Conditions: Asthma
Keywords: Bioequivalence Fluticasone Propionate
Allocation: RANDOMIZED | Intervention model: CROSSOVER
Enrollment: 24
Healthy volunteers: yes

Brief summary:
When a drug company first develops a drug, the company has to show the Food and Drug Administration (FDA) that the drug is safe and effective. If FDA concludes that the drug is safe and effective, FDA approves the drug. The company can then sell the drug, which the company does using "trade name." Only the drug company that developed the "trade name" drug is allowed to sell it. However, other drug companies can create their own version of the "trade name" drug, which usually happens after the patents for the "trade name" product run out. These drugs, often called "generic drugs," potentially will be less expensive for the patient. In order to sell generic drugs, drug companies must show that their generic version is the same as the "trade name" drug in a number of ways. For example, they generally have to show that their product is intended to be used to treat the same diseases or conditions, that it has the same label, and that the product has the same active ingredient as the "trade name" drug. The generic company also has to show that generic product is "bioequivalent" to the trade name drug, meaning that the generic product gets to the part of the body where the drug works at the same rate that the trade name drug does. How to show how much drug gets to the part of the body where it works, and how fast, depends on the type of product the drug is. The primary aim of this research study is to aid the FDA in finding methods to ensure that the versions of generic drugs that are inhaled (for example, drugs used to treat asthma) are bioequivalent to the trade name drug. As a part of the research study, pharmacokinetic (PK) studies (studies measuring drug levels in the blood over time after inhalation) will be done using three different versions of fluticasone propionate (FP, a drug routinely used in asthmatic patients) administered using a dry powder inhaler (DPI, an inhalation device that delivers the drug as a dry powder). The results from this study will help FDA ensure that generic products are the same as the trade name drugs.

Detailed description:
Formulation Development

The aim of the pharmaceutical development was to manufacture three Dry powder Inhalation (DPI) formulations containing Fluticasone Propionate in an Plastiape Monodose DPI device, which provide distinct in vitro deposition patterns on a Next Generation Impactor. NGI is a high performance cascade impactor used to characterize aerosol particles by particle size. It is desirable to develop three formulations with the same emitted dose (ED), impactor size mass (ISM) but different mass median aerodynamic diameter (MMAD). Meeting these design criteria for the formulations would help to see if pharmacokinetic parameters are sensitive to regional differences in drug deposition whilst having the dose deposited in the lung from different formulations being the same.The three formulation developed are labeled Fluticasone Propionate Drug formulation 1 through 3.

Study Procedures and scheduling

The study is comprised of 5 visits in total - a screening visit and 4 treatment visits. At least 5 days should lapse between the treatment visits.

Screening Visit During the screening visit, the inclusion and exclusion criteria will be reviewed to ensure the volunteer is appropriate for the study. The informed consent will be reviewed with the volunteer by a member of the study team and the volunteer will be encouraged to ask questions to ensure the volunteer has a good understanding of the study. If the volunteer is eligible and agrees to participate, the volunteer will be asked to sign the informed consent form prior to any study specific procedures including randomization. After the volunteer signs the informed consent, the volunteer will be interviewed and demographic data, medical history and concomitant medications will be collected and recorded. A physical examination will be performed after the vital sign measurements are obtained. A pregnancy test for female volunteers will be obtained. Spirometry testing and inhalation training will be performed by a qualified study clinician/investigator to ensure the suitability of volunteers. Laboratory tests including a Complete Blood Count (CBC), urinalysis and metabolic panel will be collected via venipuncture and processed in the lab. Screening tests will be performed within 14 days of treatment visit 1 and no later than 2 days before treatment visit 1. All screening results will be evaluated by the study clinician/investigator against the inclusion/exclusion criteria to confirm the eligibility of the volunteers.

Inhalation training: Inhalation training will be performed by a qualified study clinician at the screening visit and at each study visit. The training will be accomplished by instructions and subsequent inhalation via training devices containing empty capsules. The instructions on how to use the device correctly are similar to the Foradil Aerolizer DPI product information label. (http://www.rxlist.com/foradil-drug/medication-guide.htm )

Treatment Visits 1, 2, 3 and 4 Eligible volunteers will be asked to return for treatment visit 1. A minimum period of 5 days should lapse between the subsequent treatment visits. Each treatment visit is scheduled for 28 hours over two days. The study will be conducted at the University of Florida (UF) CRC (Clinical Research Center). It is an outpatient study and the volunteer will be asked to come back the following day for the 24 hour blood sample. The volunteer will be asked to stay in an outpatient room during the treatment visit. The same activities are carried out at the other treatment visits.

At each treatment visit, eligibility criteria will be reviewed and confirmed to ensure volunteer is appropriate for study. Changes in medical history including concomitant medications will be documented. Vital signs will be obtained. Inhalation training will be provided to the volunteers as mentioned in the section above. An IV catheter will be inserted in a vein located in the forearm region of the volunteer. The IV catheter is used to avoid multiple pricks while collecting blood samples. The IV catheter is not used for the administration of the drug. The volunteer inhales 5 times from a given inhaler during each treatment visit. Each inhaler will be used only once and by only one volunteer to ensure the volunteer's safety from infectious agents. The schedule below summarizes the procedures performed at the screening visit and a single treatment visit.

Blood sample collection Blood samples will be drawn by inserting an indwelling catheter into the volunteer's median cubital vein in the forearm region. Blood samples will be taken 15 minutes prior to the dosing of the product (pre-dose sample) and 5, 10, 15, 20, 30, 45, 60 minutes, 1.5, 2, 3, 4, 6, 8, 10, 12, 14 and 24 hours post dosing. At each time point 9ml of blood sample will be collected of which the first 1ml will be discarded and the remaining 8 ml will be stored in a vacutainer tube for plasma preparation and storage.

Arms:
arm 1: Patients first received Fluticasone Propionate Drug formulation 1, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 2, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3\*.

All Fluticasone Propionate Drug formulations were administered via Plastiape Monodose Dry Powder Inhaler, 500mcg single dose.

Mass Median Aerodynamic Diameter (MMAD) for formulation 1 is 4.5 µm. MMAD for formulation 2 is 3.8 µm MMAD for formulation 3 is 3.7 µm. MMAD for formulation 3\* is 3.7 µm. See Formulation Development in Detailed Description for details regarding differences in formulations arm 2: Patients first received Fluticasone Propionate Drug formulation 2, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3\*, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 1, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3.

All Fluticasone Propionate Drug formulations were administered via Plastiape Monodose Dry powder Inhaler, 500mcg single dose.

MMAD for formulation 1 is 4.5 µm. MMAD for formulation 2 is 3.8 µm. MMAD for formulation 3 is 3.7 µm. MMAD for formulation 3\* is 3.7 µm. See Formulation Development in Detailed Description for details regarding differences in formulations arm 3: Patients first received Fluticasone Propionate Drug formulation 3, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 1, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3\*, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 2.

All Fluticasone Propionate Drug formulations were administered via Plastiape Monodose Dry Powder Inhaler, 500mcg single dose.

MMAD for formulation 1 is 4.5 µm. MMAD for formulation 2 is 3.8 µm MMAD for formulation 3 is 3.7 µm. MMAD for formulation 3\* is 3.7 µm. See Formulation Development in Detailed Description for details regarding differences in formulations arm 4: Patients first received Fluticasone Propionate Drug formulation 3\*, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 3, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 2, after a washout period of at least 5 days, they then received Fluticasone Propionate Drug formulation 1.

All Fluticasone Propionate Drug formulations were administered via Plastiape Monodose Dry powder Inhaler, 500mcg single dose. \* (The asterisk) A different set of capsules from the same batch of formulation 3 is given to the subject to assess the within-subject variability and to ensure that the study is sufficiently powered to show bioequivalence between two replicates of the same formulation.

MMAD for formulation 1 is 4.5 µm. MMAD for formulation 2 is 3.8 µm MMAD for formulation 3 is 3.7 µm. MMAD for formulation 3\* is 3.7 µm.

Interventions: intervention 1: Fluticasone Propionate Formulation 1 intervention 2: Fluticasone Propionate Formulation 2 intervention 3: Fluticasone Propionate Formulation 3 intervention 4: Fluticasone Propionate Formulation 3*
Intervention descriptions: intervention 1: Fluticasone Propionate dry powder inhaler Formulation 1 intervention 2: Fluticasone Propionate dry powder inhaler Formulation 2 intervention 3: Fluticasone Propionate dry powder inhaler Formulation 3 intervention 4: Fluticasone Propionate dry powder inhaler Formulation 3\*

\*(The asterisk) A different set of capsules from the same batch of formulation 3 is given to the subject to assess the within-subject variability and to ensure that the study is sufficiently powered to show bioequivalence between two replicates of the same formulation.

Protocol document:
Prot_000.pdf: 
 
Protocol Version:    v20170305  
 
 
Page 1 of 14 
 
Protocol No:   v20170305  
 
Title of Study: 
Evaluation of the sensitivity of pharmacokinetics to differences in the aerodynamic particle size 
distribution of three different formulations of fluticasone propionate dry powder inhalers 
 
Principal Investigator: 
Prof. Jürgen Bulitta, Ph.D. 
College of Pharmacy 
Department of Pharmaceutics 
Center for Pharmacometrics and Systems Pharmacology 
6550 Sanger Road, Office 475, Orlando FL 32827 
Phone number: 407-313-7010 
Fax: 407-313-7030 
Email: jbulitta@cop.ufl.edu 
 
Study Team: 
Guenther Hochhaus, Ph.D. (co-principal investigator) 
Mutasim Abu-Hasan, M.D. (medically accountable investigator) 
Brandon Seay, M.D. (medically accountable investigator) 
Mongjen Chen, Ph.D. (study coordinator) 
Yuanyuan Jiao, Ph.D. (study coordinator) 
Sandra Baumstein, Pharm. D. (study coordinator) 
 
IRB Oversight: 
Institutional Review Board (IRB) 
University of Florida, Health Science Center 
FWA 00005790 
Peter Iafrate, Pharm. D. (Chair IRB-01) 
Shepard Broad Building 
1300 Center Drive, Room 130 
Gainesville, FL 32610 
Phone: (352) 273-9600 
 
Research Involving Human Subjects Committee (RIHSC): 
U.S. Food and Drug Administration 
White Oak Building 32 Room 4286 
10903 New Hampshire Ave 
Silver Spring, MD 20993 
 
Study Center: 
Department of Pharmaceutics , University of Florida 
PO Box 100494, JHMHC, 1600 SW Archer Road 
Gainesville, FL 32610-0494 
Phone: 352-273-7858  


 
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1. 
Synopsis 
 
Objectives 
The aim of the study is to determine if pharmacokinetics (PK) is sensitive to differences in the 
aerodynamic particle size distribution of three different formulations of fluticasone propionate 
(FP) dry powder inhalers (DPI). 
Methodology (Design) 
This is a single center, double-blind, single-dose (500 mcg), randomized, four period, four 
sequence cross over PK study to evaluate the local (lung) bioequivalence (BE) of 100 mcg 
fluticasone propionate formulation 1(FPF I), 100 mcg fluticasone propionate formulation 
2(FPF II) and 100 mcg fluticasone propionate formulation 3 (FPF III). FPF III will be 
replicated in the study to provide an assessment of bioequivalence when FPF III is tested against 
itself. 
During the screening visit, the inclusion and exclusion criteria will be reviewed to ensure the 
subject is appropriate for the study. The informed consent will be reviewed with the subject by 
a member of the study team and the subject will be encouraged to ask questions to ensure the 
subject has a good understanding of the study. If the subject is eligible and agrees to participate, 
the subject will be asked to sign the informed consent form prior to any study specific 
procedures. After their consent, demographic data, medical history and concomitant medications 
will be recorded. A physical examination will be performed after the vital sign measurements 
are obtained. A pregnancy test for female subjects must be negative (test repeated if the subject 
continues in the study for more than 30 days). Spirometry testing and inhalation training will be 
performed to ensure the suitability of subjects. Laboratory tests such as Complete Blood Count 
with differential, urinalysis and basic metabolic panel will be performed. Screening tests will be 
performed within 28 days of study visit 1 and no later than 2 days before study visit 1. All 
screening results will be evaluated by the study clinician/investigator against the 
inclusion/exclusion criteria to confirm the eligibility of the subjects.  
A minimum period of 5 days should lapse between the subsequent study visits. At each study 
visit, review of the eligibility criteria, documentation of any changes in medical history 
including concomitant medications, vital signs, inhalation training, insertion of the indwelling 
catheter into the forearm region of the subject for blood collection, administration of the 
formulation by inhalation, blood sampling as per protocol will be performed. The total duration 
of the study is estimated to be approximately 10 weeks.  
Number of Subjects 
The number of subjects required was calculated to be 24, which takes into account a 10% 
dropout rate to ensure 22 subjects complete the study. Subjects will include healthy males and 
females aged 18 and 50 years who meet the inclusion and exclusion criteria. 


 
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Study Medication, Dose and Mode of Administration 
On each study visit the subject will receive 5 inhalations from one of the following 3 devices: 
 
Study Drug 
Device* 
Formulation 
1 
Plastiape Monodose 
DPI 
Fluticasone Propionate Formulation 1 (FPF I) 
2 
Plastiape Monodose 
DPI 
Fluticasone Propionate Formulation 2 (FPF 
II) 
3 
Plastiape Monodose 
DPI 
Fluticasone Propionate Formulation 3 (FPF 
III) 
* Dry Powder Inhaler. 
Inclusion Criteria 
1. Healthy male or female subjects aged 18 to 50 years (inclusive).  
2. Females will be eligible only if they are currently non-lactating and demonstrate a negative 
urine pregnancy test. Female subjects must be willing to use highly effective methods of 
contraception throughout the study. A highly effective method of birth control is defined as 
one which results in a low failure rate (i.e. less than 1% per year) when used consistently 
and correctly e.g. no sexual intercourse, an intrauterine device (IUD), using contraceptive 
foam AND a condom (double-barrier). 
3. Body weight ranging from 50 to 100 kg, corresponding to a BMI of 18-29 kg/m2. 
4. Non-smoker for at least 12 months prior to study screening and a maximum smoking history 
of less than ten-pack years (i.e. the equivalent of one-pack per day for ten years). 
5. Healthy and free of significant abnormal findings as determined by medical history, physical 
examination, vital signs, laboratory tests (including serum cortisol at screening), complete 
blood count (CBC) with differential, urinalysis and basic metabolic panel. 
6. Ability to read, comprehend and sign the consent form. 
7. Ability and willingness to comply with all study procedures, discontinue and/or withhold 
medications as specified in the protocol, and attend scheduled study visits. 
8. No history of respiratory disease. 
9. Normal baseline spirometry as predicted for age, sex and height, including forced expiratory 
volume in 1 second / forced vital capacity (FEV1/FVC) ≥  0.8. 
10. Healthy and without any pre-existing medical conditions. 
 
 


 
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Exclusion Criteria 
1. Any history and/or conditions that might interfere with drug absorption, distribution, 
metabolism or excretion of FP, e.g., pre-existing lung and liver disease.  
2. Known or suspected sensitivity to Flonase (Fluticasone Propionate), Veramyst (Fluticasone 
Furoate), or related compounds in that class. 
3. Hypersensitivity to milk proteins or lactose (inactive ingredients in the formulation). 
4. Having a history and/or currently having the medical condition in the opinion of medically 
accountable investigator and hence taking any medication for the following (including but 
not limited to): 
4.1 Significant cardiac, dermatologic, gastrointestinal, hepatic, renal, hematological, 
neurological and psychiatric disease (determined by physical exam, CBC with 
differential, urinalysis, basic metabolic panel and medical history). 
4.2 Presence of glaucoma, cataracts, ocular herpes simplex or carcinoma (other than basal 
cell). 
4.3 Presence of tuberculosis and other respiratory diseases (including but not limited to 
intermittent or persistent asthma, emphysema and chronic bronchitis); or respiratory 
infection, common cold, sinusitis or ear infections. 
5. Current use of hormone replacement therapy (HRT), hormonal contraceptives and/or 
corticosteroid treatment within the last 2 months. 
6. Smoker during the last 1 year prior to study screening (self-report). 
7. Evidence of a positive pregnancy urine test for female volunteers or females who are 
pregnant or breast-feeding or are likely to become pregnant during the trial. Women of child-
bearing potential may be included in the study if, in the opinion of the investigator, they are 
taking adequate contraceptive precautions as described above. 
8. Exposure to any investigational drug within 30 days of enrolment. 
9. Subjects who are unable to demonstrate proper inhalation of the test products. 
10. Subjects who have a history of anemia. 
11. Exposure to any medication that alters CYP3A4 activity within last 2 weeks (e.g.: azole 
antifungals, rifampin). 
12. Nausea, vomiting or diarrhoea within 7 days of dosing. 
13. Subjects who have donated 1 pint (450 mL) of blood or more within the previous 8 weeks 
prior to study administration. 
14. Any history or current drug or alcohol abuse, which would interfere with the subject’s 
completion of the study and with adherence to the protocol. 
15. A subject will not be eligible for this study if he/she is an immediate family member of the 
participating investigator, sub-investigator, study coordinator, or employee of the 
participating investigator. 
16. The subject is the student of the Principal Investigator (PI). 
17. Lack of willingness to have personal study related data collected, archived and transmitted 
according to the protocol. 
 
 


 
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Pharmacokinetic Evaluation 
Blood samples will be drawn by inserting an indwelling catheter into the subject’s median 
antecubital vein in the forearm region. Blood samples will be taken within 30 minutes prior to 
the dosing of the product (pre-dose sample) and at approximately 5, 10, 15, 20, 30, 45, 60 
minutes, 1.5, 2, 3, 4, 6, 8, 10, 12, 14 and 24 hours post dosing (approximately means within 
±2 min of the scheduled time during the first hour, within ±5 min of the scheduled time for the 
1.5 to 14 h samples and within ±1 hour for the 24 h sample). The actual sampling time will be 
recorded for all samples. At each time point 0.24 ounces (7.5 mL) of blood sample will be 
collected, of which 0.033 ounces (1 mL) will be discarded in waste tubes and the remaining 0.21 
ounces (6.5 mL) will be collected in a EDTA-pretreated blood collection tube (vacutainer) for 
plasma preparation and storage. A HPLC-MS-MS method capable of quantifying FP in human 
plasma at levels down to 1.5 picogram /mL (or better) will be used. Bioanalysis will be 
performed at Worldwide Clinical Trials (Austin, TX, USA).  
Safety Evaluation 
Safety parameters will include vital signs (temperature, heart rate, respiratory rate and blood 
pressure), physical examinations, and routine clinical laboratory tests. 
Statistical Methods 
Non-Compartmental Analysis will be performed in WinNonlin using an inbuilt extra-vascular 
PK model. Standard PK metrics such as AUC(0-30min), AUC(0-t), AUC(0-inf), Cmax, tmax, terminal 
slope/half-life, CL/F, Vdss/F, AUMC(0-t), AUMC(0-inf), and MRT will be calculated for each 
individual plasma-concentration-time profile. ANOVA model will be used to construct 
univariate 90% confidence intervals for AUC(0-t) and Cmax ratios and applied to conventional 0.8 
– 1.25 average BE limits. If needed, alternative BE evaluation methodology and or BE criteria 
may be investigated. Data will also be assessed by ANOVA methods (SAS 9.2 or another 
equally suitable software package) to test for statistical differences (p<0.05). 
 
 


 
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Schedules+ 
Activity 
Screening Visit Study Visit* 
Visit Window (Days) 
2-28 Days prior 
study visit 
Day 1 
Day 2 
Subject Information 
X 
 
 
Informed Consent 
X 
 
 
Review Inclusion/Exclusion Criteria 
X 
 
 
Demographic Data 
X 
 
 
Medical History  
X 
X 
 
Concomitant Medications 
X 
X 
X 
Measure BMI 
X 
 
 
Vital Signs 
X 
X 
 
Physical Examination 
X 
 
 
Lung Function Measurements (FEV1) 
X 
 
 
Pregnancy Test** 
X 
X 
 
Laboratory Assessments 
X 
 
 
ECG 
X 
 
 
Adverse Events 
X 
X 
X 
Proper Inhalation Technique 
X 
X 
 
Inhalation of Study Formulation 
 
X 
 
Blood Sampling  
 
X 
X 
* At least 5 days (maximum 3 weeks) of washout between each study visit. 
+ A more detailed chart is given in Appendix A. 
** To be repeated if subject’s participation exceeds 30 days. 
 
 
 


 
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2. 
Glossary 
 
List of Abbreviations and Definitions of Terms 
ANOVA 
Analysis of Variance 
APSD 
Aerodynamic Particle Size Distribution 
AUC(0-t) 
Area under the concentration of analyte vs. time curve (AUC) from time zero to 
the last quantifiable concentration 
AUC(0-inf) 
AUC from time zero extrapolated to infinity 
AUMC(0-t) 
Area under the moment curve from time zero to the last observation 
AUMC(0-inf) 
Area under the moment curve when the concentration time curve is extrapolated to 
infinity 
BE 
Bioequivalence 
BMI 
Body Mass Index 
β-hCG 
Serum beta-human chorionic gonadotropin 
Cmax 
The peak or maximum concentration 
CL/F 
Ration of the total body clearance and the fraction of dose absorbed (F) 
DPI 
Dry Powder Inhaler 
ECG 
Electrocardiogram 
ED 
Emitted Dose 
FEV1 
Forced Expiratory Volume in 1 second 
FDA 
Food and Drug Administration 
FP 
Fluticasone Propionate 
FPF I 
100 mcg Fluticasone Propionate Formulation 1 
FPF II 
100 mcg Fluticasone Propionate Formulation 2 
FPF III 
100 mcg Fluticasone Propionate Formulation 3 
FPF III* 
100 mcg Fluticasone Propionate Formulation 3 replicate 
GLP 
Good Laboratory Practices 
GSD 
Geometric Standard Deviation 
ISM 
Impactor-Sized Mass 
IUD 
Intrauterine Device 
Lamda Z 
First order rate constant associated with the terminal (log-linear) elimination phase 
LC-MS/MS 
Liquid Chromatography – Mass Spectroscopy/Mass Spectroscopy 
MMAD 
Mass Median Aerodynamic Diameter 
MRT 
Mean Residence Time, is the amount of time a particle remains in a system , 
calculated as AUMC/AUC 
NCA 
Non Compartmental Analysis 
tmax 
The time of peak concentration 
VZ/F 
Volume of distribution based on the terminal phase corrected with F 
* The asterisk is used to distinguish the replicate from the original formulation. 
 
 


 
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3. 
Study Design 
 
The study comprises of 5 visits: 1 screening visit and 4 study visits. 
On each study visit day, subjects will get 5 inhalations from one of the following formulations: 
 
Study Drug 
Device 
Formulation 
1 
Plastiape Monodose DPI 
Fluticasone Propionate Formulation 1 (FPF I) 
2 
Plastiape Monodose DPI 
Fluticasone Propionate Formulation 2 (FPF II) 
3 
Plastiape Monodose DPI 
Fluticasone Propionate Formulation 3 (FPF III) 
 
The following sequences will be used in the study: 
Sequence Study Visit 1 Study Visit 2 Study Visit 3 Study Visit 4 
1 
FPF I 
FPF II 
FPF III 
FPF III* 
2 
FPF II 
FPF III* 
FPF I 
FPF III 
3 
FPF III 
FPF I 
FPF III* 
FPF II 
4 
FPF III* 
FPF III 
FPF II 
FPF I 
Note that in this design every study drug appears once in each period. Also every study drug is 
immediately followed only once by each of the other study drugs. 
 
4. 
Risk/Benefit Considerations 
Benefits: Participation in a human pharmacokinetic study like the present one cannot be of benefit 
to healthy volunteers. Nevertheless, the information form the physical examination, vital signs, ECG 
and the breathing tests will be shared with the personal physician if the subject chooses.  
Risks: The subjects are exposed to risks associated with the pharmacological properties of the 
investigational product and the study procedures. One single-dose of 500 mcg of fluticasone 
propionate rarely has side effects. Bad reactions can happen, but are very rare. Some of the bad 
reactions we know about happen in people using the drug for a long time and with larger doses (e.g., 
500 mcg of fluticasone propionate twice daily for 28 weeks, as per labeling information of the FDA-
approved ADVAIR DISKUS). These reactions are pharyngitis (sore throat), nasal congestion, 
allergic rhinitis (runny nose), oral candidiasis (yeast infections in the mouth), upper respiratory 
infection, nausea, vomiting and headache. 
Risk/Benefit Ratio: There is no direct benefit to the healthy volunteer in participating of the study. 
The subjects’ contribution to the study is of major importance to agencies like the U.S. Food and 


 
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Drug Administration (FDA) for helping them better evaluate the generic alternatives and thus 
make available cheaper and effective formulations for asthmatic subjects. 
Women of Childbearing Age: This drug will not be given to anyone who is pregnant. All women 
must take a pregnancy test before getting any study drug on this study. All woman of child bearing 
potential enrolled on this study must use effective birth control during the study. These include: 
no sexual intercourse, an IUD, using contraceptive foam AND a condom (double-barrier). You 
must notify the doctor if you become pregnant during the course of the study. 
Intravenous (IV) Catheter Insertion: During the insertion of the IV catheter, soreness or bruising 
at the insertion site can occur but is unlikely. Infection at the IV site is possible but unlikely. 
Dizziness and lightheadedness can occur during insertion of IV catheter or during the blood draws.  
 
4.1 
Adverse Events (AEs) 
An adverse reaction is defined as a response to a drug that is unintended and which occurs at doses 
normally used in man for prophylaxis, diagnosis or therapy of a disease. Since it is a single-dose 
study adverse reactions are rare but the following adverse reactions can occur with the use of 
fluticasone propionate – pharyngitis (sore throat), nasal congestion, allergic rhinitis (runny nose), 
oral candidiasis (yeast infections in the mouth) and upper respiratory infection. 
The drug is contraindicated when there is severe hypersensitivity to milk proteins (exclusion 
criteria) and as a primary treatment of status asthmatics. The latter is not an issue since healthy 
subjects are enrolled in the study. 
A very serious allergic reaction to this drug is rare. However if any symptoms of a serious allergic 
reaction, including: rash, itching/swelling (especially of the face/tongue/throat), severe dizziness, 
trouble breathing are observed, medical help will be immediately provided. 
The physicians associated with this study are: Mutasim Abu-Hasan, M.D., and Brandon Seay, 
M.D., and they will review all AEs monthly and SAEs as it occurs, for its severity and causality 
assessment, using Common Terminology Criteria for Adverse Events Version 5 (CTCAE V5).  
 
4.2 
Serious Adverse Events (SAEs) 
An adverse event or suspected adverse reaction is considered "serious" if, in the view of either the 
principal investigator or sponsor (FDA), it results in any of the following outcomes: death, a life-
threatening adverse event, inpatient hospitalization or prolongation of existing hospitalization, a 
persistent or significant incapacity or substantial disruption of the ability to conduct normal life 
functions, or a congenital anomaly/birth defect. 
All serious and unexpected adverse events will be reported to UF IRB-01within 5 working days 
using the “Adverse Event Reporting Form” (available at http://irb.ufl.edu/irb01/forms/htm) 
All serious but expected adverse events will be reported UF IRB-01 at Continuing Review on the 
“Cumulative Adverse Event Summary Table” (available at: http://irb.ufl.edu/irb01/forms.htm) 
 


 
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4.3 
Adverse Event Reporting to RIHSC 
The Principal Investigator must report to the FDA Sponsor and the RIHSC (FDA IRB) any 
unanticipated problems involving risk to human subjects or others 45 CFR 46.103(b)(5)(i). 
Anticipated adverse events may be reported to the RIHSC in the annual report for continuing 
review. However, certain adverse events must be reported to the RIHSC within 10 working days 
after identifiying the event. The RIHSC defines these adverse events as including, but not limited 
to, the following:  
• An adverse event that is not expected, i.e., not listed in the informed consent document;  
• An expected adverse event that occurs at a greater frequency or duration than expected;  
• Any adverse event that would require modification of the protocol and/or informed consent 
document.  
When reporting any adverse event to the RIHSC, the Principal Investigator should address the 
significance of the adverse event including, whether it was related to the study and whether there 
is a need to communicate information to the subjects. The Principal Investigator should discuss 
whether a change in description of risk is warranted in the protocol and consent form is needed, 
and whether a change in the study design is warranted. 
 
Selection of Study Population 
Asthma subjects typically experience a higher degree of central deposition of the inhaled drug than 
healthy subjects. Because of the skewed central/peripheral ratio towards a more central deposition, 
asthmatics are a less sensitive study population; therefore healthy volunteers will be used as they 
will be more sensitive to detect potential differences in the deposition profile between the three 
formulations. Healthy volunteers also represent a homogeneous study population for 
investigations of bioequivalence, owing to a lesser scope for variability as compared to subjects 
with asthma. Higher variability leads to lesser sensitivity in detecting differences. 
 
[Please see synopsis (above) for details on the inclusion and exclusion criteria.] 
 
5. 
Study Procedures and Scheduling 
 
The study is comprised of 5 visits in total – a screening visit and 4 study visits. At least 5 days 
(washout period, maximum 3 weeks) should lapse between the study visits. A phone call within 7 
days after the last study visit will be performed for follow-up end of the study.   
 
5.1 
Screening Visit 
During the screening visit (please see Appendix A for location), the inclusion and exclusion criteria 
will be reviewed to ensure the subject is appropriate for the study. The informed consent will be 
reviewed with the subject by a member of the study team and the subject will be encouraged to 
ask questions to ensure the subject has a good understanding of the study. If the subject is eligible 


 
Protocol Version:    v20170305  
 
 
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and agrees to participate, the subject will be asked to sign the informed consent form prior to any 
study specific procedures including randomization. After the subject signs the informed consent, 
the subject will be interviewed and demographic data, medical history and concomitant 
medications will be collected and recorded. A physical examination will be performed after the 
vital sign measurements are obtained. A pregnancy test for female subjects will be obtained. 
Spirometry testing and inhalation training will be performed by a qualified study 
clinician/investigator to ensure the suitability of subjects. Laboratory tests including a CBC with 
differential, urinalysis and basic metabolic panel will be collected via venipuncture and processed 
in the lab. Screening tests will be performed within 28 days of study visit 1 and no later than 2 
days before study visit 1. All screening results will be evaluated by the study clinician/investigator 
against the inclusion/exclusion criteria to confirm the eligibility of the subjects. Subject ID will be 
assigned sequentially during the screening visit.  
Inhalation Training: Inhalation training will be performed by a qualified study team member at 
the screening visit and at each study visit. The training will be accomplished by instructions and 
subsequent inhalation via Vitalograph AIM™ inhalation trainer. The instructions on how to use 
the device correctly are similar to the Foradil Aerolizer DPI product information label. 
(http://www.rxlist.com/foradil-drug/medication-guide.htm) 
 
5.2 
Study Visits 1, 2, 3 and 4 
Eligible subjects will be asked to return for study visit 1 (see Appendix A for location). A minimum 
period of 5 days should lapse between the subsequent study visits. Each study visit is scheduled 
for 28 hours over two days. The study will be conducted at the UF CRC (Clinical Research Center). 
It is an outpatient study and the subject will be asked to come back the following day for the 24 
hour blood sample. The subject will be asked to stay in an outpatient room during the study visit. 
The same activities are carried out at the other study visits. 
At each study visit, changes in medical history including concomitant medications will be 
documented. Vital signs will be obtained. Inhalation training will be provided to the subjects as 
mentioned in the section above. An IV catheter will be inserted in a vein located in the forearm 
region of the subject. The IV catheter is used to avoid multiple pricks while collecting blood 
samples. The IV catheter is not used for the administration of the drug. The subject inhales 5 times 
from a given inhaler during each study visit. Each inhaler will be used only once and by only one 
subject to ensure the subject’s safety from infectious agents. The schedule below summarizes the 
procedures performed at the screening visit and a single study visit.  
 
 


 
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Activity 
Screening Visit Study Visit 
Visit Window (Days) 
2-28 Days prior 
study visit 
Day 1 Day 2 
Subject Information 
X 
 
 
Informed Consent 
X 
 
 
Review Inclusion/Exclusion Criteria 
X 
 
 
Demographic Data 
X 
 
 
Medical History  
X 
X 
 
Concomitant Medications 
X 
X 
X 
Measure BMI 
X 
 
 
Vital Signs 
X 
X 
 
Physical Examination 
X 
 
 
Lung Function Measurements (FEV1) 
X 
 
 
Pregnancy Test 
X 
X 
 
Laboratory Assessments 
X 
 
 
ECG 
X 
 
 
Adverse Events 
X 
X 
X 
Proper Inhalation Technique 
X 
X 
 
Inhalation of Study Formulation 
 
X 
 
Blood Sampling for PK 
 
X 
X 
 
5.3 
Washout Period 
The washout period between study drug administrations will be at least 5 days (maximum 3 
weeks). 
 
6. 
Blood Sample Collection 
Blood samples will be drawn by inserting an indwelling catheter into the subject’s median cubital 
vein in the forearm region. Blood samples will be taken within 30 minutes prior to the dosing of 
the product (pre-dose sample) and at approximately 5, 10, 15, 20, 30, 45, 60 minutes, 1.5, 2, 3, 4, 
6, 8, 10, 12, 14 and 24 hours post dosing (approximately means within ±2 min of the scheduled 
time during the first hour, within ±5 min of the scheduled time for the 1.5 to 14 h samples and 
within ±1 hour for the 24 h sample). The actual sampling time will be recorded for all samples. At 
each time point 0.24 ounces (7.5 mL) of blood sample will be collected of which the first 0.033 
ounces (1 mL) will be discarded and the remaining 0.21 ounces (6.5 mL) will be stored in a blood 
collection tube for plasma preparation and storage. The volume of blood to be collected at the 
screening visit is 0.25 ounces (7.5 mL), and that at each study visit (from 15 min pre-dose to 24 
hours post-dose) is 4.56 ounces (135 mL). The total volume of blood to be collected in the study 
is 18.51 ounces (547.5 mL).  


 
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6.1 
Labeling 
The blood collection tubes and the cryovials containing whole blood and plasma samples 
respectively will be labeled with subject number, date and time of sampling. 
 
6.2 
Bioanalytical Method 
A HPLC-MS-MS method capable of quantifying FP in human plasma at levels down to 1.5 
picogram /mL (or better) will be used. Bioanalysis will be performed at Worldwide Clinical Trials 
(Austin, TX). 
 
7. 
Statistical Methods and Analysis of Data 
 
7.1 
Sample Size Calculation  
Using in-house and literature PK data for fluticasone propionate, the variability in the PK 
parameters Cmax and tmax were estimated to be between 20% to 40% (%CV).24,25,26 If the %CV is 
30% and the products differ by only 5%, a sample size of 40 (no dropouts) was initially thought to 
be appropriated to have 80% power to show that the products are equivalent.27 
𝑁= 2(𝑡𝛼,2𝑁−2 + 𝑡𝛽,2𝑁−2)
2 ∗[𝐶𝑉/(𝑉−𝛿)]^2 
N = total number of subjects required to be in the study 
t = the appropriate value from the t distribution 
α = the significant level 
1-β = the power, usually 80% 
CV = the coefficient of variation 
V = the bioequivalence limit (ln 1.25 = 0.223) 
δ = the difference between the products (for 5% difference, delta equals (ln(1.05) = 0.0488) 
However, due to budgetary constraints, the sample size was re-assessed and it was determined that 
the minimum number of subjects necessary to complete the study is approximately 22 subjects 
(considering 10% dropout rate, 24 subjects will be recruited). This sample size number does not 
affect the study design, which is kept as a four-way crossover (i.e., no study drug was removed 
from the study), and is sufficient to be utilized to achieve 80% power to show that the products are 
equivalent, as shown in the table below. 
 
 


 
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Case 
Test / Reference Ratio 
(point estimator) 
Intra-Individual 
Variability 
(ANOVA-CV) 
Power 
Sample Size  
(without 
dropouts) 
Dropout 
Rate 
Final Sample 
Size 
1 
0.95 
30% 
80% 
40 
~10% 
44 (previous 
sample size) 
2 
0.95 
21% 
80% 
22 
~10% 
24 subjects 
3 
1.00 
24% 
80% 
22 
~10% 
24 subjects 
4 
1.05 
22% 
80% 
22 
~10% 
24 subjects 
 
Safety Population: All subjects randomized who received at least one dose of study drugs. 
Study Population to be Analyzed: Analyses and summaries of pharmacokinetic data will be 
based on subjects who are administered all doses, complete at least 75% of blood sampling and 
follow the protocol procedures. 
Data Analysis: Once the bioanalytical data is available, the pharmacokinetic data analysis will be 
conducted under blinded conditions. Afterwards, formulation-based analysis will be conducted 
under unblinded conditions.    
 
7.2 
Non-Compartmental Analysis (NCA) 
Hypothesis of the proposed research is that the three test formulations (differing in the c/p ratio) 
will differ in AUC and Cmax estimates. Three multiple comparisons will be performed i.e. 
Reference against the reference, reference against test 1 and reference against test 2. A multiplicity 
adjustment (e.g. bonferroni type) will be made for the 3 multiple comparisons. No additional 
adjustment will be made for the use of multiple metrics. NCA will be performed in WinNonlin 
using an inbuilt extra-vascular PK model. Standard PK metrics such as AUC(0-30min), AUC(0-t), 
AUC(0-inf), Cmax, tmax, terminal slope/half-life, CL/F, Vdss/F, AUMC(0-t), AUMC(0-inf), and MRT 
will be calculated for each individual plasma-concentration-time profile. ANOVA model will be 
used to construct univariate 90% confidence intervals for AUC(0-t) and Cmax ratios and applied to 
conventional 0.8 – 1.25 average BE limits. If needed, alternative BE evaluation methodology and 
or BE criteria may be investigated. Data will also be assessed by ANOVA methods (SAS 9.2 or 
another equally suitable software package) to test for statistical differences (p<0.05). 
 
7.3 
Compartmental Analysis 
In addition to NCA analysis, compartmental data analysis methodology will be applied to the 
observed in vivo pharmacokinetic (PK) data. Compartmental PK analysis using NONMEM (or 
another equally suitable software package) will incorporate information on physiological 
processes such as pulmonary absorption, mucociliary clearance rates and dissolution rates (bulk 
and after cascade impactor separation), and cascade impactor information.  
 
 

SAP_001.pdf: 
Data analysis plan for 
V06 FINAL 
Page 1 of 3 
FDA DPI study (HHSF223201610099C) 
 
January 19, 2018 
 
 
Data analysis plan 
Pharmacokinetic Comparison of Locally Acting Orally  
Inhaled Drug Products (‘DPI study’, FDA BAA-16-00122) 
1. Purpose 
This document describes the non-compartmental data analysis plan for the above 
mentioned clinical trial. This clinical pharmacokinetic (PK) study assessed three 
fluticasone propionate DPI formulations (formulation 1, 2, 3) which have been 
characterized through in vitro methods. These formulations were compared in the present 
double-blind, randomized 4-way cross-over study; one formulation was repeated to 
evaluate intra-subject variability; this replicated formulation is named 3a and 3b.   
 
2. Objectives 
The primary aim of this research study is to aid the FDA in finding methods to ensure that 
the versions of generic drugs that are inhaled (for example, drugs used to treat asthma) 
are bioequivalent to the trade name drug. As a part of the research study, pharmacokinetic 
(PK) studies (studies measuring drug levels in the blood over time after inhalation) will be 
done using three different versions of fluticasone propionate (FP, a drug routinely used in 
asthmatic patients) administered using a dry powder inhaler (DPI, an inhalation device 
that delivers the drug as a dry powder).  
 
3. Available in vitro data and proposed normalization 
We will use the formulation with the largest Median Mass Aerodynamic Diameter (MMAD) 
as reference (Table 1).  
 
Table 1: 
Summary of FPD data averaged over the 12 and 20 month time points 
 
Formulation 
FPD (<5 µm) 
FPD (<3 µm) 
MMAD 
 
(µg) 
(µg) 
(µm) 
15-mm-016 
18.707 
9.977 
3.77 
15-mm-017 
12.230 
5.274 
4.53 
15-mm-015 
15.869 
8.599 
3.73 
 
We expect that the c/p ratio is determined by the MMAD with larger particles being 
deposited more centrally compared to smaller particles. 
 
We will use three approaches of normalizing the PK parameters: 
1) Normalization based on the ex-throat data 
2) Normalization based on fine particle dose (FPD <3 µm) data 
3) Normalization based on fine particle dose (FPD <5 µm) data 


Data analysis plan for 
V06 FINAL 
Page 2 of 3 
FDA DPI study (HHSF223201610099C) 
 
January 19, 2018 
 
 
 
Normalization factors for the ex-throat data will be calculated separately for the VCU, AIT 
and OPC throats. The average normalization factors over the three throats will be used 
for the normalization using this approach. The normalization factors from this ex-throat 
approach will be used for the primary analysis.  
For the FPD <3 µm and FPD <5 µm normalization, we will average the respective FPD 
data at the 12 and 20 month stability time-points. These two time-points include the time 
of the clinical trials performance. The 15-mm-017 formulation will be used as reference 
(i.e. normalization factor is defined as 1.0). Normalization factors using the FPD approach 
will be used as secondary analyses. 
 
4. Methods 
4.1. 
Non-compartmental pharmacokinetic analysis 
Non-compartmental analysis (NCA) will be performed as described previously.1 The NCA 
PK parameters will be calculated for each subject and study period using the formulas 
implemented in the Phoenix / WinNonlin Professional software. Whenever possible, at 
least three observations above the quantification limit will be used to estimate the terminal 
slope. The ANOVA statistics will be calculated for Cmax, AUC0-last, and AUC0-inf. 
Descriptive statistics will be calculated and reported for all PK parameters. 
The NCA PK parameters will be calculated and NCA PK results sent to the FDA 
colleagues before unblinding. Summary statistics will be calculated after subsequent 
unblinding. 
4.2. 
Statistical analysis to address objective I 
For bioequivalence statistics, the formulation with the larger MMAD will be treated as the 
reference formulation in each pairwise comparison (Table 2). 
 
Table 2: 
Pairwise comparisons while treating the replicates of formulation 15-mm-015 
separately 
 
Comparison 
Test formulation 
Reference 
formulation 
1 
15-mm-016 
15-mm-017 
2 
15-mm-015 (3a) 
15-mm-017 
3 
15-mm-015 (3b) 
15-mm-017 
4 
15-mm-015 (3a) 
15-mm-016 
5 
15-mm-015 (3b) 
15-mm-016 
6 
15-mm-015 (3a) 
15-mm-015 (3b) 
 


Data analysis plan for 
V06 FINAL 
Page 3 of 3 
FDA DPI study (HHSF223201610099C) 
 
January 19, 2018 
 
 
Bioequivalence statistics: Following the FDA guidelines 2, ANOVA statistics will be 
performed based on ln-transformed data. For each comparison, the ratio of geometric 
means and the 90% confidence interval for this ratio will be calculated. We will include 
effects for treatment, period, sequence and subject within sequence in the ANOVA. 
Equivalence statistics will be performed using the equations implemented in Phoenix / 
WinNonlin Professional or SAS.  
 
Reference 
1. Bulitta JB, Holford NHG. An Introductory Guide to Non-Compartmental 
Analysis. In: D'Agostino RB, Sullivan L, Massaro J, eds. Wiley Encyclopedia 
of Clinical Trials. Hoboken, NJ: John Wiley & Sons, Inc, 2008; 1-28. 
2. Food and Drug Administration; Center for Drug Evaluation and Research 
(CDER). Guidance for Industry Statistical Approaches to Establishing 
Bioequivalence. FDA Guideline 2001: 1-48.