
Pregnancy Reports – Still Birth Outcomes 
Cohort 
Participant 
ID 
Date of 
Initial 
Report 
Fetus 
Number 
Pregnancy 
Outcome 
(for this 
Fetus) 
Fetal 
Distress 
During 
Labor and 
Delivery? 
Delivery 
Method 
Gestational 
Age at Still 
Birth 
Size for 
Gestational 
Age 
Cord pH 
Congenital 
Anomalies? 
Autopsy 
Performed? 
If Autopsy, 
Etiology for Still 
Birth Identified? 
 
 
ddMMMyyyy 
 
 
 
 
 
 
 
 
 
 
Listing 33: 
Pregnancy Reports – Spontaneous, Elective, or Therapeutic Abortion Outcomes 
Cohort 
Participant 
ID 
Date of Initial 
Report 
Fetus 
Number 
Pregnancy 
Outcome (for this 
Fetus) 
Gestational Age at 
Termination 
Abnormality in Product of 
Conception? 
Reason for Therapeutic Abortion 
 
 
ddMMMyyyy 
 
 
 
 
 
 


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Listing 34: 
Birth Control Listing 
[Implementation Note: If participant is not of childbearing potential, indicate the reason the subject is not of childbearing potential in parentheses in the 
“Childbearing Potential” column. Sort order: Cohort, Participant ID, Birth Control Start Day.] 
Cohort 
Participant ID 
Sex 
Childbearing Potential 
Birth Control Method 
Birth Control Start Day 
Birth Control End Day 
 
 
 
 
 
ddMMMyyyy 
ddMMMyyyy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.6 
Individual Pharmacokinetic Data 
Listing 35: 
Participant Level Total Apramycin Concentrations in Plasma 
[Implementation Note: Laboratory Reported Concentration will give verbatim value reported by lab (with minimal formatting, as needed) and will use a 
character value such as PC.PCORRES. Analysis Concentrations will report the value exactly used for analysis and will use a numeric variable such as 
ADNCA.AVAL. If one or more samples are excluded from NCA, two additional columns will be provided in the PK report listing: Excluded from NCA 
(Yes/No) and Reason for Exclusion from NCA. In the actual time column, mark out of window times with one asterisk (*), mark substantially out of 
window times with two asterisks (**), and mark imputed times with three asterisks (***). Sort order: Cohort, Participant ID, Actual Time.] 
Cohort 
Participant ID 
Nominal Timea (h) 
Actual Timea (h) 
Laboratory Reported 
Concentration (µg/mL) 
Analysis Concentration 
(µg/mL) 
Used in Ke Calculations 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: BQL = Below the Quantification Limit. 
a Times are relative to time of dosing. For actual time, out of window times are indicated by an asterisk (*), substantially out of window times are indicated by two asterisks (**), and imputed times are 
indicated by three asterisks (***). 
 


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Listing 36: 
Participant Level Free Apramycin Concentrations in Plasma 
[Implementation Note: Laboratory Reported Concentration will give verbatim value reported by lab (with minimal formatting, as needed) and will use a 
character value such as PC.PCORRES. Analysis Concentrations will report the value exactly used for analysis and will use a numeric variable such as 
ADNCA.AVAL. If one or more samples are excluded from NCA, two additional columns will be provided in the PK report listing: Excluded from NCA 
(Yes/No) and Reason for Exclusion from NCA. In the actual time column, mark out of window times with one asterisk (*), mark substantially out of 
window times with two asterisks (**), and mark imputed times with three asterisks (***). Sort order: Cohort, Participant ID, Actual Time.] 
Cohort 
Participant ID 
Nominal Timea (h) 
Actual Timea (h) 
Laboratory Reported Concentration 
(µg/mL) 
Analysis Concentration 
(µg/mL) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: BQL = Below the Quantification Limit. 
a Times are relative to time of dosing. For actual time, out of window times are indicated by an asterisk (*), substantially out of window times are indicated by two asterisks (**), and imputed times are 
indicated by three asterisks (***). 


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Listing 37: 
Participant Level Free Portion of Plasma Apramycin 
[Implementation Note: In the actual time column, mark out of window times with one asterisk (*), mark substantially out of window times with two 
asterisks (**), and mark imputed times with three asterisks (***). Sort order: Cohort, Participant ID, Actual Time.] 
Cohort 
Participant ID 
Nominal Timea (h) 
Actual Timea (h) 
Free Portion (fu, % Unbound) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
a Times are relative to time of dosing. For actual time, out of window times are indicated by an asterisk (*), substantially out of window times are indicated by two asterisks (**), and imputed times are 
indicated by three asterisks (***). 
 


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Listing 38: 
Participant Specific Total Apramycin in Plasma PK Parameters  
[Implementation Note: Sort order: Cohort, Participant ID, Actual Time.] 
Cohort 
Participant 
ID 
Cmax 
(µg/mL) 
Tmax 
(h) 
AUC(0-24) 
(µg*h/mL) 
AUC(0-last) 
(µg*h/mL) 
AUC(0-∞) 
(µg*h/mL) 
Ke 
(1/h) 
t1/2 
(h) 
CL 
(L/h) 
Vd 
(L) 
Lambda Z 
Acceptance 
Criteria 
Met? 
%AUCex 
Criteria 
Met? 
 
 
 
 
 
 
 
 
 
 
 
Yes/No 
Yes/No 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 39: 
Participant Level Apramycin Concentrations in ELF 
[Implementation Note: Laboratory Reported Concentration will give verbatim value reported by lab (with minimal formatting, as needed) and will use a 
character value such as PC.PCORRES. Analysis Concentrations will report the value exactly used for analysis and will use a numeric variable such as 
ADNCA.AVAL. If one or more samples are excluded from NCA, two additional columns will be provided in the PK report listing: Excluded from NCA 
(Yes/No) and Reason for Exclusion from NCA. In the actual time columns, mark out of window times with one asterisk (*), mark substantially out of 
window times with two asterisks (**), and mark imputed times with three asterisks (***). Sort order: Cohort, Participant ID.] 
Cohort 
Participant 
ID 
Nominal 
Timea (h) 
Actual Time of 
BAL  
(h) 
Actual Time of 
Urea Collection 
(h) 
Volume of BAL 
Supernatant 
(Volume BAL 
SUP) 
(mL) 
Urea 
Concentration 
in Plasma  
(Urea Plasma) 
(µg/mL)  
Urea 
Concentration 
in BAL 
Supernatant 
(Urea BAL 
SUP) 
(µg/mL)  
Volume of 
ELFb 
(Volume ELF) 
(mL) 
Apramycin 
Concentration 
in BAL 
Supernatant 
(Apramycin 
BAL SUP) 
(µg/mL) 
Apramycin 
Concentration 
in ELFc,d 
(Apramycin 
ELF) 
(µg/mL) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: BQL = Below the Quantification Limit.  
a Times are relative to time of dosing. For actual time, out of window times are indicated by an asterisk (*), substantially out of window times are indicated by two asterisks (**), and imputed times are 
indicated by three asterisks (***). 
b Volume ELF = Volume BAL SUP * (Urea BAL SUP / Urea Plasma).  
c Apramycin ELF = Apramycin BAL SUP x (Volume BAL SUP / Volume ELF). 
d Apramycin ELF = Apramycin BAL SUP * (Urea Plasma / Urea BAL SUP). 
 


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Listing 40: 
Participant Level Apramycin Concentrations in AM 
[Implementation Note: Laboratory Reported Concentration will give verbatim value reported by lab (with minimal formatting, as needed) and will use a 
character value such as PC.PCORRES. Analysis Concentrations will report the value exactly used for analysis and will use a numeric variable such as 
ADNCA.AVAL. If one or more samples are excluded from NCA, two additional columns will be provided in the PK report listing: Excluded from NCA 
(Yes/No) and Reason for Exclusion from NCA. In the actual time column, mark out of window times with one asterisk (*), mark substantially out of 
window times with two asterisks (**), and mark imputed times with three asterisks (***). Sort order: Cohort, Participant ID.] 
Cohort 
Participant 
ID 
Nominal 
Timea (h) 
Apramycin 
Concentration in 
BAL Pellet  
(Concentration 
BAL AC) 
(µg/mL) 
Total Volume of 
BAL Pellet in 
Suspension 
(µL) 
Number of 
Alveolar Cells per 
mL BAL Volume  
(BAL AC) 
(106/mL) 
Total BAL 
Volume 
(BAL Volume) 
(mL)  
Percent of 
Macrophages in 
BAL Pellet 
(% AM) 
(%)  
Amount of 
Apramycin in 
Total Volume of 
Alveolar Cellsb 
(Apramycin 
BAL AC) 
(µg) 
Total Volume of 
AM in BAL 
Pelletc 
(Volume AM) 
(mL) 
Apramycin 
Concentration 
in AMd 
(Apramycin 
AM) 
(µg/mL) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: BQL = Below the Quantification Limit.  
a Times are relative to time of dosing.  
b Apramycin BAL AC = Concentration BAL AC * Total Volume of BAL Pellet in Suspension. 
c Volume AM = BAL AC * BAL Volume * % AM * 2.42 µL/106 cells. 
d Apramycin AM = Apramycin BAL AC / Volume AM. 
 


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Listing 41: 
Apramycin in ELF and AM to Total Apramycin in Plasma Concentration Ratios 
[Implementation Note: Sort order: Cohort, Participant ID.] 
Cohort 
Participant ID 
Timepoint 
Apramycin in ELF to Total Apramycin 
in Plasma Concentration Ratio 
Apramycin in AM to Total Apramycin in 
Plasma Concentration Ratio 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 42: 
Bronchoalveolar Lavage Cell Counts 
[Implementation Note: Sort order: Cohort, Participant ID.] 
Cohort 
Participant 
ID 
Timepoint 
Total Cell 
Count 
(103/mL) 
WBC Count 
(103/mL) 
Macrophages 
(%) 
Neutrophils 
(%) 
Eosinophils 
(%) 
Lymphocytes 
(%) 
Epithelial 
Cells          
(%) 
Other Cells 
(%) 
If other cells 
observed, 
specify 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Prot_001.pdf: 
DMID Protocol 20-0012 
Version 5.0 
Apramycin 
10 July 2023 
 
A PHASE I, OPEN-LABEL STUDY TO ASSESS LUNG 
PHARMACOKINETICS AND SAFETY OF A SINGLE DOSE OF 
APRAMYCIN ADMINISTERED INTRAVENOUSLY IN 
HEALTHY ADULT SUBJECTS 
 
 
DMID Protocol Number: 20-0012 
 
 
DMID Funding Mechanism: Phase 1 Clinical Trial Units 
 
Pharmaceutical Support: Juvabis AG 
 
IND Sponsor: Division of Microbiology and Infectious Diseases (DMID), 
National Institute of Allergy and Infectious Diseases (NIAID), 
National Institutes of Health (NIH) 
 
 
Site Principal Investigator: William B. Smith, MD 
 
DMID Clinical Project Manager: Maureen Mehigan, RN 
 
Version Number: 5.0 
 
10 July 2023 
 


DMID Protocol 20-0012 
Version 5.0 
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10 July 2023 
DMID/NIAID/NIH 
CONFIDENTIAL 
 
 
 
2 
STATEMENT OF COMPLIANCE 
The clinical trial will be carried out in accordance with Good Clinical Practice (GCP) and as 
required by the following: 
• United States (U.S.) Code of Federal Regulations (CFR) applicable to clinical trials 
(45 CFR Part 46, 21 CFR Part 50, 21 CFR Part 54, 21 CFR Part 56, and 21 CFR Part 312). 
• International Council for Harmonization of Technical Requirements for Registration of 
Pharmaceuticals for Human Use E6(R2) Good Clinical Practice (ICH E6 GCP): Integrated 
Addendum to ICH E6(R1) Guidance for Industry, published in the Federal Register (83 
Federal Register 8882 (2018)), including the latest finalized National Institutes of Health 
(NIH) Clinical Terms of Award, as applicable. 
Compliance with these standards provides public assurance that the rights, safety, and well-being 
of subjects are protected, consistent with principles that originate in the Declaration of Helsinki. 
All key personnel (all individuals responsible for the design and conduct of the trial) have 
completed Human Subjects Protection Training.  
 
 


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3 
SIGNATURE PAGE 
The signature below constitutes the approval of the protocol and its attachments and provides the 
necessary assurances that the trial will be conducted according to all stipulations of the protocol, 
including all statements regarding confidentiality, and according to local legal and regulatory 
requirements and applicable U.S. federal regulations and ICH guidelines. 
 
 
 
Site Investigator: 
 
 
William B. Smith, MD 
_________________________________________________ 
Name/Title (Print) 
 
 
Signature: 
 
_________________________________________________ 
 
Date:  
 
_________________________________________________ 
 
 


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Version 5.0 
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10 July 2023 
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CONFIDENTIAL 
 
 
 
4 
TABLE OF CONTENTS 
STATEMENT OF COMPLIANCE .................................................................................................2 
SIGNATURE PAGE .......................................................................................................................3 
TABLE OF CONTENTS .................................................................................................................4 
LIST OF TABLES .........................................................................................................................10 
LIST OF FIGURES .......................................................................................................................10 
LIST OF ABBREVIATIONS ........................................................................................................11 
PROTOCOL SUMMARY .............................................................................................................17 
1 
Key Roles ..............................................................................................................................23 
2 
Background Information and Scientific Rationale ...............................................................24 
2.1 
Background Information ...........................................................................................24 
2.2 
Apramycin.................................................................................................................24 
2.3 
Rationale for the Development of Apramycin for Human Use ................................27 
2.4 
Clinical Experience ...................................................................................................27 
 First-in-Human Single Dose Escalation Study .............................................27 
2.5 
Rationale ...................................................................................................................29 
 Study Rationale .............................................................................................29 
 Rationale for Dose Selection ........................................................................29 
2.6 
Identified and Potential Risks and Benefits Associated with the Study ...................29 
 Risks and Benefits Associated with the Study Drug, Apramycin ................29 
 Bronchoscopy and Bronchoalveolar Lavage Risks ......................................31 
 Audiometry and Otoacoustic test Risks ........................................................31 
 Venipuncture and IV Catheter Placement Risks ...........................................32 
 Additional Risks............................................................................................32 
2.7 
Known Potential Benefits .........................................................................................32 
2.8 
Risk/Benefit Ratio .....................................................................................................32 
3 
Objectives and Outcome Measures.......................................................................................34 
3.1 
Study Objectives .......................................................................................................34 
 Primary ..........................................................................................................34 
 Secondary ......................................................................................................34 


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 Exploratory ...................................................................................................34 
3.2 
Study Outcome Measures .........................................................................................34 
 Primary ..........................................................................................................34 
 Secondary ......................................................................................................34 
 Exploratory ...................................................................................................35 
4 
Study Design .........................................................................................................................36 
4.1 
Sub-studies ................................................................................................................37 
5 
Study Enrollment and Withdrawal .......................................................................................38 
5.1 
Subject Inclusion Criteria .........................................................................................38 
5.2 
Subject Exclusion Criteria ........................................................................................40 
5.3 
Treatment Assignment Procedures ...........................................................................42 
 Enrollment Procedures ..................................................................................42 
 Reasons for Withdrawal and Discontinuation of Study Drug 
Administration ..............................................................................................43 
 Handling of Withdrawals and Discontinuation of Administration ...............43 
 Lost to Follow-up ..........................................................................................44 
 Termination of Study ....................................................................................44 
6 
Study Intervention/investigational product ...........................................................................45 
6.1 
Description of Study Product ....................................................................................45 
 Acquisition ....................................................................................................45 
 Formulation, Packaging, and Labeling .........................................................46 
 Product Storage and Stability........................................................................46 
6.2 
Dosage, Preparation, and Administration of Study Intervention/ Investigational 
Products.....................................................................................................................46 
6.3 
Modification of the Study Intervention/ Investigational Products ............................47 
 Overdose .......................................................................................................47 
6.4 
Accountability Procedures for the Study Intervention/ Investigational Products .....48 
6.5 
Assessment of Subject Compliance with the Study Intervention/ Investigational 
Products.....................................................................................................................48 
6.6 
Prior and  Concomitant Medications/Treatments .....................................................49 


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6.7 
Subject Restrictions  and Precautions .......................................................................50 
 Smoking ........................................................................................................50 
 Physical Activity ...........................................................................................50 
 Food and Fluid intake ...................................................................................50 
 Alcohol, Marijuana and Illicit Drugs ............................................................50 
 Caffeinated beverages ...................................................................................51 
 Exposure to loud noise ..................................................................................51 
7 
Study Schedule .....................................................................................................................52 
7.1 
Recruitment ...............................................................................................................52 
7.2 
Screening Visit (Day -28 to Day -3) .........................................................................52 
7.3 
Check-in (Day -2) .....................................................................................................53 
7.4 
Baseline (Day -1) ......................................................................................................54 
7.5 
Inpatient Treatment Period (Days 1 to 3) .................................................................55 
 DAY 1: Administration of Study Drug – Bronchoscopy (Cohorts T1 – 
T4) .................................................................................................................55 
 DAY 2: Inpatient Follow-up – Bronchoscopy T5 ........................................57 
 DAY 3: Inpatient Follow-up .........................................................................58 
7.6 
Outpatient Follow-up Period (Days 14 and 30) ........................................................59 
 DAY 14 (± 3 days) ........................................................................................59 
 Day 30 (± 4 days) – FINAL VISIT ...............................................................59 
7.7 
Early Termination (if needed) ...................................................................................60 
7.8 
Unscheduled Visit (if needed) ..................................................................................60 
8 
Study Procedures/evaluations ...............................................................................................62 
8.1 
Clinical Procedures/Evaluations ...............................................................................62 
 Informed Consent..........................................................................................62 
 Demographics ...............................................................................................62 
 Inclusion/Exclusion Criteria .........................................................................62 
 Medical History ............................................................................................62 
 Physical Examination....................................................................................63 
 Vital Signs .....................................................................................................63 


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 12-lead Standard Electrocardiogram (ECG) .................................................64 
 Spirometry.....................................................................................................64 
8.2 
Laboratory Evaluations .............................................................................................64 
 Clinical Laboratory Evaluations (Hematology, Coagulation, Chemistry, 
and Urinalysis). .............................................................................................64 
 Viral Serology Testing ..................................................................................65 
 Pregnancy Testing .........................................................................................65 
 Serum FSH Testing .......................................................................................66 
 Urine Toxicology Screening .........................................................................66 
 Alcohol Breathalyzer Test ............................................................................66 
 Urine Cotinine Testing ..................................................................................66 
 Exploratory Biomarkers for Renal Injury .....................................................66 
 Serum Cystatin C ..............................................................................66 
 Urinary KIM-1 ..................................................................................66 
8.3 
Bronchoscopy with Bronchoalveolar Lavage ...........................................................66 
8.4 
Ear Examination and Otoacoustic Testing ................................................................67 
8.5 
Bioanalytical Assays for Pharmacokinetics (PK) .....................................................68 
 Assay for Total Apramycin in Plasma ..........................................................68 
 Protein Binding Test for Apramycin in Plasma ............................................68 
 Assays for Total Apramycin in Bronchoalveolar Lavage Components 
(ELF and AM)...............................................................................................69 
 Plasma and BAL Urea assay .........................................................................69 
 Specimen Preparation, Handling, and Shipping ...........................................69 
 Instructions for Specimens Preparation, Handling, and Storage ......69 
 Specimen Shipment ..........................................................................69 
 Long-term Storage of Plasma and BAL (ELF and AM) PK Samples ..........70 
9 
Assessment of Safety ............................................................................................................71 
9.1 
Specification of Safety Parameters ...........................................................................71 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety Parameters 71 
 Adverse Events .............................................................................................71 


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 Severity of Events .............................................................................72 
 Relationship to Study Products .........................................................72 
 Reporting Adverse Events ................................................................73 
 Serious Adverse Events ................................................................................73 
 Procedures to be Followed in the Event of Abnormal Laboratory Test 
Values or Abnormal Clinical Findings .........................................................74 
9.3 
Reporting Procedures ................................................................................................74 
 Serious Adverse Events ................................................................................74 
 Regulatory Reporting for Studies Conducted Under DMID-Sponsored 
IND ...............................................................................................................75 
 Other Adverse Events (if applicable) ............................................................76 
 Reporting of Overdose ......................................................................76 
 Reporting of Pregnancy ................................................................................76 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events .........................77 
9.5 
Halting Rules ............................................................................................................77 
 Study Halting Criteria ...................................................................................77 
9.6 
Safety Oversight........................................................................................................78 
 Safety Monitoring Committee (SMC) ..........................................................78 
10 
Clinical Monitoring...............................................................................................................79 
10.1 
Site Monitoring Plan .................................................................................................79 
11 
Statistical Considerations ......................................................................................................80 
11.1 
Study Hypotheses......................................................................................................80 
11.2 
Sample Size Considerations ......................................................................................80 
11.3 
Safety Review ...........................................................................................................80 
11.4 
Final Analysis Plan ...................................................................................................80 
 Analysis Populations .....................................................................................81 
 Demographics and Baseline Characteristics .................................................81 
 Safety Analysis .............................................................................................81 
 Adverse and Serious Adverse Events ...............................................81 
 Additional Safety Analyses...............................................................82 


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 PK Analysis ..................................................................................................82 
 Plasma PK parameters: .....................................................................82 
 Bronchoalveolar ELF and AM PK parameters: ................................83 
 Ratio of exposure parameters of Lung PK to Plasma PK: ................83 
 Missing Values and Outliers .........................................................................83 
 Analysis of ECG Parameters ........................................................................84 
 ECG Analysis and Interpretation ......................................................84 
 Categorical Analyses ........................................................................84 
 Cardiac Abnormalities: .....................................................................85 
 Analysis of Audiology Data..........................................................................85 
 Interim Reports .............................................................................................85 
12 
Source Documents and Access to Source Data/Documents .................................................86 
13 
Quality Control and Quality Assurance ................................................................................87 
14 
Ethics/Protection of Human Subjects ...................................................................................88 
14.1 
Ethical Standard ........................................................................................................88 
14.2 
Institutional Review Board .......................................................................................88 
14.3 
Informed Consent Process ........................................................................................88 
14.4 
Exclusion of Women, Minorities, and Children (Special Populations) ....................90 
14.5 
Subject Confidentiality .............................................................................................90 
14.6 
Certificate of Confidentiality ....................................................................................90 
14.7 
Study Discontinuation ...............................................................................................91 
14.8 
Future Use of Stored Specimens ...............................................................................91 
15 
Data Handling and Record Keeping .....................................................................................93 
15.1 
Data Management Responsibilities...........................................................................93 
15.2 
Data Capture Methods ..............................................................................................93 
15.3 
Types of Data ............................................................................................................93 
15.4 
Timing/Reports .........................................................................................................94 
15.5 
Study Records Retention...........................................................................................94 
15.6 
Protocol Deviations ...................................................................................................94 
16 
Publication Policy .................................................................................................................95 


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17 
Literature References ............................................................................................................96 
18 
Appendices............................................................................................................................99 
Appendix A: Schedule of Study Procedures and Evaluations .....................................................100 
Appendix B: Adverse Events Toxicity Grading Criteria .............................................................102 
Appendix C: Blood Volume Withdrawn During the Trial ..........................................................111 
 
LIST OF TABLES 
Table 1: Schedule of Events ....................................................................................................... 100 
Table 2: Toxicity Grading Tables – CLINICAL AEs ................................................................ 103 
Table 3: Toxicity Grading Tables – LABORATORY AEs ........................................................ 107 
Table 4: Toxicity Grading Tables – ECG ................................................................................... 110 
Table 5: Laboratory Samples and Estimated Total Blood Volume (mL) ................................... 111 
 
LIST OF FIGURES 
Figure 1: Description Schematic of Study Design ........................................................................ 22 
 


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LIST OF ABBREVIATIONS 
AAC(2)-Ia 
Acetyltransferase (2) subtype Ia 
AAC(3)-IV 
Acetyltransferase (3) subtype IV 
ACE 
Angiotensin-Converting Enzyme 
AE 
Adverse Event 
AII 
Angiotensin II 
ALT 
Alanine Aminotransferase 
AM 
Alveolar Macrophages 
AMR 
Alliance for Multispecialty Research 
ARA 
Angiotensin II-receptor antagonists 
APTT 
Alpha Partial Thromboplastin Time 
AP 
Alkaline Phosphatase 
AST 
Aspartate Aminotransferase 
AUC 
Area Under the Concentration – Time Curve 
AUC (0-t) 
Area under the concentration time-curve from time zero to time t 
AUC(0-24) 
Area under the concentration time-curve from time zero to 24 h 
AUC(0-∞) 
Area under the concentration time-curve from time zero to infinity 
AUC(0-last) 
Area under the concentration time-curve from time zero to the last 
concentration above the lower limit of quantitation 
β-HCG 
Beta Human Chorionic Gonadotropin 
BAL 
Bronchoalveolar Lavage 
BMI 
Body Mass Index 
BP 
Blood Pressure 
bpm 
Beats per Minute 
BUN 
Blood Urea Nitrogen 
CBC 
Complete Blood Count 
CDC 
Centers for Disease Control and Prevention 
CKD – EPI  
Chronic Kidney Disease Epidemiology collaboration equation for 
estimating GFR 
CFR 
Code of Federal Regulations 
CHEM 
Chemistry Panel 
CI 
Confidence Interval 
Cmax 
Maximum Concentration 


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CLT 
Total Clearance 
CMS 
Clinical Materials Services 
ConMed(s) 
Concomitant Medication(s) 
COAG 
Coagulation 
COX-2 
Cyclooxygenase 2 
CPM 
Clinical Project Manager 
CROMS 
Clinical Research Operations and Management Support 
CSR 
Clinical Study Report 
CTU 
Clinical Trial Unit 
ΔΔQTcF 
Time-matched, baseline-adjusted difference in QTcF interval 
dB 
Decibel 
DBP 
Diastolic Blood Pressure 
DVC 
Dynport Vaccine Company, LLC, a GDIT company 
dL 
Deciliter 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH 
DPOAE 
Distortion product otoacoustic emissions  
eCRF 
Electronic Case Report Form 
EC 
Ethics Committee 
ECG  
Electrocardiogram 
EDC 
Electronic Data Capture 
FEV1 
Forced Expiratory Volume in the first second 
FIH 
First-in Human 
FH 
Family History 
fT3 
Free triiodothyronine (T3)  
fT4 
Free thyroxine (T4) 
FVC 
Forced Vital Capacity 
ELF 
Epithelial Lining Fluid 
ET 
Early Termination 
EU 
European Union 
FDA 
Food and Drug Administration 
FH 
Family History 
FSH 
Follicle-Stimulating Hormone 


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FWA 
Federalwide Assurance 
g 
Gram(s) 
GCP 
Good Clinical Practice 
GFR 
Glomerular Filtration Rate 
eGFR 
Estimated GFR 
h 
Hour(s) 
HAP 
Hospital Acquired Pneumonia 
HBsAg 
Hepatitis B Surface Antigen 
Hct 
Hematocrit 
HCV 
Hepatitis C Virus 
HEENT 
Head, Eyes, Ears, Nose, and Throat 
HEM 
Hematology Panel 
Hgb 
Hemoglobin 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HPF 
High-Powered Field 
HIPAA 
Health Insurance Portability and Accountability Act 
HLGT 
High Level Group Term 
HR 
Heart Rate 
IATA 
International Air Transport Association 
IB 
Investigator’s Brochure 
ICF 
Informed Consent Form 
ICH 
International Council for Harmonization 
ICMJE 
International Committee of Medical Journal Editors 
IM 
Intramuscular 
IMP 
Investigational Medical Product 
IND 
Investigational New Drug  
INR 
International Normalized Ratio 
IRB 
Institutional Review Board 
IUD 
Intrauterine Devise 
IV 
Intravenous 
kg 
Kilogram(s) 


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KIM 
Kidney Injury Molecule  
LC-MS/MS 
Liquid Chromatography with Tandem Mass Spectrometry 
LLN 
Lower Limit of Normal 
LQTS 
Long QT Syndrome 
MBC 
Minimal Bactericidal Concentration 
MDMA 
3,4-methylenedioxy-methamphetamine 
MDR 
Multiple Drug Resistant 
MedDRA 
Medical Dictionary for Regulatory Activities 
MRSA 
Methicillin Resistant Staphylococcus aureus 
MM 
Medical Monitor 
µg 
Microgram(s) 
mg 
Milligram(s) 
MH 
Medical History 
MI 
Myocardial Infarction 
MBC 
Minimal Bactericidal Concentration 
MIC 
Minimal Inhibitory Concentration 
min 
Minute(s) 
mL 
Milliliter(s) 
MM 
Medical Monitor 
mmHg 
Millimeters of Mercury 
mmol 
Millimole(s) 
MOP 
Manual of Procedures 
msec 
Milliseconds(s) 
N 
Number (typically refers to the total number of subjects) 
n 
Number (typically refers to a subset of the total number of subjects) 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH 
NIH 
National Institutes of Health 
NOAEL 
No Observable Adverse Effect Level 
NSAID 
Nonsteroidal Anti-Inflammatory Drug 
OAEs 
Otoacoustic emissions 
OER 
Office of Extramural Research, NIH 
OTC 
Over the Counter 


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PAE 
post-antibiotic effect 
PBF 
Post-bronchoscopy Fever 
PD 
Pharmacodynamic(s) 
PE 
Physical Examination 
PI 
Principal Investigator 
PK 
Pharmacokinetic(s) 
POC 
Point of Contact 
Protime 
Prothrombin time 
PVG 
Pharmacovigilance Group 
QTc 
Corrected QT Interval of the ECG 
QTcF  
Corrected QT interval of the ECG using Fridericia’s Formula 
RAAS 
Renin – Angiotensin - Aldosterone System  
RNA 
Ribonucleic Acid 
rRNA 
ribosomal ribonucleic acid 
RBC 
Red Blood Cell 
RP 
Research Pharmacist 
RR 
Respiratory Rate 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SBP 
Systolic Blood Pressure 
SD 
Standard Deviation 
SDCC 
Statistical and Data Coordinating Center 
sec 
Second(s) 
SMC 
Safety Monitoring Committee 
SNHL 
Sensorineural Hearing Loss 
SOC 
System Organ Class 
SSP 
Study Specific Procedures 
STH6 
Score Threshold 
T 
Temperature 
TCA 
Tricyclic Antidepressant 
Tmax 
Time to Maximum Concentration 
TEAE 
Treatment-emergent Adverse Event 


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TSH 
Thyroid Stimulating Hormone 
UA 
Urinalysis 
ULN 
Upper Limit of Normal 
US 
United States 
UTMC 
University of Tennessee Medical Center, Knoxville 
t1/2 
Terminal Elimination Half-life 
TLF 
Tables, Listing and Figures 
TN 
Tennessee 
VAP 
Ventilator-associated Pneumonia 
Vd 
Volume of Distribution 
VS 
Vital Signs 
Vz/F 
Apparent volume of distribution 
WBC 
White Blood Cell 
WHO 
World Health Organization 
 
 


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PROTOCOL SUMMARY 
Title: 
A Phase I, Open-Label Study to Assess Lung 
Pharmacokinetics and Safety of a Single Dose of 
Apramycin Administered Intravenously in Healthy 
Adult Subjects. 
Phase: 
1 
Population: 
Approximately 20 evaluable healthy male or female 
subjects 18 to 45 years of age (both inclusive) at the 
time of dosing. 
Number of Sites: 
One (Alliance for Multispecialty Research [AMR] 
Phase 1 clinical trial unit [CTU], Knoxville, 
Tennessee [TN] [(AMR-Knoxville]). 
Study Duration:  
Approximately 10 months from study activation to 
last subject last visit. 
Subject Participation Duration: 
Up to 58 days (from Screening Visit to Final Visit). 
The trial will consist of a Screening Period of up to 
26 days as outpatient (Day -28 to Day -3); a Check-
in Day (Day -2); Baseline Day (Day -1); a 
Treatment Period of 3 days (Days 1 to 3) as 
inpatient, with dosing on Day 1 and follow-up on 
Days 2 and 3; and an out-patient Follow-up Period 
of 27 days (Days 4 to 30) with two site visits, on 
Day 14 (± 3 days) and Day 30 (± 4 days) (Final 
Visit). 
Description of Agents: 
Apramycin is a mono-substituted 2-
deoxystreptamine compound that represents a 
subclass of aminoglycoside antibiotics. Apramycin 
is provided in 20 mL vials as an aqueous solution of 
150 mg/mL apramycin base, with pH adjusted to 
5.5 to 6.0 with sulfuric acid. The filling volume is 
19 mL, and the extractable volume is 18 mL. A 
single dose of 30 mg/kg will be administered 
intravenously (IV) in a total volume of 30 mL over 
a period of 30 (± 5) min as an open label. Sterile 


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normal saline will be used to adjust the volume to 
the desired Investigational Medical Product (IMP) 
concentration.  
Objectives: 
Primary: 
• 
To assess plasma pharmacokinetic (PK) 
profile of apramycin and lung penetration of 
apramycin in epithelial lining fluid (ELF) 
and alveolar macrophages (AM) after single 
intravenous (IV) apramycin dose of 
30 mg/kg in healthy subjects.  
Secondary:  
• 
To assess the safety of single IV 
administration of 30 mg/kg apramycin in 
healthy subjects. 
• 
To assess changes in otoacoustic testing.  
Exploratory:  
• 
To assess changes in kidney function 
biomarkers. 
Outcome Measures:  
Primary: 
• Lung concentration and PK parameters of total 
apramycin: 
o ELF: Cmax, AUC(0-24), AUC(0-t), AUC(0-∞), Tmax, 
t½ 
o AM: Cmax, AUC(0-24), AUC(0-t), AUC(0-∞), Tmax, 
t½ 
• Plasma concentration and PK parameters of total 
apramycin:  
o AUC(0-∞), AUC(0-t), AUC(0-24), Cmax, Tmax, t½, 
CLT, Vd central 
• Ratio of exposure parameters of Lung PK to 
Plasma PK: 
o ratio of Cmax in ELF over Cmax in plasma  
o ratio of Cmax in AM over Cmax in plasma  
o ratio of AUC(0-24), AUC(0-t), and AUC(0-∞) in 
ELF over AUC(0-24), AUC(0-t), and AUC(0-∞) in 
plasma  
o ratio of AUC(0-24), AUC(0-t), and AUC(0-∞) in 
AM over AUC(0-24), AUC(0-t), and AUC(0-∞) in 
plasma  
 


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Secondary: 
• 
Type and incidence of treatment-emergent 
adverse events (TEAEs) and serious adverse 
events (SAEs) to Day 30 (± 4 days) after dosing. 
• 
Frequency of clinically significant TEAEs in 
physical examination (PE) findings or changes 
from baseline in vital signs (VS) and clinical 
laboratory parameters to Day 14 (± 3 days) after 
dosing. 
• 
Frequency of changes in electrocardiographic 
(ECG) intervals (QTcF, PR, QRS, HR) and 
morphology from baseline until 24 h ± 1 h after 
dosing. 
• 
Type and incidence of TEAEs related to 
auditory (cochlear) function tests (pure-tone 
audiometry and distortion product otoacoustic 
emissions [DPOAEs]) to Day 30 (± 4 days) after 
dosing.  
Exploratory: 
 
• 
Changes from baseline in urinary kidney injury 
molecule 1 (KIM-1) and serum cystatin C 
concentrations until Day 30 (± 4 days) after 
dosing. 
Description of Study Design: 
Open label study of a single dose of 30 mg/kg of 
apramycin administered intravenously (IV) over 30 
(± 5) minutes. Twenty subjects will be enrolled in 
the study to one of 5 cohorts, T1-T5, each 
corresponding to a timepoint after initiation of 
infusion at which a single fiberoptic bronchoscopy 
with bronchoalveolar lavage (BAL) is performed. 
There will be 4 subjects per cohort. Cohort T5 will 
be enrolled after plasma and lung apramycin 
concentrations and preliminary PK data analysis are 
completed in cohorts T1-T4. Enrollment and dosing 
will be determined by bronchoscopy schedule. For 
each cohort, if 2 subjects are scheduled to receive 
study drug on the same day, the dose will be 
administered sequentially at least 2 hours apart. 
Bronchoscopy with BAL will be performed under 
sedation or light anesthesia and oxygenation 
support. Apramycin concentrations after IV 
infusion will be measured in BAL and alveolar 


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20 
macrophages (AM) prepared by centrifugation of 
BAL samples. Apramycin concentrations in 
epithelial lining fluid (ELF) will be corrected for 
dilution using the urea method. BAL samples will 
be collected at the following timepoints after the 
start of the infusion in each T cohort: 30 min ± 
5 min (T1), 2 h ± 5 min (T2), 4 h ± 10 min (T3), 8 h 
± 15 min (T4) or 24 h ± 1 h (T5). The timing of T5 
is nominal; final time will be determined after 
analysis of apramycin concentrations in the BAL 
(ELF and AM) in cohorts T1-T4. 
Apramycin concentrations (total) will be measured 
in plasma samples prepared from blood samples 
collected pre-dose, and at the following timepoints 
after the start of the infusion: 30 ± 5 min, 1 h ± 
5 min, 2 h ± 5 min, 4 h ± 10 min, 8 h ± 15 min, 16 h 
± 15 min, 24 h ± 1 h, 36 h ± 1h, 48 h ± 1 h and 60 h 
± 1 h after dosing. An additional plasma PK sample 
may be collected at the end of the BAL sample 
collections if bronchoscopy with BAL is delayed. 
Free apramycin concentration will be measured at 
30 ± 5 min (immediately after the end of infusion) 
and at 36 h ± 1h.   
Urea concentration will be measured in plasma and 
time-matched BAL samples as a volume dilution 
marker of BAL ELF. 
All concentration measurements for apramycin and 
urea will be performed with validated bioanalytical 
assays. 
Safety will be evaluated by standard clinical 
assessments, VS measurements, safety clinical 
laboratory tests, and  
12-lead standard ECGs. Potential nephrotoxicity 
will be evaluated with creatinine concentrations and 
estimated glomerular filtration rate (eGFR) and 
exploratory biomarkers in plasma (Cystatin C) and 
urine (kidney injury marker 1 [KIM-1]). Potential 
ototoxic effects will be evaluated by cochlear 
otoacoustic tests (audiometry and DPOAEs). 


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Changes in assessments after administration of the 
IMP and comparison with the pre-dosing baseline 
will be reported. TEAEs and SAEs will be collected 
and graded from the time of dosing to completion 
of the study (Day 30 ± 4 days). A Safety 
Monitoring Committee (SMC) will oversee the 
conduct of the study. 
Estimated Time to Complete 
Enrollment 
Up to 6 months (from dosing of first subject to 
Final Visit of last subject). 
 
 


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Figure 1: Description Schematic of Study Design 
 
NOTE: All subjects who signed the ICF and met screening criteria will have baseline 
assessments on Day -2 and Day -1. Following a single IV dose of 30 mg/kg of apramycin, blood 
will be drawn for measuring the concentration and PK parameters of total plasma apramycin 
starting on Day 1 and finishing on Day 3 at the indicated timepoints. A single bronchoscopy with 
bronchoalveolar lavage will be performed after initiation of dosing at one of the following 
timepoints in the respective cohorts: 30 min (T1), 2 h (T2), 4 h (T3), 8 h (T4), and 24 h (T5). 
Total apramycin concentration will be measured at each BAL timepoint and PK parameters will 
be estimated in BAL fluid (containing ELF) and BAL AM. Four subjects will be assigned to 
each T cohort. Enrollment and dosing will be rolling and coordinated with the bronchoscopy 
service. For each cohort, if 2 subjects are scheduled to receive study drug on the same day, the 
second subject will be dosed at least 2 hours after the first subject. Safety assessments will be 
reported from the start of the IMP infusion on Day 1 to the last visit on Day 30, and will include 
physical examination, vital signs, ECG, clinical laboratory tests (hematology, chemistry and 
urinalysis), audiology examinations (ENT exam, audiometry, DPOAEs), and exploratory 
markers of nephrotoxicity (serum cystatin C and urinary KIM-1).   
 
 
 


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1 
KEY ROLES 
Site Principal Investigator: 
William B. Smith, MD  
AMR-Knoxville  
1928 Alcoa Highway, Suite 107 
Knoxville, TN 37920 
Office: 865-305-9100 ext.246   
Email: William.smith@amrllc.com  
 
Site Pneumonologist: 
John C. Callison, Jr., MD  
University of Tennessee Medical Center 
c/o AMR-Knoxville  
1928 Alcoa Highway, Suite 107 
Knoxville, TN 37920 
Office: 865-524-7471 
Email: jcallison@utmck.edu   
 
DMID Clinical Project Manager: 
Maureen Mehigan, RN 
DMID/NIAID/NIH/HHS 
5601 Fishers Lane, Room 7E19  
Bethesda, MD 20892-9826 
Office: 240-627-3317 
Email: mmehigan@niaid.nih.gov   
  
Statistical and Data Coordinating 
Center (SDCC): 
The Emmes Company, LLC 
401 N. Washington Street, Suite 700 
Rockville, MD 20850 
Phone: 301-251-1161 
Fax: 301-576-3558 
Email: phase1@emmes.com    
 
Bioanalytical Laboratory: 
KCAS Bioanalytical and Biomarker Services 
10830 South Lay Blair Boulevard 
Phone: 913-248-3042 
Fax: 913-248_3106 
Email: TMM@kcasbio.com   


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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
Aminoglycosides are a class of antibiotics established with the introduction of streptomycin in 
the 1940s for the treatment of tuberculosis and have defended a strong clinical presence ever 
since and until today. 
The common mode of action of aminoglycoside antibiotics is the disruption of bacterial protein 
synthesis by binding to the 16S ribosomal nucleic acid (rRNA) of the decoding site inside the 
small ribosomal subunit. Aminoglycosides are characterized by rapid and concentration-
dependent killing and demonstrate significant post-antibiotic effect (PAE). All aminoglycosides 
including apramycin are polar, poly-cationic, and highly water-soluble, a property that limits 
their ability to cross lipid-rich cellular membranes of the host cells. 
Mainly due to the common recognition of the rapid bactericidal activity of aminoglycosides - 
also at very high bacterial burden - the drug class has established itself as an important 
therapeutic component in the treatment of serious hospital infections worldwide. Despite the 
introduction of several new chemical classes of antibacterial agents over the past few decades, 
aminoglycosides still remain an extremely important therapeutic option. The World Health 
Organization (WHO) continues to classify aminoglycoside antibiotics as Essential Medicines 
(WHO 2019a) and as Critically Important Antimicrobials for Human Medicine (WHO 2019b). 
The therapeutic efficacy of standard-of-care aminoglycosides currently in clinical use is 
compromised by significant rates of antimicrobial resistance acquired by horizontal gene 
transfer. Infections with antibiotic-resistant bacteria caused an estimated 33,000 deaths in Europe 
in 2015 (Cassini 2018). According to the CDC’s Antibiotic Resistance Threats in the United 
States, 2019, more than 2.8 million antibiotic-resistant infections occur in the U.S. each year, and 
more than 35,000 people die as a result (CDC 2019). Bacterial pathogens highly resistant to 
various classes of antibiotics are estimated to account for a total of about 700,000 deaths 
annually worldwide. Without successful intervention, the burden of continued emergence and 
global spread of antimicrobial resistance is expected to multiply by the year 2050 (O’Neil 2014; 
CDC 2019). New antibiotics are needed to adequately respond to the increasing prevalence of 
drug- resistant infections. 
2.2 
Apramycin  
Apramycin is a mono-substituted 2-deoxystreptamine compound that is significantly different 
from the di-substituted 2-deoxystreptamines which include all the standard-of-care 
aminoglycoside antibiotics (gentamicin, amikacin, tobramycin, arbekacin, plazomicin and 
others) and, therefore, represents a subclass of aminoglycosides (O’Connor 1976). 


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Apramycin was originally approved as an oral and intramuscular veterinary antibiotic in the 
European Union (EU) for various indications (e.g., colibacillosis and salmonellosis in calves, 
bacterial enteritis in pigs, or Escherichia coli septicemia in poultry (EMA 1999). Its use as a 
growth-promoter in livestock has been banned in the 1980’s and it is authorized only for clinical 
veterinary targeted therapy in the EU and the USA (EU Commission 2005, FDA 2021).  
Antibacterial activity 
Apramycin is active against a wide variety of pathogenic Gram-positive and Gram-negative 
bacteria including pathogens with resistance to gentamicin and all other clinical aminoglycosides 
of relevance. The primary therapeutic spectrum comprises all relevant Enterobacterales 
including Escherichia coli, Klebsiella spp., Enterobacter spp., Serratia marcescens, Morganella 
morganii, Citrobacter freundii, and Providencia spp (Smith 2016, Juhas 2019). The activity 
against Providencia spp. is of note because of its chromosomal aac (2’)-Ia gene that causes 
intrinsic resistance to several aminoglycoside antibiotics including plazomicin, the newest 
semisynthetic aminoglycoside to receive Food and Drug Administration (FDA) market approval. 
Apramycin also has significant in vitro activity against multi-drug resistant Acinetobacter 
baumannii and Pseudomonas aeruginosa (Kang 2017, Juhas 2019). Amongst Gram-positive 
cocci, both methicillin-sensitive and resistant Staphylococcus aureus (MRSA) are sensitive to 
apramycin (Truelson 2018). The in vitro minimum inhibitory concentration (MIC) of apramycin 
against drug-susceptible clinical isolates at neutral pH tends to be about 4-fold lower than that of 
gentamicin, and about 2-fold lower than that of amikacin. The potency of apramycin at slightly 
acidic pH is similar to that of gentamicin and amikacin. The overall MIC90 of apramycin is 
8 μg/mL against highly drug- and carbapenem-resistant Enterobacterales (IB 2021, Juhas 2019).  
Apramycin treatment in mouse models of urinary tract infection (UTI), septicemia, pneumonia, 
and neutropenic thigh infection significantly reduced the bacterial burden even with bacterial 
isolates that are multidrug-, carbapenem- and plazomicin-resistant (Becker 2020, IB 2021).  
Safety Pharmacology  
In vitro and ex vivo studies showed no relevant secondary pharmacological effects on receptors 
or ion channels, which indicates that no off-target effects of apramycin are to be expected. The 
rat calcium channel N-type appeared to show full inhibition at 2 mM. This receptor interaction 
has been reported to be associated with the reversible neuromuscular blockade that can be 
commonly induced by peak aminoglycoside concentrations. In vitro investigation of effects on 
human ether-a-go-go-related gene (hERG) potassium channel assays in Chinese hamster ovary 
(CHO) cell lines showed IC50 > 40 mM. High IC50’s were also found for patch recordings of the 
Nav 1.5 sodium channel (>3 mM) in CHO cells and the Cav 1.2 channel in CHO cells (between 
1 to 3 mM), suggesting no potentially clinically significant effects (IB 2021). 
Nonclinical Assessment of Safety 
Extensive nonclinical safety core battery studies have been conducted in the rat and the Beagle 
dog to evaluate the effects of apramycin on organ systems (IB 2021). 


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Repeat-dose toxicity studies identified the kidneys as the main target organ of toxicity. 
Nephrotoxicity manifested as increases in plasma creatinine and BUN, and histopathological 
changes in the kidney, primarily tubular necrosis with or without regeneration. Nephrotoxicity 
was exposure-related, with a lower incidence and severity after the recovery period, suggesting 
potential reversibility. Nephrotoxicity appeared to correlate better with cumulative exposure than 
with total AUC (0-24) (IB 2021).  
Ototoxicity is a known risk of aminoglycosides. A repeat, 16-day dose study in guinea pigs 
showed that apramycin ototoxicity progressed more gradually than under gentamicin treatment, 
and cochlear function was partly retained even at high concentrations (430 mg apramycin/kg 
body weight). “No ototoxic effects were observed in the guinea pig model at the 152 mg/kg/day 
dose level. This would convert to a human equivalent dose (HED) of 33.3 mg/kg, a single dose 
exposure of 0.8 mMÅ~h, and a cumulated exposure of 13.7 mMÅ~h, all of which are markedly 
higher than the nephrotoxic no observed adverse effect levels (NOAELs) in both rats and dogs.” 
(Matt 2012) Recent literature data support the finding that apramycin is of lower ototoxicity than 
gentamicin (Ishikawa 2019).  
Besides nephrotoxicity and ototoxicity, no other noteworthy findings in clinical signs and 
symptoms were observed. Apramycin showed no effects on the CNS, cardiovascular, or 
respiratory systems (IB 2021).  
No secondary pharmacological effects were observed. No mutagenic and no genotoxic effects 
were observed. Therefore, no adverse effects or risks related to CNS, cardiovascular system, 
respiratory or off-target effects are to be expected. No special precautions regarding exposure to 
ultraviolet light have to be taken, as apramycin has no phototoxic potential. No signs of 
hepatotoxicity were observed in nonclinical safety pharmacology or toxicology studies. No 
fertility studies or data on embryofetal toxicity of apramycin are available to date. From 
veterinary toxicology data, no adverse effects on reproductive performance or evidence of 
maternal toxicity, fetal toxicity, or teratogenicity have been reported. Overall, the results of the 
nonclinical safety studies and toxicology studies are in line with the safety profiles reported for 
other aminoglycosides, e.g., gentamicin, amikacin and plazomicin (IB 2021). 
 
Nonclinical Pharmacokinetics 
Pharmacokinetic (PK) data for apramycin is available for mouse, rat, guinea pig and dog. In 
animal models, the PK of apramycin was equivalent to benchmark aminoglycosides tested (e.g., 
gentamicin). Aminoglycosides are minimally absorbed in the gastrointestinal tract and thus must 
be administered intravenously (IV), subcutaneously (SC), or intramuscularly (IM) in order to 
treat systemic infections. Based on the volume of distribution, apramycin showed no wide tissue 
distribution. Protein binding was low, with fractions unbound (fu) of 90% in human and animal 
plasma. Apramycin was not metabolized. No interactions with cytochrome P450 (CYP) 
isoenzymes were found. Almost all of the administered apramycin was excreted via the kidneys 
and recovered in urine. In mouse models, apramycin content in kidneys increased with 


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increasing doses and duration of treatment. At higher doses and with longer duration of 
treatment, apramycin exposure increased due to a decrease in renal excretion (IB 2021). 
2.3 
Rationale for the Development of Apramycin for Human Use 
Apramycin is a mono-substituted 2-deoxystreptamine comprising a unique bicyclic octadiose 
moiety. It represents a subclass of aminoglycoside antibiotics significantly different from the 
clinically well-established di-substituted 2-deoxstreptamines gentamicin, amikacin, tobramycin, 
arbekacin, plazomicin and others (Mingeot-Leclercq 1999). 
Based on its chemical structure, it was predicted that apramycin evades known aminoglycoside 
resistance mechanisms (Böttger & Vasella 2012). This was further confirmed by screening a 
panel of engineered E. coli strains expressing isolated aminoglycoside resistance genes under 
equipotent constitutive promoter control, which showed the evasion of almost all known 
aminoglycoside-modifying enzymes involved in resistance (aminoglycoside acetyltransferases, 
phosphotransferases, and nucleotidyltransferases). The only exception is the acetyltransferase (3) 
subtype IV (AAC(3)-IV) - a resistance gene that was previously described to confer apramycin 
resistance in animals but is of low clinical prevalence when compared to all other 
aminoglycoside resistance mechanisms (Plattner 2020). Apramycin further retains binding to 
target sites methylated by ribonucleic acid (RNA) methyltransferases. In summary, cross-
resistance to clinically approved antibiotics has been demonstrated to be minimal (Böttger 2020). 
The antibacterial potency of apramycin against wild-type and drug resistant clinical isolates, has 
been endorsed by several independent investigators in the scientific literature. An extensive data 
set on antibacterial potency of apramycin based on more than 1,200 clinical isolates has been 
generated by the IMI ENABLE project (Juhas 2019) The MIC distribution and in vivo efficacy 
of EBL-1003 against highly drug-resistant A. baumannii isolates and lung infections in mice has 
been particularly encouraging (Becker 2020). 
The aim of the clinical development program for apramycin is to allow a full assessment of 
safety and efficacy as well as ensuring that the product is rapidly made available to fulfill an 
unmet medical need in the treatment of infections due to multi-drug resistant (MDR) bacteria. 
2.4 
Clinical Experience 
 
First-in-Human Single Dose Escalation Study 
In the First-in-Human (FIH) single dose escalation trial, single doses of 0.3, 1.2, 3.6, 10.8 and 
30 mg/kg apramycin were investigated in healthy volunteers. The subjects were randomized to 
either apramycin (6 per dose level) or placebo (2 per dose level). 
Safety 
Overall, single IV doses of EBL-1003 (apramycin) up to 30 mg/kg were safe and well tolerated 
in healthy subjects in this clinical trial. 


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There were no deaths or discontinuations due to TEAEs during the study. The 2 SAEs observed 
during the study (1 in Dose Group 0.3 mg/kg: emotional disorder of moderate intensity; 1 in 
Dose Group 30 mg/kg: ligament rupture of moderate intensity) were both not related to the IMP. 
Overall, incidence of TEAEs was low; with 9 subjects (22.5%) reporting altogether 13 TEAEs, 
mostly not (9 TEAEs), or unlikely (3 TEAEs) related to the IMP. Only for 1 TEAE (mild back 
pain in Dose Group 30 mg/kg), a causal relationship to the IMP was considered possible. The 
events were mainly mild (10 TEAEs) or moderate (3 TEAEs); no TEAEs of severe intensity 
were observed. Except for 1 TEAE for which outcome could not be ascertained (SAE: torn 
ligament) as the subject was lost to follow-up, all TEAEs were recovered or resolved at the end 
of the study. 
One subject in Dose Step 1 had a TEAE of special interest (pure tone audiometry with 
sensorineural hearing loss [SNHL] >3 dB). The SNHL was not confirmed upon re-measurement 
and therefore, this TEAE/AESI was assessed as unlikely related to the study drug. 
No effects on safety laboratory parameters, vital signs, ECG parameters or exploratory 
biomarkers of renal toxicity were observed. No medically relevant effects were seen in otological 
examinations. 
EBL-1003 (apramycin) was well tolerated locally at the infusion site. 
There were 2 serious adverse events (SAEs): 
• One subject (0.3 mg/kg apramycin) had admitted himself to hospital due to acute 
emotional decompensation 10 days after treatment. This was recorded as SAE. The event 
was not related to the study treatment but to a personal crisis. The subject completed the 
entire course of the study up to the Day 90 visit. 
• One subject (30 mg/kg apramycin) was admitted to hospital after tearing the ligament of 
his right ankle joint. The event was not related to the study treatment (the subject had 
played soccer). He had completed the study up to Day 14. The subject was lost to follow-
up, so the outcome and end date of the SAE could not be verified. 
Pharmacokinetics 
In the 30 subjects who received apramycin, the plasma concentration profiles of apramycin were 
similar to those observed by other aminoglycosides, such as gentamicin, after IV infusion. 
Exposure characteristics (AUC(0-t) and Cmax) increased with increasing doses with mean values 
(arithmetic and geometric) increasing by approximately the same factor as the dose increases. 
Variability of AUC was small, with coefficients of variation <20%. Variability of Cmax was 
slightly larger. Observed mean exposure characteristics were in line with the predictions based 
on the population PK model for gentamicin (Xuan 2004) and the observed kidney function in the 
healthy volunteers. 


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2.5 
Rationale  
 
Study Rationale 
In a study of apramycin in a mouse lung infection model demonstrated ≥90% lung penetration 
and multilog CFU reduction at doses of ≥5 mg/kg for an XDR Acinetobacter baumannii  
(A baumannii) isolate with an MIC of 8 mg/L (Becker 2020). This finding warrants particular 
interest in determining the ability of apramycin to penetrate human lung tissue. It has previously 
been shown that the lung penetration of plazomicin or amikacin in mice is not a good indicator 
for the ELF targets in humans. Since apramycin has shown lung penetration in mice higher than 
that of plazomicin, it remains to be confirmed whether it also penetrates well into human ELF.  
The current Phase I lung pharmacokinetic study intends to administer apramycin in healthy 
subjects by intravenous infusion to assess primarily pulmonary drug penetration. It is the first 
step in assessing the pharmacokinetics of apramycin in the body compartments relevant for 
diseases associated with bacterial respiratory infections including hospital-associated 
pneumonia/ventilator-associated pneumonia (HAP/VAP). Secondarily, this study will continue 
assessing the overall safety and tolerability of apramycin and exploring specifically ototoxic and 
nephrotoxic effects. The effects on ECG will also be evaluated.  
 
Rationale for Dose Selection 
A single dose of 30 mg/kg had been selected for this study based on the favorable safety and 
tolerability profile observed in the FIH clinical trial in healthy subjects (IB 2021). In addition, 
based on the translational PK-PD modeling study to assess efficacy, 30 mg/kg of EBL-1003 
(apramycin) is the anticipated therapeutic dose for future clinical investigations in patients 
(Tomas Sou 2021).  
2.6 
Identified and Potential Risks and Benefits Associated with the Study 
 
Risks and Benefits Associated with the Study Drug, Apramycin 
Identified Risks 
There are no identified risks associated with apramycin to date. Single apramycin doses ranging 
from 0.3 to 30 mg/kg body weight were generally safe and well-tolerated in the FIH Phase 1 
clinical trial conducted in healthy subjects (IB 2021).  
Potential Risks  
Based on the knowledge on other aminoglycosides, the following contraindications will be 
assumed for apramycin: 
• Known hypersensitivity to aminoglycosides 
• Myasthenia gravis 
 
 


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Potential Drug  Interactions 
There are no clinical data on drug interactions with apramycin. 
Based on the knowledge on other aminoglycosides, concomitant administration of the following 
drugs should be avoided: 
• Drugs affecting renal function, e.g., renin–angiotensin aldosterone system [RAAS] 
inhibitors: angiotensin-converting enzyme [ACE]-inhibitors, angiotensin II [AII]-receptor 
antagonists, spironolactone, epleronone; non-steroidal anti-inflammatory drugs [NSAIDs] 
including cyclooxygenase 2 [COX-2] selective inhibitors). 
• Drugs with potential ototoxic effects, e.g., vancomycin, loop diuretics, quinine and 
quinidine derivatives including mefloquine. 
• Neuromuscular blocking agents. e.g., nondepolarizing (steroidal [e.g., d-tubocurarine, 
rocuronium, vecuronium, pancuronium) and benzylisoquinolinium [e.g., mivacurium, 
atracurium, cisatracurium]). 
• Other aminoglycosides, e.g., gentamicin, tobramycin, amikacin, plazomicin, 
streptomycin, neomycin, and paromomycin. 
 Use During Pregnancy and Lactation 
• Aminoglycosides pass the placenta barrier and may cause fetal harm. Therefore, 
apramycin should not be administered to pregnant women. 
• Aminoglycosides have been reported to be detected in maternal milk. Therefore, 
apramycin should not be administered to breastfeeding women. 
 Undesirable Effects 
Until clinical experience with apramycin is available, similar undesirable effects as observed for 
other aminoglycosides should be anticipated. Special attention should be paid to signs and 
symptoms of nephrotoxicity, ototoxicity, and vestibular toxicity. 
If symptoms are observed after the IMP has been administered, therapy has to be guided by the 
predominant symptoms. Subjects will have to remain under medical surveillance until all 
relevant symptoms have subsided. In case of adverse events, the investigator has to decide if the 
subject has to be withdrawn from the trial. Emergency drugs and equipment have to be available 
at the study site. 
 Overdose 
There is no specific antidote in case of apramycin overdose. The same measures as 
recommended for other aminoglycosides (treatment cessation, adequate hydration, dialysis, 
calcium salts) should be considered until further clinical data are available for apramycin 
(IB 2021). 
 Drug Abuse and Dependency 
No clinical experience is available. 


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Bronchoscopy and Bronchoalveolar Lavage Risks  
Bronchoscopy with bronchoalveolar lavage has a proven safety record in both clinical 
applications as well as human research during more than 40 years of clinical use of this 
procedure in hundreds of thousands of subjects, without any serious lung troubles and side 
effects. However, like all procedures, there are some risks associate with it (Zhang 2020). 
The most common side effect related to the procedure is transient hypoxemia, which may require 
administration of oxygen, and transient tachycardia and premature contractions, which may 
require no treatment or appropriate intervention.   
Other side effects that could be related to the procedure include nausea and dizziness, due to the 
drugs used for sedation or anesthesia, and nasal stuffiness and irritation, throat irritation, 
hoarseness and cough, due to the passage of the bronchoscope and the bronchoalveolar lavage 
(BAL).  
A post-bronchoscopy fever (PBF), typically less than 40°C / 100°F is rare and reported with 
variable rate, ranging from 0% to 10% in some series. It may occur approximately 8 h (range 4 to 
24 h) after BAL and lasts about 14 h. It is not accompanied by bacterial infection or pneumonia 
with abnormal chest X-ray. Antibiotics are not used for prevention and treatment targets the 
symptoms (drugs to lower fever, good hydration). Additional tests may be needed to guide 
treatment (Um 2004). 
Rare complications, with incidence 0.1 to 0.3%, are perforation of the bronchus with 
pneumothorax, atelectasis, and infection. Bleeding has been described for bronchoscopy with 
biopsy of the lung but is very rare (0.1% or less) if only BAL, and is related to mechanical 
trauma, especially in subjects who have a disorder of blood coagulation or take anticoagulants 
(heparin, warfarin, new anticoagulants) and non-prescription non-steroidal anti-inflammatory 
drug (such as aspirin, and ibuprofen).  
These risks are mitigated by adherence to the protocol eligibility criteria. During the 
bronchoscopy, subjects will be placed on supplemental oxygen and monitored continuously for 
oxygen saturation and ECG. Topical anesthetic, sedative or anesthetic dosages will be 
administered according to standard medical practices by experienced and hospital-certified staff. 
Pre- and post-bronchoscopy care will be provided and will include adequate nursing staff and 
standard post-sedation recovery care with direct investigator oversight. Finally, the procedure 
will be performed by experienced pulmonologists that are hospital-certified to perform this 
procedure and assisted by experienced and certified personnel in specially equipped hospital 
rooms equipped to handle emergencies.  
 
Audiometry and Otoacoustic test Risks  
Generally, there are no risks associated with the planned procedures to examine the ear and 
assess cochlear (hearing) function. 


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Venipuncture and IV Catheter Placement Risks 
Venipuncture: Venipuncture causes transient discomfort and may cause fainting. Bruising at the 
site of venipuncture may occur but can be prevented or lessened by applying pressure for several 
minutes. Infection at the site is possible but highly unlikely as aseptic technique will be used. 
IV catheter placement: An indwelling catheter may be placed in an arm vein (preferably 
antecubital) for frequent blood drawing for PK measurements. The catheter may cause phlebitis 
with signs of redness and warmth at or near the IV insertion site, and thrombophlebitis with a 
hard area palpable near the IV insertion site. These risks are minimal as the IV catheters, when 
used, are only used briefly after dosing. Careful inspection of the catheter site, including 
visualization of blood return, and withdrawal of the catheter if needed will minimize this risk. 
There is a risk of infection; however, this is a small risk as aseptic technique will be used. 
 
Additional Risks 
ECG: Possible side effects from ECG patches include a rash or minor irritation of the skin. 
Blood draws: The amount of blood drawn is about 25.7 mL at Screening Visit, 22.2 mL on  
Day -2, 188.5 mL during the inpatient period (Days 1 to 3), 22.2 mL at Day 14 Visit, 3.5 mL at 
Day 30 Visit, and 262.1 mL during the entire trial (Appendix C, Table 5). Additionally, small 
amounts of blood loss may occur if an IV catheter is used or additional blood samples are 
collected (for repeat laboratory testing, TEAE evaluation, etc.). Overall, the amount of blood that 
may be drawn during the trial is within the amount that are considered safe to be drawn during 
short or extended periods, respectively, and not excessive for the safety and PK assessment 
requirements of Phase 1 trials. However, there is a small risk that some subjects may develop 
mild symptoms of hypovolemia or anemia during the trial. These are reversible with specific 
treatments (e.g., fluid replacement, good nutrition, vitamins, or iron supplementation). 
2.7 
Known Potential Benefits 
The trial has no direct benefit for subjects participating in the trial. Knowledge gained in the trial 
could be of future benefit to public health and to individuals with infections, who might benefit if 
the study drug is licensed.  
In accordance with the Health Insurance Portability and Accountability Act (HIPAA) regulation, 
subjects will be promptly notified of any clinical test results that would have an impact on their 
health. Screening evaluations may detect previously unknown abnormalities that could be 
clinically significant. This could benefit a subject, as it may lead to earlier diagnostic evaluation 
and treatment of an underlying disorder. 
2.8 
Risk/Benefit Ratio 
This ratio cannot be calculated by available data. The risk for an SAE occurring in a subject 
treated with single doses of 30 mg/kg body weight is considered to be very low. Subjects will be 


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treated in an inpatient unit and closely monitored during the trial, so that appropriate emergency 
care can be provided immediately if an acute event occurs.  
Data collected in the clinical development of apramycin to date, support further development of 
apramycin. The benefit of a potential new addition to the drugs available for the treatment of 
antibiotic-resistant bacteria and of using a new generation aminoglycoside with lower toxicity 
than licensed preparations, outweighs the known risks of the medication and the procedures to be 
used in this trial.


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3 
OBJECTIVES AND OUTCOME MEASURES 
3.1 
Study Objectives 
 
Primary 
• To assess plasma pharmacokinetic (PK) profile of apramycin and lung penetration of 
apramycin in epithelial lining fluid (ELF) and alveolar macrophages (AM) after single 
intravenous (IV) apramycin dose of 30 mg/kg in healthy subjects. 
 
Secondary  
• To assess the safety of single IV administration of 30 mg/kg apramycin in healthy 
subjects. 
• To assess changes in otoacoustic testing.  
 
Exploratory  
• To assess changes in kidney function biomarkers. 
3.2 
Study Outcome Measures 
 
Primary 
• Lung concentration o and PK parameters of total apramycin: 
o ELF: Cmax, AUC(0-24), AUC(0-t), AUC(0-∞), Cmax, Tmax, t½ 
o AM: Cmax, AUC(0-24), AUC(0-t), AUC(0-∞), Cmax, Tmax, t½ 
• Plasma concentration and PK parameters of total apramycin:  
o AUC(0-∞), AUC(0-t), AUC(0-24), Cmax, Tmax, t½, CLT, Vd central, 
• Ratio of exposure parameters of Lung PK to Plasma PK: 
o ratio of Cmax in ELF over Cmax in plasma  
o ratio of Cmax in AM over Cmax in plasma  
o ratio of AUC (0-24), AUC(0-t), and AUC (0-∞) in ELF over AUC (0-24), AUC(0-t), and  
AUC (0-∞) in plasma  
o ratio of AUC (0-24), AUC(0-t), and AUC (0-∞) in AM over AUC (0-24), AUC(0-t), and  
AUC (0-∞) in plasma  
 
Secondary  
• Type and incidence of treatment-emergent adverse events (TEAEs) and serious AEs 
(SAEs) to Day 30 (± 4 days) after dosing. 


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• Frequency of clinically significant TEAEs in physical examination (PE) findings and of 
changes from baseline in vital signs and clinical laboratory parameters until Day 14  
(± 3 days) after dosing. 
• Frequency of changes in ECG intervals (QTcF, PR, QRS, HR), and of morphological 
changes from baseline until 24 ± 1 h after dosing. 
• Type and incidence of TEAEs related to auditory (cochlear) function tests (pure-tone 
audiometry and distortion product otoacoustic emissions [DPOAEs]) to Day 30 
(± 4 days) after dosing.  
 
Exploratory 
• Changes from baseline in kidney injury molecule 1 (KIM-1) and serum cystatin C 
concentrations until Day 30 (± 4 days) after dosing. 
 


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4 
STUDY DESIGN 
This is a Phase 1, single center, open-label, study to evaluate the plasma and lung 
pharmacokinetics (PK), safety and tolerability of a single intravenous (IV) dose of apramycin in 
twenty healthy male and female subjects 18 to 45 years of age (inclusive).  
Each subject will be enrolled to one of five cohorts (T1-T5) before dosing on Day 1. Four 
subjects will be allocated to each cohort. Each cohort corresponds to the timepoint after dosing 
when a single bronchoscopy with bronchoalveolar lavage (BAL) will be performed: 0.5 h  
(± 5 min) (Cohort T1), 2 h (± 5 min) (Cohort T2), 4 h (± 10 min) (Cohort T3), 8 h (± 15 min) 
(Cohort T4), and 24 h (± 1 h) (Cohort T5). The timing of bronchoscopy and BAL in cohort T5 is 
nominal. The final timepoint will be determined after analysis of apramycin concentrations in 
BAL. If apramycin is detectable in BAL collected from cohorts T1-T4, then T5 will be enrolled. 
If it is not detectable, then cohort T5 will not be enrolled. Blood will be collected at several 
timepoints to 60 hours after dosing (see collection schedule in Section 8.5.1) and will include 
samples that will coincide with BAL collections. The concentration and PK exposure parameters 
(Cmax and AUC(0-t)) of total apramycin in epithelial lining fluid (ELF) and alveolar macrophages 
(AM), collected through BAL, and in plasma collected at the scheduled timepoints to 8 h after 
dosing, will be evaluated from the first four cohorts, T1-T4, to determine if apramycin penetrates 
into the lung tissues and dosing of the T5 cohort is warranted. If cohort T5 is dosed, the final 
concentration and PK analysis of apramycin in BAL samples and plasma will include data from 
all T1-T5 cohorts. 
Subjects will participate in the study for approximately 58 days, including a 28-day screening 
period, consisting of an out-patient period of 26 days (Day -28 to Day -3) with 1 to 2 site visits 
and 2 inpatient days, Day -2 and Day -1, to confirm eligibility, enroll and complete baseline 
assessments; a 3-day in-patient treatment period, with treatment administered on Day 1, 
bronchoscopy with BAL done within 24 h after dosing, and in-patient follow up completed on 
Days 2 and 3; and a 27-day out-patient follow-up period with site visits on Day 14 (± 3 days) and 
Day 30 (± 4 days), the last follow up visit. It is anticipated that the duration of the study will be 
about 6 months to enroll Cohorts T1-T4 and 10 months to complete Cohort T5, if enrolled, 
following an interim data analysis to review lung PK data.  
Enrollment will be rolling and will be coordinated with the bronchoscopy service. Tentative 
reservations will be made and confirmed once subject eligibility is determined. Enrollment and 
treatment of subjects will be staggered and no more than two subjects will be dosed on a single 
day not less than 2 hours apart. Additional subjects may also be admitted to the Clinical Trial 
Unit (CTU) before dosing and may serve as back-up study subjects. A subject assigned to one of 
the T1 to T4 cohorts may be reassigned to another cohort depending on the availability of a 
bronchoscopy time slot. (See Section 5.3.1)   


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Apramycin will be reconstituted in sterile normal saline and a 30 mg/kg dose will be 
administered as a single IV infusion in a forearm vein over 30 min (± 5 min) using a syringe or 
infusion pump. (See Section 6.2 for DP preparation and administration). 
Safety monitoring during the inpatient period will include: (a) Scheduled assessments per 
protocol: a physical examination (PE), vital signs (VS; including HR, BP, RR and T); 12-lead 
standard ECG (in triplicate at scheduled timepoints on Day-1 matching planned timepoints on 
Day 1); clinical safety laboratory tests (Hematology [HEM], Coagulation [COAG], Chemistry 
[CHEM] including estimated Glomerular Filtration rate [eGFR], and Urinalysis [UA]; and 
audiology testing (middle ear examination and cochlear testing [hearing testing, consisting of 
pure-tone audiometry and DPOAEs). Exploratory plasma and urinary nephrotoxicity biomarkers 
(plasma cystatin C and urinary KIM-1) will also be measured. In addition, (b) Unscheduled 
assessments after bronchoscopy will be performed according to orders by the attending 
pulmonologist and as needed. Subjects with on-going TEAEs will be followed until resolution or 
stability of the TEAE as assessed by the PI. 
Safety Monitoring Committee (SMC) Role 
A SMC will be appointed to oversee the safe conduct of the trial, review safety data and provide 
recommendations on safety monitoring in the current and future clinical trials with apramycin. A 
scheduled interim SMC meeting will be held after all subjects in Cohorts T1-T4 complete all 
assessments. If the last cohort (T5) will be enrolled for dosing, a final SMC will be held to 
review cumulative data on all subjects. If Cohort T5 is not dosed, the interim meeting will be 
considered final.  
 If criteria for halting the trial (as listed in Section 9.5.1) are met, enrollment and dosing of new 
subjects will be suspended, and an ad hoc SMC meeting will be held to review all available 
safety data. Study procedures in subjects already enrolled and dosed will continue per schedule. 
A suspended clinical trial will resume upon recommendation by the SMC and decision by 
DMID, NIAID (the sponsor). 
4.1 
Sub-studies 
No sub-studies are planned. 


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5 
STUDY ENROLLMENT AND WITHDRAWAL 
Only subjects who meet all inclusion criteria and no exclusion criteria will be eligible for 
enrollment into the trial. No exemptions are granted on Inclusion/Exclusion Criteria in DMID-
sponsored trials. 
Twenty healthy male and female subjects, aged 18 to 45 years inclusive, will be enrolled in the 
trial.  
5.1 
Subject Inclusion Criteria 
All must be answered YES for the subject to be eligible for study participation: 
 
1. Subject reads and signs the Informed Consent Form (ICF) and agree to have 
bronchoscopy with bronchoalveolar lavage under sedation or light anesthesia and comply 
with study procedures. 
2. Healthy male or non-pregnant, non-lactating female subjects 18 to 45 years of age (both 
inclusive) at the time of dosing.  
a. Note 1: Determined by medical history (MH), medication use, physical 
examination (PE), and vital signs, clinical laboratory tests and 12-lead ECG 
within reference ranges at Screening and Day-2. (See Sections 8.1 and 8.2, 
Appendix B, Table 2, Table 3 and Table 4 and the study-specific MOP.) 
Exceptions to BP, HR and laboratory test values being with normal ranges are: 
o Abnormal HR and BP on first measurement may be repeated twice more with 
the subject resting between measurements for at least 5 min according to 
Section 8.1.6. 
o Subjects with baseline HR ≥ 45 to 50 bpm may be accepted if otherwise 
healthy adults with known history of asymptomatic bradycardia.  
o Subjects with baseline SBP up to 140 mmHg and DBP up to 90 mmHg may 
be accepted if otherwise healthy.  
o A laboratory value that is Grade 1 will be allowed if not considered to be 
clinically significant by the investigator, with the exception of ALT, AST, AP, 
BUN, urine protein, serum creatinine or estimated glomerular filtration rate 
(eGFR) <70 mL/min /1.73 m2 by the Chronic Kidney Disease Epidemiology 
collaboration (CKD-EPI) equation. 
3. 
Female subjects of childbearing potential should use highly effective methods of 
contraception from the time of screening to 30 days after dosing. 
• Note 1: A female is considered of childbearing potential unless post-menopausal 
(defined as history of ≥1 year of spontaneous amenorrhea and a FSH level 
>40 IU/L), or permanently surgically sterilized. 


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• Note 2: Highly effective contraceptive methods include: (a) surgical sterilization 
methods, such as tubal ligation, bilateral oophorectomy, salpingectomy, 
hysterectomy, or successful tubal obliteration (e.g., Essure®) with documented 
radiological confirmation test at least 90 days after the procedure, or (b) long-
acting reversible contraception, such as progestin-releasing subdermal implants, 
copper intrauterine devices (IUDs), levonorgestrel-releasing IUDs. 
• Note 3: A subject who is not sexually active and abstains from sexual intercourse 
can be enrolled and abstinence documented.   
4. Males, including vasectomized men, having sexual intercourse with women of 
childbearing potential must agree to consistent use of condoms from IMP 
administration through at least 30 days after dosing, and must also agree to not donate 
sperm during this same time period. 
• 
Note: A subject who is not sexually active and abstains from sexual intercourse 
can be enrolled and abstinence documented. 
5. BMI 18.0 to 32.0 kg/m2 (inclusive) and body weight not less than 50 kg.  
6. Subjects with normal hearing, i.e., symmetric hearing with air conduction thresholds 
no worse than 20 dB hearing loss for the frequencies 0.5-1-2-4-6-8 kHz bilaterally. 
7. Normal (reproducibility 70% or better) of distortion product otoacoustic emissions 
(DPOAEs). 
Note: Absence of DPOAEs at no more than two consecutive or non-consecutive 
DPOAEs in each ear is acceptable. 
8. Normal otoscopic findings in the ears, normal tympanic membrane mobility and 
stapedial reflex present. 
9. From the signing of the informed consent until the last follow-up visit, subjects must 
be willing to avoid exposure to loud music or noise.  
• Note: Noise avoidance to include continuous usage of earpieces at high volume, 
attending loud concerts or dance events, or using firearms or attending fireworks. 
10. Normal lung function with Forced Expiratory Volume in the first second (FEV1) 
predicted ≥ 80% and FEV1/Forced Vital Capacity (FVC) > 70%. 
11. Subjects must be willing to avoid excessive physical exercise within 48 h prior to 
dosing until discharge from the CTU on Day 3, and 24 h before each follow-up visit 
(Day 14 ± 3 days and Day 30 ± 4 days). 
12. No history of acute febrile or infectious illness for at least 7 days prior to the 
administration of the IMP.  
13. No history of lower respiratory tract infection within 4 weeks prior to screening. 


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14. Have adequate venous access for infusion and blood draws. 
5.2 
Subject Exclusion Criteria 
All must be answered NO for the subject to be eligible for study participation: 
1. Lactating females. 
Medical and surgical history: 
2. Any history of hypersensitivity to aminoglycosides. 
3. Any history of drug hypersensitivity, asthma, urticaria or other severe allergic diathesis.  
4. Any history of seasonal allergies with ongoing symptoms for more than a week prior to 
dosing requiring glucocorticoids and/or frequent use of antihistamines for treatment. 
5. Any history of a chronic condition that may increase risk to subject or interfere with 
endpoint assessment, or any unstable chronic disease.   
• Note 1: Unstable chronic disease is defined by need for frequent medical 
interventions that lead to a change in medications and/or required hospitalization, 
surgery or an invasive procedure or emergency department/urgent care visit. 
• Note 2: Any chronic disease, that has been diagnosed within 90 days of screening is 
excluded. 
6. History of any psychiatric medical condition that has required hospitalization in the last 5 
years or subject is considered psychologically unstable by the investigator. 
7. History of acute or chronic problems with hearing and/or balance in the last 24 months. 
• Note: These include but not limited to use of hearing aid, head injury leading to 
otologic damage, tumor of the head or neck, autoimmune disease of the inner ear, 
tinnitus, vestibular disease, auditory neurinoma, endolymphatic hydrops and/or 
Meniere’s disease, perilymphatic fistula, otitis media, labyrinthitis, sudden hearing 
loss, known retrocochlear hearing impairment, conductive hearing loss exceeding 10 
dB at any frequency, ear canal and/or middle ear disease including inflammation or 
effusion, pathological tympanometry. 
8. Past injury or surgery to the middle or inner ears.  
• Note: Myringotomy or tympanic tube insertion in childhood with complete healing 
and normal hearing test are excluded. 
9. Family history of hearing loss before the age of 60. 
10. Subjects who have had previous intolerance or contraindications to medications applied for 
sedation or anesthesia during bronchoscopy. 


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• Note: These include benzodiazepines or topical anesthetic agents (lidocaine or 
xylocaine) including reversal agents such as flumazenil.  
Laboratory examinations: 
11. Positive serum pregnancy test for women at screening or urine pregnancy test at check-in. 
12. Positive test for HIV antibodies, hepatitis B-virus surface antigen (HBsAg), or  
anti-hepatitis C-virus antibodies (anti-HCV). 
Prior medication: 
13.  Use of any prescription or non-prescription medication prior to the dose of IMP as 
described in Section 6.6. 
• Note: Exceptions are hormonal contraceptives, which are permitted throughout the 
study, and solitary doses of up to 1,000 mg paracetamol.  
14. Use of any investigational drug product within 30 days or 5 half-lives (whichever is longer) 
before dosing.  
15. Planned participation in a clinical research study that requires treatment with a study drug 
or blood draws or other invasive assessments during the study period (screening until final 
visit). 
Lifestyle restrictions: 
16. More than low-risk alcohol consumption (men: ≥ 24 g of pure alcohol regularly per day; 
women: ≥12 g of pure alcohol regularly per day) for the previous 3 months. 
17. Any history of alcohol or drug abuse or positive alcohol breathalyzer test. 
18. Suspicion of illicit drug use / abuse or positive urine drug screen test (See Section 6.7.4 and 
Section 8.2.5 for prohibited substances). 
19. “History of ≥10 pack-years smoking, or history of any nicotine use in the 6 months before 
check-in (Day -2) or positive urine cotinine screen at check-in. 
• Note 1: Nicotine products include cigarettes, e-cigarettes, pipe, cigar, chewing 
tobacco, nicotine patch. 
• Note 2: A positive urine cotinine at screening is allowed if negative at check-in 
(Day -2).  
20. Caffeinated beverages/foods are prohibited within 48 hours before dosing to Day 3 of the 
trial. During the follow up period, consumption is restricted to not more than 3 cups or 
equivalent per day. 
21. Judged by the investigator to have occupational noise exposure of high risk during the trial 
(e.g., construction site workers, military workers, etc.). 


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22. Blood or plasma donation of 500 mL within 3 months or more than 100 mL within 30 days 
before signing informed consent or planned donation prior to completion of this trial.  
5.3 
Treatment Assignment Procedures 
 
Enrollment Procedures 
Twenty healthy subjects who consent to participate in the trial and meet the eligibility criteria 
will be enrolled following admittance to the CTU and confirmation of eligibility. Subjects will 
be registered using Advantage eClinical®, a web-based application developed by The Emmes 
Company, LLC, the DMID Statistical and Data Coordinating Center (SDCC) for the trial. 
Reasons for screen failure or no enrollment and dosing of an eligible subject will be listed in the 
MOP and also be entered in the system. Instructions for using the enrollment module are 
included in the Advantage eClinical® User’s Guide. 
Since this is an open-label study of a single dose of IMP, subjects will be enrolled on a rolling 
basis. Four subjects will be assigned to each one of five cohorts, T1-T5 (Section 4), for lung PK 
with equal allocation (1:1:1:1:1) and registered into the Advantage eClinical®. Each cohort 
corresponds to a single Lung PK sampling timepoint. It is planned to enroll subjects into cohorts 
T1 to T4 according to availability of the bronchoscopy suite. However, on the day of 
bronchoscopy, a subject may be reassigned to another cohort than planned on the same day, 
depending on emergencies at the bronchoscopy suite at the time originally planned, or cancelled.   
Appointments for the bronchoscopy procedure will be made in advance and every effort will be 
made to admit, dose and perform bronchoscopy with BAL according to the schedule. However, 
if a scheduled bronchoscopy cannot be performed according to schedule on a dosing day, the 
following steps may be taken according to the time of cancellation before or after dosing: 
• If the procedure is cancelled before administration of study drug, an effort will be made 
to reschedule the procedure.  
• If the procedure is cancelled after administration of the study drug, bronchoscopy may be 
either rescheduled if it can be performed at a later time that can accommodate an 
available Lung PK timepoint up to 8 h after dosing or cancelled. 
o If bronchoscopy is rescheduled, the registration in the Advantage eClinical® will be 
updated to indicate the cohort that the subject completed and a future subject would 
be assigned in the registry to complete the missed Lung PK timepoint.  
o If it is not feasible to reschedule bronchoscopy, the subject will be terminated from 
participation in the study and will be replaced. The subject will be asked to remain in 
the study to complete safety and otoacoustic testing until Final Visit (Day 30 ± 4 
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appropriate early termination assessments will be completed prior to discharge from 
the CTU. (See Section 5.3.3 and Section 7.7) 
 
Reasons for Withdrawal and Discontinuation of Study Drug Administration 
A subject may withdraw from the trial at any time for any reason, without any consequences. 
A subject will be discontinued from the trial if any of the following occur before dosing: 
• Request by the subject to terminate participation. 
• Failure to receive the study drug due to difficulty initiating or maintaining an intravenous 
infusion line. 
A subject may be removed from the trial after dosing for the following reasons; however, 
whenever possible, the subject will be followed for safety per protocol to Day 30 (± 4 days): 
• Failure to adhere to protocol requirements. 
• Loss to follow-up. 
• Request of primary care provider. 
• Request of the Institutional Review Board (IRB)/Ethics Committee (EC), FDA or DMID. 
• The subject’s well-being, based on the opinion of the investigator. 
• The occurrence of an SAE or TEAE warranting withdrawal. 
• Failure to receive the entire volume of apramycin solution during the designated infusion 
time. 
 
Handling of Withdrawals and Discontinuation of Administration 
• Subjects who are withdrawn before dosing may be replaced with a subject assigned to the 
same lung PK timepoint group (T1-T5).  
• Subjects who did not receive the entire infusion volume of apramycin will be replaced 
and withdrawn from bronchoscopy and associated procedures and from PK analysis but 
will be encouraged to continue follow up for safety. 
• Subjects who withdraw after receiving the entire study treatment but before the scheduled 
bronchoscopy and associate procedures will be replaced and withdrawn from the study 
and data analysis but will be followed for safety.  
• Subjects who withdraw after receiving the entire study treatment and completed 
scheduled bronchoscopy and assessments will not be replaced unless more than 50% of 
subjects in a Lung PK timepoint cohort are withdrawn from the trial.   
• Generally, subjects who received the entire amount of the study drug but withdraw from 
the trial will be encouraged to continue follow-up (with subjects’ consent) for safety 
assessments and plasma PK sample collection.  


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• Subjects withdrawing will be asked to complete safety and otoacoustic testing according 
to the ET schedule (See Section 7.7).  
 
Lost to Follow-up 
If subjects fail to appear for a follow-up assessment, extensive effort (i.e., three documented 
contact attempts via phone calls, e-mail, etc., made on separate occasions and followed by a 
certified letter) will be made to locate or recall them, or at least to determine their health status. 
These efforts will be documented in the subjects’ records. 
 
Termination of Study 
Although DMID has every intention of completing the trial, it reserves the right to terminate the 
trial at any time for clinical or administrative reasons. In addition, the trial may be terminated or 
suspended at the request of the FDA, SMC or IRB/EC. 


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6 
STUDY INTERVENTION/INVESTIGATIONAL 
PRODUCT 
6.1 
Description of Study Product  
Product: Apramycin (EBL-1003) 
The active drug substance apramycin is chemically a monosubstituted 2-deoxystreptamine. It is 
manufactured as crystalline free base by purification of the commercially available intermediate 
apramycin sulfate (CAS 37321-09-8). Apramycin sulfate is manufactured by fermentation 
(Streptoalloteichus tenebrarius) by Shandong Qilu King-Phar Pharmaceutical Co, China and 
then recrystallized to apramycin free base and purified by Research Institutes of Sweden (RISE). 
Drug Substance is then formulated and bottled by Patheon Ferentino, Italy (a Thermo Fisher 
Scientific company) for Juvabis AG, the pharmaceutical sponsor of the study. 
The diluent for apramycin will be 0.9 % sodium chloride injection, USP. The USP grade 0.9% 
Sodium Chloride Injection, or normal saline, is a sterile, nonpyrogenic, isotonic solution; each 
mL contains sodium chloride 9 mg. It contains no bacteriostatic agent, antimicrobial agent, 
preservatives, or added buffer and is supplied only in single-dose vials. The solution may contain 
hydrochloric acid and/or sodium hydroxide for pH adjustment (pH 5.3, range 4.5 to 7.0). This 
product should be used to dilute apramycin to the desired concentration. 0.9% sodium chloride 
for injection, USP is clear in color.  
 
Acquisition 
Product: Apramycin (EBL-1003) 
Upon request by DMID, study product will be shipped for clinical labeling from a German 
location supplied by CSM (a Clinigen company) to the following address:    
DMID-Clinical Materials Services (CMS) 
Fisher BioServices 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 
Tel: 240-477-1350 
Fax: 240-477-1360 
E-mail: DMID.CMS@ThermoFisher.com    
The diluent, 0.9% Sodium Chloride Injection, USP, will make up the infusion volume (30 mL) 
and will be provided by the clinical research site. Details will be provided in the protocol-
specific Manual of Procedures (MOP). 


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The IMP will be shipped from DMID-CMS to the CTU upon request and approval by DMID. 
Details will be provided in the protocol-specific MOP. 
Ancillary supplies required for IV administration (including syringes or infusion bag, tubing and 
IV catheter and line-filter) will be supplied by the clinical research site as described in the 
protocol-specific MOP. 
 
Formulation, Packaging, and Labeling 
Product: Apramycin (EBL-1003) 
Apramycin for Infusion, the IMP, is formulated as 150 mg free base /mL in sterile aqueous 
solution, pH 5.5 to 6.0 adjusted with sulfuric acid. The study product is supplied in 20 mL glass 
vials. A volume of ≥18 mL can be withdrawn from each vial. To protect the solution from light, 
the vials are stored in folding boxes. 
The clinical research site will provide the diluent, 0.9% sodium chloride injection, USP. 
Commercial source, product information and packaging will be described in the MOP.  
Each product vial and diluent will be labeled in compliance with applicable regulatory 
requirements, including the FDA-required cautionary statement, “Caution – New drug – Limited 
by Federal (or United States) Law to Investigational Use”.  
 
Product Storage and Stability 
Product: Apramycin (EBL-1003) 
Apramycin for Infusion will be shipped refrigerated at +5 ± 3°C (35.6 to 46.4°F) and should be 
stored refrigerated at 2 to 8°C (35.6 to 46.4°F) within the packaging until time of preparation. 
Protect from freezing. The reconstituted solution must be stored at 2 to 25°C (storage at 2 to 8°C 
if possible) and used within 5 hours or discarded. If not used, it must be quarantined and 
maintained for study product accountability as per Section 6.4.  
The diluent, 0.9% NaCl for injection, USP, will be stored per manufacturing instructions. 
Product storage and stability will be described in the MOP. 
The clinical supplies storage area at the CTU will be monitored by its staff for temperature 
consistency with the acceptable storage temperature range specified in this protocol or in the 
MOP. Documentation of temperature monitoring will be maintained. 
6.2 
Dosage, Preparation, and Administration of Study Intervention/ 
Investigational Products 
The site Research Pharmacist (RP) will prepare the study product on the same day as 
administration. The study drug will be inspected for damage, contamination, discoloration, or 


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particulate matter before use. Any study drug that fails inspection will be quarantined at 
appropriate temperature and labeled ‘Do Not Use’ until further notice. The Site Principal 
Investigator (PI) or responsible person will immediately contact the DMID Clinical Project 
Manager (CPM) and study clinical team for further instructions before administering any 
additional study drug infusions. Based on the information collected, DMID and/or Juvabis AG 
will determine whether the affected study drug can be used. If it cannot be used, the CTU will 
receive specific instructions on how to return it to DMID CMS or destroy it on site.  
Preparation of the product will be performed using aseptic technique under a sterile environment 
(e.g., Biologic Safety Cabinet or laminar flow hood). Each subject will receive 30 mg/kg of 
apramycin in a volume of 30 mL as a single IV infusion over 30 (± 5) minutes in a hand or 
forearm vein using a syringe or infusion pump. Based on the subject weight, the appropriate 
weight-based apramycin for infusion dose will be calculated and the appropriate number of vials 
will be removed from storage to prepare the infusion. Sterile normal saline will be used to adjust 
the volume. 
The subjects will be admitted to the Phase 1 unit 2 days before the planned infusion. Verification 
that the subject still meets all inclusion criteria and does not have any exclusion criteria must be 
made on the morning of Day 1 prior to dosing. The site Research Pharmacist will prepare the 
infusion as described in the protocol-specific MOP. Apramycin for infusion should be 
administered as a single IV infusion over 30 min (± 5) using a syringe or infusion pump via a 
hand or forearm vein. The drug should not be administered as a bolus. The IV administration set 
must contain a 0.20 µm in-line filter. Blood for plasma PK sampling will be drawn from a 
catheter inserted in a vein in the other forearm. 
Refer to protocol-specific MOP for detailed information on apramycin dose and volume 
calculations, as well as preparation and administration of the study product and handling of 
infusion interruptions. 
6.3 
Modification of the Study Intervention/ Investigational Products  
Not applicable for single-dose study drug trial. See Study Halting Criteria, Section 9.5.1. 
Handling of infusion interruptions will be reported in the MOP. 
 
Overdose 
An overdose is defined as a dose greater than the high-dose level evaluated in the trial. All 
overdoses will be reported as a deviation; if the overdose is associated with a TEAE, then the 
TEAE will also be reported. In the event of an overdose of apramycin, the Investigator will use 
clinical judgment in treating the overdose and contact the DMID Medical Monitor (MM). There 
is no specific antidote in case of apramycin overdose. The same measures as recommended for 
other aminoglycosides (treatment cessation, adequate hydration, dialysis, administration of 


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calcium salts for neuromuscular blockade) would be considered until further clinical data are 
available for apramycin. (IB 2021) The investigator will refer to the relevant document(s) for 
detailed information regarding warnings, precautions, contraindications, TEAEs, and other 
significant data pertaining to apramycin. Such documentation may include but not be limited to 
the IB. 
6.4 
Accountability Procedures for the Study Intervention/ 
Investigational Products   
The Site PI is responsible for the distribution and disposition of the study drugs and has ultimate 
responsibility for accountability. The Site PI may delegate this responsibility to the Site RP. If 
delegated, the Site RP will be responsible for maintaining complete records and documentation 
of the study drug’s receipt, accountability, dispensation, temperature monitoring, storage 
conditions, and final disposition. Time of study drug administration to the subject must be 
recorded on the appropriate data collection form (DCF). 
All study drugs, whether administered or not, will be documented on the appropriate study drug 
accountability record or dispensing log. Used and unused apramycin vials will be retained until 
monitored. Upon completion of the trial and after the final monitoring visit, any remaining 
unused study drugs will either be returned or destroyed appropriately at the CTU as per DMID 
requirements and instructions that will be communicated to the CTU by the DMID CPM. 
Unused apramycin vials will be released for disposition per DMID requirements. DMID does not 
require used containers of study product to be maintained at the research pharmacy, except when 
the local institution’s SOP/policy mandates retaining used IV vials. If local SOPs allow 
destruction of used study product containers, the used vials can be destroyed per the site’s SOPs; 
a second staff member must observe the destruction and sign verification (two signatures) that 
the used vials were discarded. 
Any unused solution left in the IV infusion syringe or bag or the IV administration tubing after 
administration to the subject should be discarded as biohazardous waste. If a container of study 
product is unusable due to breakage, or any other reason, this explanation should also be noted 
on the Study Product Accountability Record (e.g., broken – dropped on floor). Details will be 
provided in the MOP.  
6.5 
Assessment of Subject Compliance with the Study Intervention/ 
Investigational Products  
Since each dose of apramycin will be administered by site personnel, subject compliance is not 
anticipated to be an issue. Complete information regarding any partial or interrupted dosing will 
be documented. Subjects unable to receive the full volume of apramycin infusion will be 
withdrawn from data analysis and followed up for safety as described in Section 5.3.3.   


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6.6 
Prior and Concomitant Medications/Treatments 
Medications include the following: prescription drugs, birth control hormonal preparations, non-
prescription medication, herbs, vitamins, nutritional supplements, and illicit and recreational 
substances.  
Medications taken before or after dosing will be reported as Prior Medications or Concomitant 
Medications (ConMeds), respectively.  
Prior prescription medications will be recorded at Screening Visit. All prior medications are 
not allowed during the study period with the exception of oral contraceptives, which are 
permitted throughout the study, and solitary doses of up to 1,000 mg acetaminophen 
(paracetamol). 
The following medications are prohibited for the indicated periods prior to dosing: 
• Use of any investigational drug product within 30 days or 5 half-lives (whichever is 
longer) before dosing or planned use during the study period (screening until Final Visit). 
• Use of aminoglycosides within 3 months prior to dosing. 
• Use of neuromuscular blocking agents within 1 week or 5 half-lives (whichever is longer) 
prior to dosing. 
• Use of potentially nephrotoxic medication 2 weeks prior to dosing: Renin– angiotensin-
aldosterone system [RAAS] inhibitors: angiotensin-converting enzyme [ACE]-inhibitors, 
angiotensin II [AII]-receptor antagonists (enalapril, lisinopril, ramipril captopril, 
benazepril), spironolactone, eplerenone; non-steroidal anti-inflammatory drugs [NSAIDs] 
(aspirin, ibuprofen, naproxen, indomethacin, etc.) including cyclooxygenase 2 [COX-2] 
selective inhibitors (Bextra, Celebrex, Vioxx).  
• Use of potentially ototoxic medication 3 months prior to dosing: vancomycin, loop 
diuretics (bumetanide, ethacrynic acid, furosemide), quinine and quinidine derivatives 
including mefloquine with the exception of occasional intake of quinine containing 
beverages. 
Non-prescription medications, herbs, vitamins, and nutritional supplements will not be taken 
within 15 days before dosing and during the trial. Exceptions: vitamins and OTC medications 
taken for < 48 h for the treatment of common symptoms (e.g., headache, indigestion, muscle 
pain) may be allowed if approved by the designated study clinician. 
Blood/blood products (RBCs, WBCs, platelets, and plasma) donation of 500 mL within 
3 months or more than 100 mL within 30 days before signing informed consent to this trial is not 
allowed, and it is prohibited during the course of this trial. 


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Following dosing, each new ConMed and changes to existing medications will be recorded. 
Subjects will be required not to utilize non-study medications during the trial except those 
deemed necessary by the Site PI or sub-investigator. 
Any drug (e.g., non-prescription medications, herbal supplements, vitamins, or prescription 
medications) or vaccines or blood/blood products used by the subject during the trial will be 
recorded in the subject’s source documents and on the appropriate electronic case report form 
(eCRF), and the PI or authorized study clinician (listed on FDA Form 1572) will note whether 
the use was medically indicated and immediately necessary. Any use of medications not 
authorized by the study PI or authorized clinician will be recorded as a deviation. 
6.7 
Subject Restrictions and Precautions 
From signing informed consent until end-of-study, the subjects have to follow the instructions of 
the study site. They have to adhere to the following restrictions during participation in the study. 
 
Smoking 
Smoking is prohibited during the in-patient period of the study (Day -2 to discharge on Day 3) 
and should be avoided during the out-patient follow-up period until the last visit (Day 30 ± 
4 days). 
 
Physical Activity 
Subjects will be counseled to refrain from rigorous physical activity starting 2 days before dosing 
until discharge from the CTU on Day 3, and 24 h before each follow-up visit (Day 14 ± 3 days 
and Day 30 ± 4 days).  
 
Food and Fluid intake 
Subjects will be provided food and non-alcoholic beverages by the CTU during the dosing period 
of the trial. Subjects will fast at least 6 hours prior to blood draw on days of clinical laboratory 
testing (screening, Day -2, Day 1 [pre-dose], Day 2, Day 3 and Day 14 [± 3 days]). Subjects will 
fast for at least 6 h before dosing on Day 1 and 2 h after dosing. In addition, subjects will fast 
from 4 hours before to at least 2 hours after bronchoscopy. A light nutritional broth or beverage 
may be served if bronchoscopy is scheduled 4 h to 8 h after dosing on Day 1. Water can be taken 
ad lib during those periods of fasting. Water consumption will be at least 2 L daily from the time 
of dosing to 48 h (Day 2) after dosing, and the time and amount consumed will be recorded.   
 
Alcohol, Marijuana and Illicit Drugs 
Subjects will have to agree to consume not more than 24 g  (men)/ 12 g (women) pure alcohol per 
day from Screening to enrollment (Day -2) and from the day of discharge from the CTU on 
Day 3 to Day 30 ± 4 days (final visit). Alcohol will not be served during the in-patient period 


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from Day -2 check-in to Day 3. Marijuana and illicit drugs are prohibited during the course of 
the trial. 
 
Caffeinated beverages 
Caffeinated beverages or foods containing caffeine are prohibited within 48 hours before dosing 
to Day 3 of the trial. On other days, consumption is restricted to not more than 3 cups or 
equivalent per day. 
 
Exposure to loud noise 
Subjects have to avoid exposure to loud noise (e.g., construction sites, visiting concerts or dance 
events, fireworks, listening to music or playing videogames with headphones) from screening 
until Day 30 ± 4 days (final visit).  
 


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7 
STUDY SCHEDULE 
The Schedule of Study Procedures and Evaluations is included as Appendix A.  
7.1 
Recruitment 
The subject population will be recruited from the local population surrounding the CTU, utilizing 
the CTU subject database and IRB-approved advertisements and social media. IRB-approved, 
prescreening questionnaires will be used to determine if subjects meet study requirements before 
scheduling screening visits. 
7.2 
Screening Visit (Day -28 to Day -3) 
This will be an out-patient visit to complete the following within 28 days before study drug 
dosing:  
Note: A second screening visit may be scheduled to complete Otoacoustic screening tests to 
accommodate subjects’ personal schedule. 
• Obtain informed consent. 
• Assign a study ID number to subjects who consent to participate. 
• Record demographics including age, gender, race, and ethnicity. 
• Obtain contact information. 
• Obtain height and body weight, and calculate Body Mass Index (BMI; wt [kg] / ht [m2]) 
• Take VS (supine and resting for at least 5 min systolic and diastolic BP [SBP and DBP], 
heart rate [HR], respiratory rate [RR], and oral temperature [T]). 
• Obtain Medical History (MH).   
• Review history of Prior Medications.  
• Perform complete Physical Examination (PE). 
• Obtain blood and urine samples for screening clinical laboratory tests that include a 
calculation of Glomerular Filtration Rate (GFR) by the CKD-EPI equation. 
• Obtain blood samples for viral serology (HIV antibody, HBsAg, HCV antibody).  
• Obtain serum for β-HCG pregnancy test from all women.  
• Obtain serum for FSH level from only post-menopausal women.  
• Obtain urine sample for illicit drugs and drugs of abuse (urine drug screen), and cotinine. 
• Perform alcohol breathalyzer test.  
• Obtain a single 12-lead ECG with 10-sec rhythm strip. 
• Perform spirometry. 
• ENT Examination that includes: 
o Ear Otoscopy. 
o Valsalva test. 


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o Tympanometry. 
o Stapedial Reflexes. 
• Cochlear (Hearing) Examination that includes: 
o Ear Audiometry (pure tone audiometry). 
o Distortion Product Otoacoustic Emissions (DPOAEs). 
• Counsel on use of contraception (males and females), avoidance of pregnancy (women of 
childbearing potential), avoidance of prohibited medication, illicit drugs, alcohol, 
nicotine products, vigorous exercise, and exposure to loud noises). 
• Confirm eligibility. 
Subjects who meet the eligibility criteria of the otoacoustic assessments will be contacted by 
CTU personnel and asked to return on Day -2 to complete check-in assessments and be admitted 
to the CTU if they continue to meet eligibility criteria.  
Subjects who fail screening due to a medical condition or abnormal laboratory tests including 
pregnancy test and positive tests for HIV, HBV, and HCV will be informed of the findings and 
counseled to seek medical care for further evaluation and treatment. If subjects test positive for 
HIV antibody, HBsAg, and/or HCV antibody, they will be informed that test results may be 
reported to the local health authorities according to state or local law. 
Subjects who fail the otoacoustic screening will be informed of the findings and counseled to 
seek medical care by an otolaryngologist or audiologist for further evaluation and treatment. 
Subjects who fail screening assessments on either one of these visits will not be rescreened 
unless they had an inter-current, short-term medical illness. Subjects who meet eligibility criteria 
but could not be enrolled within permissible window may be rescreened. 
7.3 
Check-in (Day -2)    
Subjects meeting all inclusion and no exclusion criteria at Screening Visit will check into the 
CTU on Day -2 and the following procedures will be performed: 
• Review inclusion/exclusion criteria to confirm the subject remains eligible for admission. 
• Update MH. 
• Update Prior Medications.  
• Obtain VS.  
• Obtain body weight. 
• Perform complete PE.  
• Obtain blood and urine samples for clinical laboratory tests that include a calculation of 
GFR by the CKD-EPI equation (for determining eligibility). 


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• For all women, a urine β-HCG pregnancy test will be done, and negative results 
confirmed before dosing. 
• Obtain urine sample for illicit drugs and drugs of abuse (urine drug screen) and cotinine. 
• Perform alcohol breathalyzer test.   
• Obtain sample for serum Cystatin-C. 
• Obtain sample for urinary KIM-1.  
• Obtain a single 12-lead ECG with 10-sec rhythm strip. 
• Perform otoacoustic assessments. 
o Ear otoscopy. 
o Audiometry (pure tone audiometry including high frequencies). 
o DPOAEs. 
Note: Day -2 otoacoustic assessments may be completed on Day -4 or Day-3 as if needed.  
• Admit eligible subjects in the CTU. 
• Counsel to avoid rigorous physical activity, encourage good hydration, avoid caffeinated 
beverages and exposure to loud noises.  
7.4 
Baseline (Day -1) 
Admitted subjects will have the following assessments: 
• Review inclusion/exclusion criteria, before 12-lead ECGs, to confirm the subject remains 
eligible for enrollment. 
• Update MH. 
• Update Prior Medications.  
• Obtain VS. 
• Perform abbreviated PE.  
• Assign subject to Lung PK timepoint cohorts (T1-T4) and enroll in Advantage 
eClinical®. 
• Obtain baseline 12-lead standard ECG with 10-sec rhythm strip in triplicate at the same 
timepoints as the projected timepoints for ECGs to be recorded before and after dosing on 
Day 1 (within 30 min before and 1 h [± 5 min], 4 h [± 10 min], and 16 h [± 15 min] after 
start of dosing).     
• Counsel to avoid rigorous physical activity, encourage good hydration, avoid caffeinated 
beverages.  
 
 


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7.5 
Inpatient Treatment Period (Days 1 to 3)  
 
DAY 1: Administration of Study Drug – Bronchoscopy (Cohorts T1 – T4) 
Before Dosing:   
• Withhold breakfast or food at least 4 hours before scheduled post-dose bronchoscopy, 
according to subject assignment into one the Lung PK cohorts (T1-T5).  
• Allow access to water.  
• Review inclusion/exclusion criteria to confirm the subject remains eligible for dosing. 
• Update MH.  
• Update Prior Medications and review any new medication. 
• Obtain VS within 30 min before dosing (baseline). 
• Perform symptom-directed PE. 
• Obtain 12-lead standard ECG with 10-sec rhythm strip in triplicate within 30 min before 
dosing.  
• Insert IV catheter for blood collection into a hand or forearm vein. 
• Insert IV line for study drug infusion into a vein in the other forearm. 
• Obtain blood and urine samples for clinical laboratory tests that include a calculation of 
GFR by the CKD-EPI equation (for baseline). 
• Obtain sample for serum Cystatin-C (for baseline). 
• Obtain sample for urinary KIM-1 (for baseline). 
• Obtain pre-dose blood (plasma) PK sample for total apramycin within 30 min before 
starting administration of study drug. 
o Measure pre-dose urea concentration in an aliquot of the blood 9plasma) PK sample 
collected within 30 min before starting administration of study drug. 
Note 1: If 12-lead standard ECGs, VS assessments, and PK blood draws to be done at the 
same timepoint before or after dosing, these procedures will be performed in this 
order: 12-lead standard ECG, VS, and PK blood draw.  
Note 2: In cohort T3, obtain ECG, VS and PK draws at the 4 h timepoint before 
administration of sedation or anesthetic for BAL. 
Note 3: CTU staff may complete pre-dosing procedures in the bronchoscopy ward instead of 
the CTU to enable timely initiation of bronchoscopy at 0.5 h (± 5 min) after initiation 
of the dosing in subjects assigned to cohort T1.   
Dosing:  
• Administer a single 30 mg/kg body weight dose of apramycin as a single IV infusion 
with a syringe or infusion pump over 30 (± 5) minutes in a forearm vein, as described in 
Section 6.2. 
• Initiate TEAE assessments (excluding laboratory events) and SAE assessments. 
• Perform focused PE (for TEAE assessments). 


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• Initiate recording of ConMeds. 
• Withhold food until 2 h after end of dosing or 2 h after end of scheduled bronchoscopy 
according to subject assignment into one of the Lung PK cohorts (T1-T5).   
• Allow access to water. 
After Dosing: 
• Bronchoscopy with BAL once per subject at the following nominal timepoints after 
starting IMP infusion according to the assigned cohort: 0.5 h (± 5 min) (T1),  
2 h (± 5 min) (T2), 4 h (± 10 min) (T3) and 8 h (T4) (± 15 min).   
o Transfer subject to the UTMC bronchoscopy suite accompanied by CTU staff. 
 Note: Subjects in T1 may start receiving study drug in the bronchoscopy suite. 
o Collect scheduled safety information before and after the procedure in the 
bronchoscopy suite and during the recovery period.  
o Monitor subject before and after the procedure per orders by the bronchoscopy and 
anesthesia services.  
o Transfer subject to the CTU from the UTMC bronchoscopy suite accompanied by 
CTU staff. 
o Collect BAL samples for the measurement of total apramycin concentration and 
estimate of PK parameters once per subject at the assigned timepoint above, as 
described in Section 8.5, corresponding to the assigned cohort: 0.5 h (± 5 min) (T1), 
2 h (± 5 min) (T2), 4 h (± 10 min) (T3) and 8 h (T4) (± 15 min) after initiation of 
dosing. Details on processing the BAL samples and preparation of ELF and AM 
samples for apramycin concentration measurements and PK analysis will be provided 
in the MOP. 
Note 1: Collect plasma samples for total apramycin concentration within 5 min of the 
corresponding BAL procedure at 0.5 h (± 5 min),  
2 h (± 5 min), 4 h (± 10 min), or 8 h (± 15 min) after initiation of the infusion. If 
collection of BAL samples is delayed, plasma samples will be collected at the 
indicated nominal timepoint (and allowable windows), as described above, and an 
additional plasma PK will be collected within 5 min after the last BAL sample.   
Note 2: Measure urea concentration at the same timepoint as the nominal timepoint of 
plasma PK or in the plasma sample that is collected within 5 min after the last BAL 
sample if BAL is delayed. 
o Obtain the bronchoscopy Procedure Record. Data to be transcribed from procedure 
record onto study CRF will be described in the MOP.  
• Check infusion site (during and end-of-infusion, and 1 h (±5 min) and 16 h (± 15 min) 
after the infusion for infusion site reactions. Details will be provided in the MOP. 
• Obtain 12-lead standard ECG with 10-sec rhythm strip in triplicate at 1 h (±5 min), 4 h 
(±10 min) and 16 h (±15 min) after dosing. Record ECG before VS and PK draws and, at 
4 h, before administration of sedation or anesthetic for BAL in cohort T3. 


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• Obtain VS at 1 h (±5 min), 4 h (±10 min) and 16 h (± 15 min) after dosing:  
o Note: More frequent monitoring will be at the PI’s discretion based on subject’s 
clinical status after bronchoscopy. 
• Obtain blood (plasma) PK samples for total apramycin at 0.5 h (±5 min, immediately at 
the end of infusion), 1 h (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h (±15 min), and 16 h 
(±15 min) after initiating the IMP infusion.  
o Note 1: An aliquot of plasma PK collected at 0.5 h (±5 min; immediately after the end 
of infusion) after starting the IMP infusion will be used for the protein binding test 
(for measurement of free apramycin).  
o Note 2: An aliquot of plasma PK collected at 0.5 h (±5 min) after starting the IMP 
infusion will be used for the measurement of urea.  
• Monitor subject closely until fully recovered per bronchoscopy instructions. 
o Provide oxygenation as needed. 
• Perform TEAE assessments (excluding laboratory events) and SAE assessments. 
• Document ConMeds. 
• Do not provide solid food until at least 2 h after bronchoscopy and the subject is able to 
swallow. 
• Encourage water intake at least 2 L daily – Document time and volume consumed. 
• 
Counsel to avoid rigorous physical activity and avoid caffeinated beverages. 
 
DAY 2: Inpatient Follow-up – Bronchoscopy T5 
For a subject assigned to lung PK timepoint group T5:  
• Perform bronchoscopy with BAL on Day 2 at 24 h (± 1 h) after dosing as described for 
subjects in cohorts T1-T4 (see Section 7.5.1, Day 1, After Dosing). 
o Collect BAL samples for PK analysis once per subject at the assigned timepoint, 24 h 
(±1 h) (T5) after initiation of dosing, as described in Section 8.5, corresponding to the 
assigned cohort T5.  
Note 1: Collect plasma sample for total apramycin concentration within 5 min of the 
corresponding BAL procedure at 24 h (± 1 h) after initiation of the infusion. If 
collection of BAL samples is delayed, plasma sample will be collected at the 
indicated nominal timepoint (and allowable windows), and an additional plasma PK 
will be collected within 5 min after the last BAL sample. (See Section 8.5.1) 
Note 2: Urea will be measured in plasma collected at the same nominal timepoint as 
the plasma PK sample or in the plasma PK sample that is collected within 5 min after 
the last BAL sample. 
• Monitor subject closely until fully recovered per bronchoscopy instructions. 
o Provide oxygenation as needed. 
For all subjects: 
• Obtain single 12-lead standard ECG with 10-sec rhythm strip at 24 h (±1 h) and 36 h  
(±1 h) after dosing. 
• Obtain VS at 24 h (±1 h) and 36 h (±1 h) after dosing. 


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• Obtain blood (plasma) PK samples for total apramycin at 24 h (±1 h) and 36 h (± 1 h) 
after dosing. 
o An aliquot of plasma PK collected at 36 h (±1 h) after starting the IMP infusion will 
be used in the protein binding test (for the measurement of free apramycin). 
• Obtain blood and urine samples for safety clinical laboratory tests that include a 
calculation of GFR by the CKD-EPI equation at 24 h (±1 h) after dosing. 
• Check on infusion site at 24 h (±1 h) and 36 h (±1 h) after dosing. 
• Perform TEAE and SAE assessments. 
• Perform symptom-directed (focused) PE for evaluation of TEAEs as needed. 
• Document ConMeds. 
• Encourage water intake at least 2 L daily – Document time and volume consumed. 
• Counsel to avoid rigorous physical activity and avoid caffeinated beverages. 
 
DAY 3: Inpatient Follow-up 
• Obtain 12-lead standard ECG with 10-sec rhythm strip at 48 h (±1 h) and 60 h (±1 h) 
after dosing. 
• Obtain VS at 48 h (±1 h) and 60 h (±1 h) after dosing. 
• Obtain blood (plasma) PK samples for total apramycin at 48 h (±1 h) and 60 h (±1 h) 
after dosing. 
• Obtain blood and urine samples for clinical laboratory tests that include a calculation of 
GFR by the CKD-EPI equation at 48 h (±1 h) after dosing. 
• Obtain blood sample for Cystatin C and urine sample for KIM-1 (exploratory biomarkers 
of renal injury) at 48 h (±1h) after dosing. 
• Perform otoacoustic assessments.  
o Ear otoscopy. 
o Audiometry (pure tone audiometry including high frequencies). 
o DPOAEs. 
Note: Day -2 otoacoustic assessments may be completed on Day 3 (+2 days) if needed.  
• Check on infusion site at 48 h (±1 h) and 60 h (±1 h).  
• Perform TEAE and SAE assessments.  
• Document ConMeds. 
• Perform complete PE.  
• Counsel on use of contraception (males and females), avoidance of pregnancy (women of 
childbearing potential), avoidance of prohibited medication, illicit drugs, alcohol, 
nicotine products, vigorous exercise, and exposure to loud noises. 
• Instruct on the next scheduled visit. 
• Discharge subject from the CTU after review of clinical laboratory tests, ECGs and other 
assessments by PI or authorized clinician and audiologist. 


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Note 1: If the time of discharge is late in the evening, due to delays in starting procedures on 
Day 1 or personal reasons (such as distance from residence), the subject may stay in the CTU 
overnight and be discharged the following morning. 
 
Note 2: If otoacoustic assessments could not be performed on Day 3, the subject will be 
discharged from the CTU on Day 3 after all other assessments are completed. Otoacoustic 
assessments will be completed up to Day 3 (+2 days) as outpatient. 
7.6 
Outpatient Follow-up Period (Days 14 and 30)  
Subjects will return to the CTU to have the following assessments: 
 
DAY 14 (± 3 days) 
• Obtain 12-lead standard ECG with 10-sec rhythm strip. 
• Obtain VS. 
• Obtain blood and urine samples for clinical laboratory tests that include a calculation of 
GFR by the CKD-EPI equation.  
• Obtain blood sample for Cystatin C and urine sample for KIM-1 (exploratory biomarkers 
of renal injury). 
• Perform otoacoustic assessments. 
o Ear otoscopy. 
o Audiometry (pure tone audiometry including high frequencies). 
o DPOAEs. 
• Check on infusion site.  
• Perform TEAE and SAE assessments. 
• Document ConMeds. 
• Perform a complete PE. 
• Counsel on use of contraception (males and females), avoidance of pregnancy (women of 
childbearing potential), avoidance of sperm donation, avoidance of prohibited 
medication, illicit drugs, alcohol, nicotine products, vigorous exercise, and exposure to 
loud noises. 
• Instruct on the next scheduled visit. 
 
Day 30 (± 4 days) – FINAL VISIT 
• Perform TEAE and SAE assessments. 
• Document ConMeds. 
• Obtain blood sample for Cystatin C and urine sample for KIM-1 (exploratory biomarkers 
of renal injury). 
• Perform symptom-directed (focused PE) for evaluation of TEAEs as needed. 


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• Perform otoacoustic assessments. 
o Ear otoscopy. 
o Audiometry (pure tone audiometry including high frequencies). 
o DPOAEs.  
• Discharge subject from the study.  
• Follow up on-going TEAEs including otoacoustic abnormalities according to PI 
instruction to resolution or until stable. 
7.7 
Early Termination (if needed) 
• Obtain VS. 
• Obtain weight. 
• Check infusion site. 
• Perform complete PE.  
• Perform TEAE and SAE assessments. 
• Update ConMeds. 
• Obtain single 12-lead standard ECG with 10-sec rhythm strip. 
• Obtain blood and urine for clinical laboratory tests that include a calculation of 
Glomerular Filtration Rate (GFR) by the CKD-EPI equation. 
• Obtain blood sample for Cystatin C and urine sample for KIM-1 (exploratory biomarkers 
of renal injury).  
• Obtain blood PK sample for total apramycin if ET occurs within 24 h of dosing. 
• Perform otoacoustic assessments (ear otoscopy, audiometry [pure tone audiometry 
including high frequencies], and  DPOAEs). 
• Counsel on use of contraception (males and females), avoidance of pregnancy (women of 
childbearing potential), avoidance of sperm donation. 
7.8 
Unscheduled Visit (if needed) 
A subject may return to the clinic for an unscheduled visit at any time after discharge on Day 3. 
The following activities at a minimum will be performed:  
• Perform TEAE and SAE assessments. 
• Update ConMeds. 
• Obtain VS. 
• Perform symptom-directed (focused) PE, if applicable. 
• Obtain blood and/or urine for clinical safety laboratory tests that include a calculation of 
GFR by the CKD-EPI equation, if applicable. 


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• Obtain single 12-lead standard ECG with 10-sec rhythm strip as needed for the 
evaluation of a TEAE, if applicable. 
• Perform otoacoustic assessments, if applicable (Ear otoscopy, Audiometry [pure tone 
audiometry including high frequencies], and DPOAEs). 


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8 
STUDY PROCEDURES/EVALUATIONS 
8.1 
Clinical Procedures/Evaluations 
 
Informed Consent 
The informed consent form (ICF) will be approved by the reviewing IRB/EC and executed 
before performing any study-related activities. 
Informed consent will be obtained for all subjects participating in the trial before performing any 
screening assessments. Subjects may withdraw consent at any time. Participation in the trial may 
be terminated at any time without the subject’s consent as determined by the Investigator. 
 
Demographics 
Demographic information (date of birth, gender, ethnicity and race) will be recorded on the 
subject’s source documents and eCRF at Screening Visit. Name, address, phone number, and 
emergency contact information will be documented in the source documents only. 
 
Inclusion/Exclusion Criteria 
Eligibility screening of healthy subjects will be completed within 28 days before study drug 
dosing and will be documented on the subject’s source documents and eCRF. Review of 
inclusion and exclusion criteria will be performed on Day -2 to determine eligibility for 
admission, on Day -1, before 12-lead ECG, to determine eligibility for enrollment, and on Day 1 
pre-dose to confirm the subject remains eligible for dosing. 
Screening failures and the reason for failure to meet the study eligibility requirements will be 
documented in the source documents and entered into the study database. 
 
Medical History 
For subjects enrolled in the trial, the medical history (MH) will be obtained by direct interview 
of the subject and recorded on the subject’s source document and eCRF. The MH will capture 
the subject’s current disease processes, past disease processes, history of hospitalization, history 
of surgery, allergies, and prior medications (see Section 6.6). Subjects will be queried regarding 
a history of significant medical disorders of the head, eyes, ears, nose, throat (HEENT), mouth, 
skin, and the cardiovascular, gastrointestinal, renal, urological, nervous, hematological, 
lymphatic, endocrine, musculoskeletal, and genital/reproductive systems. A history of hearing 
abnormalities or hearing loss, especially following administration of aminoglycoside antibiotics 
or other medications, kidney disorders, cardiovascular disorders, including MI and angina, 
cardiac arrhythmias including Long QT Syndrome (LQTS), syncope due to cardiac arrhythmias 
or unexplained, psychiatric illness, and substance abuse will be specifically solicited. 


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Occupational history of exposure to loud noise will be obtained. Family history of hearing loss, 
especially on the maternal side, renal disorders, and sudden death will be obtained. The MH will 
be obtained at Screening Visit and updated upon admission to the CTU at check-in (Day -2), on 
Day -1, and before dosing on Day 1. After start of study drug dosing, any worsening of pre-
dosing MH or new symptoms will be evaluated and reported as TEAEs. 
 
Physical Examination 
A complete PE – except genital, breast, and rectal exams – will be performed at the Screening 
Visit, check-in visit (Day -2), Day 3, and Day 14 (± 3 days) after dosing, or ET Visit, and will 
assess general appearance, HEENT, heart, lungs, abdomen, skin, musculoskeletal system, and 
lymph nodes, and a standard neurological exam. 
An abbreviated PE will be performed at Day -1. An abbreviated PE differs from a complete PE 
in that the abdomen and neurological system are not evaluated.  
A symptom-directed (focused) PE may be performed before dosing on Day 1 and at any time 
after dosing on Day 1 to the last visit (Day 30 ± 4 days) for evaluation of TEAEs. 
Height and weight will be measured, and BMI calculated, at the Screening Visit. Weight only 
will be measured on Day -2, and Day 14 (±3 days), or ET.  
Refer to the protocol-specific MOP for further details. The findings of each examination will be 
recorded on the subject’s source documents and eCRF. Any new findings on examination or 
worsening of existing conditions after dosing are to be reported as TEAEs.  
 
Vital Signs 
VS include resting (measured after supine for at least 5 min) systolic blood pressure (SBP) and 
diastolic BP (DBP), heart rate (HR), respiratory rate (RR) and oral temperature (T). VS will be 
measured at the Screening Visit, on Day -2, on Day -1, on Day 1 (within 0.5 h [30 min] before 
dosing and at 0.5 h ±5 min, 1 h ±5 min, 4 h ±10 min, and 16 h ±15 min after dosing); on Day 2 
(at 24 h ±1 h and 36 h ±1 h after dosing); on Day 3 (at 48 h ±1 h and 60 h ± 1 h after dosing); 
and on Day 14 (± 3 days), or ET. Normal references ranges are shown in Appendix B, Table 2, 
and exceptions for enrollment in Section 5.1, Inclusion Criterion #2. 
VS that are considered aberrant due to an error in measurement may be repeated. At Screening 
and after dosing, an abnormal VS measurement may be repeated up to two more times at rest, 
within 5 min of each other. If the second measurement is abnormal, it will be reported at the 
highest grade of the two measurements and the subject will be excluded (if at Screening Visit) or 
the event reported as a TEAE and graded for severity per Appendix B, Table 2 (if after dosing). 
If the second measurement is normal, a third measurement will be taken at least after 5 min at 
rest. If the third measurement is still normal, the subject is eligible (if at Screening Visit) or there 
is no TEAE (if after dosing); if it is abnormal, the subject will be excluded (if at Screening Visit) 


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or a TEAE will be reported at the highest assessed grade (between first and third measurements) 
and graded for severity per Appendix B, Table 2 (if after dosing).  
 
12-lead Standard Electrocardiogram (ECG) 
Single 12-lead standard ECG and 10-sec rhythm strip will be obtained at the Screening Visit, on 
Day -2, at 24 ±1 h and 36 ± 1 h (Day 2) and 48 h ±1 h and 60 h ±1 h (Day 3) after dosing, and on 
Day 14 (± 3 days), or ET.  
Triplicate 12-lead standard ECGs will be obtained over a period of 5 minutes on Day -1 (at the 
same 4 timepoints as projected to be taken on Day 1), and on Day 1 pre-dose (within 30 min) and 
at 1 h ±5 min, 4 h ±10 min and 16 h ± 15 min after dosing.  
ECGs will be performed after the subject rests quietly in a supine position for at least 5 min. The 
ECGs will be reviewed by the PI or a designated clinician (listed on FDA Form 1572). ECGs 
will be analyzed for PR, QRS and QT intervals, ventricular rate (based on RR intervals), and for 
morphological abnormalities. The QT interval will be corrected by the Fridericia (QTcF) 
formula, recorded and analyzed. To be eligible for participation, the QTcF interval must be 
within protocol reference range criteria at Screening and Day -2 and there must be no clinically 
significant ECG abnormalities after enrollment on Day -1 and before dosing on Day 1. If the site 
PI identifies clinically significant changes in the ECGs after enrollment in Day -1 and before 
dosing on Day 1, the subject would not be eligible to receive study drug and should be 
withdrawn and replaced.  ECG PR and QTcF intervals after dosing will be reported as TEAEs if 
they meet toxicity grading criteria (See Appendix B, Table 4). If a question regarding ECG 
interpretation arises, the study investigators will have the ECG reviewed by a cardiologist. 
 
Spirometry 
Spirometry will be performed to evaluate FEV1 and FEV1/FVC at Screening. A subject will be 
eligible if predicted FEV1 is ≥ 80% and FEV1/FVC is > 70 %. 
8.2 
Laboratory Evaluations 
Venipuncture schedule and blood volumes are shown in Appendix A and Appendix C, 
respectively.  
 
Clinical Laboratory Evaluations (Hematology, Coagulation, Chemistry, and 
Urinalysis). 
Blood and urine samples for clinical laboratory tests will be collected at the Screening Visit and 
on Day -2 to determine eligibility, and on Day 1, pre-dose, to determine (Baseline). Clinical 
laboratory tests will also be collected on Day 2 (24 h ± 1 h after dosing), Day 3 (48 h ± 1 h after 
dosing), and on Day 14 (± 3 days) or ET. Subjects must be fasting at least for 6 h before any 
blood draw for clinical laboratory assessments. These tests will include: 


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• HEM: Hgb, Hct, RBC, platelet count, and WBC count with absolute differential count. 
• COAG: INR with prothrombin time, alpha partial thromboplastin time. 
• CHEM: electrolytes (sodium, potassium, chloride, total carbon dioxide [CO2]), calcium, 
magnesium, creatinine, with estimation of GFR by the CKD-EPI equation, blood urea 
nitrogen (BUN), glucose (fasting), total bilirubin, direct bilirubin, alanine 
aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AP), 
total protein, albumin, TSH, free T4, and free T3. 
o The CKD-EPI equation, expressed as a single equation, is GFR = 141 × min(Scr/κ, 
1)α × max(Scr/κ, 1)-1.209 × 0.993Age × 1.018 [if female] _ 1.159 [if black], where 
Scr is serum creatinine, κ is 0.7 for females and 0.9 for males, α is  
-0.329 for females and -0.411 for males, min indicates the minimum of Scr/κ or 1, 
and max indicates the maximum of Scr/κ or 1 (Levey et al 2009). 
• UA: Routine dipstick testing of clean-catch urine for blood, protein, and glucose  
o If urine dipstick is abnormal, urine microscopy will be performed. 
Clinical laboratory tests at the Screening Visit and Day -2 should be in the normal reference 
range with exceptions. (See Section 5.2, Inclusion Criteria #2 and Appendix B, Table 3). 
Laboratory values that are outside the range of eligibility but are thought to be due to an acute 
condition or due to collection or laboratory error may be repeated once, preferably within 24 h or 
as soon as subject is available.   
Laboratory values will be transferred to Advantage eClinical®.  
Abnormal safety laboratory values after dosing will be graded for severity per Appendix B, 
Table 3 and Table 4 . Abnormal values within the allowable Grade 1 range per Appendix B 
noted at screening or baseline will only be considered TEAEs if they deteriorate after study drug 
dosing to Grade 2 or higher. 
 
Viral Serology Testing 
Serological testing for HIV antibody, HBsAg, and HCV antibody will be performed at Screening 
Visit. These tests must be negative for study eligibility. In cases where a false-positive result is 
suspected, confirmatory testing (e.g., polymerase chain reaction) may be performed. 
 
Pregnancy Testing 
In all women, a serum β-HCG pregnancy test will be done at the Screening Visit. A urine 
pregnancy test will be done upon check in on Day -2 and results must be negative for dosing 
with study drug.   


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Serum FSH Testing 
A serum FSH level for confirmation of post-menopausal status in female subjects will be 
measured at the initial Screening Visit only.   
 
Urine Toxicology Screening 
A urine toxicology screen will be performed at Screening Visit and on Day -2 to detect the 
presence of amphetamines, cocaine, barbiturates, benzodiazepines, opiate, 
methylenedioxymethamphetamine (MDMA), methadone, marijuana (THC), methamphetamine, 
tricyclic antidepressants (TCAs), and phencyclidine. Results must be negative for study 
eligibility. Urine creatinine will be measured as part of the profile to assess quality of collected 
sample.  
 
Alcohol Breathalyzer Test 
A breathalyzer test for alcohol will be performed at Screening and Check-in (Day -2). Results 
must be negative for study eligibility. 
 
Urine Cotinine Testing  
Detection of urine cotinine, for recent tobacco products consumption, will be performed at the 
Screening Visit, and on Day -2. A positive urine cotinine at screening is allowed but results must 
be negative on Day -2 for study eligibility for enrollment. 
 
Exploratory Biomarkers for Renal Injury 
The following tests will be performed on indicated days for exploratory purposes, not to 
determine eligibility:  
 
Serum Cystatin C 
The test will be performed in enrolled subjects on Day -2, pre-dose on Day 1 (Baseline), Day 3 
(at 48 h ± 1 h after start of dosing), Day 14 (± 3 days), and Day 30 (± 4 days), or ET.  
 
Urinary KIM-1  
The test will be performed in enrolled subjects on Day -2, pre-dose on Day 1 (Baseline), Day 3 
(at 48 h ±1 h after start of dosing), Day 14 (± 3 days), and Day 30 (± 4 days), or ET.  
8.3 
Bronchoscopy with Bronchoalveolar Lavage 
Bronchoscopy with bronchoalveolar lavage is performed to obtain fluid samples from a lung 
segment following dosing with apramycin to measure the accumulation of the drug in the lung. 
The procedure will be performed in the Pulmonology Department of the UTMC, which is 
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pulmonologists and support staff. The procedure will be done in eligible and enrolled subjects 
who were assigned to cohorts T1-T5, at the following nominal timepoints: 0.5 h ±5 min (T1), 2h 
±5 min (T2), 4 h ± 10 min (T3), 8 h ± 15 min (T4) and 24 h (nominal) ±1 h (T5) after the start of 
apramycin infusion. (The exact timing of T5 will be determined after review of BAL apramycin 
concentrations in cohorts T1 to T4.) Subjects in each cohort will have only one procedure done 
during the study.  
Subjects, who had previously been informed of the procedure and provided informed consent, 
will be taken to the bronchoscopy suites by CTU staff, will review the procedure with the 
operator and confirm understanding prior to the procedure. The subject will be monitored during 
the procedure by VS, pulse oximetry and will receive oxygen as needed. Sedation or light 
anesthesia will be induced by appropriate intravenous medications. The procedure will be 
performed according to standard protocol and a study-specific protocol (SSP) for the collection 
of BAL samples for PK analysis. BAL samples will be stored on ice and then transferred into test 
tubes to the CTU research lab for measurement of total and differential cell counts and for 
centrifugation and processing into BAL supernatant (containing ELF) and BAL pellet 
(containing AM) samples. After the procedure, the subject will be observed in the recovery room 
for side effects and will be released by the pulmonologist to the care of the CTU staff when 
stable. The subject will continue to be monitored closely until discharge from the CTU. 
Procedure records will be obtained by the CTU staff.   
8.4 
Ear Examination and Otoacoustic Testing 
Ear exams and otoacoustic tests will be performed by licensed and experienced audiologists at 
the Audiology section of the ENT department of the UTMC. The following tests will be 
performed at the indicated study visits. These tests may be performed at the screening visit, or at 
a second screening visit due to time restrictions on the initial visit. 
• 
Ear otoscopy, Valsalva test, Tympanometry and Stapedial Reflexes (Screening visit). 
• 
Ear otoscopy only (Day -2 [or Day -4 or Day -3 if needed], Day 3 (+2 days), Day 14 
(± 3 days) and Day 30 (± 4 days), or ET).  
• 
Audiometry. 
o Pure tone audiometry at standard frequencies, 0.5 to 8 kHz (Screening visit).  
o Pure tone audiometry at standard (0.5 to 8 kHz) and higher frequencies (Day -2 [or 
Day -4 or Day -3 if needed], Day 3 (+2 days), Day 14 (± 3 days), and Day 30 
(± 4 days), or ET. 
• 
Distortion product otoacoustic emissions (DPOAEs) at Screening Visit, and on Day -2 
[or Day -4 or Day -3 if needed], Day 3 (+2 days), Day 14 (± 3 days), and Day 30 
(± 4 days), or ET. 
Note 1: On Day 3, ear otoscopy, audiometry and DPOAEs can be done at any time. 


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Note 2: If otoacoustic assessments could not be performed on Day 3, the subject will be 
discharged from the CTU on Day 3 after all other assessments are completed. Otoacoustic 
assessments will be completed up to Day 3 (+2 days) as outpatient. 
Audiology records will be obtained by the CTU staff and results will be recorded as described in 
the MOP.  
8.5 
Bioanalytical Assays for Pharmacokinetics (PK)  
The following LC-MS/MS methods will be validated before subject enrollment starts for 
determination of concentration of apramycin in plasma and bronchoalveolar lavage (BAL) 
samples: Total and Free apramycin in plasma, total apramycin in BAL supernatant (BAL SUP, 
containing ELF), and total apramycin in BAL pellet (BAL AM). In addition, a validated LC-
MS/MS method will be used for the measurement of urea in plasma and BAL SUP.  
 
Assay for Total Apramycin in Plasma 
A single dose of apramycin will be administered by 30 (± 5) min IV infusion.  Blood (plasma) 
samples for assay of total apramycin concentration and estimate of PK parameters will be 
collected at the following study days and timepoints: On Day 1 within 30 min before dosing, and 
0.5 h (±5 min, immediately at end of infusion), 1 h (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h 
(±15 min), and 16 h (±15 min) after dosing; on Day 2 at 24 h (± 1 h) and 36 h (± 1 h) after 
dosing; and on Day 3 at 48 h (± 1 h) and 60 h (± 1 h) after dosing, or ET if occurs within 24 h of 
dosing.  
Plasma samples for total apramycin concentration should be collected within 5 min of the 
corresponding BAL procedure at 0.5 h (± 5 min, immediately after the end of infusion), 2 h  
(± 5 min), 4 h (± 10 min), 8 h (± 15 min) and 24 h (± 1 h) after the start of the infusion. If BAL is 
delayed, blood (plasma) for apramycin will be collected at the nominal timepoint (including 
windows) and a second sample will be collected within 5 min of the BAL sample.  
Blood samples will be obtained promptly after each corresponding 12-lead standard ECG 
timepoint and VS assessment. 
Sample collections will be scheduled for the nominal time point and actual collection times 
recorded in source documents. If bronchoscopy/BAL is delayed, an additional sample for plasma 
apramycin assay should be collected within 5 min after collection of the last BAL sample, to be 
used for the calculation of Lung PK / Plasma PK ratios.  
 
Protein Binding Test for Apramycin in Plasma 
Protein binding of apramycin in plasma will be measured at the following timepoints: 0.5 h  
(± 5 min; immediately at the end of infusion), and 36 h (± 1 h) after the start of the infusion. The 


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assay will be performed in plasma PK aliquots collected at the matching time points as described 
in Section 8.5.1. A separate blood collection is not required. 
 
Assays for Total Apramycin in Bronchoalveolar Lavage Components (ELF and 
AM)  
During bronchoscopy with bronchoalveolar lavage (BAL), samples will be obtained at the 
following timepoints overall during the study: 0.5 h (± 5 min), 2 h (± 5 min), 4 h (± 10 min), 8 h 
(±15) min and 24 h (± 1 h) after the start of the infusion. Each subject will have bronchoscopy 
with BAL at one timepoint only. At each time point, BAL samples will be collected from 4 
subjects assigned to one of the cohorts T1-T5. Each BAL collection will be centrifuged for the 
separation of the supernatant (BAL SUP containing ELF) and the pellet (BAL AM) components, 
and total apramycin will be measured in BAL SUP and BAL AM. Due to dilution during BAL, 
the concentration of apramycin in ELF will be corrected using the urea method (Tenero 2013; 
Gottfried 2017; Rizk 2018). Formulae for the calculations will be provided in the SAP. 
 
Plasma and BAL Urea assay  
Urea will be assayed in plasma samples collected at baseline (as part of the total apramycin 
plasma PK) and at 0.5 h (± 5 min, immediately after the end of infusion), 2 h (± 5 min), 4 h 
(± 10 min), 8 h (± 15 min) and 24 h (± 1 h) after the start of the infusion), and in BAL SUP 
samples corresponding to plasma PK timepoints. If BAL is delayed and a second plasma sample 
is collected within 5 min of the last BAL sample, plasma urea will be measured only in the 
second sample.  
 
Specimen Preparation, Handling, and Shipping 
 
Instructions for Specimens Preparation, Handling, and Storage 
Details regarding the specimen preparation, handling, and storage of bioanalytical samples for 
the measurement of plasma total and free apramycin, total apramycin in BAL SUP (containing 
ELF) and BAL AM, plasma urea, and urea in BAL SUP (ELF) will be described in the protocol-
specific MOP.  
Blood samples and BAL samples left after all routine clinical laboratory testing and 
pharmacokinetic assays are completed will not be stored indefinitely or used for purposes other 
than those described but will be disposed of per DMID (sponsor) guidance. No genetic testing 
will be done on collected blood samples and BAL samples.  
 
Specimen Shipment 
Specimen shipment will occur at intervals during the trial following all applicable International 
Air Transport Association (IATA) requirements and according to the specifics for storage 


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temperature and documentation as detailed in the central clinical laboratory manual and protocol-
specific MOP, as appropriate. 
All specimens for clinical screening and safety laboratory evaluations will be transported from 
the CTU to the local clinical laboratory. 
Plasma and Bronchoalveolar BAL SUP (containing ELF) and BAL AM samples for 
bioanalytical assays will be shipped from the CTU to DMID-CMS at: 
Fisher BioServices 
c/o DMID Clinical Materials Services (CMS) 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 
Phone: 240-477-1350  
Fax: 240-477-1360 
Email: DMID.CMS@thermofisher.com   
Plasma and Bronchoalveolar BAL SUP (containing ELF) and AM samples will then be provided 
by DMID-CMS to the bioanalytical lab, KCAS Inc., at:  
Test Materials Management (TMM) 
KCAS Bioanalytical and Biomarker Services  
10830 S. Clay Blair Boulevard 
Olathe, KS, 66061 
Telephone: 913-248-3006 
Fax: 913-248-3106 
Email: TMM@kcasbio.com  
Further information will be provided in the study-specific MOP.   
 
Long-term Storage of Plasma and BAL (ELF and AM) PK Samples 
Any residual plasma and BAL samples (BAL SUP [ELF] and BAL AM) collected for the 
measurement of total apramycin concentration may be stored at the DMID CMS long-term and 
used for future exploratory research if collected from subjects who consented to its use for this 
purpose. (See Section 14.8)  


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9 
ASSESSMENT OF SAFETY 
Regulatory requirements including FDA regulations and ICH Guidelines for GCP set forth safety 
monitoring and reporting responsibilities of sponsors and investigators to ensure the safety and 
protection of human subjects participating in clinical trials. 
Responsibilities: 
Investigators participating in this clinical trial are responsible for and will: 
• Evaluate subject safety including assessment of treatment-emergent adverse events 
(TEAEs) for seriousness, severity, and causality (relatedness to study drug). 
• Notify DMID of treatment-emergent serious AEs (SAEs) within 24 h of site awareness. 
• Provide detailed written reports, including necessary documentation requested by DMID 
or IRB/EC, promptly following immediate initial reports. 
• Inform the IRB/EC of SAEs and TEAEs as required by applicable regulatory 
requirements. 
9.1 
Specification of Safety Parameters 
Safety will be assessed by the timing, frequency, causality, and severity of: 
1. Treatment-emergent AEs and SAEs occurring from time of dosing through Final Visit 
(Day 30 ± 4 days), or ET. 
2. Clinical laboratory TEAEs occurring from time of first dose through Day 14 (± 3 days),  
or ET. 
3. 12-lead standard ECGs performed in the 24-h period after dosing, on Day 3, and Day 14 
 (± 3 days), or ET. 
4. Audiology (Ear examination and otoacoustic testing) TEAEs at Day 3 (+ 2 days), Day 14 
(± 3 days) and Day 30 (± 4 days), or ET.  
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
 
Adverse Events 
Definitions 
ICH E6 defines an AE as any untoward medical occurrence in a subject or clinical investigation 
subject who was administered a pharmaceutical product, regardless of its causal relationship to 
the product. An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease temporally associated with use of the product, 


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and are described as treatment-emergent AEs (TEAEs). The occurrence of a TEAE may come to 
the attention of study personnel during study visits and interviews, or by a subject presenting for 
medical care. 
The FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether it is considered drug-related or not. 
Any systemic medical condition, VS and ECG measurement, and clinical safety lab test value 
that is present on Day 1 prior to dosing will be considered a baseline finding for the purpose of 
data analysis. For ear and otoacoustic testing, baseline comprises assessments performed on  
Day -2. However, if the condition increases in severity or frequency after IMP administration at 
any time during the trial, it will be recorded as a TEAE. 
Any medical condition that is reported after screening but before study drug administration will 
be evaluated and reported as MH update.  
All TEAEs will be graded for severity according to Appendix B, Table 2, Table 3, and Table 4 
and for relationship to the study drug.  
 
Severity of Events 
Intensity of TEAEs will be graded as follows, unless otherwise specified in Appendix B: 
Mild: Require minimal or no treatment; do not interfere with the subject’s daily activities. 
Moderate: Result in a low level of inconvenience or concern with therapeutic measures; may 
cause some interference with functioning. 
Severe: Interrupt a subject’s usual daily activity and may require systemic drug therapy or other 
treatment; are usually incapacitating. 
 
Relationship to Study Products 
TEAEs and treatment-emergent SAEs will be assessed by the investigator to determine 
relationship to the study drug, using the following two terms. In a clinical trial, the study drug 
must always be suspect.   
• Related: There is a reasonable possibility that the study drug caused the treatment-
emergent AE/SAE. 
• Not Related: There is not a reasonable possibility that the study drug caused the 
treatment-emergent AE/SAE. 
The investigator will provide an assessment of association or relationship of each treatment-
emergent AE/SAE to the study drug based on: 
• Temporal relationship of the TEAE/SAE to study drug dosing. 


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• Whether an alternative etiology has been identified. 
• Biological plausibility. 
• Existing therapy and/or ConMeds. 
 
Reporting Adverse Events 
TEAEs will be captured on the appropriate subject’s source document and eCRF. Information 
collected for TEAEs includes event description, time of onset, investigator assessment of 
severity and relationship to the study drug, date of resolution of the event, seriousness, and 
outcome. 
All TEAEs will be documented from the time of study drug dosing through the time of last 
assessment (e.g., Day 14 ± 3 days for clinical safety labs and ECGs) or Final Visit (Day 30 ± 
4 days), or ET. All TEAEs including abnormal safety laboratory test results will be followed to 
resolution or until considered stable in the clinical judgment of the study investigator. Evaluation 
of TEAEs may require unscheduled visits and clinical and laboratory investigations, according to 
the clinical judgment of the Site PI and study physicians. 
 
Serious Adverse Events 
An SAE is any treatment-emergent AE that meets at least one of the following criteria: 
• Death. 
• Life-threatening AE*. 
• Inpatient hospitalization or prolongation of existing hospitalization. 
• Persistent or significant disability or incapacity, or substantial disruption of the ability to 
conduct normal life function. 
• Congenital anomaly/birth defect. 
• Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered an SAE when, based upon appropriate medical 
judgment, they may jeopardize the subject and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition. Examples of such 
medical events include allergic bronchospasm requiring intensive treatment in an 
emergency room or at home, blood dyscrasias or convulsions that do not result in 
inpatient hospitalization, or the development of drug dependency or drug abuse. 
*A TEAE is considered “life-threatening” if, in the view of either the investigator or DMID, its 
occurrence places the subject at immediate risk of death. It does not include a TEAE that, had it 
occurred in a more severe form, might have caused death. 
SAEs will be: 
• Assessed for severity and relationship to study product and alternate etiology by an 
authorized study physician (listed on FDA Form 1572). 


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• Recorded on the appropriate SAE data collection form and eCRF. 
• Followed through resolution. 
• Reviewed and evaluated by the SMC (periodic review unless related), DMID, and the 
IRB. 
 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings 
A licensed study clinician (listed on FDA Form 1572) will determine seriousness, severity, and 
causality of abnormal laboratory values; provide a medical evaluation of TEAEs; and classify 
TEAEs based upon medical judgment. 
Abnormal laboratory values or clinical findings for all enrolled subjects after dosing will be 
assessed using the toxicity scales in Appendix B. Out of range values and findings noted at 
screening or baseline, but allowable for enrollment and dosing if within Grade 1, will only be 
considered TEAEs if their severity increases to Grade 2 or higher. For abnormalities noted from 
the time of study drug dosing, any Grade 1 or higher laboratory abnormality listed on the toxicity 
Table 3 in Appendix B will be entered in the database as a TEAE. Safety laboratory results that 
are reported as TEAEs will be evaluated by the CTU clinician as clinically significant (CS) or 
not clinically significant (NCS).  Abnormal laboratory values, performed as part of HEM, 
COAG, CHEM, or UA but not listed in this toxicity table will be evaluated by the study 
clinicians, recorded in the source document and, if clinically significant, considered TEAEs and 
graded according to the criteria in Section 9.2.1. 
Protocol-specific laboratory normal range values are included in the study-specific MOP and 
toxicity grades for evaluation of TEAEs are included in Appendix B, Table 3. 
Gross blood in urine that is confirmed due to menses is not a TEAE (but is for all other reasons). 
9.3 
Reporting Procedures 
 
Serious Adverse Events 
SAEs will be followed until resolution even if this extends beyond the study-reporting period.  
Resolution of an SAE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic. 
 
All SAEs will be: 
• Recorded on the appropriate SAE report form and sent to DMID Pharmacovigilance 
Group (PVG). 
• Entered into the appropriate subject source document and eCRF in Advantage eClinical® 
• Reported to the IRB. 


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• Reviewed and followed to resolution or stability by an authorized study physician (listed 
on FDA Form 1572). 
• Collected on each subject until Day 30 ± 4 days (Final Visit), or ET Visit. 
Any TEAE that meets a protocol-defined serious criterion will be submitted immediately (within 
24 h of site awareness) on an SAE report form to DMID PVG: 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support (CROMS) 
6500 Rock Spring Drive, Suite 650 
Bethesda, MD 20817, USA 
SAE Hot Line: 1-800-537-9979 (US) or 1-301-897-1709 (outside US) 
SAE FAX Number: 1-800-275-7619 (US) or 1-301-897-1710 (outside US) 
SAE Email Address: PVG@dmidcroms.com  
In addition to the SAE report form, selected SAE data fields will also be entered into Advantage 
eClinical®. Please see the protocol-specific MOP for details regarding this procedure. 
Other supporting documentation of the SAE may be requested by DMID PVG and will be 
provided as soon as possible. 
The DMID Medical Monitor (MM) and DMID CPM will be notified of the SAE by the DMID 
PVG. The DMID MM will review and assess the SAE for regulatory reporting and potential 
impact on subject safety and protocol conduct. 
At any time after completion of the trial, if the Site PI or appropriate sub-investigator becomes 
aware of an SAE that is suspected to be related to study drug, the Site PI or appropriate sub-
investigator will report the event to the DMID PVG. 
 
Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND 
Following notification from the investigator, DMID, the Investigational New Drug (IND) 
sponsor, will report any suspected TEAE that is both serious and unexpected. DMID will report 
an AE as a suspected AE only if there is evidence to suggest a causal relationship between the 
drug and the TEAE. DMID will notify FDA and all participating investigators (i.e., all 
investigators to whom DMID is providing drug under its IND(s) or under any PI’s IND(s)) in an 
IND safety report of potential serious risks from clinical trials or any other source, as soon as 
possible, but in no case later than 15 calendar days after DMID determines that the information 
qualifies for reporting as specified in 21 CFR Part 312.32. DMID will also notify FDA of any 
unexpected fatal TEAE or suspected life-threatening TEAE as soon as possible, but in no case 


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later than 7 calendar days after DMID’s initial receipt of the information. Relevant follow up 
information to an IND safety report will be submitted as soon as the information is available. 
Upon request from FDA, DMID will submit to FDA any additional data or information that the 
agency deems necessary, as soon as possible, but in no case later than 15 calendar days after 
receiving the request.  
All serious events designated as “not related” to the study drug will be reported to the FDA at 
least annually in a summary format. 
 
Other Adverse Events (if applicable) 
 
Reporting of Overdose 
An overdose is defined as a dose greater than the high-dose level evaluated in the trial as 
described in Section 6.3.1 of the protocol. All overdoses will be reported; if the overdose is 
associated with a TEAE, then the TEAE will also be reported. In the event of an overdose of 
study drug, the investigator will use clinical judgment in treating the overdose and contact the 
DMID MM. There is no specific antidote in case of apramycin overdose. The same measures as 
recommended for other aminoglycosides (treatment cessation, adequate hydration, dialysis, 
administration of calcium salts for symptoms of neuromuscular blockade) should be considered 
until further clinical data are available for apramycin. The investigator will refer to the relevant 
document(s) for detailed information regarding warnings, precautions, contraindications, TEAEs, 
and other significant data pertaining to apramycin. Such documentation may include but not be 
limited to the IB. 
 
Reporting of Pregnancy 
Pregnancies that occur in female subjects during the trial will be reported via Advantage 
eClinical® on a pregnancy report form. With the subject’s permission, all protocol-required 
venous blood samples will be obtained, and the subject will continue to be followed for safety 
until Day 30 ± 4 days (Final Visit). Efforts will be made to follow all pregnancies reported 
during the trial to pregnancy outcome, as described in the protocol-specific MOP (e.g., delivery, 
spontaneous abortion, or therapeutic abortion), pending the subject’s permission.  
A female subject who participates in the trial and becomes pregnant will be asked to inform 
study personnel of a pregnancy occurring 30 days after Final Visit. For all reported pregnancies, 
subjects will be asked to provide pregnancy outcome upon delivery or pregnancy termination to 
the CTU.  
Serious adverse outcomes of pregnancy affecting the mother and/or fetus or neonate (e.g., 
spontaneous abortion, congenital anomaly(ies) in fetus or child, late fetal death, or reports of 
adverse drug reactions in a newborn/neonate that is fatal, life-threatening, resulting in persistent 


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or significant disability/incapacity or resulting in or prolonging hospitalization) will be 
documented in a SAE form and reported to the CROMS PVG within 24 h of clinical site 
awareness of the events. 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events 
Treatment-emergent SAEs and AEs will be followed until resolution or until considered stable in 
the clinical judgment of the investigator. Resolution of an AE is defined as the return to 
pretreatment status or stabilization of the condition in the clinical judgment of the study 
investigator, with the expectation that it will remain chronic. 
Follow-up procedures, evaluations, and outcomes will be recorded in the subject's source 
document and eCRF. 
9.5 
Halting Rules 
 
Study Halting Criteria 
Systemic or Lab Criteria: 
• One subject has an SAE that is determined to be related to the study product. OR 
• Two or more subjects experience Grade 2 or higher related TEAE (laboratory or 
systemic) that is coded in the same HLGT per MedDRA classification, and if AE is not 
resolved within 48 hours through Day 4. Resolution is defined as return to a baseline 
status. Note: If the event starts on Day 4, the review period will be extended to Day 5. 
Note: VS abnormalities will be considered part of a systemic disorder or an organ-
specific condition, as described in Appendix B: Adverse Events Toxicity Grading 
Criteria, in order to be included among the Study Halting Criteria. 
Ototoxicity Criteria: 
• Two or more subjects experience Grade 2 or higher ototoxic change of pure-tone 
audiometry through Day 30 ± 4 days that is confirmed by repeat testing, generally within 
24 h, and assessed as related to IMP.  
• A single subject experiences significant ototoxic change in DEOAEs, characterized by 
decrease in reproducibility to lower than 70% at three consecutive frequencies where 
responses were previously obtained, through Day 30 (± 4 days) and is confirmed by 
repeat testing, generally within 24 h: 
If the predefined criteria for the halting rules are met, the SMC will review the study data and 
provide guidance on how to proceed. 
A halted study will resume upon recommendation by the Sponsor (DMID).  


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9.6 
Safety Oversight 
 
Safety Monitoring Committee (SMC) 
Safety oversight will be conducted by a SMC, which is an independent group of experts that 
monitors subject safety and advises DMID. SMC members will be separate and independent of 
study personnel participating in the trial and will not have scientific, financial, or other conflicts 
of interest related to the trial. The SMC will consist of a minimum of three members with 
appropriate expertise to contribute to the interpretation of the data from the trial. 
The SMC will meet as follows: 
• Organizational meeting (before study initiation). 
• Ad hoc meeting. 
o When study halting criteria are met. 
o At the request of DMID to review a potential safety concern identified by either the 
Site PI or DMID MM. 
• Scheduled meeting. 
o The SMC will review cumulative, interim safety data when available after completion 
of all subjects in cohorts T1 to T4. 
o The SMC will review final safety data after completion of all subjects in all cohorts, 
including T5, when available after database lock.  
Procedures for SMC reviews/meetings will be defined in the SMC charter. The SMC will review 
safety data and provide recommendations to DMID, NIAID on resumption or stopping of a 
temporary halted trial, and safety monitoring in future clinical trials with apramycin.  


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10 
CLINICAL MONITORING 
10.1 
Site Monitoring Plan 
Site monitoring will be conducted to ensure that human subject protections, study and laboratory 
procedures, study intervention administration, and data collection processes are of high quality 
and meet DMID, GCP/ICH, and regulatory guidelines, when appropriate. Site visits may be 
conducted by an authorized representative of DMID or other regulatory agencies to inspect study 
data, subjects’ medical records, and eCRFs in accordance with ICH guidelines, GCP, and the 
respective local and national government regulations and guidelines. 
The investigator will permit authorized representatives of DMID and the respective local and 
national health authorities to inspect facilities and records relevant to the trial, if needed. 
A separate monitoring plan developed by DMID will describe protocol-specific items to be 
monitored. 
Site visits will be made at standard intervals defined by DMID and may be made more frequently 
as directed by DMID. Monitoring visits will include, but not be limited to, review of regulatory 
files, accountability records, subjects’ source documents, eCRFs, ICFs, clinical and laboratory 
reports, and protocol and GCP compliance. Site monitors will have access to the CTU, study 
personnel, and all study documentation according to the DMID-approved site monitoring plan. 
Study monitors will meet with the Site PI to discuss any problems and actions to be taken and 
document visit findings and discussions. 
 


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11 
STATISTICAL CONSIDERATIONS 
11.1 
Study Hypotheses 
The objectives of the study are to obtain lung and plasma PK and safety data for apramycin. 
There are no formal hypotheses being tested in this Phase 1 trial. 
11.2 
Sample Size Considerations 
Enrollment of 20 subjects assigned into 5 cohorts, T1-T5 (T5 will undergo bronchoscopy with 
BAL after completion of cohorts at the 24 h nominal timepoint, with the optimum timepoint to 
be selected after completion of T1-T4), with 4 subjects per cohort is based on clinical experience 
and judgment and should provide adequate clinical information to meet the objectives of the 
study. Power calculations have not been performed. 
11.3 
Safety Review 
An SMC will be appointed to oversee the safe conduct of the trial. A scheduled SMC meeting 
will be held after all subjects in timepoint cohorts T1 to T4 complete the trial to review safety 
data. If criteria for halting the trial (as listed in Section 9.5.1) are met, an ad hoc SMC meeting 
will be held to review all available safety data and to make recommendations about the dosing of 
all further subjects in the trial. A statistician may present data in a closed session of an SMC 
meeting. If group T5 will enroll, a final SMC meeting will be held to review all data and advise 
on safety monitoring in future clinical trials of apramycin. 
11.4 
Final Analysis Plan 
The ICH Guidance E9 (Statistical Principles for Clinical Trials) will be followed for all statistical 
content. Summaries of frequencies and percentages will be presented for categorical data, and 
summary statistics such as minimum, median, mean, SD, and maximum values will be presented 
for continuous data. Details of plasma and BAL SUP (containing ELF) and BAL AM apramycin 
PK, lung PK/plasma apramycin exposure ratios, safety, ECG interval measurements (QT, PR, 
QRS, and HR), and ECG morphology data analyses and presentations will be described in the 
Statistical Analysis Plan (SAP) and accompanying Tables, Listings, and Figures (TLF) 
templates. The SAP for PK and safety data will be prepared and finalized by the SDCC before 
analysis of the PK data after completion of cohorts T1-T4. A final analysis containing safety data 
and PK data from cohorts T1-T4 and cohort T5, if enrolled, will be performed by the SDCC after 
final data lock and included in the clinical study report (CSR). Any change from originally 
planned statistical analyses will be reported in the CSR.   


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Analysis Populations 
The safety population will include all subjects who received any amount of the IMP. The per 
protocol safety population subset will include all subjects who received a complete dose of study 
drug (apramycin) and analyzed as treated.  
The lung PK population will consist of all the subjects who received a complete dose of 
apramycin, underwent BAL at the assigned sampling timepoint with BAL return volume 
adequate for analysis and have at least one quantifiable drug concentrations separately for BAL 
SUP (containing ELF) and BAL AM.  
The plasma PK analysis population will consist of all subjects who received a complete dose of 
apramycin and have at least one quantifiable post-dosing plasma drug concentration measured.  
The PK analysis subset will be based on the lung and plasma PK population, which includes all 
subjects who completed the lung PK and plasma PK part of the trial without any protocol 
violations that would likely affect the PK results and who have evaluable plasma PK and BAL 
PK concentration data for apramycin from which at least a subset of the designated PK 
parameters can be determined. If any subjects are found to be noncompliant with respect to 
dosing or have incomplete data, a decision to include them in the analysis will be made on a 
case-by-case basis.  
 
Demographics and Baseline Characteristics 
Subject demographics and baseline characteristics will be summarized. The number of subjects 
who enroll in the trial, and the number and percentage of subjects who complete each 
assessment, will be presented. The percentage of subjects who withdraw from the trial or 
discontinue the study drug, and reasons for withdrawal or discontinuation, will be summarized. 
 
Safety Analysis 
 
Adverse and Serious Adverse Events 
TEAEs will be coded using the latest version of the Medical Dictionary for Regulatory 
Activities® (MedDRA). All AEs occurring after study drug dosing will be summarized using 
frequency counts and percentages. The following summaries will be presented for treatment-
emergent AEs and SAEs:  
• Overall (i.e., regardless of severity or relationship to treatment). 
• By severity grade (mild, moderate, or severe). 
• By relationship to study drug. 


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• By MedDRA level hierarchy (system organ class [SOC], higher level group term 
[HLGT] and preferred term [PT]). 
Unless otherwise specified, at each level of subject summarization in reporting the incidence of 
TEAEs, a subject will be counted once if the subject reported one or more TEAEs. If more than 
one occurrence of a TEAE is reported, the TEAE of the worst severity or the worst-case 
relationship assessment will be summarized.  
 
Additional Safety Analyses 
Descriptive summary statistics (mean, SD, median, minimum, and maximum) for clinical 
laboratory data, 12-lead standard ECG parameters, and VS at admission and each applicable 
post-dosing visit, including changes from baseline values, will be calculated. Baseline values 
will be the last values recorded before administration of study drug. For change-from-baseline 
summaries, subjects with an undefined change from baseline, due to missing data, will be 
excluded. Clinical significance of abnormalities will be indicated in the listings. 
 
PK Analysis  
PK parameters will be estimated for total plasma apramycin, and for ELF and AM total 
apramycin by noncompartmental analysis (NCA) methods using Phoenix® WinNonlin® version 
8.0 or higher.  
 
Plasma PK parameters: 
When evaluable, estimated plasma PK parameters will include: 
• AUC(0-∞): Area under the concentration time-curve from time zero to infinity.  
• AUC(0-t): Area under the concentration time-curve from time zero to time t. 
• AUC (0-24): Area under the concentration time-curve from time zero to 24 h. 
• Cmax: Maximum observed concentration. 
• Tmax: Time of maximum observed concentration. 
• Ke: Elimination rate constant.  
• t1/2: Terminal elimination half-life. 
• CLT: Total clearance. 
• Vd: Volume of distribution. 
Other PK parameters may be calculated, as appropriate.  
The free portion (fu, % unbound) of plasma apramycin will be calculated from the ratio of free to 
total apramycin concentration in plasma PK samples obtained at 0.5 h (± 5 min, immediately 
after the end of infusion) and 36 h (± 1 h) after the start of infusion. 


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Bronchoalveolar ELF and AM PK parameters: 
When evaluable, estimated PK parameters from BAL ELF and BAL AM will include: 
• Cmax: Maximum observed concentration.  
• AUC(0-24): Area under the concentration time-curve from time zero to 24 h.  
• AUC(0-t): Area under the concentration time-curve from time zero to the last 
concentration. above the lower limit of quantitation.  
• AUC(0-∞): Area under the concentration time-curve from time zero to infinity. 
• Tmax: Time of maximum observed concentration.  
• t½: Terminal elimination half-life. 
The concentration of apramycin in ELF will be calculated using the urea method (Tenero 2013, 
Gottfried 2017, Rizk 2018). The formulae for calculating ELF concentration will be provided in 
the SAP.  
 
Ratio of exposure parameters of Lung PK to Plasma PK: 
When evaluable, the following exposure ratios of total apramycin will be calculated: 
• ratio of Cmax in ELF over Cmax in plasma. 
• ratio of Cmax in AM over Cmax in plasma. 
• ratio of AUC(0-24), AUC(0-t) and AUC(0-∞)  in ELF over AUC(0-24), AUC(0-t) and AUC(0-∞)  in 
plasma. 
• ratio of AUC(0-24), AUC(0-t) and AUC(0-∞) in AM over AUC(0-24), AUC(0-t) and AUC(0-∞)  in 
plasma. 
The results of PK parameters for apramycin in plasma, ELF, AM, and the exposure (Cmax and 
AUC) ratios of lung/plasma (total and free) will be listed by subject, and summarized with 
descriptive statistics including n, mean, SD, coefficient of variation, median, minimum, 
maximum, geometric mean and geometric SD at each timepoint.  
Graphical presentations of concentration vs. time profiles will be provided for apramycin and 
will include individual subject and mean concentration profiles. Semi-log concentration profiles 
will be provided for individual subjects. The ratios of lung PK exposure / plasma PK exposure vs 
time profiles for total plasma apramycin will be presented graphically for the ELF and AM 
components of BAL samples.  
 
Missing Values and Outliers 
All attempts will be made to collect all data per protocol. No imputation will be performed for 
missing values. Outliers will not be excluded from the primary analyses. Outliers identified 
during the PK analysis will be discussed in the analysis report. 


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Analysis of ECG Parameters  
 
ECG Analysis and Interpretation 
For replicate ECGs recorded on Day -1 and Day 1, the ∆∆QTcF, calculated as difference 
between mean change-from-baseline QTcF values at each timepoint on Day 1 after treatment 
with apramycin relative to time-matched ECG recording before dosing in Day -1, will be 
evaluated to quantify the QTcF prolongation. For single ECG recordings after dosing, ∆QTcF, 
will be calculated as difference between mean change-from- baseline QTcF values recorded at 
the pre-dose timepoints on Day -1 and Day 1.  
QTcF corrections will be calculated using Fridericia’s formula: 
• 
QTcF=QT/RR0.33 
Other endpoints to be evaluated include HR (bpm), PR (msec), QRS (msec), and cardiac 
abnormalities (morphologies, rhythm, and conduction).   
All statistical analyses and reporting of ECG data will be conducted on the mean of the replicate 
intervals within a timepoint on Day -1 and Day 1. Descriptive statistics including two-sided 90% 
CI will be generated for observed, change-from-baseline for all ECGs recorded in the study, and 
time-matched change-from-baseline ECG intervals for replicate ECGs recorded on Day -1 and 
Day 1. 
 
Categorical Analyses 
QTcF intervals will be categorized and tabulated for each treatment. The following criteria will 
be used to specify noteworthy QTcF intervals:  
• QTcF >450 msec  
• QTcF >480 msec  
• QTcF >500 msec  
• QTcF increase ≥30 msec from baseline 
• QTcF increase ≥60 msec from baseline 
Categorical analysis of PR, QRS, and RR intervals will also be performed. The following criteria 
will be used to specify noteworthy non-QTc intervals (PR, QRS, and RR intervals) and HR:   
• PR change-from-baseline >25% resulting in PR >200 msec  
• QRS change-from-baseline >25% resulting in QRS >120 msec 
• HR change-from-baseline >25% decrease resulting in HR <50 bpm 


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• HR change-from-baseline >25% increase resulting in HR >100 bpm 
The number and percent of subjects in each category will be calculated. Comparisons across 
timepoints may be conducted if sufficient counts are observed.  
 
Cardiac Abnormalities: 
The morphological waveform analysis of the ECG data at each timepoint will be utilized for the 
summary of new onset cardiac abnormalities for treatment. Results will be summarized in 
frequency tables with counts and percentages for both number of subjects and number of 
timepoints. 
 
Analysis of Audiology Data 
Descriptive summary statistics (mean, SD, median, minimum, and maximum) of ear audiometric 
and otoacoustic variables at baseline and each applicable post-dosing visit, including changes 
from baseline values, will be calculated. Baseline values will be the last values recorded before 
administration of study drug. For change-from-baseline summaries, subjects with an undefined 
change from baseline, due to missing data, will be excluded. Clinical significance of 
abnormalities will be indicated in the listings.  
All events occurring after study drug dosing will be summarized using frequency counts and 
percentages. The following summaries will be presented for each event:  
• Overall (i.e., regardless of severity or relationship to treatment). 
• By severity grade (mild, moderate, or severe). 
Detailed analysis plans will be documented in the SAP.  
Results of the safety and PK evaluations will be reported in the CSR.  
 
Interim Reports 
Plasma and BAL (ELF and AM) apramycin concentration and PK data from cohorts T1 to T4 
will be analyzed and summarized.  
Interim safety data will be presented to the SMC.  
  


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12 
SOURCE DOCUMENTS AND ACCESS TO SOURCE 
DATA/DOCUMENTS 
The CTU will maintain appropriate medical and/or research records for the trial, in compliance 
with ICH E6 GCP, Section 4.9, and regulatory and institutional requirements for the protection 
of confidentiality of subjects. As part of participating in a DMID-sponsored, DMID-affiliated, or 
manufacturer-sponsored trial, the CTU will permit authorized representatives of DMID, to 
include The Emmes Company, LLC (the SDCC), DynPort Vaccine Company, LLC (DVC), and 
regulatory agencies to review (and, when required by applicable law, to copy) clinical records for 
the purposes of quality assurance reviews, audits, and evaluation of the study safety and 
progress. 
Forms for use as source documents will be derived from eCRFs and will be provided by the 
SDCC and CTU. Additional source data are all information, original records of clinical findings, 
observations, or other activities in a clinical trial necessary for the reconstruction and evaluation 
of the trial. Examples of these original documents and data records include, but are not limited 
to, hospital records, clinical and office charts, laboratory notes, memoranda, pharmacy 
dispensing records, recorded data from automated instruments, copies or transcriptions certified 
after verification as being accurate and complete, microfiches, photographic negatives, microfilm 
or magnetic media, x-rays, ECG print-outs, and subject files and records kept at the pharmacy, 
laboratories, and medico-technical departments involved in the trial. 
 


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13 
QUALITY CONTROL AND QUALITY ASSURANCE 
Following a written DMID-accepted site quality management plan, the CTU is responsible for 
conducting routine quality assurance and quality control activities to internally monitor study 
progress and protocol compliance. The Site PI will provide direct access to the CTU, source 
data/documents, and reports for monitoring and auditing by DMID, and inspection by local and 
regulatory authorities. The Site PI will ensure all study personnel are appropriately trained and 
applicable documentations are maintained on site.  
The SDCC will implement quality control procedures beginning with the data entry system and 
generate data quality control checks that will be run on the database. Any missing data or data 
anomalies will be communicated to the CTU for clarification and resolution. 


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14 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 
Ethical Standard 
The PI will ensure that the trial is conducted in full conformity with principles of the Belmont 
Report: Ethical Principles and Guidelines for the Protection of Human Subjects of Research of 
the National Commission for the Protection of Human Subjects of Biomedical and Behavioral 
Research (April 18, 1979) and codified in 45 CFR 46, 21 CFR 50 and 56, and ICH E6; 
62 Federal Regulations 25692 (1997), if applicable. The PI’s institution will hold a current 
Federal Wide Assurance (FWA) issued by the Office of Human Research Protection for 
federally funded research. 
14.2 
Institutional Review Board 
The CTU will provide for the review and approval of this protocol and associated ICFs by an 
appropriate IRB/EC listed on the FWA. Any amendments to the protocol or consent materials 
will also be approved before they are used, unless change is for the safety of the subject. Only 
those IRB members who are independent of the investigators and DMID will provide an opinion 
on study-related matters. Verification of IRB approval of the protocol and the written ICF will be 
transmitted by the investigator or designee before shipment of the study drug. No deviations 
from or changes to the protocol will be initiated without prior approval of an appropriate 
amendment unless change is for the safety of the subject. 
14.3 
Informed Consent Process 
The written consent document will embody the elements of informed consent as described in the 
Declaration of Helsinki and will adhere to the ICH Harmonized Tripartite Guideline for GCP. 
Informed consent will be obtained before any protocol-specified procedures or interventions are 
carried out, and in accordance with 21 CFR 50.25 and 45 CFR 46. Information will be presented 
both orally and in written form. 
An investigator or designee will describe the protocol to potential subjects face-to-face. The ICF 
may be read to the subjects, but, in any event, the investigator shall give the subjects ample 
opportunity to inquire about details of the trial and ask any questions before signing the ICF. 
Study staff will inform subjects that the trial involves research, and explain the purpose of the 
trial, those aspects of the trial that are experimental, any expected benefits, all possible risks 
(including a statement that the particular treatment or procedure may involve risks to the subject 
or to the embryo or fetus, if the subject is or may become pregnant, that are currently 
unforeseeable), the expected duration of the subject’s participation in the trial, the procedures of 
the research study, including all invasive procedures, and the probability for assignment to 


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treatment cohorts. Subjects will be informed that they will be notified in a timely manner if 
information becomes available that may be relevant to their willingness to continue participation 
in the trial. They will also be informed of alternative procedures that may be available, and the 
important potential benefits and risks of these available alternative procedures. Subjects will 
receive an explanation as to whether any compensation and any medical treatments are available 
if injury occurs, and, if so, what they consist of, or where further information may be obtained. 
Subjects will be informed of the anticipated financial expenses, if any, to the subject for 
participating in the trial, as well as any anticipated prorated payments, if any, to the subject for 
participating in the trial. They will be informed of whom to contact (e.g., the Investigator) for 
answers to any questions relating to the research project. Information will also include the 
foreseeable circumstances and/or reasons under which the subject’s participation in the trial may 
be terminated. Subjects will be informed that participation is voluntary and that they are free to 
withdraw from the trial for any reason at any time without penalty or loss of benefits to which 
the subject is otherwise entitled. 
Neither the investigator, nor the trial staff, will coerce or unduly influence a subject to participate 
or continue to participate in the trial. The extent of the confidentiality of the subjects’ records 
will be defined, and subjects will be informed that applicable data protection legislation will be 
followed. Subjects will be informed that the monitor(s), auditor(s), IRB, DMID, and regulatory 
authority(ies) will be granted direct access to the subject’s original medical records for 
verification of clinical trial procedures and/or data without violating the subject’s confidentiality, 
to the extent permitted by applicable laws and regulations, and that, by signing a written ICF, the 
subject is authorizing such access. Subjects will be informed that records identifying the subject 
will be kept confidential, and, to the extent permitted by the applicable laws and/or regulations, 
will not be made publicly available and, if the results of the trial are published, the subject’s 
identity will remain confidential. 
ICFs will be in a language fully comprehensible to the prospective subjects. Informed consent 
shall be documented using a written consent form approved by the IRB and signed and dated by 
the subject and the person who conducted the informed consent discussion. The signature 
confirms that the consent is based on information that has been provided and all questions have 
been answered to the prospective subject’s satisfaction. Each subject’s signed ICF will be kept 
on file by the investigator for possible inspection by regulatory authorities and/or DMID and 
regulatory compliance persons. The subject will receive a copy of the signed and dated written 
ICF and any other written information provided to the subjects and will receive copies of any 
signed and dated ICF updates and any amendments to the written information provided to 
subjects. 


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14.4 
Exclusion of Women, Minorities, and Children (Special Populations) 
Children aged < 18 years will be excluded from participation because insufficient data are 
available in adults to judge potential risk in children, and as a Phase 1 trial, there is no known 
benefit.   
Neither women nor minorities will be routinely excluded from participation in the trial. Subjects 
will be recruited without regard to gender or race. It is expected that race and gender 
distributions in the trial will approximate the proportion to their numbers within the community.   
Women of childbearing potential will be included but will be repeatedly counseled to use 
effective measures (Section 5.1) to avoid becoming pregnant from the time of screening until 
Day 30 (± 4 days) (last visit) after the dose of study drug is received because the effects of the 
study drug on the unborn fetus are not known.    
14.5 
Subject Confidentiality 
Subject confidentiality is held strictly in trust by the participating investigators, their staff, and 
DMID and its agents. This confidentiality is extended to cover testing of biological samples, and 
also clinical information related to participating subjects. 
The study protocol, documentation, data, and all other information generated will be held in 
strict confidence. No information concerning the trial or data will be released to any 
unauthorized third party without prior written approval from DMID. This information and data 
will not be used by the Site PI or other study personnel for any purpose other than conducting the 
trial. These restrictions do not apply to: (1) information which becomes publicly available 
through no fault of the Site PI or other study personnel; (2) information which is necessary to 
disclose in confidence to an IRB solely for the evaluation of the trial; (3) information which is 
necessary to disclose in order to provide appropriate medical care to a subject; or (4) study 
results which may be published as described in Section 16. 
The study monitors or other authorized representatives of DMID may inspect all documents and 
records required to be maintained by the Site Investigator, including, but not limited to, medical 
records (office, clinic or hospital) and pharmacy records for the subjects in the trial. The CTU 
will permit access to such records. 
14.6 
Certificate of Confidentiality 
To protect privacy, we have received a Certificate of Confidentiality.  With this Certificate, the 
researchers cannot be forced to release information that may identify the research subject, even 
by a court subpoena, in any federal, state, or local civil, criminal, administrative, legislative, or 
other proceedings. The researchers will use the Certificate to resist any demands for information 
that would identify the subject, except as explained below. 


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The Certificate cannot be used to resist a demand for information from personnel of the United 
States Government that is used for auditing or evaluation of federally funded projects, like this 
study, or for information that must be released in order to meet the requirements of the Federal 
Food and Drug Administration (FDA). 
A Certificate of Confidentiality does not prevent the subject from voluntarily releasing 
information about themselves or their involvement in this research. If any person or agency 
obtains a written consent to receive research information, then the researchers may not use the 
Certificate to withhold that information.   
The Certificate of Confidentiality does not prevent the researchers from reporting without the 
subject’s consent, information that would identify the subject as a participant in the research 
project regarding matters that must be legally reported, including child and elder abuse, sexual 
abuse, or wanting to harm themselves or others.   
The release of individual private information or specimens for other research will only occur if 
consent was obtained from the individual to whom the information, document, or biospecimen 
pertains, or for the purposes of other research that is in compliance with applicable Federal 
regulations governing the protection of human subjects in research. 
14.7 
Study Discontinuation 
DMID has the right to terminate the trial or the CTU’s participation at any time. Reasons for 
terminating the trial may include, but are not limited to: 
• Incidence or severity of TEAEs indicates a potential health hazard. 
• Data recording is inaccurate or incomplete. 
• Investigator does not adhere to the protocol or applicable regulatory guidelines in 
conducting the trial. 
If the trial is discontinued, subjects who have signed the ICF and received the study drug will 
continue to be followed for safety for the duration of the trial. No further study treatments will be 
administered to other subjects. 
14.8 
Future Use of Stored Specimens 
Blood (plasma) and BAL fluid samples (ELF and AM) will be collected as outlined in the 
protocol. Subjects will be given a choice during the informed consent process to have their 
residual linked samples stored indefinitely for future research, have their residual samples de-
linked from any subject information and stored indefinitely, or have their residual linked samples 
destroyed at completion of the study. The use of long-term stored linked samples will be 


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conducted under the restrictions regarding confidentiality, as outlined in the preceding 
paragraphs. 
Only coded specimens will be sent to the sponsor with the code identifiers maintained by the 
principal investigator and The Emmes Company, LLC, the NIAID SDCC. Any future research 
studies will utilize only the residual long-term stored specimens from subjects consenting to 
future use, which will be stored at the DMID CAR. Residual, coded samples from the plasma PK 
and BAL (ELF and AM) PK assays may also be used for future exploratory research.  
Residual clinical samples will be available upon the completion of the study; however, future use 
clinical samples may be requested from DMID and shipped from the DMID CMS at any time. 
The samples will not be sold or used directly for production of any commercial product. No 
human genetic tests will be performed on the samples.  Each sample will be encoded (labeled) 
only with a barcode and a unique tracking number to protect subject confidentiality. 
There are no benefits to subjects in the collection, storage and subsequent use of their specimens 
for future research. Reports about future research done with subjects’ samples will NOT be kept 
in their health records. 
Subjects may be given the option to decide if they want their samples to be used for future 
research or have their samples destroyed at the end of this trial. The subject’s decision can be 
changed at any time by notifying the study doctors or nurses in writing. However, if the subject 
originally consents to future use and subsequently changes his/her decision, any data from a 
previously collected sample may still be used for this research. 


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15 
DATA HANDLING AND RECORD KEEPING 
The Site PI is responsible to ensure the accuracy, completeness, legibility, and timeliness of 
reported data. All data collection forms will be completed legibly to ensure accurate 
interpretation of data. Black or blue ink is required to ensure clarity of reproduced copies. When 
making changes or corrections, cross out the original entry with a single line, and initial and date 
the change. Do not erase, overwrite, or use correction fluid or tape on the original. 
Data collection forms will be provided by the SDCC and the CTU will use them to develop the 
CTU’s source documents to record and maintain data for each subject enrolled in the trial. Data 
reported in the eCRF derived from source documents will be consistent with the source 
documents or the discrepancies will be explained. 
DMID and/or its designee will provide guidance to investigators and other study personnel on 
making corrections to the data collection forms, source documents and eCRFs. 
15.1 
Data Management Responsibilities 
All source documents and laboratory reports will be reviewed by the clinical team and data entry 
staff, who will ensure that they are accurate and complete. TEAEs will be graded, assessed for 
severity and causality, and reviewed by the Site PI or designee. 
Data collection is the responsibility of the clinical trial staff at the CTU under the supervision of 
the Site PI. During the trial, the investigator will maintain complete and accurate documentation 
for the trial. 
The Emmes Company, LLC will serve as the SDCC for the trial, and will be responsible for data 
management, quality review, analysis, and reporting of the study data. 
15.2 
Data Capture Methods 
Clinical data (including, but not limited to treatment-emergent AE/SAEs, ConMeds, MH, PE, 
clinical laboratory data, and ECG data [ECG intervals and interpretations]) will be entered into a 
21 CFR 11-compliant Internet Data Entry System provided by the SDCC. The data system 
includes password protection and internal quality checks (e.g., automatic range checks) to 
identify data that appear inconsistent, incomplete, or inaccurate. Clinical data will be entered 
directly from the source documents by CTU study personnel.  
15.3 
Types of Data 
Data for the trial will include clinical and laboratory safety assessments, plasma and urine renal 
toxicity markers, 12-lead standard ECG interval measurements and interpretations, audiology 
assessments and interpretations, bronchoscopy procedure reports including pre- and post-


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bronchoscopy records, plasma PK and lung PK (ELF and AM) bioanalytical assay results and 
corrected ELF and AM concentration data, and concentration of urea in plasma, ELF and AM at 
each specified timepoint.  
15.4 
Timing/Reports 
A final CSR will be prepared after all safety, 12-lead standard ECG, and plasma and lung PK 
data are available. See Section 9.6.1 and Section 11.4.8 for additional reporting requirements.  
15.5 
Study Records Retention 
Study files and ICFs will be maintained for 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 study drug. These documents will be retained for a longer period, 
however, if required by local regulations. No records will be destroyed without the written 
consent of DMID, if applicable. It is DMID’s responsibility to inform the investigator when 
these documents no longer need to be retained. 
15.6 
Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, GCP, or protocol-
specific MOP requirements. The noncompliance may be either on the part of the subject, the 
investigator, or the CTU staff. Corrective actions for protocol deviations are to be developed by 
the CTU and implemented promptly. 
These practices are consistent with ICH E6: 
4.5 Compliance with Protocol, Sections 4.5.1, 4.5.2, and 4.5.3 
5.1 Quality Assurance and Quality Control, Section 5.1.1 
5.20 Noncompliance, Sections 5.20.1, and 5.20.2. 
It is the responsibility of the Site PI/study staff to use continuous vigilance to identify and report 
deviations within 5 working days of identification of the protocol deviation, or within 5 working 
days of the scheduled protocol-required activity. All deviations will be promptly reported to 
DMID via SDCC’s Advantage eClinical®. 
All protocol deviations, as defined above, will be addressed in subject source documents. A 
completed copy of the DMID Protocol Deviation Form will be maintained in the regulatory file, 
as well as in the subject's source document. Protocol deviations will be sent to the local IRB/EC 
per their guidelines. The Site PI/study staff is responsible for knowing and adhering to their IRB 
requirements. 


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16 
PUBLICATION POLICY 
Following completion of the study, the lead Principal Investigator is expected to publish the 
results of this research in a scientific journal. All investigators funded by the NIH must submit or 
have submitted for them to the National Library of Medicine’s PubMed Central 
(http://www.ncbi.nlm.nih.gov/pmc/) an electronic version of their final, peer-reviewed 
manuscripts upon acceptance for publication, to be made publicly available no later than 
12 months after the official date of publication. The NIH Public Access Policy ensures the public 
has access to the published results of NIH funded research.  It requires investigators to submit 
final peer-reviewed journal manuscripts that arise from NIH funds to the digital archive PubMed 
Central upon acceptance for publication. Further, the policy stipulates that these papers must be 
accessible to the public on PubMed Central no later than 12 months after publication. 
Refer to: 
• NIH Public Access Policy, http://publicaccess.nih.gov/   
• NIH Office of Extramural Research (OER) Grants and Funding, 
http://grants.nih.gov/grants/oer.htm   
As of January 2018, all clinical trials supported by the NIH must be registered on 
ClinicalTrials.gov no later than 21 days after the enrollment of the first subject. Results of all 
clinical trials supported by the NIH, generally, need to be submitted no later than 12 months 
following the primary completion date. A delay of up to 2 years is available for trials that meet 
certain criteria and have applied for certification of delayed posting. As part of the result posting, 
a copy of this protocol (and its amendments) and a copy of the Statistical Analysis Plan will be 
posted on ClincialTrials.gov. For this trial the responsible party is DMID/NIAID/NIH, which 
will register the trial and post results.   
Refer to: 
• Public Law 110-85, Section 801, Clinical Trial Databases 
• 42CFR11 
• NIH NOT-OD-16-149 
 


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17 
LITERATURE REFERENCES 
1. WHO Model Lists of Essential Medicines, June 2019. (WHO 2019a) Accessed in:   
https://www.who.int/groups/expert-committee-on-selection-and-use-of-essential-
medicines/essential-medicines-lists  
 
2. Critically important antimicrobials for human medicine, 6th revision. Geneva: World Health 
Organization; 2019. License: CC BY-NC-SA 3.0 IGO. (WHO 2019b) Accessed in:  
https://apps.who.int/iris/bitstream/handle/10665/312266/9789241515528-eng.pdf 
 
3. Cassini A, Högberg LD, Plachouras D, Quattrocchi A, Hoxha A, Simonsen GS, et al.; Burden of 
AMR Collaborative Group. Attributable deaths and disability-adjusted life-years caused by 
infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: a 
population-level modelling analysis. Lancet Infect Dis. Lancet Infect Dis. 2019;19(1):56-66. 
 
4. O’Neill, J. (2014). Antimicrobial Resistance: Tackling a Crisis for the Future Health and 
Wealth of Nations. HM Government. Accessed in: 
https://wellcomecollection.org/works/rdpck35v 
 
5. CDC. Antibiotic Resistance Threats in the United States, 2019. Atlanta, GA: U.S. Department of 
Health and Human Services, CDC (CDC 2019); 2019.  Accessed in: 
https://www.cdc.gov/drugresistance/pdf/threats-report/2019-ar-threats-report-508.pdf 
 
6. O’Connor S, Lam LKT, Jones ND, Chaney MO. Apramycin, a unique aminocyclitol antibiotic. L 
Org Chem 1976; 41 (12): 2087-2092.  https://doi.org/10.1021/jo00874a003  
 
7. Apramycin. Summary Report (2). The European Agency for the Evaluation of Medicinal 
Products, Veterinary Medicines Unit. January 1999. (EMA 1999). Accessed in: 
https://www.ema.europa.eu/en/documents/mrl-report/apramycin-summary-report-2-committee-
veterinary-medicinal-products_en.pdf 
 
8. EU Commission Press Release IP/05/1687. Ban on antibiotics as growth promoters in animal feed 
enters into effect. Brussels, 22 December 2005. Accessed in: 
http://europa.eu/rapid/pressrelease_IP-05-1687_en.htm 
 
9. FDA Drugs with Veterinary Feed Directive (VFD) Marketing Status (FDA 2021). Accessed in: 
 https://www.fda.gov/animal-veterinary/development-approval-process/drugs-veterinary-feed-
directive-vfd-marketing-status 
  
10. Investigator’s Brochure, EBL-1003 (apramycin). V8.0, Juvabis AG, 28 January 2021 
 
11. Smith KP, Kirby JE. Evaluation of apramycin activity against carbapenem-resistant and -
susceptible strains of Enterobacteriaceae. Diagn. Microbiol. Infect. Dis., 2016 Dec; 86, 439-441 
 


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12. Juhas M, Widlake E, Teo J , et al. In vitro activity of apramycin against multidrug-, carbapenem- 
and aminoglycoside-resistant Enterobacteriaceae and Acinetobacter baumannii. J Antimicrob 
Chemother 2019; 74: 944–952. doi:10.1093/jac/dky546 
 
13. Kang AD, Smith. KP, Eliopoulos GM, Berg AH, McCoy C, Kirby JE. Invitro Apramycin 
Activity against multidrug-resistant Acinetobacter baumannii and Pseudomonas aeruginosa. 
Diagn. Microbiol. Infect. Dis., 2017 Jun; 88(2) 188-191. 
 
14. Truelson KA, Brennan-Krohn T, Smith KP, Kirby JE. Evaluation of apramycin activity against 
methicillin-resistant, methicillin-sensitive, and vancomycin-intermediate Staphylococcus aureus 
clinical isolates. Diagn Microbiol Infect Dis 2018 Oct; 92(2):168-171. 
doi:10.1016/j.diagmicrobio.2018.05.018.  
 
15. Becker K, Aranzana-Climent V, Cao S, Nilsson A, Shariatgorji R, Haldimann K, Bjorn Platzack 
B, Hughes D, Per E. Andren PE, Bottger EC, Friberg LE, Hobbie SN, on behalf of the ENABLE 
consortium. Efficacy of EBL-1003 (apramycin) against Acinetobacter baumannii lung infections 
in mice. Clinical Microbiology and Infection 2021 Sept; 27(9):1315-1321.  
https://doi.org/10.1016/j.cmi.2020.12.004  
 
16. Matt T, Ng CL, Sha SH, Akbertenov R, Shcherbakov D, Meyer M, et al. Dissociation of 
antibacterial activity and aminoglycoside ototoxicity in the 4-monosubstituted 2- 
deoxystreptamine apramycin. Proc. Natl. Acad. Sci. U S A, 2012 Jul 3;109(27):10984-9. doi: 
10.1073/pnas.1204073109 
 
17. Ishikawa M, García-Mateo N, Čusak A, López-Hernández I, Fernández-Martínez M, Müller M, 
et al. Lower ototoxicity and absence of hidden hearing loss point to gentamicin C1a and 
apramycin as promising antibiotics for clinical use. Sci Rep. 2019 Feb 20;9(1):2410. 
 
18. Mingeot-Leclercq, M-P, Glupczynski Y, and Tulkens PM. Aminoglycosides: Activity and 
Resistance. Antimicrob Agents Chemother. 1999 Apr; 43(4): 727–737. DOI: 
10.3390/ijms21176133 
 
19. Plattner M, Gysin M, Haldimann K, Becker K, Hobbie SN. Epidemiologic, Phenotypic, and 
Structural Characterization of Aminoglycoside-resistance Gene aac(3)-IV. Int J Mol Sci 2020 
Aug 25; 21(17): 6133. DOI: 10.3390/ijms21176133 
 
20. Böttger EC, Crich D. Aminoglycosides: Time for Resurrection of a Neglected Class of 
Antibacterials? ACS Infect Dis. 2020 Feb 14; 6(2): 168–172.  doi:10.1021/acsinfecdis.9b00441. 
 
21. Sou T, Hansen J, Liepinsh E, et al. Model-Informed Drug Development for Antimicrobials: 
Translational PK and PK/PD Modeling to Predict an Efficacious Human Dose for Apramycin. 
Clinical Pharmacology and Therapeutics 2021; 109(4); 1063-1073. doi:10.1002/cpt.2104 
 
22. Zhang W, Yi Huang Y, Helmers R . Bronchoalveolar Lavage. Chapter 14 in Flexible 
Bronchoscopy, Book Editor(s): Ko-Pen Wang MD, Atul C. Mehta MD, FACP, FCCP, J. Francis 
Turner Jr. MD, FACP, FCCP, FCCM 4th edition, 08 May 2020, Wiley Online Library | 
https://doi.org/10.1002/9781119389231  


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23. Um SW, Choi CM, Lee CT et al. Prospective analysis of clinical characteristics and risk factors 
of post-bronchoscopy fever. Chest 2004; 125: 945–52. 
 
24. Dezube R. Bronchoscopy. Merch Manuals, May 2021  
Accessed at https://www.merckmanuals.com/professional/pulmonary-disorders/diagnostic-and-
therapeutic-pulmonary-procedures/bronchoscopy  
 
25. Levey AS, Stevens LA. Estimating GFR using the CKD Epidemiology Collaboration (CKD-EPI) 
creatinine equation: more accurate GFR estimates, lower CKD prevalence estimates, and better 
risk predictions. Am J Kidney Dis. 2010; 55(4):622-627.  
https://doi.org/10.1053/j.ajkd.2010.02.337  
 
26. Tenero D, Bowers G, Rodvold KA, Patel A, Kurtinecz K, Dumont E, Tomayko J, Patel P. 
Intrapulmonary Pharmacokinetics of GSK2251052 in Healthy Volunteers. Antimicrob Agents 
Chemother 2013; 57(7): 3334-3339 | DOI: 10.1128/AAC.02483-12 
 
27. Gotfried MH, Horn K, Garrity-Ryan L, Villano S, Tzanis E, Chitra S, Manley A, Tanaka SK, 
Rodvold KA. Comparison of omadacycline and tigecycline pharmacokinetics in the plasma, 
epithelial lining fluid, and alveolar cells of healthy adult subjects. Antimicrob Agents Chemother 
2017; 61:e01135-17. https://doi.org/10.1128/AAC.01135-17 
 
28. Rizk ML, Rhee EG, Jumes PA, Gotfried MH, Zhao T, Mangin E, Bi S, Chavez-Eng CM, Zhang 
Z, Butterton JR. Intrapulmonary pharmacokinetics of relebactam, a novel β-lactamase inhibitor, 
dosed in combination with imipenem-cilastatin in healthy subjects. Antimicrob Agents 
Chemother 2018; 62:e01411-17. https://doi.org/10.1128/AAC.01411-17 
 


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APPENDICES


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Appendix A: Schedule of Study Procedures and Evaluations 
Table 1: Schedule of Events 
Procedures and Evaluations 
Out- 
patient 
In- 
patient 
In- 
patient 
Treatment Period (In-patient) 
Follow-up  
(out-patient) 
Early 
Termina-
tion 
Unsche-
duled 
Day -28  
to -3 
Day -2 
Day -1 
Day 1 
Day 2 
Day 3 
Day 14 
 (± 3 
days) 
Day 30 
(±4 
days) 
Screening 1 
Check-
in 
Baseline 
Pre-
dose 
0 h 
(dose) 
0.5 h 
(±5 
min) 
1 h  
(±5 
min) 
2 h  
(±5 
min) 
4 h  
(±10 
min) 
8 h 
(±15 
min) 
16 h 
(±15 
min) 
24 h 
(±1 
h) 
36 h  
(±1  
h) 
48 h 
(±1 
h) 
60 h 
(±1 
h) 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion and exclusion criteria 
X 2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Review of Inclusion and exclusion criteria 3 
 
X 3 
X 3 
X 3 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history (including family history of auditory loss, 
renal disorders and sudden death) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical History update 4 
 
X 4 
X 4 
X 4 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Prior medication 
X 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Concomitant medication 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Complete Physical examination (PE) 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
 
X 
 
Abbreviated PE 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Symptom-directed (Focused) PE 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
Vital signs (BP, HR, RR, T) 5 
X 
X 
X 
X 
 
X 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
Height, Weight, BMI calculation 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Weight 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
Spirometry (FEV1 and FEV1/FVC) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12-lead ECG (single recording) 6 
X 
X 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
X 
 
X 
X 
12-lead ECG (triplicate recording)7 
 
 
X 7 
X 7 
 
 
X 7 
 
X 7 
 
X 7 
 
 
 
 
 
 
 
 
Viral Serology 8 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Alcohol breathalyzer test 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine drug toxicity and cotinine tests 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical laboratory tests 9 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
Calculation of glomerular filtration rate (CKD-EPI) 10 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
 
Urinalysis 11 
X 
X 
 
X 
 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
Serum β-HCG pregnancy test (females only) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serum FSH (post-menopausal females only) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine pregnancy test (females only) 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood sample for exploratory biomarkers 12 
 
X 
 
X 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
X 
 
Urine sample for exploratory biomarkers 13 
 
  X 
 
X 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
X 
 
Ear otoscopy, Valsalva test, tympanometry, stapedial 
reflexes14 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Ear otoscopy14 
 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
 
Ear audiometry and otoacoustic tests 14 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
 
Admission 15 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Enrollment and Assignment to T cohorts 16 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Discharge 17 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
IV administration of apramycin 18 
 
 
 
 
X-------
21
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Check of infusion site 
 
 
 
 
X 
X 
X 
 
 
 
X 
X 
X 
X 
X 
X 
 
X 
 
Treatment- emergent adverse events (TEAE) and serious 
AEs (SAEs) 19 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Blood (plasma) sampling for apramycin PK 20 
 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
X 
 
Protein Binding Test 21 
 
 
 
 
 
X 
 
 
 
 
 
 
X 
 
 
 
 
 
 
Administration of sedative or anesthetic medication 22 
 
 
 
 
 
X 
 
X 
X 
X 
 
X 
 
 
 
 
 
 
 
Bronchoscopy /BAL (for ELF and AM PK) 23 
 
 
 
 
 
X 
 
X 
X 
X 
 
X 
 
 
 
 
 
 
 
Oxygenation support (TBC) 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
 
 
 
 
Drinking schedule 24 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
X 
 
Counseling 25 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
1. 
Screening including Ear -Nose-Throat (ENT)/Audiology assessments are planned to be completed in a single Screening Visit. If schedule cannot be accommodated, 
ENT/Audiology assessments will be completed on a second visit within the screening period. 
2. 
Inclusion and exclusion criteria regarding ENT/Audiology assessments to be reviewed by audiologist. 
3. 
Review of inclusion and exclusion criteria on Day -2 to determine eligibility for admission; on Day -1, before 12-lead ECG, to determine eligibility for enrollment; and 
on Day 1 pre-dose to confirm the subject remains eligible for dosing. 
4. 
Medical history update to include pre-dose events.  
5. 
Vital signs include systolic and diastolic BP, heart rate, respiratory rate and oral temperature. 
6. 
12-lead ECG single recording with 10 sec rhythm strip.   
7. 
12-lead ECG: triplicate safety ECGs with 10 sec rhythm strips on Day -1 at the same timepoints as the projected timepoints for ECGs to be recorded on Day 1, and on 
Day 1 within 30 min of dosing, and at 1h (± 5 min), 4h (± 10 min) and 16 h (± 15 min) after starting the apramycin dose.  Record ECG before VS and PK draws and, at 
4 h post-dosing, before administration of sedation or anesthetic for BAL in cohort T3.  
8. 
Hepatitis B surface antigen (HBsAg), hepatitis virus C antibody (HCV) and human immunodeficiency virus (HIV) antibody. 
9. 
Clinical laboratory tests under fasting conditions: hematology, coagulation, and clinical chemistry panels and thyroid stimulating hormone (TSH) & free T3/free T4. (See 
Section 8.2.1) 
10. The CKD-EPI equation, expressed as a single equation, is GFR = 141 × min(Scr/κ, 1)α × max(Scr/κ, 1)-1.209 × 0.993Age × 1.018 [if female] _ 1.159 [if black], where 
Scr is serum creatinine, κ is 0.7 for females and 0.9 for males, α is -0.329 for females and -0.411 for males, min indicates the minimum of Scr/κor 1, and max indicates 
the maximum of Scr/κ or 1 [Levey 2009].(Formula will be used and result will be included in the lab report) 
11. Urinalysis (UA) by dipstick. If abnormal for blood, protein and glucose, perform microscopic UA. The results of microscopic UA supersede those of UA by dipstick. UA 
may be postponed in a menstruating female. Abnormal UA in a menstruating female is not graded as a TEAE; the UA should be completed after the end of menstruation. 
12. Cystatin C in serum. 
13. Kidney-injury molecule 1 (KIM-1) in urine. 
14. Ear otoscopy, Valsalva test, tympanometry, stapedial reflexes, and pure-tone audiometry with standard frequencies and distortion product otoacoustic emissions 
(DPOAEs) at screening. Ear otoscopy, and pure-tone audiometry including high frequencies and DPOAEs at check-in (Day -2 [or Day -4 or Day -3 if needed]) and at 
Days 3 (+2 days), 14 (± 3 days), and 30 (± 4 days), or ET. 
15.  Subjects will be admitted to the ward on Day -2 after confirmation of eligibility. 
16. Assignment to Lung PK timepoint cohorts (T1 – T5) and Enrollment to be performed before 12-lead ECG on Day -1. – Subjects with ECGs on Day-1 and pre-dose on 
Day 1 with clinically significant abnormalities according to the PI should not be eligible for receiving study drug and should be withdrawn and replaced. 
17. Subjects may be discharged from the CTU after Day 3 collection of the 60-h plasma PK sample. 
18.  Duration of apramycin infusion is 30 ± 5 min. 
19. Adverse events from the start of dosing to the end of the trial are considered treatment-emergent adverse events (TEAE) or serious AEs (SAE). 
20. Blood samples for plasma PK are taken pre-dose, and at 30 (± 5) minutes (end of infusion) and 1 h (±5 min), 2 h (±5 min), 4 h (±10 min), 8 h (±15 min), 16 h (±15 min), 
24 h (±1 h), 36 h (±1 h), 48 h (±1 h) and 60 h (±1 h) after start of the infusion - Blood samples for plasma PK are taken at the nominal timepoints. If collection of BAL is 
delayed, plasma PK will be collected at the nominal timepoint and again within 5 min after collection of the last BAL sample. In that case, total apramycin concentration 
will be measured in both samples, but only the drug concentration in the delayed plasma PK sample will be used to calculate the Lung PK/Plasma PK ratio.  
21. A protein binding test for the measurement of free apramycin will be performed using aliquots of plasma PK samples collected at 0.5 h (± 5 min) and 36 h (± 1 h) after 
start of infusion.  
22. Type of sedation or light anesthesia medication for bronchoscopy/BAL procedure and local anesthetic TBD by the anesthesia and bronchoscopy protocols. 
23. Each subject is assigned to a single bronchoscopy/BAL procedure. Lung PK samples (for measurement of total apramycin concentration) after initiation of infusion and 
corresponding cohorts (T) are: (Cohort T1), 30 min (±5 min); (Cohort T2), 2 h (±5 min); (Cohort T3), 4 h (±10 min); (Cohort T4), 8h (± 15 min); and (Cohort T5), 24 h 
(± 1 h). Lung PK timepoints are aligned with plasma PK samples collected for the measurement of total apramycin concentration at 30 min (±5 min), 2 h (±5 min), 4 h 
(±10 min), 8 h (±15 min) and 24 h (±1 h) after start of apramycin infusion, If BAL is delayed, plasma sample will be collected at the nominal timepoint and again within 
5 minutes after the last BAL sample collection. Urea will be measured at the same timepoints in plasma and BAL (ELF) samples.  
24. Subjects to be kept hydrated water starting 30 min before until 24 h after start of infusion (document time and volume of water intake). 
25. Provide counseling at the end of each encounter with the subject. Counsel on use of contraception (males and females), avoidance of pregnancy 
(women of childbearing potential), avoidance of sperm donation, and avoidance of prohibited medications, illicit drugs, alcohol, nicotine products, vigorous exercise, and 
exposure to loud noises. 


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Appendix B: Adverse Events Toxicity Grading Criteria 
ABBREVIATIONS: Abbreviations utilized in the Tables: 
ADL = Activities of Daily Living 
 
CTU = Clinical Trial Unit 
ULN = Upper Limit of Normal 
IV    = Intravenous 
ESTIMATING SEVERITY GRADE 
For abnormalities NOT found elsewhere in the Toxicity Tables, use the scale below to 
estimate grade of severity: 
GRADE 1 
Mild 
Events require minimal or no treatment; do not interfere with the 
subject’s daily activities.  
GRADE 2 
Moderate 
Events result in a low level of inconvenience or concern with 
therapeutic measures; may cause some interference with 
functioning.  
GRADE 3 
Severe 
Events interrupt a subject’s usual daily activity and may require 
systemic drug therapy or other treatment; are usually 
incapacitating.  
SERIOUS OR LIFE-THREATENING AEs 
Clinical events considered to be serious or life-threatening include, but are not limited to 
seizures, coma, tetany, diabetic ketoacidosis, disseminated intravascular coagulation, diffuse 
petechiae, paralysis, acute psychosis, and severe depression. 
COMMENTS REGARDING THE USE OF THESE TABLES 
• Standardized and commonly used toxicity tables (Division of AIDS, National Cancer 
Institute [NCI] Common Toxicity Criteria [CTC], and World Health Organization 
[WHO]) have been adapted for use by the Division of Microbiology and Infectious 
Diseases (DMID) and modified to better meet the needs of subjects in DMID trials. 
• For parameters not included in the following Toxicity Tables, the CTU will refer to the 
“Guide for Estimating Severity Grade” located above. 
• Criteria are generally grouped by body system. 
Some protocols may have additional protocol specific grading criteria, which will supersede 
the use of these tables for specified criteria. 
 
 


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Note 1:   No recent hot or cold beverages or smoking. A protocol should select either 0C or 0F for inclusion 
Note 2:  Assume awake and in supine position for 5 min at rest. For TEAE, measurements at least 3 times with 2 
concordant results (See Section 8.1.6) 
Note 3:   Abnormal HR and BP on first measurement may be repeated twice more with the subject resting between 
measurements for at least 5 min (See Section 8.1.6). 
Note 4:   Exceptions to screening BP and HR reference range are: 
(a) Subjects with baseline HR ≥ 45 to 50 bpm may be accepted if they do not have symptomatic bradycardia, 
including syncope, heart disease, ECG abnormalities, or history of syncope or use of medication  
(b) Subjects with baseline SBP up to 140 mmHg and DBP up to 90 mmHg may be accepted if they do not have 
MH of symptomatic hypertension, including paroxysmal hypertension, heart disease, renal disease, CNS 
disease, visual abnormalities or evidence of peripheral vascular disease, ECG abnormalities, use of 
medication, use of illicit drugs or FH of hypertension  
Note 5:   Isolated/individual abnormalities of VS would not be considered toward halting criteria. Abnormalities of 
VS will be described as “increased X” or “decreased X” (X = HR, BP, RR, temperature) if asymptomatic, 
transient and not associated with a systemic or organ-specific disorder and coded by MedDRA within the 
System Organ Class (SOC) “Investigations.” These abnormalities will be graded per criteria in this table, 
but not considered in determining whether study stopping criteria have been met. On the other hand, 
abnormalities of VS that are either secondary to systemic or organ-specific clinical syndrome or primary 
disorders will be coded in the appropriate SOC (e.g., “cardiac disorders”, “respiratory disorders”, 
“immunological disorders”, etc). These abnormalities will be considered in determining whether stopping 
criteria have been met. 
 
Table 2: Toxicity Grading Tables – CLINICAL AEs 
Clinical AEs 
Reference range 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
VITAL SIGNS 1 
 
 
 
 
Fever - °C 
Fever - °F 
36.1 - 37.2 1 
97.0 - 99.0 1 
37.3 - 38.4 
99.1 - 101.1 
38.5 - 38.9 
101.2 - 102.0 
>38.9 
>102.0 
Tachycardia - bpm 
50 - 100 2,3,4,5 
101 - 115 
116 - 130 
>130 or ventricular 
dysrhythmias 
Bradycardia – bpm 
45 - 49 
40 - 44 
<40 
Hypertension (systolic) 
- mmHg 
130/89 2,3,4,5 
131 - 150 
151 - 160 
>160 
Hypertension (diastolic) 
- mmHg 
90 - 95 
96 - 100 
>100 
Hypotension (systolic) - 
mmHg 
85 - 88 
80 - 84 
 <80 
Tachypnea – breaths 
per min 
10 - 20 2,3,4,5 
21 - 25 
26 - 30 
>30 


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Table 2: Toxicity Grading Tables – CLINICAL AEs 
Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
CARDIOVASCULAR 
Arrhythmia 
 
Asymptomatic or transient 
signs; no medical 
intervention required. 
Recurrent and/or persistent 
signs; symptomatic medical 
intervention required. 
Hemorrhage 
Estimated blood loss 
<100 mL. 
Estimated blood loss 
>100mL; no transfusion 
required. 
Blood transfusion required. 
RESPIRATORY 
Cough  
Transient cough no treatment 
required. 
Persistent cough treatment 
required. 
Interferes with daily 
activities. 
Bronchospasm, Acute 
Transient bronchospasm no 
treatment required; FEV1 71-
80% of predicted peak flow. 
Requires treatment; 
normalizes with 
bronchodilator; FEV1 60-
70% of predicted peak flow. 
No normalization with 
bronchodilator; FEV1 <60% 
of predicted peak flow. 
Dyspnea   
Does not interfere with usual 
and social activities. 
Interferes with usual and 
social activities; no treatment. 
Prevents usual and social 
activities OR requires 
treatment. 
EAR AND LABYRINTH DISORDERS  
Hearing Impairment 
 
(Adult enrolled on a 
Monitoring Program (on a 1, 
2, 4, 3, 6, and 8 kHz 
audiogram) 
Threshold shift of 15 - 25 dB 
averaged at 2 contiguous test 
frequencies in at least one 
ear. 
Threshold shift of >25 dB 
averaged at 2 contiguous test 
frequencies in at least one 
ear. 
Threshold shift of >25 dB 
averaged at 3 contiguous test 
frequencies in at least one 
ear; therapeutic intervention 
indicated. 
Tinnitus 
Mild symptoms intervention 
not indicated. 
Moderate symptoms; limiting 
instrumental ADL. 
Severe symptoms; limiting 
self-care ADL. 
Vertigo  
Mild symptoms. 
Moderate symptoms; limiting 
instrumental ADL. 
Severe symptoms; limiting 
self-care ADL. 
GASTROINTESTINAL   
Nausea  
No interference with normal 
activity. 
Some interference with 
normal activity. 
Prevents daily activities. 
Vomiting  
No interference with activity 
OR 1-2 episodes in a 24-h 
period. 
Some interference with 
activity OR >2 episodes in a 
24-h period. 
Prevents daily activity OR 
requires medical intervention. 


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Table 2: Toxicity Grading Tables – CLINICAL AEs 
Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Diarrhea  
2-3 loose or watery stools in 
a 24-h period. 
4-5 loose OR watery stools in 
a 24-h period. 
6 or more loose or watery 
stools in a 24-h period OR 
requires IV hydration OR 
requires medical intervention. 
Oral Discomfort / Dysphagia 
Mild discomfort; no difficulty 
swallowing. 
Some limits on eating 
/drinking. 
Eating/talking very limited; 
unable to swallow solid 
foods. 
LOCAL IV CATHETER REACTION 
IV site reaction 
Not applicable. 
Erythema with associated 
symptoms (e.g., edema, pain, 
induration, phlebitis). 
Ulceration or necrosis; severe 
tissue damage; operative 
intervention indicated. 
SYSTEMIC REACTIONS 
Anaphylaxis ** 
-- 
-- 
Symptomatic bronchospasm, 
with or without urticaria; 
parenteral intervention 
indicated; allergy-related 
edema or angioedema; 
hypotension. 
**Definition: A disorder characterized by an acute inflammatory reaction resulting from the release of histamine and histamine-
like substances from mast cells, causing a hypersensitivity immune response. Clinically, it presents with breathing difficulty, 
dizziness, hypotension, cyanosis, and loss of consciousness, and may lead to death. 
Allergic Reaction  
Pruritus without rash. 
Localized urticaria OR 
requires oral therapy. 
Generalized urticaria OR 
angioedema OR anaphylaxis 
OR requires epinephrine 
Hypersensitivity (including 
drug fever) 
Transient flushing or rash; 
temperature 38.0-38.4°C 
(100.4-101.1°F). 
Rash; flushing; urticaria; 
dyspnea; temperature 38.5 - 
38.9°C (101.2 – 102.0°F). 
Symptomatic 
bronchospasm with or 
without urticaria; parenteral 
medication indicated; 
allergy-related edema or 
angioedema; hypotension; 
temperature >38.9°C 
(>102.0°F).  
Headache  
No interference with activity. 
Repeated use of non-narcotic 
pain reliever for more than 24 
h OR some interference with 
activity. 
Significant; any use of 
narcotic pain reliever OR 
prevents daily activity OR 
requires triptans. 
Fatigue  
No interference with activity. 
Some interference with 
activity. 
Significant; prevents daily 
activity. 


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Table 2: Toxicity Grading Tables – CLINICAL AEs 
Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Myalgia 
No interference with activity. 
Some interference with 
activity. 
Significant; prevents daily 
activity. 
SKIN 
Mucocutaneous 
Erythema, pruritus. 
Diffuse, maculo-papular rash, 
dry desquamation. 
Vesiculation OR moist 
desquamation OR ulceration. 
Pruritus 
No or minimal interference 
with usual social and 
functional activities.  
Greater than minimal 
interference with usual social 
and functional activities.  
Inability to perform usual 
social and functional daily 
activities. 
ALL OTHER CONDITIONS 
Illness or clinical AE (as 
defined according to 
applicable regulations) 
Require minimal or no 
treatment; does not interfere 
with the subject’s daily 
activities. 
Result in a low level of 
inconvenience or concern 
with therapeutic measures; 
may cause some interference 
with functioning. 
Interrupt a subject’s usual 
daily activity and may require 
systemic drug therapy or 
other treatment; are usually 
incapacitating. 
 
 


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Table 3: Toxicity Grading Tables – LABORATORY AEs 
Laboratory AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Blood, serum, or plasma * 
HEMATOLOGY 
 
 
 
Hemoglobin decrease, female – 
g/dL 
9.6 – 11.0 
8.0 – 9.5 
<8.0 
Hemoglobin decrease, male – 
g/dL 
11.0 – 12.9 
8.5 – 10.9 
<8.5 
WBC increase – x103/ µL 
>10.8 – 15.0 
>15.0 – 20.0 
>20.0 
WBC decrease – x103/ µL 
2.4 – <3.4 
1.4 – <2.4 
<1.4 
Neutrophils decrease – x103/ µL 
1.0 – <1.4 
0.75 – <1.0 
<0.75  
Lymphocytes decrease – x103/ µL 
<0.7 –0.4 
<0.4 – 0.3 
<0.30 
Monocytes increase – x103/ µL 
>0.9 – 2.0 
>2.0 – 3.0 
>3.0 
Eosinophils increase – x103/ µL 
>0.4 – 0.75 
>0.75 – 1.0 
>1.0 
Basophils increase – x103/ µL 
>0.2 – 0.5 
>0.5 – 0.8 
>0.8 
Platelets decrease – x103/ µL 
90 – <150 
55 – <90 
<55 
COAGULATION 
 
 
 
PT INR ratio 
>1.2 – 1.8 
>1.8 – 2.1 
>2.1 
Prothrombin Time (PT) - seconds 
>12.0 – 15.0 
>15.0 – 18.6 
>18.6 
Activated Partial Thromboplastin 
Time (APTT) - seconds 
>33.0 – 54.0 
>54.0 – 75.0 
>75.0 
CHEMISTRY 
 
 
 
Sodium decrease – mmol/L 
130 - 133 
124 –129 
<124 
Sodium increase – mmol/L 
145 - 150 
151 – 156 
>156 
Potassium increase – mmol/L 
5.3 – 6.0 
6.1 – 6.5 
>6.5 
Potassium decrease – mmol/L 
3.4– 3.0  
2.5 – 2.9 
<2.5 
Total Carbon Dioxide (CO2) 
increase – mmol/L 
30 - 35 
36 - 37 
>37 
Total Carbon Dioxide (CO2) 
decrease – mmol/L 
17 - 19 
14 - 16 
<14 
Calcium decrease – mg/dL 
7.8 – 8.6 
7.0 – 7.7 
<7.0 
Calcium increase – mg/dL 
10.3 – 11.4 
11.5 – 12.5 
>12.5 
Magnesium decrease – mg/dL 
1.2 – 1.5 
1.1 – 0.9 
<0.9 


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Table 3: Toxicity Grading Tables – LABORATORY AEs 
Laboratory AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Blood urea nitrogen (BUN) 
increase, 18-39Y - mg/dL 
21 - 58 
59 – 120 
>120 
Blood urea nitrogen (BUN) 
increase, 40-59Y – mg/dL  
25 - 58 
59 – 120 
>120 
Glucose decrease, fasting – 
mg/dL 
55 - 69 
40 – 54 
<40 
Glucose increase, fasting – mg/dL 
100 – 160 
161 – 250 
>250 
Creatinine increase, male – 
mg/dL 
1.28 – 1.90 
1.91 – 3.80 
>3.80 
Creatinine increase, female – 
mg/dL 
0.58 – 1.50 
1.51 – 3.00 
>3.00 
Direct bilirubin 
0.41 – 0.70 
0.71 – 1.20 
>1.20 
Total bilirubin (serum) increase – 
mg/dL (with other LFTs in the 
normal range) 
1.3 – 2.0 
2.1 – 2.5 
>2.5 
Total bilirubin (serum) increase – 
mg/dL (accompanied by a >3 x 
ULN increase in ALT or AST) 
1.3 – 1.7 
1.8 – 2.4 
>2.4 
Total protein decrease – g/dL 
5.1 – 5.9 
4.6 – 5.0 
<4.6 
Albumin, decrease, Male, 18-30Y 
- g/dL 
4.0 – 3.0 
2.0– 2.9 
<2.0 
Albumin decrease, Female,  
18-30Y – g/dL 
3.8 – 3.0 
2.0– 2.9 
<2.0 
Albumin decrease, Male,  
31-50Y – g/dL 
3.9 – 3.0 
2.0– 2.9 
<2.0 
Albumin decrease, Female,  
31-50Y – g/dL 
3.7 – 3.0 
2.0– 2.9 
<2.0 
AST increase – U/L 
41 – 80 
81 – 120 
>120 
ALT increase, male – U/L 
45– 88 
89 - 132 
>132 
ALT increase, female – U/L 
33 – 64 
65 – 96 
>96 
Alkaline phosphatase (AP) 
increase, males 18-20Y – U/L 
126 – 250 
251 – 375 
>375 
Alkaline phosphatase (AP) 
increase, females, 18-20Y – U/L 
107 – 250 
251 – 375 
>375 
Alkaline phosphatase (AP) 
increase, 21-150Y – U/L 
122 – 250 
251 – 375 
>375 


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Table 3: Toxicity Grading Tables – LABORATORY AEs 
Laboratory AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Urine  
URINALYSIS by 
Dipstick 
 
 
 
Protein 
1+ 
2+ 
>2+ 
Blood (occult) 
1+ 
2+ 
>2+ 
Glucose 
1+ 
2+ 
>2+ 
URINE 
MICROSCOPY 
 
 
 
Red blood cells (RBC) per HPF 
3-10 
11-30 
>30 and/or gross blood 
White blood cells (WBC) per 
HPF 
6-10 
11-30 
>30 and/or symptomatic 
urogenital infection 
Bacteria (microscopic) 
few 
moderate 
many 
Note 1:  With the exception of AST, ALT, AP, bilirubin, electrolytes, urine protein, BUN and creatinine, which should be within 
reference range, lab values of other analytes in the grade 1 range are acceptable for enrollment if (a) they are not 
considered to be clinically significant by the investigator and (b) there is no cluster of abnormal labs that combined are 
suggestive of an underlying disorder. 
Note 2: Other Exceptions to screening laboratory tests’ normal reference ranges are: 
a. 
Racially based low total WBC or neutrophil counts up to toxicity Grade 1 are allowed, but toxicity Grades 2 or 3 are 
exclusionary. 
b. 
Labs performed as part of a panel but not listed above are to be recorded in the database. If abnormal, they are not 
exclusionary and are not to be graded per Toxicity table, however, the investigator would make a clinical decision 
about their clinical significance and, if clinically significant, they will be graded according to the criteria in 
Section 9.2.1 . (Examples include mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean 
corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), mean platelet volume (MPV), and 
nucleated red blood cell count (NRBC CT), which are included in a CBC with differential.)  
Note 3: If a subject was accepted into the trial with a laboratory value of an analyte that overlaps with values used for grading 
Grade 1 laboratory abnormalities, a TEAE will be reported if the on-study value of the same analyte increases to 
Grade 2 or higher. 
Note 4: If the dipstick UA is abnormal, a microscopic UA will be performed, and the results will supersede the results of the 
dipstick UA. 
Note 5:  Menstruating females with a positive dipstick UA or microscopic UA may be retested following cessation of menses. 
Note 6: Isolated laboratory values that are outside the range of eligibility but are thought to be due to an acute condition or due 
to collection or laboratory error may be repeated once. 
 
 


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Table 4: Toxicity Grading Tables – ECG 
ECG interval 
abnormality 
Reference 
range 
Grade 1 
Grade 2 
Grade 3 
QTcF interval 
prolonged (msec): 
• 
Male 
• 
Female 
•  ≤ 450 msec 
•  ≤ 470 msec 
Asymptomatic,  
QTcF  
• 451  ̶  479 msec  
• 471  ̶  479 msec 
Asymptomatic,  
QTcF 480-500 msec  
OR  
increase in interval 30-
59msec above baseline 
Asymptomatic,  
QTcF >500 msec  
OR 
increase in interval 
≥60 msec above baseline 
PR interval prolonged 
(msec) 
≤210 msec 
211-250 msec 
>250 msec 
Type II 2nd degree AV 
block  
OR  
ventricular pause >3.0 sec 
Note 1: Events will be coded as treatment-emergent SAEs if there are life-threatening associated symptoms or signs (arrhythmia, 
CHF, hypotension, syncope, TdP, etc.). 
Note 2: If a male subject was accepted into the trial with a QTcF value that overlaps with values used for grading Grade 1 QTcF 
prolongation, a TEAE will be reported if the QTcF value is higher than the baseline value. 


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Appendix C: Blood Volume Withdrawn During the Trial 
Table 5: Laboratory Samples and Estimated Total Blood Volume (mL) 
Study Periods 
Out-
patient 
In-
patient  
In-
patient 
In-patient Dosing Period (Days 1 – 3) 
Out-patient 
ET 
Study Visit 
Screen 
Check-
in 
Baseline 
Dosing 
Follow-up 
Discharge 
Follow-
up 
Final 
Study Day a 
-28 to -3 
-2 
-1 
1 
2 (24 h) 
2 (36 h) 
3 (48 h) 
3 (60 h) 
14 (±3 
days) 
30 (±4 
days) 
HEMATOLOGY 1 
4.0 
4.0 
 
4.0 
4.0 
4.0 
4.0 
4.0 
4.0 
 
4.0 
CHEMISTRY and serum β-HCG and FSH 1 
8.5 
8.5 
 
8.5 
8.5 
8.5 
8.5 
8.5 
8.5 
 
8.5 
COAGULATION 1 
2.7 
2.7 
 
2.7 
2.7 
2.7 
2.7 
2.7 
2.7 
 
2.7 
TSH, Free T4, Free T3 1 
3.5 
3.5 
 
3.5 
3.5 
3.5 
3.5 
3.5 
3.5 
 
3.5 
Viral Serology (HIV, HBsAg, HCV) 2 
7.0 
 
 
 
 
 
 
 
 
 
 
Exploratory Marker (serum Cystatin) 3 
 
3.5 
 
3.5 
 
 
3.5 
 
3.5 
3.5 
3.5 
PK 4 
 
 
 
56.0 
8.0 
8.0 
8.0 
8.0 
 
 
8.0 
Total volume/visit 
25.7 
22.2 
0 
78.2 
26.7 
26.7 
30.2 
26.7 
22.2 
3.5 
30.2 
Cumulative total volume 
25.7 
47.9 
47.9 
126.1 
152.8 
179.5 
209.7 
236.4 
258.6 
262.1 
 
a Study Days shown correspond to days in each study period. For a view of the cumulative numbering of study days, please refer to Section 7. 
1 Clinical Safety blood tests (HEM, CHEM, COAG, TSH, fT4, fT3) are drawn at Screening Visit, on Day -2, and on Days 1, 2, 3 and 14 (± 3 days), or ET. Serum pregnancy test 
in all women is drawn at Screening. FSH is drawn only at Screening in post-menopausal women. 
2 Viral serology tests are drawn at Screening. 
3 Serum Cystatin test is drawn on Day -2, Day 3 (at 48 h) and Day 14 (± 3 days). 
4 PK plasma samples are drawn on Day 1 (30 min before dosing and at 0.5, 1, 2, 4, 8, 16 h after dosing); on Day 2 (24 and 36 h after dosing) and on Day 3 (48 and 60 h after 
dosing), or ET if occurs within 24 h of dosing.