Microbiome Effect of Omadacycline on Healthy Volunteers
Phase: PHASE1
Conditions: Healthy Volunteers
Sponsor: University of Houston

Microbiome Effect of Omadacycline on Healthy Volunteers 
Jinhee Jo, Anne J Gonzales-Luna, Khurshida Begum, Chris Lancaster, M Jahangir 
Alam, and Kevin W. Garey 
University of Houston College of Pharmacy 
NCT number: Pending 
August 30, 2023 
 
 

 
Background and summary:  Omadacycline is a novel aminomethylcycline antibiotic approved 
for the treatment of community-acquired bacterial pneumonia and acute skin and skin structure 
infections.  In clinical trials, a remarkably low number of cases of Clostridiodies difficile infection 
(CDI) cases were observed in patients given omadacycline vs. comparators.  In collaboration 
with the University of Houston College of Pharmacy, Dr. Garey and his research team have 
shown excellent in vitro susceptibility to a large number of C. difficile isolates.  The clinical 
question that arises from this work is whether omadacycline should be considered the drug of 
choice in patients at risk for CDI?  An effective antibiotic that does not increase the likelihood of 
CDI should have potent in vitro activity to C. difficile and also have a favorable microbiome 
disruption profile that minimizes dysbiosis. The purpose of this study is to define the microbiome 
effect of omadacycline.  We will do this by conducting a phase I, healthy volunteer study.  With a 
unique spectrum of activity that should limit destruction to the host gut microbiome, 
omadacycline is poised to become the drug of choice for patients at risk for CDI.   
Specific aim 1.  To identify microbiome changes associated with omadacycline vs. 
moxifloxacin.       
Sub-aim 1.1. Quantitate changes over time on host fecal microbiota biomass of specific microbial 
families (Eubacteria and 5 different microbiota groups; Bacteroides, C. coccoides, C. leptum, 
Enterobacteriaceae, and Prevotella) using qPCR in healthy subjects given a 10-day course of 
omadacycline or comparators (moxifloxacin and oral vancomycin) 
Sub-aim 1.2.  Assess relative abundances of resident taxa, microbiota diversity, and taxonomic 
composition using high-throughput sequencing in health subjects given a 10-day course of 
omadacycline or comparators (moxifloxacin and oral vancomycin) 
 
Significance:  Using advanced microbiologic genomics analysis, this will be the defining study 
to assess the microbiome effects of omadacycline on the human gut microbiome in direct 
comparison to moxifloxacin and vancomycin.   
 
 

Methods: 
General overview:  The goal of this study is to assess microbiome changes associated with 
omadacycline vs. comparators (moxifloxacin or oral vancomycin).  Using a phase I study 
design, healthy volunteers aged 18-40 years will be given a 10-day course of either 
omadacycline 450 mg days 1 and 2 followed by 300 mg PO once daily, moxifloxacin 400 mg 
once daily, or oral vancomycin 125 mg given four times daily.  Stool will be collected at baseline, 
daily during therapy, and at two follow-up time periods (days 13-14 and days 30-32).  DNA will 
be extracted from stool and used for qPCR biomass and microbial metagenomic experiments.  
Results from the study will provide definitive data on the microbiome effects of omadacycline 
versus comparator antibiotics. 
Overview of clinical trial, inclusion and exclusion, and sample collection:  A 10-day, oral course 
of omadacycline, moxifloxacin, or vancomycin will be randomly assigned to healthy volunteers 
aged 18-40. All volunteers will be considered healthy based on medical history, none will have a 
history of cardiovascular, gastrointestinal, hepatic, or renal disease.  Subjects will not have 
taken an antibiotic for at least three months prior to enrollment.  Patients will not have taken a 
probiotic for at least a month prior to enrollment and during the entire study period.  After 
screening for inclusion and exclusion and signing an informed consent, fecal samples will be 
collected at baseline prior to dosing, and on days 1 to 10.  Follow-up stool samples will be 
collected between days 13-14 and days 30-32. Instructions on collection will be provided to all 
volunteers. 
Safety assessment:  See appendix 1 
Sample size: Eight subjects, each will be randomized to receive either omadacycline, 
moxifloxacin, or vancomycin (total subjects = 24). 
Drug supply:  Omadacycline: Subjects will receive 3 tablets on day 1 and day 2 (450 mg) 
followed by 2 tablets (300 mg) for days 3 through 10.  Thus, each subject will require 22 tablets.  
Total numbers of tablets required is 176 tablets (22 tablets per subjects X 8 subjects = 176). 
Comparator antibiotics:  Comparator antibiotics will be purchased from a local distributor.  
Moxifloxacin 400 mg tablets will be taken once per day for a total of 10 days.  Thus, 10 tablets 
per subject will be required for a total of 80 tablets.  Vancomycin 125 mg capsules will be taken 
four times daily for a total of 10 days.  Thus, 40 capsules per subjects will be required for a total 
of 320 capsules. 
Missed dose:  If a dose of medication is missed, it will be given as soon as possible anytime up 
to 8 hours prior to the next scheduled dose. If less than 8 hours remains before the next dose, 
then dosing will occur at the next scheduled dose. A double dose will not be given to 
compensate for a missed dose. Subjects given vancomycin will follow the same protocol except 
a cut off time of three hours will be chosen.  If the patient vomits following oral dosing and any 
tablet is visible, then another dose will be taken immediately. If the tablet is not visible, then the 
next scheduled dose will to be taken as planned. 
Clinical trial supplies and sample collection:  Volunteers will be instructed to collect the first stool 
sample passed each day into appropriate collection containers.  The Collection accessory for 
use with OMNIgene GUT (DNA Genotek, Ottawa, CA) will be used for all DNA studies while the 

Fisher Scientific commode collection container will be used for microbiologic studies.  Samples 
will be brought to the research lab within 12 hour of collection.   
Laboratory procedures: For each sample, weight and consistency (Bristol Stool chart) will be 
recorded.   Approximately 500 mg of sample will be aliquoted into the OMNIgene GUT OMR-
200 tube (DNA Genotek, Ottawa, CA) and stored via the manufacturer’s directions.  The 
remaining fecal samples will be aliquoted into appropriate storage containers and stored at -80C 
until analysis. 
Stool DNA extraction:  Prior to DNA extraction, the OMNIgene stool tube will be vortexed 
vigorously for at least 10 seconds.  DNA will be extracted from the OMNIgene stool tube by 
aliquoting 300 μL of the tube sample using the AnaPrep system DNA extractor (AnaPrep 2, 
BioChain Instruments, Newark CA) and the following protocol.  Approximately 300 mg stool will 
be dissolved in 250 μL of 1X PBS, homogenized for 60 seconds in a bead homogenizer (Bead 
Blaster24, Benchmark) for cell lysis, and added to 250 uL of lysis buffer BL2B with RNase A 
(Bacterial DNA Extraction Kit, BioChain Inc.).  Approximately 400 uL of lysed bacteria will be 
mixed and loaded into the AnaPrep system and gDNA will be extracted using 400 microliter 
input volume and 100 uL elution volume.  DNA will be eluted in 100μL of buffer and 
concentration measured using a Nanodrop 2000 (Fisher Scientific, Waltham, MA).  If the DNA 
concentration is less than 10 μg/mL, the extraction will be repeated. A 50 μL portion of the 
extracted DNA will be used for qPCR analysis and the remainder used for 16S rDNA amplicon 
generation.  
Methods for specific aim 1 
Quantitative PCR (qPCR) analysis: Ethanol precipitation of the extracted DNA will be done to 
eliminate inhibitors.  The reconstituted DNA will be checked for purity and concentration using a 
Nanodrop 2000 (Fisher Scientific, Waltham, MA).  Sample DNA will then be diluted with PCR 
grade water to 5 ng/μL. The DNA levels of bacterial groups will be assessed using the PCR 
primers/conditions used in the ridinilazole phase II study (Table 1) and published references [1-
4]. Using the Viia 7 (Fisher Scientific, Waltham, MA), qPCR will be performed on each sample in 
triplicate in a final volume of 20 μL containing 25 ng DNA template, primers at 0.3 μM, and 
SYBR Green 2x Master Mix (QIAGEN), with a FAM-tagged probe at 0.25 μM for Eubacteria. 
Threshold cycle values will be converted to copies per ng of DNA using a standard curve. 
Standards will be prepared by performing PCR using species specific primers on appropriate 
bacterial strains or DNA from normal stool. The PCR products will be cloned using Invitrogen 
TOPO PCR Cloning Kit (Fisher Scientific, Waltham, MA), and verified by sequencing at the 
University of Houston Core Facility. A Basic Local Alignment Search Tool (BLAST) search will 
be performed to identify the closest matching database sequence (Table 2). A range of 10-fold 
serially diluted plasmid standard DNA (5 X 108 to 500 copies) will be run on each qPCR plate in 
triplicate. Standard curve R2 values will be calculated for standards.  Copies per gram of stool 
will be calculated, accounting for initial sample DNA concentrations and stool weights. The 
change in bacterial levels (Δlog10 copies/ gram stool) from entry level to each available 
successive time-point will be determined for each participant and median changes calculated  
High-throughput metagenomics sequencing:  Microbiome characterization will be performed by 
sequencing the V4 region of the 16S rRNA gene followed by extensive bioinformatics analysis 
related to microbial composition, diversity, and community structure. Microbial DNA extraction, 
16S rRNA gene-based next-generation sequencing (using the Illumina MiSeq), quality filtering, 

and microbiome-based analysis will be performed [5].  Each sample will be amplified using a 
barcoded primer, which yields a unique sequence identifier tagged onto each individual sample 
library. Illumina-based sequencing will yield > 15,000 reads per sample.  Operational taxonomic 
units (OTUs) will be defined as >99% identity. To analyze differences between treatment groups 
and across timepoints, the OTU table will be normalized to the lowest number of sequences per 
sample, then consolidated by summing to species level and to each successively higher 
taxonomic level. The alpha diversity parameters Chao and Shannon (based on number of 
OTUs) and Phylogentic Diversity (PD) (based on phylogenetic relationships), will be generated 
in QIIME or R; alpha diversity is widely regarded as an indication of microbiota health. For beta 
diversity, which describes the relationships between samples, weighted Unifrac distance 
matrices will be generated in QIIME.  
Statistical analyses.  In general, the healthy volunteer analyses will assess changes in total 
and species-specific microbiota between groups and over time as well as changes in microbial 
diversity.  For both qPCR and high-throughput sequencing data, significance differences 
between treatment groups at each time point will be calculated using Mann-Whitney U test while 
within each treatment arm, significance of differences between time-points will be assessed by 
the Wilcoxon Signed Rank test.  Analyses will be performed using SAS (SAS Institute, Cary NC) 
or R on log-transformed data. For high-throughput sequencing data, the LEfSe (Linear Effect 
Size) algorithm [6] will be used to identify significant differences in microbiota composition 
between baseline and each study time point, followed by Wilcoxon Signed Rank tests on 
identified taxa. The Benjamin-Hochberg procedure will be used to control the false discovery 
rate at 0.10 [7]. The MaAsLin algorithm will be used to find associations of taxa with treatment. 
For beta diversity, principal co-ordinates analyses on weighted Unifrac distance matrices 
generated in QIIME will be performed in the vegan package in the statistical program R. Finally, 
using the methods described by Burdet at al, Shannon diversity (a measure of alpha diversity) 
will be compared at each time point for subjects that receive moxifloxacin or comparators.[8]  
For qPCR data, to analyze whether trajectories across time differed between treatment groups, 
repeated measures models with time and group as categorical fixed factors [9] will be used on 
log-transformed data, with diagnostics to assess the impact of potential influential points.  
Deliverables: Results from this study will be presented at a major scientific meeting (ID-week, 
ECCMID, for example and submitted to a top tier medical journal (Clinical Infectious Diseases / 
Journal of Antimicrobial Chemotherapy, for example). 
Budget (total costs):  $204,725 (detailed budget attached) 
Timeline:  1-year 
Budget justification:    
Dr. Gonzales-Luna will be the lead for all day to day activities on the grant.  She will also be 
responsible for the healthy volunteer study with Dr. Lancaster.  Drs. Begum and Alam will be 
responsible for all qPCR and metagenomics work in the study. Dr. Gonzales-Luna will also be 
responsible for analyses, generation of study reports, and first draft of the study manuscript. Dr. 
Garey will have oversight of the entire project as Principle Investigator.  $77,781 in salary 
support is requested.  
Supply costs are calculated $125 qPCR costs per sample and 16S rRNA (microbiome) costs of 
$125/sample.  qPCR includes six relevant microbiota species.  $28,800 and $38,400 are 

requested for sequencing costs for the healthy volunteer.  $10,000 is requested for clinical 
supplies and recruitment requirements for the phase I study along with study drug 
(omadacycline). Travel budget of $2500 is requested to present the results of the study at a 
major scientific meeting.  Finally, $47,244 is requested for university indirect costs (mandatory 
per university policy).   
 
 

Detailed budget: 
  
Personnel 
 Costs 
Kevin Garey, PI 
$11,290  
M Jahangir Alam, Co-I 
$16,318  
Khurshida Begum, Co-I 
$16,318  
Anne J. Gonzales-Luna, Co-I 
$33,855  
  
  
Total Salary 
$77,781  
Material and Supplies 
  
  
Experiments 
  
Phase I clinical study supplies 
$10,000  
Healthy volunteer microbiome studies 
  
qPCR 
$28,800  
  
Metagenomics 
$38,400  
Travel (scientific meeting) 
$2,500  
Total Materials & Supplies 
$79,700  
Total Direct Costs 
$157,481  
  
  
IDC Calculation 
  
IDC Base 
Direct costs 
$157,481  
Total IDC 
30% 
$47,244  
Total Costs 
$204,725  
 
 

References: 
1. 
Bernhard AE, Field KG. A PCR assay To discriminate human and ruminant feces on the 
basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. Appl 
Environ Microbiol 2000; 66(10): 4571-4. 
2. 
Louie TJ, Cannon K, Byrne B, et al. Fidaxomicin preserves the intestinal microbiome 
during and after treatment of Clostridium difficile infection (CDI) and reduces both toxin 
reexpression and recurrence of CDI. Clin Infect Dis 2012; 55 Suppl 2: S132-42. 
3. 
Matsuki T, Watanabe K, Fujimoto J, et al. Development of 16S rRNA-gene-targeted group-
specific primers for the detection and identification of predominant bacteria in human 
feces. Appl Environ Microbiol 2002; 68(11): 5445-51. 
4. 
Bartosch S, Fite A, Macfarlane GT, McMurdo ME. Characterization of bacterial 
communities in feces from healthy elderly volunteers and hospitalized elderly patients by 
using real-time PCR and effects of antibiotic treatment on the fecal microbiota. Appl 
Environ Microbiol 2004; 70(6): 3575-81. 
5. 
Fadrosh DW, Ma B, Gajer P, et al. An improved dual-indexing approach for multiplexed 
16S rRNA gene sequencing on the Illumina MiSeq platform. Microbiome 2014; 2(1): 6. 
6. 
Segata N, Izard J, Waldron L, et al. Metagenomic biomarker discovery and explanation. 
Genome Biol 2011; 12(6): R60. 
7. 
Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful 
approach to multiple testing. Journal of the Royal Statistical Society Series B 
(Methodological) 1995; 57(1):289±300. 
8. 
Burdet C, Nguyen TT, Duval X, et al. Impact of Antibiotic Gut Exposure on the Temporal 
Changes in Microbiome Diversity. Antimicrob Agents Chemother 2019; 63(10). 
9. 
Diggle PJ, Liang K-L, Zeger SL. Analysis of Longitudinal Data Oxford, UK: Oxford 
University Press;1996. 
 

 
Table 1.  DNA primers used for qPCR to quantify changes in major components of microbiota 
 
 
Group 
Primer name 
Sequence (5’-3’) 
Reference 
Bacteroides 
Bac303F 
GAA GGT CCC CCA CAT TG 
Bernhard et al, 2000 (10) 
Bac708R 
CAA TCG GAG TTC TTC GTG 
Clostridium coccoides 
Erec482R 
GCT TCT TAG TCA RGT ACC G 
Louie et al, 2012 (11) 
Eub338F 
ACT CCT ACG GGA GGC AGC 
Clostridium leptum 
Sg-clept-F 
GCA CAA GCA GTG GAG T 
Matsuki et al, 2002 (12) 
Sg-clept-R 
CTT CCT CCG TTT GTC AA 
Enterobacteriaceae 
Eco1457F 
CAT TGA CGT TAC CCG CAG AAG AAG C 
Bartosch et al, 2004 (13) 
Eco1652 
CTC TAC GAG ACT CAA GCT TGC 
Eubacteria 
8F 
AGT TTG ATC CTG GCT CAG 
Jiang et al, 2009 (14) 
 
515R 
GWA TTA CCG CGG CKG CTG 
 
338P 
FAM GCT GCC TCC CGT AGG AGT BHQ1 
Prevotella 
CFB286F 
GTA GGG GTT CTG AGA GGA 
Louie et al, 2012 (11) 
CFB719R 
AGC TGC CTT CGC AAT CGG 
 
 
 

Table 2. Sequences of plasmid standards for qPCR 
 
 
 
Primer Set 
Referenced 
Organism 
Length 
Match 
Sequence 
Bacteroides 
Bacteroides sp. 
HGA0257 16S 
ribosomal RNA 
gene, partial 
sequence 
419 
99% 
GAAGGTCCCCCACATTGGAACTGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAAT
ATTGGTCAATGGGCGAGAGCCTGAACCAGCCAAGTAGCGTGAAGGATGACTGCCCTATGGGTTG
TAAACTTCTTTTATAAAGGAATAAAGTCGGGTATGGATACCCGTTTGCATGTACTTTATGGATAAG
GATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATCCGAGCGTTATCCGGATTTAT
TGGGTTTAAAGGGAGCGTAGATGGATGTTTAAGTCAGTTGTGAAAGTTTGCGACTCAACCGTAAA
ATTGCAGTTGATACTGGATATCTTGAGTGCAGTTGAGGCAGGCGGAATTCGTGGTGTAGCGGTG
AAATGCTTAGATATCACGAAGAACTCCGATTG 
Clostridium 
coccoides 
Blautia coccoides 
strain DSM 29138 
16S ribosomal 
RNA gene, 
139 
 
95% 
 
ACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCAACGC
CGCGTGAGTGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGAAAGAAAATGACGGTACCT
GACTAAGAAGC 
Clostridium 
leptum 
 
Clostridium leptum 
16S rRNA gene, 
strain DSM 753T 
Clostridium leptum 
ATCC 29065 
241 
 
90% 
 
GCACAAGCAGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACAT
CGAGTGACGAACATAGAGATATGTTCTTCCTTCGGGACACGAAGACAGGTGGTGCATGGTTGTC
GTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCATTAGTTGC
TACGCAAGAGCACTCTAATGAGACTGCCGTTGACAAAACGGAGGAAG 
Entero-
bacteriaceae 
 
Escherichia coli 
JJ1887, complete 
genome 
 
190 
 
100% 
 
CATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAG
GGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTAAGTCAGATG
TGAAATCCCCGGGCTCAACCTGGGAACTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAG 
Eubacteria 
 
Escherichia coli 
str. K-12 substr. 
MG1655 strain K-
12 16S ribosomal 
RNA, complete 
sequence 
(EUBACTERIA) 
 
517 
 
100% 
 
AGTTTGATCATGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAACGGTAAC
AGGAAGAAGCTTGCTTCTTTGCTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGAAACTGCCT
GATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAACGTCGCAAGACCAAAGAGG
GGGACCTTCGGGCCTCTTGCCATCGGATGTGCCCAGATGGGATTAGCTAGTAGGTGGGGTAACG
GCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACA
CGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGC
AGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGA
GTAAAGTTAATACCTTTGCTCATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAG
CAGCCGCGGTAATAC  
Prevotella 
 
Prevotella 
denticola gene for 
16S ribosomal 
RNA, partial 
sequence, strain: 
JCM 8528 
 
446 
 
86% 
 
GTAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACACGGTCCAAACTCCTACGGGA
GGCAGCAGTGAGGAATATTGGTCAATGGGCGAGAGCCTGAACCAGCCAAGTAGCGTGAAGGAT
GACTGCCCTATGGGTTGTAAACTTCTTTTATAAAGGAATAAAGTCGGGTATGGATACCCGTTTGCA
TGTACTTTATGAATAAGGATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATCCGA
GCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGATGGATGTTTAAGTCAGTTGTGAAAGTTT
GCGGCTCAACCGTAAAATTGCAGTTGATACTGGATATCTTGAGTGCAGTTGAGGCAGGCGGAATT
CGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCT 

Appendix 1  Safety assessments. 
Adverse Events (AE), Serious Adverse Events (SAE), and Serious Adverse Drug Events (SADE):  
Definitions and Reporting Procedures:  
Adverse Events (AE)  
Definition: an adverse event is defined as any event that was not present at baseline.  
Criteria for Assessing Severity  
The Investigator will evaluate the comments of the subject and the response to antibiotic treatment 
in order to judge the nature and severity of the adverse event. Severity refers to the intensity of 
discomfort/impairment of health and will be assessed according to the following criteria:  
Mild: Awareness of sign, symptom, or event, but easily tolerated  
Moderate: Discomfort enough to interfere with usual activity and may warrant intervention  
Severe: Incapacitating with inability to do usual activities or significantly affects clinical status and 
warrants intervention  
To clarify the difference between the terms "serious" and "severe", which are not synonymous, the 
following note is provided:  
The term "severe" is often used to describe the intensity (severity) of a specific event (as in mild, 
moderate, or severe myocardial infarction); the event itself, however, may be of relatively minor 
medical significance (such as severe headache). This is not the same as "serious", which is based 
on patient/event outcome or action criteria usually associated with events that pose a threat to a 
patient's life or functioning. Seriousness (not severity) serves as a guide for defining regulatory 
reporting obligations.  
Criteria for Assessing Causality  
The question of the relationship of an adverse event to study drug will be determined by the 
Investigator after thorough consideration of all facts that are available. Assessment of causality is 
based on considering associative connections (time or place), pharmacological explanations, 
previous knowledge of the drug, presence of characteristic clinical or pathological phenomena, 
exclusion of other causes, and/or absence of alternative explanations. The causal relationship of 
an adverse event to study drug will be assessed according to the following criteria:  
The Investigator will be responsible for determining the relationship between the administration of 
study drug and the occurrence of an AE as not suspected or suspected.  
Not suspected: The temporal relationship of the adverse event to study drug administration makes 
a causal relationship unlikely, or other drugs, therapeutic interventions, or underlying conditions 
provides a sufficient explanation for the observed event.  
Not related: Temporal relationship to study drug administration is missing or implausible, or there 
is evidence of another cause.  
Unlikely related: Temporal relationship to study drug administration makes a causal relationship 
improbable; and other drugs, chemicals, or underlying disease provide plausible explanations.  

Suspected: The temporal relationship of the adverse event to study drug administration makes a 
causal relationship possible, and other drugs, therapeutic interventions, or underlying conditions 
do not provide a sufficient explanation for the observed event.  
Possibly related: Reasonable time sequence to administration of study drug, but the event could 
also be explained by concurrent disease of other drugs or chemicals. Information on drug 
withdrawal may be lacking or unclear.  
Definitely related: Plausible time relationship to study drug administration; event cannot be 
explained by concurrent disease or other drugs or chemicals. The response to withdrawal of the 
drug (dechallenge) should be clinically plausible. The event must be definitive pharmacologically 
or phenomenologically, using a satisfactory re-challenge procedure if necessary.  
Reporting of Adverse Events  
All adverse events from the time of randomization will be recorded in the Adverse Events section of 
the case report form. All adverse events will be described in terms of duration, severity, possible 
relation to study medications and treatment. Previously documented side effects of each medication 
will be listed in each medication’s respective section.  
All adverse events will be followed until resolution; or for 30-days following the last dose of study drug 
or up until the last protocol-specified study visit, whichever occurs later.  
If the adverse event meets any of the criteria for Serious Adverse Events, the event will also be 
documented on a separate SAE Form.  
Serious Adverse Event (SAE)  
 
Definition: serious adverse events are defined as the following  
 
• Death  
 
• Life Threatening  
 
• Requiring prolonged hospitalization (greater than one week)  
 
• Permanently disabling  
 
• Causing cancer  
 
• Congenital anomaly/ birth defect  
 
• Any other clinically significant event  
 
Reporting of SAE  
In the event of a Serious Adverse Event, information will be collected and recorded on the Serious 
Adverse Event section of the case report form.  
 
Serious Adverse Events will be reported from the time informed consent is obtained through 30-days 
following the last dose of study drug or to the last protocol-specified study visit; whichever occurs later. 
In addition, any SAE occurring after the observation period will be promptly reported if causal 
relationship to study drug is suspected. Serious adverse events will be followed until the event 
resolves or the event or its sequelae stabilize.  
Serious Adverse Event (SAE) Reporting to IRB: The Principal Investigator will be responsible for 
reporting any SAE thought to be directly attributable to the study medication to the IRB.