Clinical trial NCT03866434
Conditions: Healthy Volunteers
Sponsor: Shire
Start date: 2019-02-26; Completion date: 2019-04-10
Enrollment: 20 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
This Phase 1, open-label, single-sequence, non-randomized, multiple-dose, crossover pharmacokinetic study is a single site study in the United States and will be conducted to assess the effect of a CYP1A2 inducer (omeprazole 40 mg once daily \[QD\]) on the pharmacokinetics of anagrelide (1 mg) when administered concurrently in healthy participants.

Detailed description:

Arms: arm 1: Participants will receive 1 milligram (mg) of SPD422 (Anagrelide hydrochloride) (2\*0.5 mg) capsule orally on Day 1 in fasted state (10 hours prior to and until 4 hours following administration of anagrelide), followed by 40 mg of Omeprazole capsule orally once daily (prior to breakfast) on Days 2 to Day 7, followed by 1 mg of Anagrelide in fasted state in combination with Omeprazole 40 mg on Day 8.
Interventions: intervention 1: SPD422 intervention 2: Omeprazole
Intervention descriptions: intervention 1: Participants will receive 1 mg of SPD422 (2\*0.5 mg) capsule orally on Day 1 and 8 in fasted state. intervention 2: Participants will receive 40 mg of Omeprazole orally once daily on Days 2 - 8.

Protocol documents:
Prot_000.pdf: 
PROTOCOL: SPD422-113
TITLE:
A Phase 1, Open-label, Single-sequence, Non-randomized, Crossover, 
Drug-Drug Interaction Study to Evaluate the Effect of Omeprazole on 
the Pharmacokinetics of SPD422 (anagrelide hydrochloride) in Healthy 
Adult Subjects
DRUG:
SPD422, Anagrelide Hydrochloride
IND:
Non-IND
EUDRACT NO.:
Non-EUDRACT
SPONSOR: 
Shire  300 Shire Way, Lexington, MA 02421 USA
PRINCIPAL/
COORDINATING
INVESTIGATOR:
PROTOCOL 
HISTORY:
Original Protocol: 04Oct2018
This document contains confidential and proprietary information of Shire and is disclosed pursuant to confidentiality 
and non-disclosure obligations. This information should be used solely for the purposes for which it was provided 
and should not be copied, shared with, or disclosed to any third party without the express written consent of Shire.








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PRODUCT QUALITY COMPLAINTS
Investigators are required to report investigational product quality complaints to Shire within 
24 hours. This includes any instances wherein the quality or performance of a Shire product 
(marketed or investigational) does not meet expectations (eg, inadequate or faulty closure, 
product contamination) or that the product did not meet the specifications defined in the 
application for the product (eg, wrong product such that the label and contents are different 
products). For instructions on reporting AEs related to product complaints, see Section 8.
Please use the information below as applicable to report the Product Quality Complaint:
Origin of Product Quality Complaint
E-mail Address
North and South America
European Union and Rest of World
Telephone numbers (provided for reference if needed):
Shire, Lexington, MA (USA)


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TABLE OF CONTENTS
PROTOCOL SIGNATURE PAGE .................................................................................................2
EMERGENCY CONTACT INFORMATION................................................................................3
ADDITIONAL CONTACT INFORMATION................................................................................4
PRODUCT QUALITY COMPLAINTS..........................................................................................5
TABLE OF CONTENTS.................................................................................................................6
LIST OF TABLES.........................................................................................................................10
LIST OF FIGURES .......................................................................................................................10
ABBREVIATIONS .......................................................................................................................11
STUDY SYNOPSIS ......................................................................................................................12
STUDY SCHEDULE(S) ...............................................................................................................18
1.
BACKGROUND INFORMATION.......................................................................................22
1.1
Indication and Current Treatment Options...............................................................22
1.2
Product Background.................................................................................................22
1.2.1
Clinical Information...............................................................................................22
2.
STUDY OBJECTIVES AND PURPOSE ..............................................................................24
2.1
Rationale for the Study.............................................................................................24
2.2
Study Objectives ......................................................................................................24
2.2.1
Primary Objectives.................................................................................................24
2.2.2
Secondary Objectives.............................................................................................24
3.
STUDY DESIGN ...................................................................................................................25
3.1
Study Design and Flow Chart ..................................................................................25
3.1.1
Dose selection, rationale and administration .........................................................25
3.1.2
Screening Period and Day -1 .................................................................................25
3.1.3
Treatment Period (Day 1-9)...................................................................................26
3.1.4
Follow-up...............................................................................................................26
3.1.5
Primary and Secondary Endpoints.........................................................................27
3.2
Duration and Study Completion Definition .............................................................27
3.3
Sites and Regions .....................................................................................................28
4.
STUDY POPULATION.........................................................................................................29
4.1
Inclusion Criteria......................................................................................................29
4.2
Exclusion Criteria.....................................................................................................30
4.3
Restrictions...............................................................................................................32
4.4
Reproductive Potential .............................................................................................32
4.4.1
Female Contraception ............................................................................................32
4.4.2
Male Contraception................................................................................................33
4.5
Discontinuation of Subjects .....................................................................................33


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4.5.1
Reasons for Discontinuation..................................................................................33
4.5.2
Subjects “Lost to Follow-up” Prior to Last Scheduled Visit.................................34
5.
PRIOR AND CONCOMITANT TREATMENT ...................................................................35
5.1
Prior Treatment ........................................................................................................35
5.2
Concomitant Treatment............................................................................................35
5.2.1
Permitted Treatment...............................................................................................35
6.
INVESTIGATIONAL PRODUCT ........................................................................................36
6.1
Identity of Investigational Product...........................................................................36
6.1.1
Blinding the Treatment Assignment......................................................................36
6.1.2
Commercially Obtained Investigational Product...................................................36
6.2
Administration of Investigational Product(s)...........................................................36
6.2.1
Allocation of Subjects to Treatment ......................................................................36
6.2.2
Dosing....................................................................................................................36
6.2.3
Unblinding the Treatment Assignment..................................................................37
6.3
Labeling, Packaging, Storage, and Handling ...........................................................37
6.3.1
Labeling .................................................................................................................37
6.3.2
Packaging...............................................................................................................37
6.3.3
Storage ...................................................................................................................37
6.4
Drug Accountability.................................................................................................38
6.5
Subject Compliance..................................................................................................39
6.6
Retention of Bioavailability and Bioequivalence Testing Samples.........................39
7.
STUDY PROCEDURES........................................................................................................40
7.1
Study Schedule.........................................................................................................40
7.1.1
Screening Period....................................................................................................40
7.1.1.1
Screening Failure....................................................................................40
7.1.1.2
Rescreening of Subjects .........................................................................41
7.1.2
Treatment Period....................................................................................................41
7.1.2.1
Admission to the Clinical Research Center (Day -1).............................41
7.1.2.2
Treatment Period (Day 1 through Day 9)...............................................41
7.1.3
Follow-up...............................................................................................................42
7.1.4
Additional Care of Subjects after the Study ..........................................................42
7.2
Study Evaluations and Procedures ...........................................................................42
7.2.1
Demographic and Other Baseline Characteristics .................................................42
7.2.2
Safety .....................................................................................................................42
7.2.2.1
Medical and Medication History............................................................42
7.2.2.2
Physical Examination (Including Height and Weight) ..........................43
7.2.2.3
Adverse Event Collection.......................................................................43
7.2.2.4
Vital Signs..............................................................................................43


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7.2.2.5
Clinical Laboratory Evaluations.............................................................45
7.2.2.6
Pregnancy Test .......................................................................................47
7.2.2.7
Drug and Alcohol Screen .......................................................................47
7.2.2.8
Serology Screen......................................................................................47
7.2.2.9
Electrocardiogram ..................................................................................47
7.2.3
Pharmacokinetic Procedures..................................................................................48
7.2.3.1
Blood Sample Collection and Handling Procedures..............................48
7.2.3.2
Shipment of Pharmacokinetic Samples..................................................49
7.2.3.3
Plasma Drug Assay Methodology..........................................................49
8.
ADVERSE AND SERIOUS ADVERSE EVENTS ASSESSMENT ....................................51
8.1
Definition of Adverse Events, Period of Observation, Recording of Adverse 
Events.......................................................................................................................51
8.1.1
Severity Categorization..........................................................................................51
8.1.2
Relationship Categorization...................................................................................52
8.1.3
Outcome Categorization ........................................................................................52
8.1.4
Clinical Laboratory and Other Safety Evaluations ................................................52
8.1.5
Pregnancy...............................................................................................................53
8.1.6
Abuse, Misuse, Overdose, and Medication Error..................................................53
8.2
Serious Adverse Event Procedures ..........................................................................54
8.2.1
Reference Safety Information................................................................................54
8.2.2
Reporting Procedures.............................................................................................54
8.2.3
Serious Adverse Event Definition .........................................................................55
8.2.4
Serious Adverse Event Collection Time Frame.....................................................55
8.2.5
Serious Adverse Event Onset and Resolution Dates .............................................55
8.2.6
Fatal Outcome........................................................................................................56
8.2.7
Regulatory Agency, Institutional Review Board, Ethics Committee, and Site 
Reporting................................................................................................................56
9.
DATA MANAGEMENT AND STATISTICAL METHODS...............................................57
9.1
Data Collection.........................................................................................................57
9.2
Clinical Data Management.......................................................................................57
9.3
Data Handling Considerations..................................................................................57
9.4
Statistical Analysis Process......................................................................................57
9.5
Planned Interim Analysis, Adaptive Design, and Data Monitoring Committee......57
9.6
Sample Size Calculation and Power Considerations................................................58
9.7
Study Population ......................................................................................................58
9.8
Pharmacokinetic and Pharmacodynamic Analyses..................................................58
9.8.1
Pharmacokinetic Analysis......................................................................................58
9.8.1.1
Statistical Analysis of Pharmacokinetic Parameters ..............................59


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9.9
Safety Analyses........................................................................................................60
9.10
Other Analyses .........................................................................................................60
10. SPONSOR’S AND INVESTIGATOR’S RESPONSIBILITIES...........................................61
10.1
Sponsor’s Responsibilities .......................................................................................61
10.1.1 Good Clinical Practice Compliance.......................................................................61
10.1.2 Public Posting of Study Information......................................................................61
10.1.3 Submission of Summary of Clinical Study Report to Competent Authorities 
of Member States Concerned and Ethics Committees...........................................61
10.1.4 Study Suspension, Termination, and Completion..................................................61
10.2
Investigator’s Responsibilities .................................................................................62
10.2.1 Good Clinical Practice Compliance.......................................................................62
10.2.2 Protocol Adherence and Investigator Agreement..................................................62
10.2.3 Documentation and Retention of Records.............................................................63
10.2.3.1
Case Report Forms.................................................................................63
10.2.3.2
Recording, Access, and Retention of Source Data and Study 
Documents..............................................................................................63
10.2.3.3
Audit/Inspection.....................................................................................64
10.2.3.4
Financial Disclosure...............................................................................64
10.3
Ethical Considerations..............................................................................................64
10.3.1 Informed Consent...................................................................................................64
10.3.2 Institutional Review Board or Ethics Committee..................................................65
10.4
Privacy and Confidentiality......................................................................................65
10.5
Study Results/Publication Policy .............................................................................66
11. REFERENCES .......................................................................................................................68
12. APPENDICES ........................................................................................................................69
Appendix 1
Protocol History .....................................................................................70


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LIST OF TABLES
Table 1:
Schedule of Assessments...............................................................................18
Table 2:
Schedule of Assessments – Detail of Treatment Period Day 1.....................20
Table 3:
Schedule of Assessments – Detail of Treatment Period Day 8.....................21
Table 4:
Volume of Blood to Be Drawn from Each Subject.......................................50
LIST OF FIGURES
Figure 1:
Study Design Flow Chart ..............................................................................27
Figure 2:
Procedures for Screening Vital Signs (Blood Pressure – Pulse) –
Healthy Subjects Only...................................................................................45


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04 Oct 2018
ABBREVIATIONS
AE
adverse event
AUC
area under the curve
-hCG
beta-human chorionic gonadotropin
BID
twice daily
BMI
body mass index
CI
confidence interval
CRA
clinical research associate
CRF
case report form
CRC
clinical research center
CRO
contract research organization
EC 
ethics committee
ECG
electrocardiogram
EM
extensive metabolizer
ET
essential thrombocythemia
EU
European Union
FDA
Food and Drug Administration
GCP
good clinical practice
HBsAG
hepatitis B surface antigen
HCV
hepatitis C virus antibody
HIV
human immunodeficiency virus
ICH
International Conference on Harmonisation
IM/PM
intermediate/poor metabolizer
IP
investigation product
IRB
institutional review board
IRT
interactive response technology
PK
pharmacokinetics
QD
once daily
SAE
serious adverse event
SAP
statistical analysis plan
TEAEs
treatment emergent adverse events
TSH 
thyroid stimulating hormone
T4
thyroxine
T3
triiodothyronine
US
United States


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STUDY SYNOPSIS
Protocol number: SPD422-113
Drug: Anagrelide Hydrochloride
Title of the study: A Phase 1, Open-label, Single-sequence, Non-randomized, Crossover, Drug-Drug Interaction  
Study to Evaluate the Effect of Omeprazole on the Pharmacokinetics of SPD422 (anagrelide hydrochloride) in 
Healthy Adult Subjects
Number of subjects:
Approximately 35 subjects will be screened to ensure the enrollment and completion of approximately 20 subjects.
Investigator(s): 
Site(s) and Region(s): 
Study period (planned):
Feb 2018 – May 2018
Clinical phase:
Phase 1
Objectives:
Primary: The primary objective of this study is to assess the effect of a CYP1A2 inducer (omeprazole) on the 
pharmacokinetics (PK) of anagrelide when administered concurrently.
Secondary: The secondary objective of this study is to assess the safety and tolerability of anagrelide, omeprazole, 
and co-administered anagrelide plus omeprazole.
Rationale:
CYP1A2 inducers may modify the pharmacokinetics of anagrelide and its metabolites when given concurrently. In 
this study the effect of a commonly used CYP1A2 inducer (omeprazole) on the PK of anagrelide will be assessed.
Investigational product, dose, and mode of administration:

SPD422 anagrelide hydrochloride 1 mg administered orally as two 0.5 mg capsules

Omeprazole 40 mg administered orally as a single 40 mg capsule once daily (QD) for 7 days
Methodology:
This is a Phase 1, open-label, single-sequence, non-randomized, multiple-dose, crossover pharmacokinetic study to 
assess the effect of a CYP1A2 inducer (omeprazole 40 mg) on the pharmacokinetics of anagrelide (1 mg) when 
administered concurrently in healthy subjects. Up to 35 male and female subjects, aged 18-45 years will be screened 
to ensure the enrollment and completion of approximately 20 subjects. Subjects who discontinue the study will not 
be replaced. 
The study includes a screening period of up to 28 days, a treatment period (9 days) and a follow-up phone call 
(7±2 days after the last dose of investigational drug is administered). The maximum duration of study participation 
for any subject is 46 days, if the maximum screening, treatment and follow-up durations are used.
Following the screening visit, subjects who meet the protocol specific inclusion and exclusion criteria will return to 
the Clinical Research Center (CRC) on Day -1 to reconfirm eligibility criteria for participation in the study. Subjects 
who continue to meet the eligibility criteria will be admitted to the CRC on Day -1.
Subjects will be confined at the CRC from Day -1 until after completion of the 24 hour post dose assessments on the 
morning of Day 9. Administration of study drug on Days 1-8 should be at the same time each morning for each 
subject during the study. 
Study procedures, safety and pharmacokinetic assessments will be collected and reported at scheduled time points 
during the study.


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Day 1: Anagrelide 1 mg, (administered orally as two 0.5 mg capsules) will be administered to all study subjects at 
approximately 8:00 in the morning on Day 1. Subjects are required to fast for 10 hours prior to and until 4 hours 
following administration of anagrelide on Day 1. 
Days 2-7: Omeprazole 40 mg will be administered orally QD on Days 2-7 to all subjects. Omeprazole will be 
administered at approximately 8:00 in the morning, prior to breakfast.
Day 8: Anagrelide 1 mg and omeprazole 40 mg will be administered concurrently to all subjects at approximately 
8:00 in the morning on Day 8. Subjects are required to fast for 10 hours prior to and until 4 hours following 
administration of anagrelide and omeprazole on Day 8.
Day 9: Subjects will be discharged from the CRC on Day 9 following the completion of all study procedures and 
assessments. 
Subjects who discontinue early from the study will complete the Early Termination assessments.

Safety Assessments include but are not limited to adverse events, physical examinations, 12-lead 
electrocardiograms (ECG), vital signs (blood pressure and pulse) and safety laboratory tests (hematology, 
chemistry, and urinalysis).

Pharmacokinetic assessments:

Omeprazole: Trough and serial blood samples for PK analysis will be collected for the determination of 
plasma omeprazole concentrations. Trough samples will be collected predose on Days 6 and 7. Serial 
samples will be collected on Day 8 starting at predose and up to 24 hours post dose. The samples will be 
collected according to the Schedule of Assessments.

Anagrelide: Serial blood samples for PK analysis will be collected for the determination of plasma 
concentrations of anagrelide, 3-hydroxy-anagrelide and RL603 on Day 1 and Day 8 starting at predose and 
up to 12 hours post dose. The blood samples will be collected according to the Schedule of Assessments. 
A follow up telephone call (or in-person follow-up contact at the investigator’s discretion) will take place 7± 2 days 
following the subject’s last dose of investigational product to collect information on any ongoing or new adverse 
events (AEs), serious adverse events (SAEs) or concomitant medications, as appropriate.
Inclusion Criteria:
The subject will not be considered eligible for the study without meeting all of the criteria below.
Subjects cannot be enrolled before all inclusion criteria (including test results) are confirmed.
1.
Has given, personally signed, and dated informed consent to participate in the study, in accordance with the 
International Conference on Harmonization Good Clinical Practice Guideline E6 (1996) and applicable 
regulations, before completing any study-related procedures.
2.
Age 18-45 years inclusive at the time of consent. The date of signing informed consent is defined as the 
beginning of the Screening Period. This inclusion criterion will be assessed only at the Screening Visit.
3.
Male, or non-pregnant, non-lactating female who agrees to comply with any applicable contraceptive 
requirements of the protocol or females of non-childbearing potential. A female of non-childbearing potential 
(defined as a female who is post-menopausal [amenorrhea for at least 12 consecutive months], has had a 
hysterectomy, bilateral tubal ligation, bilateral oophorectomy or bilateral salpingectomy.
4.
Satisfactory medical assessment with no clinically significant or relevant abnormal findings  as determined by 
medical/surgical history, physical examination, vital signs, 12-lead electrocardiogram, and clinical laboratory 
evaluation (hematology, biochemistry, thyroid function, and urinalysis) that are likely to interfere with the 
subject’s participation  or ability to complete the study as assessed by the investigator.




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
History of alcohol or other substance abuse within the last year.
7.
A subject's alcohol consumption that fulfils one of the following: (Note: One alcohol unit=1 beer 
[12 oz]=1 wine [5 oz]=1 liquor [1.5 oz])

Has consumed alcohol within 2 days prior to the first dose of investigational product.

Male subjects who consume more than 3 units* of alcohol per day. 

Female subjects who consume more than 2* units of alcohol per day. 
8.
Positive screening test results for alcohol, drugs of abuse, or pregnancy (females of childbearing potential only) 
at the Screening Visit or Day –1.
9.
A positive human immunodeficiency virus (HIV) antibody screen, hepatitis B surface antigen (HBsAG) or 
hepatitis C virus antibody (HCV) screen.
10. Use of tobacco in any form (eg, smoking or chewing) or other nicotine-containing products in any form (eg, 
gum, patch) within 30 days prior to the first dose of investigational product and during the in-house stay at the 
CRC.
11. A positive urine cotinine test that is ≥50 ng/mL at either the Screening Visit or on Day -1.
12. Routine consumption of more than 2 units of caffeine per day or subjects who experience caffeine-withdrawal 
headaches or have a history of caffeine-withdrawal headaches. (One caffeine unit is contained in the following 
items: one 6-oz cup of coffee, two 12-oz cans of cola, one 12-oz cup of tea, and three 1-oz chocolate bars. 
Decaffeinated coffee, tea, or cola are not considered to contain caffeine.)
13. Consumption of grapefruit, Seville oranges, and/or products containing these items within 7 days prior to the 
first dose of investigational product.
14. Donation of blood or blood products (egg, plasma, or platelets) within 60 days prior to the first dose of 
investigational product.
15. Known or suspected intolerance or hypersensitivity to the investigational products (ie, anagrelide or 
omeprazole) or closely related compounds, or any of the stated ingredients.
16. Within 30 days prior to the first dose of investigational product:

Have used an investigational product (if elimination half-life is <6 days, otherwise 5 half-lives).

Have been enrolled in a clinical study (including vaccine studies) that, in the Investigator’s opinion, 
may impact this Shire-sponsored study.
17. Prior screen failure, participation, or enrollment in this study.
18. History of sensitivity to heparin or heparin-induced thrombocytopenia. 
Maximum duration of subject involvement in the study:

Planned duration of screening period: 28 days

Planned duration of treatment period: 9 days

Planned duration of follow-up: 7 ±2 days
Endpoints and statistical analysis:
Endpoints and statistical analysis:
Three analysis populations are defined for this study: the enrolled, safety, and pharmacokinetic sets:
The enrolled set consists of all subjects for whom an enrollment number has been assigned. Usually, these are the 
subjects who meet the study inclusion/exclusion criteria and sign the informed consent form.


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The safety set consists of subjects who are administered at least 1 dose of investigational product (anagrelide or 
omeprazole) and have at least 1 post-dose safety assessment.
The pharmacokinetic set consists of subjects who receive at least 1 dose of study drug and have evaluable PK data 
(defined as complete concentration-time profile to obtain meaningful estimates of PK parameters) available for 
1 dose regimen. The PK analyses will be based on this population.
Primary endpoint:
SPD422 will be administered under fasting conditions on Days 1 and 8. Omeprazole will be administered prior to 
breakfast on Days 2-7 and under fasting conditions on Day 8. PK evaluations of anagrelide concentrations will be 
performed on Day 1 and Day 8, and PK evaluations of omeprazole concentrations will be following 
coadministration with anagrelide on Day 8.
Pharmacokinetic parameters will be determined from the plasma concentration-time data for anagrelide and 
omeprazole by non-compartmental analysis. The pharmacokinetic parameters will include, but not be limited to:
AUC0-¥: Area under the curve extrapolated to infinity, calculated using the observed value of the last non-zero 
concentration (anagrelide and metabolites)
AUC0-t:: Area under the curve from time 0 to the last time point of sample collection (anagrelide and metabolites)
AUC0-tau: Area under the concentration-time curve from time zero to the end of the dosing interval (24 hours post 
dose) (omeprazole only)
CL/F: Total body clearance for extravascular administration.
Cmax: Maximum concentration occurring at tmax
Cmin: The minimum concentration observed over the dosing interval (omeprazole only)
t½: Terminal half-life
tmax: Time of maximum observed concentration sampled during a dosing interval
Vz/F:Volume of distribution associated with the terminal slope following extravascular administration.
The primary pharmacokinetic parameters will include the Cmax, AUC0-t and AUC0-∞ for anagrelide and its active 
metabolite, 3-OH-anagrelide (BCH24426).
The geometric mean relative bioavailability and 90% confidence limits for the effect of omeprazole will be 
evaluated (eg, AUCanagrelide+omeprazole/AUCanagrelide).
Because the CYP2C19-mediated metabolism of omeprazole is subject to genetic polymorphism (approximately 
2/3 subjects are CYP2C19 extensive metabolizers (EMs) with lower omeprazole AUC and 1/3 subjects are 
CYP2C19 intermediate/poor metabolizers IMs/PMs with higher omeprazole AUC), the geometric mean relative 
bioavailability and 90% confidence interval (CI) will be calculate for (1) all subjects, (2) EMs, and (3) IMs/PMs. 
The observed omeprazole AUC0-tau will be used to define the CYP2C19 metabolizer status of subjects, with AUC0-tau
<5,200 ng•h/mL (<15 µmol/L) classified as CYP2C19 EM and AUC0-tau ≥5,200 ng•h/mL (≥15 µmol/L) classified as 
CYP2C19 IM/PM.
Secondary endpoint:
Safety will be assessed for the following evaluations for anagrelide alone, omeprazole alone, and anagrelide and 
omeprazole combination treatments:
Number, severity, seriousness, and causality of treatment emergent adverse events (TEAEs)
Changes in physical examination, vital signs, electrocardiogram (ECGs), and clinical laboratory results 
(hematology, chemistry, and urinalysis) from baseline to post-baseline time points.
Sample Size Justification:
Approximately 20 subjects will be enrolled. Subjects who discontinue will not be replaced.


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Sample size estimation is to achieve a desired precision of the effect of interest, targeting the width of the 90% 
confidence interval.
From a previous crossover study (SPD422-110), the within-subject coefficient variation in anagrelide 
log-transformed AUC was estimated to be 0.132. Allowing for a slightly larger within-subject CV of 0.15, a sample 
size of 11 subjects is required to estimate the mean difference between anagrelide + omeprazole and anagrelide 
alone in the log-transformed AUC with an error margin of at most ±0.2231 at 90% confidence with a probability of 
90%. Thus, if the true geometric mean treatment ratio is 0.6, the lower and upper 90% confidence bounds will be 
estimated to be within 0.48 and 0.75 (exp[ln(0.6) – 0.2231] and exp[ln(0.6) + 0.2231]) with 90% probability. 
Therefore, with an overall sample size of 20 subjects, anticipated 13 to 14 CYP2C19 EM subjects and 6 to 7 
CYP2C19 IM/PM subjects, this statistical power will be met for the overall population and the CYP2C19 EM 
population. The confidence bounds for the smaller population of CYP2C19 IM/PM subjects will be slightly wider.
Statistical Methodology for Pharmacokinetic Endpoint(s):
The following analyses will be conducted on the Pharmacokinetic Analysis Set. All statistical analyses will be 
presented for the CYP2C19 EM group, the CYP2C19 IM/PM group, and overall.
Summary statistics (number of observations, mean, standard deviation, coefficient of variation, median, maximum, 
minimum, and geometric mean) will be determined for all pharmacokinetic parameters and presented by treatment. 
Plasma concentrations of anagrelide, 3-hydroxy-anagrelide, RL603, and omeprazole at each nominal sampling time 
will also be summarized by treatment using descriptive statistics.
The log-transformed pharmacokinetic parameters will be compared between the 2 treatments (anagrelide plus 
omeprazole and anagrelide alone) using an analysis of variance model for a single-sequence crossover design with 
treatment and subject as fixed effects will be used. The magnitude of the effect of omeprazole on the 
pharmacokinetic profile of anagrelide and its metabolites will be evaluated by the point estimate and ninety percent 
(90%) confidence intervals for the treatment difference on the log-transformed parameters back-calculated to the 
original scale for the comparisons of Cmax, AUC0 t, and AUC0 ∞ (test treatment=anagrelide plus omeprazole, 
reference treatment=anagrelide alone).
Statistical Methodology for Safety Endpoint(s):
The safety endpoints will be summarized with descriptive statistics for the safety set.
Relevant safety endpoints as well as their changes from baseline will be summarized. Baseline is defined as the last 
assessment prior to the first dose of investigational product. Potentially clinically important findings will also be 
summarized or listed. The potentially clinically important values will be defined in the statistical analysis plan 
(SAP).
Adverse events will be coded using the Medical Dictionary for Regulatory Activities. The number of events, 
incidence, and percentage of treatment-emergent adverse events will be calculated overall, by system organ class, by 
preferred term, and by treatment group. Treatment emergent adverse events will be further summarized by severity 
and relationship to investigational product. Adverse events related to investigational product, AEs leading to 
withdrawal, SAEs, and deaths will be similarly summarized/listed.
Safety will be assessed by TEAEs, new findings from physical examination, evaluation of blood pressure, heart rate,
new findings from 12-lead ECG, and clinical laboratory test results (biochemistry, hematology, and urinalysis).
Clinical laboratory tests, vital signs, and ECG findings will be summarized by treatment group and visit. Potentially 
clinically important findings will also be summarized or listed.


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Protocol SPD422-113
SPD422, Anagrelide hydrochloride
04 Oct 2018
STUDY SCHEDULE(S)
Table 1:
Schedule of Assessments
Screening 
Period
Day -28 to 
Day -2
Treatment 
Period
Early 
Termination a Follow Up 
Phone 
Contact 7± 
2 days b
Day -1
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Day 8
Day 9
Informed Consent
X
In-Patientc
X
X
X
X
X
X
X
X
X
X
Inclusion/Exclusion Criteria
X
X
Demographics
X
Medical & Medication History
X
Physical Examination
X
X
X
X
X
Height, weight and calculate BMId
X
Vital Signs (blood pressure and 
pulse)e
X
X
X
X
X
X
12-lead ECG e
X
X
X
X
X
X
Drugs of abuse and alcohol screen
X
X
Urine cotinine
X
X
HIV, HBsAg, and HCV screen
X
Serum Pregnancy (all females)
X
X
X
X
Biochemistry, hematology, and 
urinalysis
X
X
X
X
X
X
TSH and T4 
X
Omeprazole PK Sample Collectionf
X f
X f
X
X
Anagrelide PK Sample Collection
X
X
Administration of omeprazoleg, 
X
X
X
X
X
X
Xi
Administration of anagrelide h, i
X i
X i
Concomitant Medications
X
X
X
X
X
X
X
X
X
X
X
X
X


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Protocol SPD422-113
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04 Oct 2018
Table 1:
Schedule of Assessments
Screening 
Period
Day -28 to 
Day -2
Treatment 
Period
Early 
Termination a Follow Up 
Phone 
Contact 7± 
2 days b
Day -1
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Day 8
Day 9
Adverse events/serious adverse 
events
X
X
X
X
X
X
X
X
X
X
X
X
X
BMI=body mass index; ECG=electrocardiogram; HIV=human immunodeficiency virus, HBsAG=hepatitis B surface antigen, HCV=hepatitis C virus; CBP=childbearing 
potential, TSH=thyroid stimulating hormone, T4=thyroxine, PK=pharmacokinetic
a An attempt to perform the early termination assessments and procedures will be made for any subject who withdraws or is removed from the study.
b A follow up telephone contact (or in-person follow-up contact at the investigator’s discretion) will take place 7± 2 days following the subject’s last dose of anagrelide.
c Subjects will remain in-patient at the Clinical Research Center (CRC) starting on Day -1 until discharged following the 24 hour post dose PK sample and safety assessments 
on the morning of Day 9.
d Height to be measured without shoes and weight to be taken in light clothes
e Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs. Vital sign assessments during the screening visit only include 
blood pressure, pulse, respiratory rate and temperature.
f Omeprazole PK sample collections on Days 6 and Day 7 are to be taken at pre-dose only.
g Omeprazole is to be administered prior to breakfast.
h Anagrelide should be taken by the subject at 8:00 am. If more than 1 subject is receiving anagrelide on the same day, the dosing interval between subjects may be a stagger of 
3 minutes between dose administrations.
i Subjects are required to fast for approximately 10 hours prior to and until 4 hours post dose (after PK and safety assessments) on Days 1 and 8


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Protocol SPD422-113
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04 Oct 2018
Table 2:
Schedule of Assessments – Detail of Treatment Period Day 1
Study 
Procedure/Relative
to IP
Administration
Administration 
of 
Anagrelidea
Anagrelide 
PK Sample 
Collection
Physical 
Examination
Vital Signsb
(BP and 
pulse)
12-lead 
ECGb
Biochemistry, 
hematology, 
and 
urinalysis
Concomitant 
Medications
Adverse 
Events/Serious 
Adverse Events
Pre-dosec
X
X
X
X
X
X
0
Xd
X
X
0.5 hours
X
X
X
1 hour
X
X
X
X
1.5 hours
X
X
X
2 hours
X
X
X
X
2.5 hours
X
X
X
3 hours
X
X
X
4 hours
X
X
X
X
8 hours
X
X
X
X
12 hour
X
X
X
X
X
X
X
ECG=electrocardiogram; PK=pharmacokinetic; IP=Investigation Product
a The subjects should be administered anagrelide starting at 8:00 am.
b Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs. 
c Pre-dose assessments should be performed within 30 minutes prior to dose.
d Subjects are required to fast for approximately 10 hours prior to and until 4 hours post-dose (after PK and safety assessments) on Day 1 and 8


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Protocol SPD422-113
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04 Oct 2018
Table 3:
Schedule of Assessments – Detail of Treatment Period Day 8
Study 
Procedure/Relative 
to IP 
Administration
Administration 
of Anagrelide 
&
Omeprazolea
Anagrelide 
PK 
Sample 
Collection
Omeprazole 
PK Sample 
Collection
Physical 
Examination
Vital 
Signsb
(BP 
and 
pulse)
12-
lead 
ECGb
Biochemistry,
hematology, 
and 
urinalysis
Concomitant 
Medications
Adverse 
Events/Serious 
Adverse 
Events
Pre-dosec
X
X
X
X
X
X
X
0
Xd
X
X
0.5 hours
X
X
X
X
1 hour
X
X
X
X
X
1.5 hours
X
X
X
X
2 hours
X
X
X
X
X
2.5 hours
X
X
X
X
3 hours
X
X
X
X
4 hours
X
X
X
X
X
8 hours
X
X
X
X
X
12 hour
X
X
X
X
X
X
24 hour
X
X
X
X
X
X
X
ECG=electrocardiogram; PK=pharmacokinetic; IP=Investigation Product
a The subjects should be administered anagrelide and omeprazole concurrently starting at 8:00 am.
b Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs. 
c Pre-dose assessments should be performed within 30 minutes prior to dose.
d Subjects are required to fast for approximately 10 hours prior to and until 4 hours post-dose (after PK and safety assessments) on Days 1 and Day 8.


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Protocol SPD422-113
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04 Oct 2018
1.
BACKGROUND INFORMATION
1.1
Indication and Current Treatment Options 
Anagrelide is a selective inhibitor of megakaryocyte growth and development, which is used to 
lower the high platelet count in patients with essential thrombocythemia (ET). The effect of 
anagrelide on the platelet count of patients with ET is highly variable, requiring careful dose 
titration to lower platelet counts to ≤600 x 109/L and optimally to ≤450 x 109/L. Treatment with 
Anagrelide is typically initiated at 1 mg/day (administered as 0.5 mg twice daily [BID]) for one 
week, followed by careful titration to individually subject-optimized dose regimen balancing 
lowered platelet counts and tolerability.
1.2
Product Background
Anagrelide (AGRYLIN®; XAGRID®; Shire Pharmaceuticals Group PLC), licensed in 45 territories 
including the European Union (EU), is an orally active quinazoline derivative and the only 
platelet-lowering agent showing selectivity for megakaryocytes. Anagrelide is believed to inhibit 
TPO/c-mpl receptor-mediated events in megakaryocytes, leading to reduced maturation of the 
megakaryocytes and ultimately to decreased platelet numbers. Clinical studies in subjects 
support a hypothesis of dose-related reduction in platelet production resulting from a decrease in 
megakaryocyte hypermaturation. 
Anagrelide selectively lowers platelets due to inhibitory effects due to megakaryocytopoiesis. 
The action of anagrelide has recently been determined to suppress expression of the key 
transcription factors. GATA-1 and FOG-1, downstream of TPO/c-mpl receptor-mediated events. 
These factors are required at all stages of megakaryocytopoiesis from differentiation to 
maturation. Anagrelide downregulates expression of GATA-1 and FOC-1 without altering 
TPO/c-mpl-mediated signaling, an action independent of cAMP PDE III inhibition. Anagrelide 
does not interfere with erythropoietin-stimulated erythropoiesis, or leukocytopoiesis (Ahluwalia 
et al., 2010). 
Refer to the investigator’s brochure provided under separate cover for more information.
1.2.1
Clinical Information
Anagrelide administered to healthy subjects during Phase 1 clinical studies demonstrated a 
consistent safety profile at 1mg/day dosing. In studies SPD422-107 and SPD422-110, there were 
no serious adverse events. Treatment-emergent events were primarily mild, and the most 
common events (>10%) were dizziness, postural dizziness, headache, feeling hot, procedural site 
pain, and palpitations. There were no clinically significant electrocardiogram (ECG) changes 
reported in either study. In healthy subjects receiving a single dose of anagrelide 2.5mg (study 
SPD422-111, Thorough QT study), 73% of subjects reported headache, and >20% of subjects 
reported dizziness and nausea. One subject was withdrawn from the study due to 
treatment-emergent T-wave inversion on ECG at the 2.5 mg dose. In that study, the high dose of 
anagrelide 2.5 mg rapidly increased mean heart rate during the first 1.5 hours after 
administration, with the largest mean time-matched increase of +29.1 bpm occurring at 2 hours 
after administration. 


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Protocol SPD422-113
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04 Oct 2018
Additionally, anagrelide 2.5 mg increased the QTcNi (patient-specific corrected QT) and QTcF 
(Fridericia corrected QT) with an upper 2-sided 90% confidence interval (CI) exceeded 10msec 
at 1 and 1.5 hours (values of 15.7 msec and 13.6 msec, respectively) for QtcNi and at 1 hour for 
QTcF (value of 12.7 msec). These changes corresponded to the time of tmax for anagrelide and its 
active metabolite, 3-hydroxy-anagrelide, and the magnitude of the change at these time points 
met the definition for a positive thorough QT study. However, the QTcNi and QTcF values were 
increased only when the heart rate was increasing rapidly at 0.5-1.5 hours after administration of 
anagrelide 2.5mg, and the interpretation of the increased QTcNi and QTcF may be confounded 
by the rapid increase in heart rate and the corresponding QT-RR hysteresis that occurs when the 
heart rate changes rapidly.
Anagrelide is metabolized via cytochrome P450 type 1A2 (CYP1A2) to form its active 
metabolite, 3-hydroxy-anagrelide (also known as BCH24426) which undergoes both renal 
excretion and further metabolism via CYP1A2 to form the inactive metabolite, RL603. 
Administration of anagrelide with a CYP1A2 inducer could potentially increase the clearance of 
anagrelide as well as increase both formation rate and clearance of the active metabolite. Shire 
has been requested by the European Medicines Agency to assess the effect of the CYP1A2 
inducer omeprazole on the pharmacokinetics (PK) of anagrelide when given concurrently.
Omeprazole is metabolized by CYP2C19, an important polymorphically expressed enzyme. 
Based on the incidence of this genetic polymorphism in the general population, approximately 
2/3 of the subjects in this study are expected to be CYP2C19 extensive metabolizers (EMs), and 
the remaining 1/3 of the subjects in this study will be CYP2C19 intermediate/poor metabolizers 
(IMs/PMs). The CYP2C19 IMs/PMs will have higher omeprazole plasma concentrations leading 
to a greater CYP1A2 induction than the CYP2C19 EMs. It is important to evaluate the effect of 
omeprazole on the pharmacokinetic profile of anagrelide and its active metabolite in EMs and 
IMs/PMs as well as overall (Rost et al., 1992; Rost and Roots, 1994;Nousbaum et al., 1994). The 
area under the curve (AUC) of omeprazole will be used to classify subjects into the CYP2C19 
EM or IM/PM categories in this study (Rost et al., 1992). Increased knowledge and 
understanding of the effect of the CYP1A2 inducer omeprazole on the pharmacokinetic profile 
of anagrelide and the varying impact of CYP2C19 polymorphisms on the magnitude of 
omeprazole's CYP1A2 induction effect on the pharmacokinetic profile of anagrelide are helpful 
in optimizing personalized anagrelide drug therapy.
Always refer to the latest version of the SPD422 anagrelide hydrochloride investigator’s 
brochure for the overall risk/benefit assessment and the most accurate and current information 
regarding the drug metabolism, pharmacokinetics, efficacy, and safety of SPD422 anagrelide 
hydrochloride.


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Protocol SPD422-113
SPD422, Anagrelide hydrochloride
04 Oct 2018
2.
STUDY OBJECTIVES AND PURPOSE
2.1
Rationale for the Study
The European Medicines Agency requested that Shire assess the effect of the CYP1A2 inducer 
omeprazole on the pharmacokinetics (PK) of anagrelide when given concurrently to provide 
additional guidance to the prescribing physician on this potential drug-drug interaction. CYP1A2 
inducers may modify the pharmacokinetics of anagrelide and its metabolites when given 
concurrently. In this study the effect of a commonly used CYP1A2 inducer (omeprazole) on the 
pharmacokinetics of anagrelide will be assessed.
2.2
Study Objectives
2.2.1
Primary Objectives
The primary objective of this study is to assess the effect of a CYP1A2 inducer (omeprazole) on 
the pharmacokinetics of anagrelide when administered concurrently.
2.2.2
Secondary Objectives
The secondary objective of this study is to assess the safety and tolerability of anagrelide, 
omeprazole, and co-administered anagrelide plus omeprazole.


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Protocol SPD422-113
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04 Oct 2018
3.
STUDY DESIGN
3.1
Study Design and Flow Chart
This is a Phase 1, open-label, single-sequence, non-randomized, multiple-dose, crossover 
pharmacokinetic study to assess the effect of a CYP1A2 inducer (omeprazole 40 mg once daily 
[QD]) on the pharmacokinetics of anagrelide (1 mg) when administered concurrently in healthy 
subjects. Up to 35 healthy male and female subjects, aged 18-45 years will be screened to ensure 
enrollment and completion of 20 subjects. Subjects who discontinue from the study will not be 
replaced. The study design is shown in the study schema in Figure 1. 
3.1.1
Dose selection, rationale and administration
Therapeutic doses of omeprazole typically range from 20 to 40 mg once-daily for most 
indications, but may be as high as 60 mg once-daily for pathological hypersecretory conditions. 
Omeprazole is safe and well tolerated in this dose range. Omeprazole 40 mg QD produces 
substantial CYP1A2 induction with 1 week of dosing (Rost et al., 1992; Rost and Roots, 1994; 
Nousbaum et al., 1994).
Anagrelide 1 mg is a typical starting dose for treating ET patients, and it is sufficient to provide a 
robust PK profile for both parent anagrelide and the 3-hydroxy metabolite in healthy subjects at a 
safe and tolerable single-dose (SPD422-103, SPD422-104, SPD422-109, SPD422-110). Higher 
single-doses of anagrelide 2.5 mg have led to substantial increases in heart rate (mean increase 
+29 bpm) observed in a thorough QTc study (Troy et al., 2014), therefore this drug-drug 
interaction study will use the 1mg dose which is below 2.5 mg.
For each study subject, dosing should occur at approximately the same time point in the morning 
on Days 1 through Day 8. A previous study (SPD422-109) demonstrated that food delays the 
onset of absorption and slows the rate of absorption of anagrelide, with a 20% increase in mean 
anagrelide AUC and 29% decrease in mean 3-hydroxy-anagrelide AUC. This effect is not 
considered clinically important. In general, clinical pharmacology studies are conducted with 
fasted administration to minimize the potential confounding effect of administration with a meal 
on the interpretation of the study results. Therefore, the subjects are required to fast for 
approximately 10 hours prior to and until 4 hours following administration of anagrelide on the 
primary pharmacokinetic sampling days (Days 1 and 8). Omeprazole will be administered at 
8:00 in the morning, prior to breakfast on Days 2-7.
3.1.2
Screening Period and Day -1
The maximum duration of the screening period is 28 days before the administration of the first 
dose of anagrelide on Day 1. At the initial visit to the clinical research center (CRC) the subject 
will first give informed consent. This will be the date that he/she enters the screening period. 
After giving consent, the subject’s eligibility will be evaluated against the inclusion and 
exclusion criteria and they will undergo the procedures outlined in Table 1.


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Protocol SPD422-113
SPD422, Anagrelide hydrochloride
04 Oct 2018
Following the screening visit, subjects who meet the protocol specific inclusion and exclusion 
criteria will return to the CRC on Day -1 to reconfirm eligibility criteria for participation in the 
study. Subjects who continue to meet the eligibility criteria will be admitted to the CRC on 
Day -1.
3.1.3
Treatment Period (Day 1-9)
Subjects will be admitted to the CRC after eligibility is confirmed on Day -1. Subjects will be 
confined at the CRC from Day -1 until after completion of the 24 hour post dose assessments on 
the morning of Day 9. Administration of study drug on Days 1-8 should be at the same time each 
morning for each subject during the study. 
Study procedures, safety and pharmacokinetic assessments will be collected and reported at 
scheduled time points during the study. The timing of these assessments are described in Table 1, 
Table 2, and Table 3.
Day 1: Anagrelide 1 mg, (administered orally as two 0.5 mg capsules) will be administered to all 
study subjects at approximately 8:00 in the morning on Day 1. Subjects are required to fast for 
10 hours prior to and until 4 hours following administration of anagrelide on Day 1.
Days 2-7: Omeprazole 40 mg will be administered orally QD on Days 2-7 to all subjects. 
Omeprazole will be administered at approximately 8:00 in the morning, prior to breakfast.
Day 8: Anagrelide 1 mg and omeprazole 40 mg will be administered concurrently to all subjects 
at approximately 8:00 in the morning on Day 8. Subjects are required to fast for 10 hours prior to 
and until 4 hours following administration of anagrelide and omeprazole on Day 8.
Day 9: Subjects will be discharged from the CRC on Day 9 following the completion of all study 
procedures and assessments.
Subjects who discontinue early from the study will complete the Early Termination assessments.

Safety Assessments include but are not limited to assessment of adverse events, physical 
examinations, 12-lead ECG, vital signs (blood pressure and pulse) and safety laboratory 
tests (hematology, chemistry, and urinalysis).

Pharmacokinetic assessments:
o
Omeprazole: plasma concentrations of omeprazole
o
Anagrelide: plasma concentrations of anagrelide, 3-hydroxy-anagrelide and 
RL603
3.1.4
Follow-up
A follow up telephone call (or in-person follow-up contact at the investigator’s discretion) will 
take place 7± 2 days following the subject’s last dose of investigational product to collect 
information on any ongoing or new adverse events (AEs), serious adverse events (SAEs) or 
concomitant medications, as appropriate.




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Protocol SPD422-113
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04 Oct 2018
The study will be completed in approximately 3 months.
The Study Completion Date is defined as the date the final subject, across all sites, completes 
their final protocol-defined assessment. Please note that this includes the follow-up visit or
contact, whichever is later. The Study Completion Date is used to ascertain timing for study 
results posting and reporting.
3.3
Sites and Regions
This study will be conducted at a single center Clinical Research Center in the United States.


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Protocol SPD422-113
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04 Oct 2018
4.
STUDY POPULATION
Each subject must participate in the informed consent process and provide written informed 
consent/assent before any procedures specified in the protocol are performed.
4.1
Inclusion Criteria
The subject will not be considered eligible for the study without meeting all of the criteria below.
Subjects cannot be enrolled before all inclusion criteria (including test results) are confirmed.
1. Has given, personally signed, and dated informed consent to participate in the study, in 
accordance with the International Conference on Harmonization Good Clinical Practice 
Guideline E6 (1996) and applicable regulations, before completing any study-related 
procedures.
2. Age 18-45 years inclusive at the time of consent. The date of signing informed consent is 
defined as the beginning of the Screening Period. This inclusion criterion will be assessed 
only at the Screening Visit.
3. Male, or non-pregnant, non-lactating female who agrees to comply with any applicable 
contraceptive requirements of the protocol or females of non-childbearing potential. A 
female of non-childbearing potential (defined as a female who is post-menopausal 
[amenorrhea for at least 12 consecutive months], has had a hysterectomy, bilateral tubal 
ligation, bilateral oophorectomy or bilateral salpingectomy.
4. Satisfactory medical assessment with no clinically significant or relevant abnormal findings  
as determined by medical/surgical history, physical examination, vital signs, 12-lead 
electrocardiogram, and clinical laboratory evaluation (hematology, biochemistry, thyroid 
function, and urinalysis) that are likely to interfere with the subject’s participation  or ability 
to complete the study as assessed by the investigator.
5. An understanding, ability, and willingness to fully comply with study procedures and 
restrictions.
6. Body mass index (BMI) between 18.5 and 30.0 kg/m² inclusive; assessed only at the 
screening visit.
7. Able to swallow (multiple capsules or tablets at 1 time or consecutively at 1 time) all 
investigational product.
8. Healthy as determined by the investigator on the basis of screening evaluations.




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04 Oct 2018

History of symptomatic or clinically meaningful orthostatic hypotension or syncope, 
as assessed by the investigator.

History of controlled or uncontrolled hypertension or a systolic blood pressure 
≥140 mmHg or diastolic blood pressure ≥90 mmHg at the Screening Visit or Day -1.

Subject has any history of seizure disorder.

History or presence of known structural cardiac abnormalities, syncope, cardiac 
conduction problems (PR interval >220 ms, second or third-degree heart block, 
bundle branch block [except congenital right bundle branch block], or prolonged QTc 
interval) or exercise-related cardiac events.

History of alcohol or other substance abuse within the last year.
7. A subject's alcohol consumption that fulfils one of the following: (Note: One alcohol 
unit=1 beer [12 oz]=1 wine [5 oz]=1 liquor [1.5 oz])

Has consumed alcohol within 2 days prior to the first dose of investigational product.

Male subjects who consume more than 3 units* of alcohol per day. 

Female subjects who consume more than 2* units of alcohol per day. 
8. Positive screening test results for alcohol, drugs of abuse, or pregnancy (females of 
childbearing potential only) at the Screening Visit or Day –1.
9. A positive human immunodeficiency virus (HIV) antibody screen, hepatitis B surface antigen 
(HBsAG) or hepatitis C virus antibody (HCV) screen.
10. Use of tobacco in any form (eg, smoking or chewing) or other nicotine-containing products 
in any form (eg, gum, patch) within 30 days prior to the first dose of investigational product 
and during the in-house stay at the CRC.
11. A positive urine cotinine test that is ≥50 ng/mL at either the Screening Visit or on Day -1.
12. Routine consumption of more than 2 units of caffeine per day or subjects who experience 
caffeine-withdrawal headaches or have a history of caffeine-withdrawal headaches. (One 
caffeine unit is contained in the following items: one 6-oz cup of coffee, two 12-oz cans of 
cola, one 12-oz cup of tea, and three 1-oz chocolate bars. Decaffeinated coffee, tea, or cola 
are not considered to contain caffeine.)
13. Consumption of grapefruit, Seville oranges, and/or products containing these items within 
7 days prior to the first dose of investigational product.
14. Donation of blood or blood products (egg, plasma, or platelets) within 60 days prior to the 
first dose of investigational product.
15. Known or suspected intolerance or hypersensitivity to the investigational products 
(ie, anagrelide or omeprazole) or closely related compounds, or any of the stated ingredients.


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16. Within 30 days prior to the first dose of investigational product:

Have used an investigational product (if elimination half-life is <6 days, otherwise 
5 half-lives).

Have been enrolled in a clinical study (including vaccine studies) that, in the 
Investigator’s opinion, may impact this Shire-sponsored study.
17. Prior screen failure, participation, or enrollment in this study.
18. History of sensitivity to heparin or heparin-induced thrombocytopenia. 
4.3
Restrictions
1. Subjects should refrain from strenuous physical exercise 48 hours prior to admission to the 
CRC and during the in-house stay at the CRC.
2. Subjects should refrain from alcohol 48 hours prior to admission to the CRC and during the 
in-house stay at the CRC.
3. Subjects should refrain from foods or beverages containing caffeine/xanthine 48 hours prior 
to admission to the CRC and during the in-house stay at the CRC.
4. On Day 1 and Day 8 subjects will be required to fast for approximately 10 hours prior to 
administration of investigational product and continuing through 4 hours after administration. 
In addition, water intake will be restricted (except for water to administer the dose) starting 
4 hours prior and continuing up to 2 hours after administration of the investigational product. 
5. Subjects will be required to follow standardized meal schedules and eat the meals provided 
by the site while housed in the CRC. No outside food or beverages (including gum, mints, 
etc.) will be permitted. Menus will be identical for all subjects at the CRC. Copies of the 
menus will be provided to the sponsor for approval prior to the start of the study. While 
confined, the total daily nutritional composition should be approximately 50% carbohydrate, 
35% fat, and 15% protein. The daily caloric intake per subject should not exceed 
approximately 3200 kcal.
4.4
Reproductive Potential
4.4.1
Female Contraception
Sexually active females of childbearing potential should be using an acceptable form of 
contraception. Females of childbearing potential must be advised to use acceptable 
contraceptives throughout the study period and for 30 days following the last dose of 
investigational product. Females of childbearing potential who are not currently sexually active 
must agree to use acceptable contraception, as defined below, if they become sexually active 
during the period of the study and 30 days following the last dose of investigational product. 
Female subjects should be either:

Postmenopausal (12 consecutive months of spontaneous amenorrhea and age 51 years)


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
Surgically sterile (having undergone one of the following surgical acts: hysterectomy, 
bilateral tubal ligation, bilateral oophorectomy or bilateral salpingectomy) and at least 
6 weeks post-sterilization, or

Females of childbearing potential with a negative urine and/or serum -hCG pregnancy 
test at screening and at Day -1. Females of childbearing potential must agree to abstain 
from sexual activity that could result in pregnancy or agree to use acceptable methods of 
contraception.
Acceptable methods of contraception are:

Intrauterine devices plus condoms

Double-barrier methods (eg, condoms and diaphragms with spermicidal gel or foam)
4.4.2
Male Contraception
Not Applicable.
4.5
Discontinuation of Subjects
A subject may withdraw from the study at any time for any reason without prejudice to their 
future medical care by the physician or at the institution. The investigator or sponsor may 
withdraw the subject at any time (eg, in the interest of subject safety). The investigator is 
encouraged to discuss withdrawal of a subject from investigational product with the medical 
monitor when possible.
If investigational product is discontinued, regardless of the reason, the early withdrawal 
evaluations listed in Table 1 are to be performed as completely as possible. Whenever possible, 
all discontinued subjects should also undergo the protocol-specified follow-up. Comments 
(spontaneous or elicited) or complaints made by the subject must be recorded in the source 
documents. The reason for termination and date of stopping investigational product must be 
recorded in the case report form (CRF) and source documents.
Subjects who discontinue will not be replaced. 
4.5.1
Reasons for Discontinuation
The reason for withdrawal must be determined by the investigator and recorded in the subject’s 
medical record and on the CRF. If a subject is withdrawn for more than 1 reason, each reason 
should be documented in the source document and the most clinically relevant reason should be 
entered on the CRF.
Reasons for discontinuation include but are not limited to:

Adverse event

Protocol deviation

Withdrawal by subject


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
Lost to follow-up

Other 
4.5.2
Subjects “Lost to Follow-up” Prior to Last Scheduled Visit
A minimum of 3 documented attempts must be made to contact any subject lost to follow-up at 
any time point prior to the last scheduled contact (office visit or telephone contact). At least 1 of 
these documented attempts must include a written communication sent to the subject’s last 
known address via courier or mail (with an acknowledgement of receipt request) asking that they 
return to the site for final safety evaluations and return any unused investigational product.


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5.
PRIOR AND CONCOMITANT TREATMENT
5.1
Prior Treatment
Prior treatment includes all treatment (including but not limited to herbal treatments, vitamins, 
non-pharmacological treatment such as psychotherapy, as appropriate)  received within 30 days 
(or pharmacokinetic equivalent of 5 half-lives, whichever is longer) of the date of first dose of 
investigational product. Prior treatment information must be recorded on the appropriate CRF 
page.
5.2
Concomitant Treatment 
Concomitant treatment refers to all treatment taken between the dates of the first dose of 
investigational product and the end of the follow-up period, inclusive. Concomitant treatment 
information must be recorded on the appropriate CRF page.
5.2.1
Permitted Treatment
Subjects should refrain from taking any medications during the course of the study. Any 
medication which is considered necessary for the subject’s safety and well-being may be given at 
the discretion of the investigator. The administration of all medications (including investigational 
products) must be listed on the appropriate CRF page.


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6.
INVESTIGATIONAL PRODUCT
6.1
Identity of Investigational Product
The test product is commercial anagrelide, which will be provided as 0.5mg capsules. Additional 
information is provided in the SPD422 anagrelide hydrochloride investigator’s brochure.
6.1.1
Blinding the Treatment Assignment
Not applicable.
6.1.2
Commercially Obtained Investigational Product
Commercially available omeprazole 40 mg will be sourced by the site or Clinical Research 
Center.
6.2
Administration of Investigational Product(s)
6.2.1
Allocation of Subjects to Treatment
This is an open-label, non-randomized study.
Screening numbers are assigned to all subjects as they consent to take part in the study. The 
screening number is assigned to subjects according to the sequence of presentation for study 
participation. This will be a 4-digit number starting at 0001.
A 4-digit subject number, starting at 1001, will be allocated immediately prior to dosing after 
eligibility has been determined. If a subject number is allocated incorrectly, the study monitor 
must be notified as soon as the error is discovered. Once a subject number has been assigned, 
that number must not be used again if, for example, a subject is withdrawn from the study. For 
enrolled subjects, the subject number will be the identifying number used throughout the CRF.
6.2.2
Dosing
After satisfying all screening and entry criteria, anagrelide and/or omeprazole will be 
administered to all subjects according to the following schedule:
Day 1: Anagrelide 1 mg, (administered orally as two 0.5 mg capsules) will be administered to 
all subjects at approximately 8:00 in the morning on Day 1. Subjects are required to fast for 
10 hours prior to and until 4 hours following administration of anagrelide on Day 1.
Days 2-7: Omeprazole 40 mg (administered orally, QD) will be administered to all subjects on 
Days 2-7. Omeprazole will be administered at approximately 8:00 in the morning, prior to 
breakfast.
Day 8: Anagrelide 1 mg and omeprazole 40 mg will be administered concurrently to all subjects 
at approximately 8:00 in the morning on Day 8. Subjects are required to fast for 10 hours prior to 
and until 4 hours following administration of anagrelide and omeprazole on Day 8.


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6.2.3
Unblinding the Treatment Assignment
Not Applicable.
6.3
Labeling, Packaging, Storage, and Handling
6.3.1
Labeling
Labels containing study information and pack identification are applied to the investigational 
product(s) container.
In addition to the commercial label, all investigational product is labeled with a minimum of the 
protocol number, batch number and/or packaging reference, directions for use, the statements 
“For clinical trial use only” and/or “CAUTION: New Drug – Limited by Federal (or US) Law to 
Investigational Use, and the sponsor’s name and address. 
Additional labels may, on a case-by-case basis, be applied to the investigational product in order 
to satisfy local or institutional requirements, but must not:

Contradict the clinical study label

Obscure the clinical study label

Identify the study subject by name
Additional labels may not be added without the sponsor’s prior full agreement.
6.3.2
Packaging
Investigational product, anagrelide 0.5 mg capsules will be provided in the manufacturer’s 
original package. In addition to the commercial label, each bottle will have supplemental label 
that will contain but will not be limited to: protocol number, batch number and/or packaging 
reference, directions for use, the statements “For clinical trial use only” and/or “CAUTION: New 
Drug – Limited by Federal (or US) Law to Investigational Use, and the sponsor’s name and 
address.
Changes to sponsor-supplied packaging prior to dosing may not occur without full agreement in 
advance by the sponsor.
6.3.3
Storage
The investigator has overall responsibility for ensuring that investigational product is stored in a 
secure, limited-access location. Limited responsibility may be delegated to the pharmacy or 
member of the study team, but this delegation must be documented. Investigational products are 
distributed by the pharmacy or nominated member of the study team.
Investigational product must be stored in accordance with labeled storage conditions. 
Temperature monitoring is required at the storage location to ensure that the investigational 
product is maintained within an established temperature range. 


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The investigator is responsible for ensuring that the temperature is monitored throughout the 
duration of the study and that records are maintained; the temperature should be monitored 
continuously by using either an in house system, a mechanical recording device such as a 
calibrated chart recorder, or by manual means, such that both minimum and maximum 
thermometric values over a specific time period can be recorded and retrieved as required. Such 
a device (ie, certified min/max thermometer) would require manual resetting upon each 
recording. The sponsor must be notified immediately upon discovery of any excursion from the 
established range. Temperature excursions will require site investigation as to cause and 
remediation. The sponsor will determine the ultimate impact of excursions on the investigational 
product and will provide supportive documentation as necessary. Under no circumstances should 
the product be dispensed to subjects until the impact has been determined and the product is 
deemed appropriate for use by the sponsor.
The sponsor should be notified immediately if there are any changes to the storage area of the 
investigational product that could affect the integrity of the product(s), eg, fumigation of a 
storage room.
6.4
Drug Accountability
Investigators will be provided with sufficient amounts of the investigational product to carry out 
this protocol for the agreed-upon number of subjects. The investigator or designee will 
acknowledge receipt of the investigational product, documenting shipment content and 
condition. Accurate records of all investigational product dispensed, used, returned, and/or 
destroyed must be maintained as detailed further in this section.
The investigator has overall responsibility for administering/dispensing investigational product.
Where permissible, tasks may be delegated to a qualified designee (eg, a pharmacist) who is 
adequately trained in the protocol and who works under the direct supervision of the investigator. 
This delegation must be documented in the applicable study delegation of authority form.
The investigator or his/her designee (as documented by the investigator in the applicable study 
delegation of authority form) will administer the investigational product only to subjects 
included in this study following the procedures set out in the study protocol. Each subject will be 
given only the investigational product carrying his/her treatment assignment. All administered 
medication will be documented on the CRFs and/or other investigational product record.
No investigational product stock or returned inventory from a Shire-sponsored study may be 
removed from the site where originally shipped without prior knowledge and consent by the 
sponsor. If such transfer is authorized by the sponsor, all applicable local, state, and national 
laws must be adhered to for the transfer.
At the end of the study, or as instructed by the sponsor, all unused stock, and empty/used 
investigational product packaging are to be sent to a nominated contractor on behalf of the 
sponsor. Investigational products being returned to the sponsor’s designated contractors must be 
counted and verified by clinical site personnel and the sponsor (or designated contract research 
organization [CRO]). For unused supplies where the original supplied tamper-evident feature is 
verified as intact, the tamper-evident feature must not be broken and the labeled amount is to be 
documented in lieu of counting. 


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Shipment return forms, when used, must be signed prior to shipment from the site. Validated 
electronic return systems (ie, interactive response technology [IRT]) do not require a shipment 
form. Returned investigational products must be packed in a tamper-evident manner to ensure 
product integrity. Contact the sponsor for authorization to return any investigational product 
prior to shipment. Shipment of all returned investigational product must comply with local, state, 
and national laws.
Based on entries in the site drug accountability forms, it must be possible to reconcile 
investigational products delivered with those used and returned. All investigational products 
must be accounted for and all discrepancies investigated and documented to the sponsor’s 
satisfaction.
6.5
Subject Compliance
Compliance must be assessed by observation of dosing by the investigator or designee. In 
addition, the CRC personnel should perform a hand-and-mouth check (mouth check is only 
required for oral dosing) of the subject to assure the investigational product has been ingested. 
The investigator/nominated person will record details on the drug accountability log(s) and/or 
source documents. In addition, details of the dosing time (time, date, dose level) will be captured 
in the appropriate CRF.
6.6
Retention of Bioavailability and Bioequivalence Testing Samples
Not Applicable.


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7.
STUDY PROCEDURES
7.1
Study Schedule
See Table 1, Table 2, and Table 3 for study procedures.
The following “priority order” will be in effect when more than 1 procedure or assessment is 
required at a particular time point.

Spontaneous or solicited AE reporting

Electrocardiogram 

Vital signs

Pharmacokinetic blood sampling

Clinical laboratory tests

Physical examination
NOTE: Blood sampling for pharmacokinetic evaluation must be performed at the precise 
protocol-scheduled time. Actual sampling time(s) must be accurately recorded in the source 
document and appropriate CRF.
7.1.1
Screening Period 
Screening procedures must be completed within 28 days prior to receiving the first dose of 
investigational product. All screening assessments and procedures are to be performed by the 
principal investigator or a qualified designee. See Table 1 for a complete list of screening 
procedures to be performed.
Written, signed, and dated informed consent from each subject prior to the performance of any 
study-related procedures must be obtained by the principal investigator or a designee. A copy of 
the signed informed consent form must be given to the subject for their records.
7.1.1.1
Screening Failure
A screen failure is a subject who has given informed consent and failed to meet the inclusion 
and/or met at least 1 of the exclusion criteria and has not been enrolled or administered 
investigational product(s).
For purposes of data collection, all subjects who give consent to the study but are not enrolled 
will be reported as screen failures even if they were otherwise fully eligible for the study (for 
example, alternates/reserve subjects).


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7.1.1.2
Rescreening of Subjects
Subjects who fail to meet all inclusion/exclusion criteria will not be permitted to be rescreened 
for the study at any point.
Eligible subjects who meet all inclusion/exclusion criteria but are unable to participate in the 
study due to scheduling conflicts/timing may be rescreened based on investigator discretion and 
sponsor approval should their availability to participate fall outside the screening window. In 
these cases, a new screening number must be assigned for each subject who is rescreened and a 
new informed consent form must be signed.
7.1.2
Treatment Period
7.1.2.1
Admission to the Clinical Research Center (Day -1)
Following the screening visit, eligible subjects will return to the CRC on Day -1 of the study. See 
Table 1 for a list of procedures and assessments to be completed upon admission to the CRC. 
Subjects who successfully complete the pre-admission procedures and assessments will be 
admitted to the CRC on Day -1 and assigned a subject number on Day 1 as described in 
Section 6.2.1. Eligible subjects will be confined to the CRC from the morning of Day -1 until 
completion of all procedures and assessments on Day 9.
7.1.2.2
Treatment Period (Day 1 through Day 9)
Subjects will be admitted to the CRC after eligibility is confirmed on Day -1. Subjects will be 
confined at the CRC from Day -1 until after completion of the 24 hour post dose assessments on 
the morning of Day 9. Administration of study drug on Days 1-8 should be at the same time each 
morning for each subject during the study. 
Study procedures, safety and pharmacokinetic assessments will be collected and reported at 
scheduled time points during the study. Subjects will undergo assessments and procedures as 
specified by visit in Table 1, Table 2, and Table 3.

Day 1: Anagrelide 1 mg, (administered orally as two 0.5 mg capsules) will be 
administered to study subjects at approximately 8:00 in the morning on Day 1. Subjects 
are required to fast for 10 hours prior to and until 4 hours following administration of 
anagrelide on Day 1. The procedures, assessments and time points for Day 1 are 
presented in Table 2.

Days 2-7: Omeprazole 40 mg will be administered orally QD on Days 2-7. Omeprazole 
will be administered at approximately 8:00 in the morning, prior to breakfast.

Day 8: Anagrelide 1 mg and omeprazole 40 mg will be administered concurrently in a 
fasted state (10 hours prior to and until 4 hours following administration of anagrelide 
and omeprazole) at approximately 8:00 in the morning on Day 8. The procedures, 
assessments and time points for Day 8 are presented in Table 3. 

Day 9: Subjects will be discharged from the CRC on Day 9 following the completion of 
all study procedures and assessments.


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Subjects who discontinue early from the study will complete the Early Termination assessments 
in Table 1.
7.1.3
Follow-up 
A follow up telephone call (or in-person follow-up contact at the investigator’s discretion) will 
take place 7 ±2 days following the subject’s last dose of investigational product to collect 
information on any ongoing or new AEs, SAEs or concomitant medications, as appropriate. All 
AEs and SAEs that are not resolved at the time of this contact will be followed to closure (see 
Section 8.1).
7.1.4
Additional Care of Subjects after the Study
No aftercare is planned for this study.
7.2
Study Evaluations and Procedures
7.2.1
Demographic and Other Baseline Characteristics
Demographic characteristics such as age, sex, weight, height and body mass index will be 
collected according to Table 1.
7.2.2
Safety
The name and address of each third-party vendor (eg, clinical laboratory) used in this study will 
be maintained in the investigator’s and sponsor’s files.
Actual safety assessment times will be monitored and recorded. The sponsor’s expectation is that 
the investigator will ensure that every effort is made to perform all assessments at the precise 
protocol-scheduled time. Any safety assessment that deviates from the scheduled assessment 
time set forth in the protocol by more than 15 minutes will be considered a protocol deviation.
7.2.2.1
Medical and Medication History
A complete medical and medication history, as well as demographic information, will be 
performed at the screening visit/time points described in Table 1 by a qualified licensed 
physician, physician’s assistant, or a nurse practitioner. The medical history will be reviewed and 
recorded, including:

Date of birth

Sex

Race and ethnicity

Recent ingestion of medication (30 days prior to entering the screening period)

History of respiratory, cardiovascular, renal, gastrointestinal, hepatic, endocrine, 
hematological, neurological, psychiatric, and other diseases

Smoking habits 


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7.2.2.2
Physical Examination (Including Height and Weight)
A complete physical examination will be performed at the time points described in Table 1,
Table 2, and Table 3 by a qualified licensed physician, physician’s assistant, or nurse 
practitioner.
The physical examination will include a review of the following body systems:

General appearance

Skin

Head, eyes, ears, nose, and throat

Spine/neck/thyroid

Musculoskeletal

Respiratory

Cardiovascular

Neurological

Abdomen (including liver and kidneys)
Abnormalities identified at the screening visit will be documented in the subject’s source 
documents and on the medical history CRF. Changes after the screening visit that are considered 
clinically significant will be captured as AEs on the AE CRF page, as deemed by the 
investigator.
7.2.2.3
Adverse Event Collection
At each study visit, subjects will be questioned in a general way to ascertain if AEs have 
occurred since the previous visit (eg, “Have you had any health problems since your last visit?”). 
Adverse events are collected from the time informed consent is signed. (Please refer to Section 8, 
Adverse and Serious Adverse Events Assessment.)
7.2.2.4
Vital Signs
Blood Pressure and Pulse Rate
Blood pressure and pulse rate will be measured at times specified in Table 1, Table 2, and
Table 3 of this protocol. Additional blood pressure and pulse rate measurements may be 
performed, as determined by the investigator, in order to ensure appropriate monitoring of 
subject safety and accurate recording of vital sign measurements. Any changes from baseline 
which are deemed clinically significant by the investigator are to be recorded as an AE.
The same method for obtaining blood pressure measurement (auscultatory or oscillometric) 
should be used throughout the study for all subjects (and documented). In addition, the 
conditions of vital sign measurements should be controlled and as consistent as possible during 
the study, in order to minimize external variability of the readings. 


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It is advised that measurements be collected at a comfortable room temperature with little to no 
background noise, using the same (appropriately sized) cuff placed at the same location of the 
same arm during the study. The bladder deflation rate should be deflated (calibrated for 
oscillometric method or manually by auscultatory method) at a rate of 2-3 mmHg/s (and the first 
and last audible sounds recorded as systolic and diastolic pressure) after at least 5 minutes of rest 
in the assumed position.
The cuff should have a bladder length that is 80% and a width that is at least 40% of arm 
circumference (a length-to-width ratio of 2:1). 
The subject should be asked to remove all clothing that covers the location of cuff placement. 
The subject should be instructed to relax as much as possible for at least 5 minutes prior to 
collection. The subject should remain quiet during this time and through the measurement.
The subject should be lying comfortably, with the legs uncrossed. The arm should be supported 
with a pillow, such that the middle of the cuff on the upper arm is at the level of the right atrium 
(approximately halfway between the bed and the level of the sternum).One reading (supine 
systolic blood pressure/diastolic blood pressure-heart rate) should be taken at each timepoint. 
The use of automated devices for measuring pulse rate is acceptable, although, when done 
manually, pulse rate will be measured in the brachial/radial artery for at least 30 seconds. When 
the timing of these measurements coincides with a blood collection, blood pressure and pulse 
rate should be obtained prior to the nominal time of the blood collection.
At the screening visit, blood pressure should be compared between both arms. When there is a 
consistent inter-arm difference confirmed over 3 consecutive measurements (>10 mmHg), the 
arm with the higher blood pressure should be used for inclusion at screening and the last 
measurement recorded in the CRF. The same (right or left) arm with the higher blood pressure 
will be used throughout the study.
For details on blood pressure and pulse procedures for healthy subjects, see Figure 2.




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The following clinical laboratory assessments will be performed:
Biochemistry
Blood samples (8.5 mL) for serum biochemistry will be collected into a gel separator tube at the 
time points described in Table 1, Table 2, and Table 3. The following parameters will be 
assessed: 
Sodium
Phosphorus
-hCG a
Potassium
Total protein
Glucose 
Total CO2 (Bicarbonate)
Blood urea nitrogen
Albumin
Creatinine
Aspartate transaminase 
Calcium
Alanine transaminase 
Chloride
Gamma glutamyl transferase 
Thyroid stimulating hormone (TSH)
Alkaline phosphatase 
Thyroxine (T4 total) 
Total bilirubin 
Triiodothyronine (T3) 
Uric acid
a-hCG=beta-human chorionic gonadotropin. Females only.
Hematology
Blood samples (4 mL) for hematology will be collected into a ethylenediaminetetraacetic acid 
tube at the time points described in Table 1, Table 2, and Table 3. The following parameters will 
be assessed:
Hemoglobin
Total neutrophils (absolute)
Hematocrit
Eosinophils (absolute)
Red blood cells
Monocytes (absolute)
Platelet count
Basophils (absolute)
White blood cell count; total and differential
Lymphocytes (absolute)


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Urinalysis
A urine sample for urinalysis will be collected at the time points described in Table 1, Table 2,
and Table 3. The following parameters will be assessed:
pH
Blood
Nitrites
Glucose
Ketones
Leukocyte esterase
Protein
Bilirubin
Specific gravity
Microscopic examination will be conducted if protein and/or blood is/are detected during 
urinalysis. At a minimum the microscopic examination will consist of red blood cells, white 
blood cells, casts, and bacteria. 
7.2.2.6
Pregnancy Test
For all female subjects (regardless of reproductive potential status), a serum pregnancy test will 
be performed at the visits specified in Table 1; or if pregnancy is suspected; or on withdrawal of 
the subject from the study.
7.2.2.7
Drug and Alcohol Screen
A urine screen for drugs of abuse and alcohol will be performed at the time points described in 
Table 1. Additional drug and alcohol screens may be performed at the investigator’s discretion.
Urine samples are to be tested for amphetamines, barbiturates, benzodiazepines, cannabinoids, 
cocaine, methadone, opiate metabolite, and phencyclidine.
Results of urine drug and alcohol screens will be reviewed and verified by the study monitor, but 
will not be collected in the CRF database.
Any positive result for drugs of abuse or alcohol at screening or on Day -1 will exclude the 
subject from further participation in the study.
7.2.2.8
Serology Screen
At the screening visit, a blood sample of approximately 8.5 mL will be drawn into a serum 
separator tube to test for the presence of HIV, hepatitis B surface antigen (HBsAg), and hepatitis 
C virus (HCV) antibody.
The test results must be confirmed negative prior to enrollment in the study. If a test result is 
positive, the subject will be excluded from entering the study. Results of the virology screen will 
be reviewed and verified by the study monitor, but will not be collected in the CRF database.
7.2.2.9
Electrocardiogram
Twelve-lead ECGs will be performed at the times specified in Table 1, Table 2, and Table 3
using the standard methods/equipment at the CRC.


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The following parameters will be recorded on the appropriate CRF page: heart rate, PR, RR, 
QRS, and QT intervals. The QTcB and QTcF will be derived from the data in the database. The 
investigator’s assessment of the ECG tracing as normal or abnormal must be documented, and if 
abnormal, his/her determination of whether the abnormality is clinically significant or not will be 
documented on the tracing and recorded in the CRF.
The subject should be asked to remove all clothing that covers the location of lead placement. 
The subject must be resting in the supine position for at least 5 minutes prior to collecting the 
ECG.
In some cases, it may be appropriate to repeat abnormal ECGs to rule out improper lead 
placement as contributing to the ECG abnormality. It is important that leads are placed in the 
same positions each time in order to achieve precise ECG recordings. 
One complete recording, including a 10-second rhythm strip, should be taken at each time point. 
It should be immediately assessed as a valid recording and if not valid, it should be repeated. 
Invalid recordings will not be entered in the CRF. The ECG collected predose on Day 1 will 
serve as the subject’s baseline ECG.
7.2.3
Pharmacokinetic Procedures
The name and address of the bioanalytical laboratory(ies) for this study will be maintained in the 
investigator’s files at the/each site and in the Trial Master File at the sponsor.
Actual pharmacokinetic blood sample collection times versus time of dosing will be 
monitored. The sponsor’s expectation is that the investigator will ensure that every effort is 
made to collect all pharmacokinetic blood samples at the precise protocol scheduled time. 
Pharmacokinetic blood collection must not deviate from the nominal collection time set forth in 
the protocol by more than 5 minutes from samples drawn within 4 hours post dose or by more
than 15 minutes for samples drawn beyond 4 hours post dose. Samples drawn outside these 
parameters will be considered a protocol deviation. 
7.2.3.1
Blood Sample Collection and Handling Procedures
Blood samples will be collected at the time specified in Table 1, Table 2, and Table 3 to measure 
plasma concentrations of anagrelide, its active metabolite, BCH24426 (3-hydroxy-anagrelide), 
RL603, and omeprazole. Other potential metabolites may also be determined as appropriate.
A full description of the PK blood collection, handling, storage, and shipping can be found in the 
laboratory manual.


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Plasma sample tubes for bioanalysis must be freezer-safe and identified with freezer-safe labels 
provided by the CRC. The labels will contain the following information:

Study number (SPD422-113)

Subject identifier

Analyte (anagrelide or omeprazole)

Nominal day (Day 1, Day 6…)

Nominal time

Matrix identifier (plasma)

Split (primary or backup or aliquot 1 or aliquot 2)
7.2.3.2
Shipment of Pharmacokinetic Samples
All pharmacokinetic samples should be placed in a cryo-box, double-bagged to contain leaks and 
packed with a sufficient quantity of dry ice to ensure that they remain frozen for at least 72 hours 
to allow for delays in shipment. All applicable shipping regulations must be followed. Shipments 
should be scheduled so that no samples arrive on the weekend and should be shipped 
Monday-Wednesday only. Samples should be transported to ensure that they arrive at the 
bioanalytical laboratory between the hours of 9:00 AM and 4:00 PM. The recipient and primary 
Shire contact must be notified by telephone or e-mail when the samples are shipped and they 
must be provided with the shipment tracking number.
Full directions for shipment of all PK samples, (along with the corresponding documentation) 
can be found in the laboratory manual provided under separate cover.
Pharmacokinetic samples will be stored nominally at -80°C for anagrelide and -20°C for 
omeprazole at YBS prior to and after analysis until their disposal is authorized by Shire.
7.2.3.3
Plasma Drug Assay Methodology
Plasma sample analysis will be performed according to the relevant Standard Operating 
Procedures at the contract bioanalytical lab.
Plasma concentrations will be measured using the most current validated bioanalytical methods. 
In addition, selected plasma samples may be used to investigate incurred sample reproducibility 
(full details will be described in the bioanalytical study plan). The presence of other metabolites 
or artifacts may be monitored or quantified as appropriate. Raw data will be stored in the archive 
of the designated bioanalytical contract laboratory. 


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Table 4:
Volume of Blood to Be Drawn from Each Subject
Assessment
Sample Volume (mL)
Number of 
Samples
Total Volume (mL)
Anagrelide Pharmacokinetic samplesa
6a
20
120
Omeprazole Pharmacokinetic samples
2
13
26
HBsAg, HIV, HCV
8.5
1
8.5
Safety
Biochemistry and -hCGb
8.5
5
42.5
Hematology
4
5
20
Thyroid function
5
1
5
Total mL
222
-hCG=beta-human chorionic gonadotropin; HBsAg=hepatitis B surface antigen; HCV=hepatitis C virus; HIV=human 
immunodeficiency virus
a If a catheter is used, the first mL is to be discarded; then take 4 mL into appropriate tube for pharmacokinetic sample. A total 
of 5 mL of blood drawn has been used in determination of sample volume.
b -hCG testing for females only.
During this study, it is expected that approximately 222 mLs of blood will be drawn from all 
subjects, regardless of sex.
Note: The amount of blood to be drawn for each assessment is an estimate. The amount of blood 
to be drawn may vary according to the instructions provided by the manufacturer or laboratory 
for an individual assessment; however, the total volume drawn over the course of the study 
should be approximately 222 mLs. When more than 1 blood assessment is to be done at the time 
point/period, if they require the same type of tube, the assessments may be combined.


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8.
ADVERSE AND SERIOUS ADVERSE EVENTS ASSESSMENT
8.1
Definition of Adverse Events, Period of Observation, Recording of Adverse Events
An AE is any untoward medical occurrence in a clinical investigation subject administered a 
pharmaceutical product and that does not necessarily have a causal relationship with this 
treatment. An AE can therefore be any unfavorable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease temporally associated with the use of a medicinal 
(investigational) product, whether or not related to the medicinal (investigational) product 
(International Conference on Harmonisation [ICH] Guidance E2A 1995).
All AEs are collected from the time the informed consent is signed until the defined follow-up 
period stated in Section 7.1.3. This includes events occurring during the screening phase of the 
study, regardless of whether or not investigational product is administered. Where possible, a 
diagnosis rather than a list of symptoms should be recorded. If a diagnosis has not been made, 
then each symptom should be listed individually. All AEs should be captured on the appropriate 
AE pages in the CRF and in source documents. In addition to untoward AEs, unexpected 
benefits outside the investigational product indication should also be captured on the AE CRF.
All AEs must be followed to closure (the subject’s health has returned to his/her baseline status 
or all variables have returned to normal), regardless of whether the subject is still participating in 
the study. Closure indicates that an outcome is reached, stabilization achieved (the investigator 
does not expect any further improvement or worsening of the event), or the event is otherwise 
explained. When appropriate, medical tests and examinations are performed so that resolution of 
event(s) can be documented.
8.1.1
Severity Categorization
The severity of AEs must be recorded during the course of the event including the start and stop 
dates for each change in severity. An event that changes in severity should be captured as a new 
event. Worsening of pre-treatment events, after initiation of investigational product, must be 
recorded as new AEs (for example, if a subject experiences mild intermittent dyspepsia prior to 
dosing of investigational product, but the dyspepsia becomes severe and more frequent after first 
dose of investigational product has been administered, a new AE of severe dyspepsia [with the 
appropriate date of onset] is recorded on the appropriate CRF).
The medical assessment of severity is determined by using the following definitions:
Mild:
A type of AE that is usually transient and may require only minimal treatment or 
therapeutic intervention. The event does not generally interfere with usual 
activities of daily living.
Moderate:
A type of AE that is usually alleviated with specific therapeutic intervention. The 
event interferes with usual activities of daily living, causing discomfort but poses 
no significant or permanent risk of harm to the research subject.
Severe:
A type of AE that interrupts usual activities of daily living, or significantly affects 
clinical status, or may require intensive therapeutic intervention.


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8.1.2
Relationship Categorization
A physician/investigator must make the assessment of relationship to investigational product for 
each AE. The investigator should decide whether, in his or her medical judgment, there is a 
reasonable possibility that the event may have been caused by the investigational product. If 
there is no valid reason for suggesting a relationship, then the AE should be classified as “not 
related”. Otherwise, if there is any valid reason, even if undetermined or untested, for suspecting 
a possible cause-and-effect relationship between the investigational product and the occurrence 
of the AE, then the AE should be considered “related”. The causality assessment must be 
documented in the source document.
The following additional guidance may be helpful:
Term
Relationship Definition
Related
The temporal relationship between the event and the administration of the 
investigational product is compelling and/or follows a known or suspected response 
pattern to that product, and the event cannot be explained by the subject’s medical 
condition, other therapies, or accident.
Not Related
The event can be readily explained by other factors such as the subject’s underlying 
medical condition, concomitant therapy, or accident and no plausible temporal or 
biologic relationship exists between the investigational product and the event.
8.1.3
Outcome Categorization
The outcome of AEs must be recorded during the course of the study on the CRF. Outcomes are 
as follows:

Fatal

Not Recovered/Not Resolved

Recovered/Resolved

Recovered/Resolved With Sequelae

Recovering/Resolving

Unknown
8.1.4
Clinical Laboratory and Other Safety Evaluations
A change in the value of a clinical laboratory, vital sign, or ECG assessment can represent an AE 
if the change is clinically relevant or if, during treatment with the investigational product, a shift 
of a parameter is observed from a normal value to an abnormal value, or a further worsening of
an already abnormal value. When evaluating such changes, the extent of deviation from the 
reference range, the duration until return to the reference range, either while continuing treatment 
or after the end of treatment with the investigational product, and the range of variation of the 
respective parameter within its reference range, must be taken into consideration.


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If, at the end of the treatment phase, there are abnormal clinical laboratory, vital sign, or ECG 
values which were not present at the pretreatment value observed closest to the start of study 
treatment, further investigations should be performed until the values return to within the 
reference range or until a plausible explanation (eg, concomitant disease) is found for the 
abnormal values.
The investigator should decide, based on the above criteria and the clinical condition of a 
subject, whether a change in a clinical laboratory, vital sign, or ECG parameter is clinically 
significant and therefore represents an AE.
8.1.5
Pregnancy
All pregnancies are to be reported from the time informed consent is signed until the defined 
follow-up period stated in Section 7.1.3.
Any report of pregnancy for any female study participant must be reported within 24 hours to the 
Shire Global Safety Department using the Shire Investigational and Marketed Products 
Pregnancy Report Form. A copy of the Shire Investigational and Marketed Products Pregnancy 
Report Form (and any applicable follow-up reports) must also be sent to the CRO/Shire medical 
monitor using the details specified in the emergency contact information section of the protocol. 
The pregnant female study participant must be withdrawn from the study.
Every effort should be made to gather information regarding the pregnancy outcome and 
condition of the infant. It is the responsibility of the investigator to obtain this information within 
30 calendar days after the initial notification and approximately 30 calendar days post partum.
Pregnancy complications such as spontaneous abortion/miscarriage or congenital abnormality 
are considered SAEs and must be reported using the Shire Clinical Study Serious Adverse Event 
and Non-serious AEs Required by the Protocol Form. Note: An elective abortion is not 
considered an SAE.
In addition to the above, if the investigator determines that the pregnancy meets serious criteria, 
it must be reported as an SAE using the Shire Clinical Study Serious Adverse Event and 
Non-serious AEs Required by the Protocol Form as well as the Shire Investigational and 
Marketed Products Pregnancy Report Form. The test date of the first positive serum/urine -hCG 
test or ultrasound result will determine the pregnancy onset date.
8.1.6
Abuse, Misuse, Overdose, and Medication Error
Abuse, misuse, overdose, or medication error (as defined below) must be reported to the sponsor 
according to the SAE reporting procedure whether or not they result in an AE/SAE as described 
in Section 8.2. Note: The 24-hour reporting requirement for SAEs does not apply to reports of 
abuse, misuse, overdose, or medication errors unless these result in an SAE. 
The categories below are not mutually exclusive; the event can meet more than 1 category.


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
Abuse – Persistent or sporadic intentional intake of investigational product when used for 
a non-medical purpose (eg, to alter one’s state of consciousness or get high) in a manner 
that may be detrimental to the individual and/or society

Misuse – Intentional use of investigational product other than as directed or indicated at 
any dose (Note: this includes a situation where the investigational product is not used as 
directed at the dose prescribed by the protocol)

Overdose – Intentional or unintentional intake of a dose of investigational product higher 
than the protocol-prescribed dose

Medication Error – An error made in prescribing, dispensing, administration, and/or use 
of an investigational product. For studies, medication errors are reportable to the sponsor 
only as defined below.
Medication errors should be collected/reported for all products under investigation.
The administration and/or use of an expired investigational product should be considered as a 
reportable medication error.
8.2
Serious Adverse Event Procedures
8.2.1
Reference Safety Information
The reference for safety information for this study is the investigator’s brochure which the 
sponsor has provided under separate cover to all investigators.
The reference for safety information for omeprazole (PRILOSEC®) is the FDA-approved package 
insert which the sponsor has provided under separate cover to all investigators
8.2.2
Reporting Procedures
All initial and follow-up SAE reports must be reported by the investigator to the Shire Global 
Safety Department and the CRO/Shire medical monitor within 24 hours of the first awareness of 
the event. Note: The 24-hour reporting requirement for SAEs does not apply to reports of abuse, 
misuse, overdose, or medication errors (see Section 8.1.6) unless they result in an SAE.
The investigator must complete, sign, and date the Shire Clinical Study Serious Adverse Event 
and Non-serious AEs Required by the Protocol Form and verify the accuracy of the information 
recorded on the form with the corresponding source documents (Note: Source documents are not 
to be sent unless requested) and fax or e-mail the form to the Shire Global Safety Department. A 
copy of the Shire Clinical Study Serious Adverse Event and Non-serious AEs Required by the 
Protocol Form (and any applicable follow-up reports) must also be sent to the CRO/Shire 
medical monitor using the details specified in the emergency contact information section of the 
protocol.


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8.2.3
Serious Adverse Event Definition
A serious adverse event (SAE) is any untoward medical occurrence (whether considered to be 
related to investigational product or not) that at any dose:

Results in death

Is life-threatening. Note: The term “life-threatening” in the definition of “serious” refers 
to an event in which the subject was at risk of death at the time of the event; it does not 
refer to an event which hypothetically might have caused death if it was more severe.

Requires inpatient hospitalization or prolongation of existing hospitalization. Note: 
Hospitalizations that are the result of elective or previously scheduled surgery for 
pre-existing conditions and have not worsened after initiation of treatment should not be 
classified as SAEs. For example, an admission for a previously scheduled ventral hernia 
repair would not be classified as an SAE; however, complication(s) resulting from a 
hospitalization for an elective or previously scheduled surgery that meet(s) serious 
criteria must be reported as SAE(s).

Results in persistent or significant disability/incapacity

Is a congenital abnormality/birth defect

Is an important medical event. Note: 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 1 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.
8.2.4
Serious Adverse Event Collection Time Frame
All SAEs (regardless of relationship to study) are collected from the time the subject signs the 
informed consent until the defined follow-up period stated in Section 7.1.3 and must be reported 
to the Shire Global Safety Department and the CRO/Shire medical monitor within 24 hours of 
the first awareness of the event.
In addition, any SAE(s) considered “related” to the investigational product and discovered by the 
investigator at any interval after the study has completed must be reported to the Shire Global 
Safety Department within 24 hours of the first awareness of the event.
8.2.5
Serious Adverse Event Onset and Resolution Dates
The onset date of the SAE is defined as the date the event meets serious criteria. The resolution 
date is the date the event no longer meets serious criteria, the date the symptoms resolve, or the
event is considered chronic. In the case of hospitalizations, the hospital admission and discharge 
dates are considered the onset and resolution dates, respectively.


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In addition, any signs or symptoms experienced by the subject after signing the informed consent 
form, or leading up to the onset date of the SAE, or following the resolution date of the SAE, 
must be recorded as an AE, if appropriate.
8.2.6
Fatal Outcome
Any SAE that results in the subject’s death (ie, the SAE was noted as the primary cause of death) 
must have fatal checked as an outcome with the date of death recorded as the resolution date. For 
all other events ongoing at the time of death that did not contribute to the subject’s death, the 
outcome should be considered not resolved, without a resolution date recorded.
For any SAE that results in the subject’s death or any ongoing events at the time of death, unless 
another investigational product action was previously taken (eg, drug interrupted, reduced, 
withdrawn), the action taken with the investigational product should be recorded as “dose not 
changed” or “not applicable” (if the subject never received investigational product). The 
investigational product action of withdrawn should not be selected solely as a result of the 
subject’s death.
8.2.7
Regulatory Agency, Institutional Review Board, Ethics Committee, and Site 
Reporting
The sponsor is responsible for notifying the relevant regulatory authorities of related, unexpected 
SAEs.
In addition the sponsor is responsible for notifying active sites of all related, unexpected SAEs 
occurring during all interventional studies across the anagrelide program. 
The investigator is responsible for notifying the local IRB, local EC, or the relevant local 
regulatory authority of all SAEs that occur at his or her site as required.


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9.
DATA MANAGEMENT AND STATISTICAL METHODS
9.1
Data Collection
The investigators’ authorized site personnel must enter the information required by the protocol 
on the CRF. A study monitor will visit each site in accordance with the monitoring plan and 
review the CRF data against the source data for completeness and accuracy. Discrepancies 
between source data and data entered on the CRF will be addressed by qualified site personnel. 
When a data discrepancy warrants correction, the correction will be made by authorized site 
personnel. Data collection procedures will be discussed with the site at the site initiation visit 
and/or at the investigator’s meeting. Once a subject is randomized, it is expected that site 
personnel will complete the CRF entry within approximately 3 business days of the subject’s 
visit.
9.2
Clinical Data Management
Data are to be entered into a clinical database as specified in the CRO’s data management plan. 
Quality control and data validation procedures are applied to ensure the validity and accuracy of 
the clinical database.
Data are to be reviewed and checked for omissions, errors, and values requiring further 
clarification using computerized and manual procedures. Data queries requiring clarification are 
to be communicated to the site for resolution. Only authorized personnel will make corrections to 
the clinical database, and all corrections are documented in an auditable manner.
9.3
Data Handling Considerations
Not applicable.
9.4
Statistical Analysis Process
The study will be analyzed by the sponsor or its agent.
The statistical analysis plan (SAP) will provide the statistical methods and definitions for the 
analysis of the pharmacokinetic, pharmacodynamic, and safety data, as well as describe the 
approaches to be taken for summarizing other study information such as subject disposition, 
demographics and baseline characteristics, investigational product exposure, and prior and 
concomitant medications. The SAP will also include a description of how missing, unused and 
spurious data will be addressed.
To preserve the integrity of the statistical analysis and study conclusions, the SAP will be 
finalized prior to database lock.
All statistical analyses will be performed using SAS(SAS Institute, Cary, NC 27513) 
version 9.4 or higher.
9.5
Planned Interim Analysis, Adaptive Design, and Data Monitoring Committee
There is no planned interim analysis, adaptive design, or Data Monitoring Committee in this 
study. 


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9.6
Sample Size Calculation and Power Considerations
Approximately 20 subjects will be enrolled. Subjects who discontinue will not be replaced.
Sample size estimation is to achieve a desired precision of the effect of interest, targeting the 
width of the 90% confidence interval.
From a previous crossover study (SPD422-110), the within-subject coefficient variation in 
anagrelide log-transformed AUC was estimated to be 0.132. Allowing for a slightly larger 
within-subject CV of 0.15, a sample size of 11 subjects is required to estimate the mean 
difference between anagrelide + omeprazole and anagrelide alone in the log-transformed AUC 
with an error margin of at most ±0.2231 at 90% confidence with a probability of 90%. Thus, if 
the true geometric mean treatment ratio is 0.6, the lower and upper 90% confidence bounds will 
be estimated to be within 0.48 and 0.75 (exp[ln(0.6) – 0.2231] and exp[ln(0.6) + 0.2231]) with 
90% probability. Therefore, with an overall sample size of 20 subjects, anticipated 13 to 
14 CYP2C19 EM subjects and 6 to 7 CYP2C19 IM/PM subjects, this statistical power will be 
met for the overall population and the CYP2C19 EM population. The confidence bounds for the 
smaller population of CYP2C19 IM/PM subjects will be slightly wider.
9.7
Study Population
The enrolled set consists of all subjects for whom an enrollment number has been assigned. 
Usually, these are the subjects who meet the study inclusion/exclusion criteria and sign the 
informed consent form.
The safety set consists of all subjects who have taken at least 1 dose of investigational product 
(anagrelide or omeprazole) and have at least 1 post-dose safety assessment.
The pharmacokinetic set consists of subjects who receive at least 1 dose of study drug and has at 
least 1 measureable post-dose plasma concentration. The PK analyses will be based on this 
population.
9.8
Pharmacokinetic and Pharmacodynamic Analyses
9.8.1
Pharmacokinetic Analysis
All the pharmacokinetic analyses will be performed using the pharmacokinetic set.
Pharmacokinetic parameters will be determined from the plasma concentration-time data for 
anagrelide and its metabolites and omeprazole by non-compartmental analysis and all 
calculations will be based on the actual time since dose. The pharmacokinetic parameters will 
include, but not be limited to:
AUC0-
Area under the curve extrapolated to infinity, calculated using the observed 
value of the last non-zero concentration (anagrelide and metabolites)
AUC0-t
Area under the curve from time 0 to the last time point of sample collection 
(anagrelide and metabolites)


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AUC0-tau
Area under the concentration-time curve from time zero to the end of the 
dosing interval (24 hours post dose) (omeprazole only)
CL/F
Total body clearance for extravascular administration.
Cmax
Maximum concentration occurring at tmax
Cmin
The minimum concentration observed over the dosing interval (omeprazole 
only)
t½
Terminal half-life
tmax
Time of maximum observed concentration sampled during a dosing interval
Vz/F
Volume of distribution associated with the terminal slope following 
extravascular administration.
The primary pharmacokinetic parameters will include the Cmax, AUC0-t and AUC0-∞for 
anagrelide and its active metabolite, 3-OH-anagrelide (BCH24426).
The geometric mean relative bioavailability and 90% confidence limits for the effect of 
omeprazole will be evaluated (eg, AUCanagrelide+omeprazole/AUCanagrelide).
Because the CYP2C19-mediated metabolism of omeprazole is subject to genetic polymorphism 
(approximately 2/3 subjects are CYP2C19 extensive metabolizers with lower omeprazole AUC 
and 1/3 subjects are CYP2C19 intermediate/poor metabolizers with higher omeprazole AUC), 
the geometric mean relative bioavailability and 90% CI will be calculate for (1) all subjects, 
(2) EMs, and (3) IMs/PMs
The observed omeprazole AUC0-tau will be used to define the CYP2C19 metabolizer status of 
subjects, with AUC0-tau <5,200 ng•h/mL (<15 µmol/L) classified as CYP2C19 EM and AUC0-tau
≥5,200 ng•h/mL (≥15 µmol/L) classified as CYP2C19 IM/PM.
9.8.1.1
Statistical Analysis of Pharmacokinetic Parameters
The following analyses will be conducted on the Pharmacokinetic Analysis Set. All statistical 
analyses will be presented for the CYP2C19 EM group, the CYP2C19 IM/PM group, and 
overall.
Summary statistics (number of observations, mean, standard deviation, coefficient of 
variation, median, maximum, minimum, and geometric mean) will be determined for all 
pharmacokinetic parameters and presented by treatment. Plasma concentrations of anagrelide, 
3-hydroxy-anagrelide, RL603, and omeprazole at each nominal sampling time will also be 
summarized by treatment using descriptive statistics.
The log-transformed pharmacokinetic parameters will be compared between the 2 treatments 
(anagrelide plus omeprazole and anagrelide alone) using a 2-factor analysis of variance model 
for a single-sequence crossover design with fixed factors for treatment and subject. The 
magnitude of the effect of omeprazole on the pharmacokinetic profile of anagrelide and its 
metabolites will be evaluated by the point estimate and ninety percent (90%) confidence 
intervals for the treatment difference on the log-transformed parameters back-calculated to the 
original scale for the comparisons of Cmax, AUC0-t, and AUC0- ∞(test treatment=anagrelide plus 
omeprazole, reference treatment=anagrelide alone).


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9.9
Safety Analyses
The safety endpoints will be summarized with descriptive statistics for the safety set. 
Relevant safety endpoints as well as their changes from baseline will be summarized. Baseline is 
defined as the last assessment prior to the first dose of investigational product. Potentially 
clinically important findings will also be summarized or listed. The potentially clinically 
important values will be defined in the SAP.
Adverse events will be coded using the Medical Dictionary for Regulatory Activities. The 
number of events, incidence, and percentage of treatment-emergent adverse events will be 
calculated overall, by system organ class, by preferred term, and by treatment group. 
Treatment-emergent adverse events will be further summarized by severity and relationship to 
investigational product. Adverse events related to investigational product, AEs leading to 
withdrawal, SAEs, and deaths will be similarly summarized/listed.
Safety will be assessed by treatment emergent adverse events (TEAEs), new findings from 
physical examination, evaluation of blood pressure, heart rate , new findings from 12-lead ECG, 
and  clinical laboratory test results (biochemistry, hematology, and urinalysis). 
Clinical laboratory tests, vital signs, and ECG findings will be summarized by treatment group 
and visit. Potentially clinically important findings will also be summarized or listed.
9.10
Other Analyses
No other analyses are planned in this study.


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10.
SPONSOR’S AND INVESTIGATOR’S RESPONSIBILITIES
This study is conducted in accordance with current applicable regulations, ICH, EU Directive 
2001/20/EC and its updates, and local ethical and legal requirements.
The name and address of each third-party vendor (eg, CRO) used in this study will be maintained 
in the investigator’s and sponsor’s files, as appropriate.
10.1
Sponsor’s Responsibilities
10.1.1
Good Clinical Practice Compliance
The study sponsor and any third party to whom aspects of the study management or monitoring 
have been delegated will undertake their assigned roles for this study in compliance with all 
applicable industry regulations, ICH Good Clinical Practice (GCP) Guideline E6 (1996), EU 
Directive 2001/20/EC, as well as all applicable national and local laws and regulations.
Visits to sites are conducted by representatives of the study sponsor and/or the company 
organizing/managing the research on behalf of the sponsor to inspect study data, subjects’ 
medical records, and CRFs in accordance with current GCP and the respective local and 
inter/national government regulations and guidelines. Records and data may additionally be 
reviewed by auditors or by regulatory authorities.
The sponsor ensures that local regulatory authority requirements are met before the start of the 
study. The sponsor (or a nominated designee) is responsible for the preparation, submission, and 
confirmation of receipt of any regulatory authority approvals required prior to release of 
investigational product for shipment to the site.
10.1.2
Public Posting of Study Information
The sponsor is responsible for posting appropriate study information on applicable websites. 
Information included in clinical study registries may include participating investigators’ names 
and contact information.
10.1.3
Submission of Summary of Clinical Study Report to Competent Authorities 
of Member States Concerned and Ethics Committees
The sponsor will provide a summary of the clinical study report to the competent authority of the 
member state(s) concerned as required by regulatory requirement(s) and to comply with the 
Community guideline on GCP. This requirement will be fulfilled within 6 months of the end of 
the study completion date for pediatric studies and within 1 year for non-pediatric studies as per 
guidance.
10.1.4
Study Suspension, Termination, and Completion
The sponsor may suspend or terminate the study, or part of the study, at any time for any reason. 
If the study is suspended or terminated, the sponsor will ensure that applicable sites, regulatory 
agencies and IRBs/ECs are notified as appropriate. 


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Additionally, the discontinuation of a registered clinical study which has been posted to a 
designated public website will be updated accordingly.
10.2
Investigator’s Responsibilities
10.2.1
Good Clinical Practice Compliance
The investigator must undertake to perform the study in accordance with ICH GCP Guideline E6 
(1996), EU Directive 2001/20/EC, and applicable regulatory requirements and guidelines.
It is the investigator’s responsibility to ensure that adequate time and appropriately trained 
resources are available at the site prior to commitment to participate in this study. The 
investigator should also be able to estimate or demonstrate a potential for recruiting the required 
number of suitable subjects within the agreed recruitment period.
The investigator will maintain a list of appropriately qualified persons to whom the investigator 
has delegated significant study-related tasks, and shall, upon request of the sponsor, provide 
documented evidence of any licenses and certifications necessary to demonstrate such 
qualification. Curriculum vitae for investigators and sub-investigators are provided to the study 
sponsor (or designee) before starting the study.
If a potential research subject has a primary care physician, the investigator should, with the 
subject’s consent, inform them of the subject’s participation in the study.
Agreement with the final clinical study report is documented by the signed and dated signature 
of the principal investigator, in compliance with Directive 2001/83/EC as amended by Directive 
2003/63/EC and ICH Guidance E3 (1995).
10.2.2
Protocol Adherence and Investigator Agreement
The investigator and any co-investigators must adhere to the protocol as detailed in this 
document. The investigator is responsible for enrolling only those subjects who have met 
protocol eligibility criteria. Investigators are required to sign an investigator agreement to 
confirm acceptance and willingness to comply with the study protocol.
If the investigator suspends or terminates the study at their site, the investigator will promptly 
inform the sponsor and the IRB/EC and provide them with a detailed written explanation. The 
investigator will also return all investigational product, containers, and other study materials to 
the sponsor. Upon study completion, the investigator will provide the sponsor, IRB/EC, and 
regulatory agency with final reports and summaries as required by (inter)national regulations.
Communication with local IRBs/ECs, to ensure accurate and timely information is provided at 
all phases during the study, may be done by the sponsor, applicable CRO, investigator, or, for 
multicenter studies, the coordinating principal investigator, according to national provisions, and 
will be documented in the investigator agreement.


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10.2.3
Documentation and Retention of Records
10.2.3.1
Case Report Forms
Case report forms are supplied by the CRO and should be handled in accordance with 
instructions from the sponsor.
The investigator is responsible for maintaining adequate and accurate medical records from 
which accurate information is recorded onto CRFs, which have been designed to record all 
observations and other data pertinent to the clinical investigation. Case report forms must be 
completed by the investigator or designee as stated in the site delegation log.
All data will have separate source documentation; no data will be recorded directly onto the 
CRF.
All data sent to the sponsor must be endorsed by the investigator.
The clinical research associate (CRA)/study monitor will verify the contents against the source 
data per the monitoring plan. If the data are unclear or contradictory, queries are sent for 
corrections or verification of data.
10.2.3.2
Recording, Access, and Retention of Source Data and Study Documents
Original source data to be reviewed during this study will include, but are not limited to: 
subject’s medical file, and original clinical laboratory reports.
All key data must be recorded in the subject’s medical records.
The investigator must permit authorized representatives of the sponsor; the respective national, 
local, or foreign regulatory authorities; the IRB/EC; and auditors to inspect facilities and to have 
direct access to original source records relevant to this study, regardless of media.
The CRA/study monitor (and auditors, IRB/EC, or regulatory inspectors) may check the CRF 
entries against the source documents. The consent form includes a statement by which the 
subject agrees to the monitor/auditor from the sponsor or its representatives, national or local 
regulatory authorities, or IRB/EC, having access to source data (eg, subject’s medical file, 
appointment books, original laboratory reports, X-rays, etc.).
These records must be made available within reasonable times for inspection and duplication, if 
required, by a properly authorized representative of any regulatory agency (eg, the US FDA, 
EMA, UK Medicines and Healthcare products Regulatory Agency) or an auditor.
Essential documents must be maintained according to ICH GCP requirements and may not be 
destroyed without written permission from the sponsor.


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10.2.3.3
Audit/Inspection
To ensure compliance with relevant regulations, data generated by this study must be available 
for inspection upon request by representatives of, for example, the US FDA (as well as other US 
national and local regulatory authorities), the EMA, the Medicines and Healthcare products 
Regulatory Agency, other regulatory authorities, the sponsor or its representatives, and the 
IRB/EC for each site.
10.2.3.4
Financial Disclosure
The investigator is required to disclose any financial arrangement during the study and for 1 year 
after, whereby the outcome of the study could be influenced by the value of the compensation for 
conducting the study, or other payments the investigator received from the sponsor. The 
following information is collected: any significant payments from the sponsor or subsidiaries 
such as a grant to fund ongoing research, compensation in the form of equipment, retainer for 
ongoing consultation or honoraria; any proprietary interest in investigational product; any 
significant equity interest in the sponsor or subsidiaries as defined in 21 CFR 54 2(b) (1998).
10.3
Ethical Considerations
10.3.1
Informed Consent
It is the responsibility of the investigator to obtain written informed consent from all study 
subjects prior to any study-related procedures including screening assessments. All consent 
documentation must be in accordance with applicable regulations and GCP. Each subject or the 
subject’s legally authorized representative, as applicable, is requested to sign and date the subject 
informed consent form or a certified translation if applicable, after the subject has received and 
read (or been read) the written subject information and received an explanation of what the study 
involves, including but not limited to: the objectives, potential benefits and risk, inconveniences, 
and the subject’s rights and responsibilities. A copy of the informed consent documentation (ie, a 
complete set of subject information sheets and fully executed signature pages) must be given to 
the subject or the subject’s legally authorized representative, as applicable. This document may 
require translation into the local language. Signed consent forms must remain in each subject’s 
study file and must be available for verification at any time.
The principal investigator provides the sponsor with a copy of the consent form that was 
reviewed by the IRB/EC and received their favorable opinion/approval. A copy of the IRB/EC’s 
written favorable opinion/approval of these documents must be provided to the sponsor prior to 
the start of the study unless it is agreed to and documented (abiding by regulatory guidelines and 
national provisions) prior to study start that another party (ie, sponsor or coordinating principal 
investigator) is responsible for this action. Additionally, if the IRB/EC requires modification of 
the sample subject information and consent document provided by the sponsor, the 
documentation supporting this requirement must be provided to the sponsor.


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10.3.2
Institutional Review Board or Ethics Committee
For sites outside the EU, it is the responsibility of the investigator to submit this protocol, the 
informed consent document (approved by the sponsor or their designee), relevant supporting 
information and all types of subject recruitment information to the IRB/EC for review, and all 
must be approved prior to site initiation.
The applicant for an EC opinion can be the sponsor or investigator for sites within the EU; for 
multicenter studies, the applicant can be the coordinating principal investigator or sponsor, 
according to national provisions.
Responsibility for coordinating with IRBs/ECs is defined in the clinical trial agreement.
Prior to implementing changes in the study, the sponsor and the IRB/EC must approve any 
revisions of all informed consent documents and amendments to the protocol unless there is a 
subject safety issue.
Investigational product supplies will not be released until the sponsor has received written 
IRB/EC approval of and copies of revised documents.
For sites outside the EU, the investigator is responsible for keeping the IRB/EC apprised of the 
progress of the study and of any changes made to the protocol, but in any case at least once a 
year; for sites within the EU, this can be done by the sponsor or investigator, or, for multicenter 
studies, it can be done by the coordinating principal investigator, according to national 
provisions. The investigator must also keep the local IRB/EC informed of any serious and 
significant AEs.
10.4
Privacy and Confidentiality
All US-based sites and laboratories or entities providing support for this study, must, where 
applicable, comply with HIPAA of 1996. A site that is not a covered entity as defined by HIPAA 
must provide documentation of this fact to the sponsor.
The confidentiality of records that may be able to identify subjects will be protected in 
accordance with applicable laws, regulations, and guidelines.
After subjects have consented to take part in the study, the sponsor and/or its representatives’ 
reviews their medical records and data collected during the study. These records and data may, in 
addition, be reviewed by others including the following: independent auditors who validate the 
data on behalf of the sponsor; third parties with whom the sponsor may develop, register, or 
market SPD422 (anagrelide); national or local regulatory authorities; and the IRB(s)/EC(s)
which gave approval for the study to proceed. The sponsor and/or its representatives accessing 
the records and data will take all reasonable precautions in accordance with applicable laws, 
regulations, and guidelines to maintain the confidentiality of subjects’ identities.


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Subjects are assigned a unique identifying number; however, their initials and date of birth may 
also be collected and used to assist the sponsor to verify the accuracy of the data (eg, to confirm 
that laboratory results have been assigned to the correct subject).
The results of studies – containing subjects’ unique identifying number, relevant medical 
records, and possibly initials and dates of birth – will be recorded. They may be transferred to, 
and used in, other countries which may not afford the same level of protection that applies within 
the countries where this study is conducted. The purpose of any such transfer would include: to 
support regulatory submissions, to conduct new data analyses to publish or present the study 
results, or to answer questions asked by regulatory or health authorities.
10.5
Study Results/Publication Policy
Shire will endeavor to publish the results of all qualifying, applicable, and covered studies 
according to external guidelines in a timely manner regardless of whether the outcomes are 
perceived as positive, neutral, or negative. Additionally, Shire adheres to external guidelines (eg, 
Good Publication Practices 2) when forming a publication steering committee. The purpose of 
the publication steering committee is to act as a non-commercial body that advises or decides on 
dissemination of scientific study data in accordance with the scope of this policy.
All publications relating to Shire products or projects must undergo appropriate technical and 
intellectual property review, with Shire agreement to publish prior to release of information. The 
review is aimed at protecting the sponsor’s proprietary information existing either at the 
commencement of the study or generated during the study. To the extent permitted by the 
publisher and copyright law, the principal investigator will own (or share with other authors) the 
copyright on his/her publications. To the extent that the principal investigator has such sole, joint 
or shared rights, the principal investigator grants the sponsor a perpetual, irrevocable, 
royalty-free license to make and distribute copies of such publications.
The term “publication” refers to any public disclosure including original research articles, review 
articles, oral presentations, abstracts and posters at medical congresses, journal supplements, 
letters to the editor, invited lectures, opinion pieces, book chapters, electronic postings on 
medical/scientific websites, or other disclosure of the study results, in printed, electronic, oral or 
other form.
Subject to the terms of the paragraph below, the investigator shall have the right to publish the 
study results, and any background information provided by the sponsor that is necessary to 
include in any publication of study results, or necessary for other scholars to verify such study 
results. Notwithstanding the foregoing, no publication that incorporates the sponsor’s 
confidential information shall be submitted for publication without the sponsor’s prior written 
agreement to publish, and shall be given to the sponsor for review at least 60 days prior to 
submission for publication. If requested in writing by Shire, the institution and principal 
investigator shall withhold submission of such publication for up to an additional 60 days to 
allow for filing of a patent application.


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If the study is part of a multicenter study, the first publication of the study results shall be made 
by the sponsor in conjunction with the sponsor’s presentation of a joint, multicenter publication 
of the compiled and analyzed study results. If such a multicenter publication is not submitted to a 
journal for publication by the sponsor within an 18-month period after conclusion, abandonment, 
or termination of the study at all sites, or after the sponsor confirms there shall be no multicenter 
study publication of the study results, an investigator may individually publish the study results 
from the specific site in accordance with this section. The investigator must, however, 
acknowledge in the publication the limitations of the single-site data being presented.
Unless otherwise required by the journal in which the publication appears, or the forum in which 
it is made, authorship will comply with the International Committee of Medical Journal Editors 
(ICMJE) current standards. Participation as an investigator does not confer any rights to 
authorship of publications.


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11.
REFERENCES
Ahluwalia, M., Donovan, H., Singh, N., Butcher, L. and Erusalimsky, J. D. 2010. Anagrelide 
represses GATA-1 and FOG-1 expression without interfering with thrombopoietin 
receptor signal transduction. J Thromb Haemost, 8, 2252-61.
Nousbaum, J., Berthou, F., Carlhant, D., Riche, C., Robaszkiewicz, M. and Gouerou, H. 1994. 
Four-week treatment with omeprazole increases the metabolism of caffeine. American 
Journal of Gastroenterology, 89, 371-5.
Rost, K., Brosicke, H., Brockmoller, J., Scheffler, M., Helge, H. and Roots, I. 1992. Increase of 
cytochrome P4501A2 activity by omeprazole: Evidence by the 13C-[N-3-methyl]-
caffeine breath test in poor and extensive metabolizers of S-mephenytoin. Clin 
Pharmacol Ther, 52, 170-180.
Rost, K. and Roots, I. 1994. Accelerated caffeine metabolism after omeprazole treatment is 
indicated by urinary metabolite ratios: Coincidence with plasma clearance and breath test. 
Clinical Pharmacology & Therapeutics, 55, 402-411.
Troy, S., Parks, V., Purkayastha, J., Gossart, S., Goodman, D., Achenbach, H., et al. 2014. Effect 
of anagrelide on cardiac repolarization in healthy volunteers: a randomized, double-blind, 
placebo- and positive-controlled, thorough QT study. Pharmacology Research & 
Perspectives, 2, e00080.


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12.
APPENDICES



SAP_001.pdf: 
Shire  
                                                       CONFIDENTIAL 
Page 1 of 31 
Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
08 May 2019 
 
STATISTICAL ANALYSIS PLAN 
SPD422, Anagrelide Hydrochloride 
PHASE 1 
A Phase 1, Open-label, Single-sequence, Non-randomized, Crossover, 
Drug-Drug Interaction Study to Evaluate the Effect of Omeprazole on 
the Pharmacokinetics of SPD422 (anagrelide hydrochloride) in Healthy 
Adult Subjects 
PROTOCOL IDENTIFIER: SPD422-113 
 
Study Sponsor(s): 
Shire 
300 Shire Way, Lexington, MA 02421 USA 
Author: 
 
 
 
Protocol: 
2018 OCT 04 Original 
SAP Version #: 
1.0 
SAP Date: 
2019 MAY 08 
Status: 
Final 
 
 
 
 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
08 May 2019 
REVISION HISTORY 
Version  
Issue Date 
Summary of Changes 
1.0 
08MAY2019 
Original final version 
 
 
 
 
 
 
 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
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TABLE OF CONTENTS 
REVISION HISTORY ..................................................................................................... 2 
TABLE OF CONTENTS ................................................................................................. 3 
ABBREVIATIONS ........................................................................................................... 6 
1. 
INTRODUCTION ................................................................................................. 8 
2. 
OBJECTIVES, ESTIMANDS, AND ENDPOINTS .......................................... 8 
2.1 
Objectives ........................................................................................................... 8 
2.1.1 Primary Objective ....................................................................................... 8 
2.1.2 Secondary Objective ................................................................................... 8 
2.2 
Endpoints ............................................................................................................ 8 
2.2.1 Primary Endpoints ...................................................................................... 8 
2.2.2 Secondary Endpoints .................................................................................. 8 
3. 
STUDY DESIGN ................................................................................................... 8 
3.1 
General Description ........................................................................................... 8 
3.2 
Randomization ................................................................................................... 9 
3.3 
Blinding .............................................................................................................. 9 
3.4 
Sample Size and Power Considerations ............................................................. 9 
4. 
STATISTICAL ANALYSIS SETS .................................................................... 10 
4.1 
Enrolled Set ...................................................................................................... 10 
4.2 
Safety Set ......................................................................................................... 10 
4.3 
Pharmacokinetic Set ......................................................................................... 10 
5. 
STUDY SUBJECTS ............................................................................................ 11 
5.1 
Disposition of Subjects .................................................................................... 11 
5.2 
Demographic and Other Baseline Characteristics ............................................ 11 
5.3 
Medical History ................................................................................................ 12 
5.4 
Prior Medications ............................................................................................. 12 
5.5 
Concomitant Medications ................................................................................ 12 
5.6 
Exposure to Investigational Product ................................................................ 13 
5.7 
Measurements of Treatment Compliance ........................................................ 13 
5.8 
Protocol Deviations .......................................................................................... 13 


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6.
EFFICACY ANALYSES .................................................................................... 13 
7.
SAFETY ANALYSIS .......................................................................................... 14 
7.1 
Adverse Events ................................................................................................. 14 
7.2 
Clinical Laboratory Data .................................................................................. 15 
7.3 
Vital Signs ........................................................................................................ 18 
7.4 
Electrocardiogram (ECG) ................................................................................ 18 
7.5 
Other Safety Data ............................................................................................. 19 
8.
PHARMACOKINETIC ANALYSIS ................................................................ 19 
8.1 
Drug Concentration .......................................................................................... 19 
8.2 
Handling Below Limit of Quantitation Values ................................................ 20 
8.3 
Pharmacokinetic Parameters ............................................................................ 20 
8.4 
Statistical Analysis of Pharmacokinetic Data .................................................. 20 
9.
OTHER ANALYSES .......................................................................................... 21 
10.
INTERIM ANALYSIS/ DATA MONITORING COMMITTEE ................... 23 
11.
DATA HANDLING CONVENTIONS ............................................................. 23 
11.1 
General Data Reporting Conventions .............................................................. 23 
11.2 
Definition of Baseline ...................................................................................... 23 
11.3 
Repeated or Unscheduled Assessments of Safety Parameters ......................... 23 
11.4 
Handling of Missing, Unused, and Spurious Data ........................................... 23 
11.4.1 Missing Date Information for Prior or Concomitant Medications 
(Therapies/Procedures) ............................................................................. 23 
11.4.1.1 Incomplete Start Date ................................................................. 24 
11.4.1.2 Incomplete Stop Date ................................................................. 24 
11.4.2 Missing Date Information for Adverse Events ......................................... 25 
11.4.2.1 Incomplete Start Date ................................................................. 25 
11.4.2.2 Incomplete Stop Date ................................................................. 25 
11.4.3 Missing Severity Assessment for Adverse Events .................................... 26 
11.4.4 Missing Relationship to Investigational Product for Adverse Events ...... 26 
11.4.5 Character Values of Clinical Laboratory Variables .................................. 26 


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12. 
ANALYSIS SOFTWARE .................................................................................. 26 
13. 
CHANGES TO ANALYSIS SPECIFIED IN PROTOCOL ........................... 26 
14. 
REFERENCES .................................................................................................... 27 
15. 
APPENDICES ..................................................................................................... 28 
15.1 
Schedule of Activities ...................................................................................... 28 
 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
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ABBREVIATIONS 
AE 
adverse event 
ANOVA 
analysis of variance 
AUC 
area under the curve 
AUC0-∞ 
area under the curve extrapolated to infinity 
AUC0-t 
area under the curve from time 0 to the last time point of sample 
collection 
AUC0-tau 
area under the curve from time 0 to the end of the dosing interval (24 
hours post dose) 
BLQ 
below limit of quantitation 
BMI 
body mass index 
CI 
confidence interval 
CL/F 
total body clearance 
Cmax 
maximum concentration 
Cmin 
minimum concentration observed over the dosing interval 
Ctrough 
Plasma concentration observed at the end of a dosing interval  
CV 
coefficient of variation 
ECG 
electrocardiogram 
eCRF 
electronic case report form 
EM 
extensive metabolizer 
IM/PM 
intermediate metabolizers/poor metabolizer 
LLOQ 
lower limit of quantification 
MedDRA 
Medical Dictionary for Regulatory Activities 
MR 
metabolite ratio 
PCI 
potentially clinically important 
PK 
pharmacokinetic(s) 
PT 
preferred term  
Q1 
25th Percentile 
Q3 
75th Percentile 
QD 
once daily 
QTcB 
QT interval corrected for heart rate using Bazett’s formula 
QTcF 
QT interval corrected for heart rate using Fridericia’s formula 
SAP 
statistical analysis plan 
SD 
standard deviation  
SOC 
system organ class 
t½ 
terminal half-life 
TEAE   
treatment-emergent adverse event 
TFL 
tables, figures, and listings 
tmax 
time of maximum observed concentration sampled during a dosing 
interval 


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Vz/F 
volume of distribution associated with the terminal slope following 
extravascular administration 
WHO 
World Health Organization 


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1. 
INTRODUCTION  
This statistical analysis plan (SAP) provides a technical and detailed elaboration of the 
statistical analyses of pharmacokinetic (PK) and safety data described in the original 
study protocol dated 04 Oct 2018. Specifications for tables, figures, and listings (TFLs) 
are contained in a separate document (SPD422-113 TFL Shells). 
2. 
OBJECTIVES, ESTIMANDS, AND ENDPOINTS 
2.1 
Objectives 
2.1.1 Primary Objective 
The primary objective of this study is to assess the effect of a CYP1A2 inducer 
(omeprazole) on the PK of anagrelide when administered concurrently. 
2.1.2 Secondary Objective 
The secondary objective of this study is to assess the safety and tolerability of anagrelide, 
omeprazole, and co-administered anagrelide plus omeprazole. 
2.2 
Endpoints 
2.2.1 Primary Endpoints 
The primary endpoints will be maximum concentration (Cmax) and area under the curve 
from time 0 to the last time point of sample collection (AUC0-t) and area under the curve 
extrapolated to infinity (AUC0-∞) of anagrelide and 3-hydroxyanagrelide compared 
between anagrelide administered alone and anagrelide administered after omeprazole 40 
mg once daily (QD) for 7 days. 
2.2.2 Secondary Endpoints 
Safety and tolerability will be assessed throughout the study by routine recording of 
electrocardiograms (ECGs), vital signs, safety laboratory assessments, and adverse events 
(AEs). 
3. 
STUDY DESIGN 
3.1 
General Description 
This is a Phase 1, open-label, single-sequence, non-randomized, multiple-dose, crossover 
PK study to assess the effect of a CYP1A2 inducer (omeprazole 40 mg QD) on the PK of 
anagrelide (1 mg) when administered concurrently in healthy subjects. Up to 35 healthy 


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male and female subjects, aged 18 to 45 years will be screened to ensure enrollment and 
completion of 20 subjects. Subjects who discontinue from the study will not be replaced. 
The study design is shown in the study schema in Figure 1. 
Figure 1: Study Design Flow Chart 
3.2 
Randomization 
Not applicable. 
3.3 
Blinding 
Not applicable. 
3.4 
Sample Size and Power Considerations 
 Up to 35 healthy male and female subjects, aged 18 to 45 years will be screened to 
ensure enrollment and completion of 20 subjects. Subjects who discontinue will not be 
replaced. 


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Sample size estimation is to achieve a desired precision of the effect of interest, targeting 
the width of the 90% confidence interval (CI). 
From a previous crossover study (SPD422-110), the within-subject coefficient variation 
(CV) in anagrelide log-transformed AUC was estimated to be 0.132. Allowing for a 
slightly larger within-subject CV of 0.15, a sample size of 11 subjects is required to 
estimate the mean difference between anagrelide + omeprazole and anagrelide alone in 
the log­transformed AUC with an error margin of at most ±0.2231 at 90% confidence 
with a probability of 90%. Thus, if the true geometric mean treatment ratio is 0.6, the 
lower and upper 90% confidence bounds will be estimated to be within 0.48 and 0.75 
(exp[ln(0.6) – 0.2231] and exp[ln(0.6) + 0.2231]) with 90% probability. Therefore, with 
an overall sample size of 20 subjects, anticipated 13 to 14 CYP2C19 extensive 
metabolizer (EM) subjects and 6 to 7 CYP2C19 intermediate/poor metabolizer (IM/PM) 
subjects, this statistical power will be met for the overall population and the CYP2C19 
EM population. The confidence bounds for the smaller population of CYP2C19 IM/PM 
subjects will be slightly wider. 
4. 
STATISTICAL ANALYSIS SETS 
4.1 
Enrolled Set 
The Enrolled Set consists of all subjects for whom an enrollment number has been 
assigned. Usually, these are the subjects who meet the study inclusion/exclusion criteria 
and sign the informed consent form. 
4.2 
Safety Set   
The Safety Set consists of all subjects who have taken at least 1 dose of investigational 
product (anagrelide or omeprazole) and have at least 1 post-dose safety assessment.  
4.3 
Pharmacokinetic Set  
The PK Set consists of subjects who receive at least 1 dose of investigational product 
(anagrelide or omeprazole) and have at least 1 measurable post-dose plasma 
concentration. Each subject’s data (eg, dosing records, AEs, sample collection records, 
etc.) will be reviewed for inclusion/exclusion from the descriptive statistics and statistical 
analysis on a case-by-case basis at the discretion of the Pharmacokineticist. 


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5. 
STUDY SUBJECTS 
5.1 
Disposition of Subjects 
The number of subjects who were included in each defined analysis set (Enrolled, Safety, 
and PK) will be summarized overall.  
The number and percentage of subjects who completed the study or prematurely 
discontinued will be presented overall, along with primary reasons for discontinuation, as 
recorded on the study completion/early termination page of the electronic case report 
form (eCRF). All enrolled subjects who prematurely discontinued the study will be listed 
in the subject disposition listing along with reasons for discontinuation.  
Subject disposition summary and listings will be based on the Enrolled Set. 
5.2 
Demographic and Other Baseline Characteristics 
Descriptive summaries of demographic and baseline characteristics will be generated for 
the Safety Set and the PK Set.  
The observed omeprazole AUC0-tau will be used to define the CYP2C19 metabolizer 
status of subjects, with AUC0-tau <5,200 ng•h/mL (<15 μmol/L) classified as CYP2C19 
EM and AUC0-tau ≥5,200 ng•h/mL (≥15 μmol/L) classified as CYP2C19 IM/PM. The 
summaries of demographic and baseline characteristics for the PK Set will be presented 
by the CYP2C19 metabolizer status (EMs and IMs/PMs) and overall. 
The following demographic and baseline characteristics will be summarized in the 
following order in the tables: age (years), sex, ethnicity, race, weight (kg), height (cm), 
and body mass index (BMI, kg/m2). 
Continuous variables, such as age, weight, height, and BMI will be summarized using 
descriptive statistics including number of subjects, mean, standard deviation (SD), 
median, minimum, and maximum. Categorical variables such as sex, ethnicity and race 
will be summarized by reporting the number and percentage of subjects in each category. 
All summarized values will be taken from the latest available assessments at the 
Screening Visit. 
Demographic and baseline characteristics will be listed for the Safety Set. 


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5.3 
Medical History 
Medical history will be collected at the Screening visit and will be coded using Medical 
Dictionary for Regulatory Activities (MedDRA) Version 21.1. A listing will be provided 
using the Safety Set. 
Medical history will be summarized by system organ class (SOC) and preferred term 
(PT) for the Safety Set. 
5.4 
Prior Medications  
Prior medications will be coded using the World Health Organization (WHO) Drug 
Dictionary Enhanced Version dated 01 September 2018. 
Prior medication is defined as any medication with the start date prior to the date of first 
dose of investigational product. 
Prior medication usage will be summarized by the number and proportion of subjects 
receiving each medication by PT and overall for the Safety Set. Multiple medication 
usage by a subject in the same category will be counted only once. 
All prior medications will be listed for the Safety Set. 
5.5 
Concomitant Medications  
Concomitant medications will be coded using the WHO Drug Dictionary Enhanced 
Version dated 01 September 2018. 
Concomitant medication is defined as any medication with a start date prior to the date of 
the first dose of investigational product and continuing after the first dose of 
investigational product or with a start date between the dates of the first and last doses of 
investigational product, inclusive. Any medication with a start date after the date of the 
last dose of investigational product will not be considered a concomitant medication. 
Concomitant medication usage will be summarized by the number and proportion of 
subjects receiving each medication by PT, by treatment group and overall, for the Safety 
Set. Multiple medication usage by a subject in the same category will be counted only 
once. Concomitant medications will be assigned to the treatment(s) with which they are 
concomitant. A medication is concomitant with a treatment if the subject received that 
medication any time between the time when that treatment was received and when the 
next treatment was received, or in the case of the last treatment, until the end of the 


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follow-up period. Therefore, a medication may be concomitant with more than one 
treatment. 
All concomitant medications will be listed for the Safety Set. 
5.6 
Exposure to Investigational Product 
Exposure to investigational product for the Safety Set will be summarized in terms of 
treatment duration, which is calculated as the number of days from the date of first dose 
of investigational product taken to the date of the last dose of investigational product 
taken, inclusively. Descriptive statistics (number of subjects, mean, standard deviation, 
minimum, median, and maximum) will be presented to describe the exposure to 
investigational product. 
A listing will be created by subject number and visit giving the date and time of dose 
administration for the Safety Set. 
5.7 
Measurements of Treatment Compliance  
This is a study in which the investigational products will be administered from Day 1 to 
Day 8 at the study site only (in-house confinement treatment period). All compliance 
related data will be included in the exposure data listing. No separate treatment 
compliance summary or data listing will be provided. 
5.8 
Protocol Deviations 
Protocol deviations will be recorded by the site separately from the clinical database. 
Protocol deviations will be classified as major or minor per the agreed protocol deviation 
management plan. The Shire study team will review the protocol deviations and their 
classification throughout the study and before database lock. 
Decisions of the review will include categorization of major and minor protocol 
deviations. 
Confirmed major and minor protocol deviations will be documented in the protocol 
deviation tracker for the study. Major/minor protocol deviations will be summarized by 
category for the Safety Set. Major/minor protocol deviations will be listed for the 
Enrolled Set. 
6. 
EFFICACY ANALYSES 
Not applicable. 


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7. 
SAFETY ANALYSIS  
The safety analysis will be performed using the Safety Set. Safety parameters include 
AEs, clinical laboratory results, vital signs, and ECG results. For each safety parameter, 
the last value collected prior to dosing on Day 1 will be used as baseline for all analyses 
of that safety parameter.  
7.1 
Adverse Events  
Adverse events will be coded using MedDRA Version 21.1. 
An AE (classified by PT) will be considered a treatment-emergent AE (TEAE) if it starts 
on or after dosing on Day 1, or if it starts before dosing on Day 1 but increases in severity 
on or after dosing on Day 1 through the end of the study. If more than 1 AE with the 
same PT is reported before dosing on Day 1, then the AE with the greatest severity will 
be used as the benchmark for comparison to the AEs occurring after dosing on Day 1 
under the PT. 
Treatment most recently taken when the AE occurred will be used for the summaries: 
• Any AE that started (or increased in severity) on or after the administration of 
anagrelide on Day 1, but before the administration of omeprazole on Day 2 will 
be assigned to anagrelide alone; 
• Any AE that started (or increased in severity) on or after the administration of 
omeprazole on Day 2, but before the administration of anagrelide on Day 8 will 
be assigned to omeprazole alone; 
• Any AE that started (or increased in severity) on or after the administration of 
anagrelide on Day 8 will be assigned to anagrelide with omeprazole. 
An overall summary of the number of subjects with TEAEs as well as the number of 
events will be presented by treatment group, including the number and percentage of 
subjects with any TEAEs, serious TEAEs, TEAEs related to investigational product, 
serious TEAEs related to investigational product, TEAEs leading to discontinuation of 
investigational product, serious TEAEs leading to discontinuation of investigational 
product, TEAEs leading to discontinuation from study, serious TEAEs leading to 
discontinuation from study, severe TEAEs, TEAEs leading to hospitalization, and TEAEs 
leading to death. 


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The number and percentage of subjects reporting TEAEs, as well as the number of 
events, in each treatment group and overall will be tabulated by SOC and PT, and by 
SOC, PT, and maximum severity. The TEAEs considered related to investigational 
product will also be summarized by SOC and PT. If more than 1 AE occurs with the 
same PT, after the same treatment for the same subject, then the subject will be counted 
only once for that treatment group and PT using the most severe and most related 
occurrence for the summarization by severity and by relationship to investigational 
product. Summaries by SOC and PT will present SOC sorted alphabetically and PT 
within SOC by descending incidence. 
The TEAEs and related TEAEs will be summarized by PT by descending frequency. 
Serious TEAEs, TEAEs leading to discontinuation of investigational product and serious 
TEAEs leading to death, will be summarized by SOC, PT, and treatment group.  
7.2 
Clinical Laboratory Data 
Descriptive statistics for clinical laboratory values (in SI units) and changes from baseline 
at each assessment time point as well as shift tables from baseline to each visit for 
quantitative variables will be presented by treatment group for the following clinical 
laboratory parameters: 
Hematology 
Hemoglobin, hematocrit, red blood cells, platelet count, white blood cell 
count – total and differential, total neutrophils (absolute), eosinophils 
(absolute), monocytes (absolute), basophils (absolute) and lymphocytes 
(absolute). 
Biochemistry 
Sodium, potassium, glucose, blood urea nitrogen, creatinine, calcium, 
chloride, phosphorus, total protein, total CO2 (bicarbonate), albumin, 
aspartate transaminase, alanine transaminase, gamma glutamyl 
transferase, alkaline phosphatase, total bilirubin, and uric acid. 
Urinalysis 
pH, glucose, protein, blood, ketones, bilirubin, nitrites, leukocyte 
esterase, and specific gravity. 
Clinical laboratory test values are potentially clinically important (PCI) if they meet 
either the low or high PCI criteria listed in Table 1. The number and percentage of 
subjects with post-baseline PCI values will be tabulated overall. The percentages will be 
calculated relative to the number of subjects with available baseline values and at least 
1 post-baseline assessment. The numerator is the total number of subjects with at least 


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1 post-baseline PCI value. A supportive listing of subjects with post-baseline PCI values 
will be provided including the subject number, baseline, and post-baseline values.  
Table 1: 
Criteria for Potentially Clinically Important Laboratory Tests 
Parameter 
Classification  
Criteria: SI Units (Conventional Units) 
Hematology 
Hemoglobin 
LOW and DECREASE 
<100 g/L (10g/dL) AND 
Decrease of ≥ 20 g/L (2.0 g/dL) from baseline value   
HIGH 
>200 g/L (20g/dL) 
Hematocrit 
LOW and DECREASE 
≤0.6 × LLN AND 
Decrease of ≥0.06 L/L (6.0%) from baseline value 
HIGH 
>1.3 × ULN 
Red blood cells count 
LOW 
<3 × 1012/L 
HIGH 
>7.5 × 1012/L 
Platelet count 
LOW 
<0.6 × LLN OR <100 × 109/L (100 × 103/µL) 
HIGH 
>1.5 × ULN OR >500 × 109/L (500 × 103/µL) 
White blood cell count 
LOW 
<0.5 × LLN OR <3.0 × 109/L (3 × 103/µL) 
HIGH 
>2 × ULN OR >16.0 × 109/L (16 × 103/µL) 
Neutrophils  
LOW 
<1.5 × 109/L (1.5 × 103/µL) OR < 40% 
HIGH 
>6.2 × 109/L (6.2 × 103/µL) OR > 70 % 
Eosinophils  
HIGH 
>0.5 × 109/L (0.5 × 103/µL) OR > 10.0% 
Monocytes 
HIGH 
>1.1 × 109/L (1.1 × 103/µL) OR >11 % 
Basophils 
HIGH 
>0.2 × 109/L (0.2 × 103/µL) OR > 2% 
Lymphocytes 
LOW 
<0.8 × 109/L (0.8 × 103/µL) OR < 22 % 
HIGH 
> 4.0 × 109/L (4.0 × 103/µL) OR > 44 % 
Biochemistry 
Sodium 
LOW 
>5 mmol/L (5 mEq/L) below LLN 
HIGH 
>5 mmol/L (5 mEq/L) above ULN 
Potassium 
LOW and DECREASE 
Below LLN AND  
Decrease of >0.5 mmol/L (0.5 mEq/L) from 
baseline value 
HIGH and INCREASE 
Above ULN AND 
Increase of > 0.5 mmol/L (0.5 mEq/L) from 
baseline value 
Glucose (fasting) 
LOW 
≤4.2 mmol/L 
HIGH 
≥6.7 mmol/L 
Blood urea nitrogen 
HIGH 
>1.5 × ULN 
Creatinine 
HIGH and INCREASE 
>150 µmol/L AND 
Increase > 30% from baseline value 
Calcium 
LOW and DECREASE 
Below LLN AND 
Decrease of ≥0.25 mmol/L (1.0 mg/dL) from 
baseline value 


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Parameter 
Classification  
Criteria: SI Units (Conventional Units) 
HIGH and INCREASE 
Above ULN AND 
Increase of ≥ 0.25 mmol/L (1.0 mg/dL) from 
baseline value 
Phosphorus 
LOW 
>0.162 mmol/L (0.5 mg/dL) below LLN 
HIGH 
>0.162 mmol/L (0.5 mg/dL) above ULN 
Total protein 
LOW and DECREASE 
Below LLN AND 
Decrease of ≥20 g/L (2.0 g/dL) from baseline value 
HIGH and INCREASE 
Above ULN AND 
Increase of ≥20 g/L (2.0 g/dL) from baseline value 
Albumin 
LOW and DECREASE 
Below LLN AND 
Decrease of ≥10 g/L (1.0 g/dL) from baseline value 
HIGH and INCREASE 
Above ULN AND 
Increase of ≥10 g/L (1.0 g/dL) from baseline value 
Aspartate transaminase 
HIGH 
>2 × ULN 
Alanine transaminase 
HIGH 
>2 × ULN 
Gamma glutamyl 
transferase 
HIGH 
>1.5 × ULN 
Alkaline phosphatase 
HIGH 
>1.5 × ULN 
Total bilirubin 
HIGH 
>1.5 × ULN 
Uric acid (with normal 
diet) 
LOW and DECREASE 
Below LLN AND 
Decrease of >0.119 mmol/L (2.0 mg/dL) from 
baseline value 
Triiodothyronine 
LOW 
<0.922 nmol/L 
HIGH 
>2.765 nmol/L 
Thyroxine 
LOW 
<57.92 nmol/L 
HIGH 
>140.28 nmol/L 
Thyroid stimulating 
hormone 
LOW 
<0.5 μU/L 
HIGH 
>5.0 μU/L 
Uric acid (with normal 
diet) 
HIGH and INCREASE 
Above ULN AND 
Increase of >0.119 mmol/L (2.0 mg/dL) from 
baseline value 
Urinalysis 
Glucose 
HIGH 
≥1+ 
Protein 
HIGH 
≥2+ 
Blood 
HIGH 
≥2+ 
Ketones 
HIGH 
≥2+ 
Bilirubin 
HIGH 
Positive 
Nitrites 
HIGH 
Positive 
Leukocyte esterase 
HIGH 
Positive 
Abbreviations: LLN=lower limit of normal value provided by the laboratory, ULN=upper limit of 
normal value provided by the laboratory 
 
All laboratory data will be listed for the Safety Set. 


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7.3 
Vital Signs 
Descriptive statistics for vital signs (systolic and diastolic blood pressure and pulse rate) 
and their changes from baseline at each post-baseline visit and at end of study will be 
presented by treatment group. 
Vital sign values will be considered PCI if they meet both the observed value criteria and 
the change from baseline criteria listed in Table 2. The number and percentage of subjects 
with PCI post-baseline values will be tabulated by time point and treatment group. The 
percentages will be calculated relative to the number of subjects with baseline and 
available assessment at that time point in that treatment group. The numerator is the total 
number of subjects with at least 1 PCI post-baseline vital sign value at that time point in 
that treatment group. A supportive listing of subjects with post-baseline PCI values will 
be provided including the subject number, baseline, and post-baseline PCI values.  
Table 2:  
Criteria for Potentially Clinically Important Vital Signs 
Parameter 
Classification  
Criteria 
Sitting and Supine 
Systolic BP (mmHg) 
LOW and DECREASE 
≤90 and decrease of ≥20 from baseline value 
HIGH and INCREASE 
≥140 and increase of ≥20 from baseline value   
Diastolic BP 
(mmHg) 
LOW and DECREASE 
≤50 and decrease of ≥15 from baseline value 
HIGH and INCREASE 
≥90 and increase of ≥15 from baseline value 
Pulse rate (bpm) 
LOW and DECREASE 
≤45 and decrease of >15 from baseline value 
HIGH and INCREASE 
≥100 and increase of >15 from baseline value 
Temperature 
(regardless of method) 
LOW 
<35°C or <95°F 
 
HIGH 
>38.3°C or >100.9°F 
Respiratory rate 
(breaths/min) 
LOW 
<10 
HIGH 
>25 
Abbreviations: BP=blood pressure, bpm=beats per minute. 
 
All vital signs data will be listed for the Safety Set. 
7.4 
Electrocardiogram (ECG) 
Descriptive statistics for ECG variables (heart rate, PR, RR, QRS, QT, and QTc intervals) 
and their changes from baseline at each time point will be presented by treatment group. 
QTc interval will be calculated using both Bazett (QTcB=QT/(RR)1/2) and Fridericia 
(QTcF=QT/(RR)1/3) corrections; and if RR is not available, it will be replaced with 
60/(heart rate) in the correction formula. The ECG interpretation at each time point will 
be summarized by treatment group. A shift table from baseline to each time point for 
qualitative ECG results will be presented by treatment group.  


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Electrocardiogram variable values will be considered PCI if they meet or exceed the 
upper limit values listed in Table 3. The number and percentage of subjects with 
post­baseline PCI values will be tabulated by treatment and time point. The percentages 
will be calculated relative to the number of subjects with available non-PCI baseline and 
available assessment at that time point in that treatment group. The numerator is the total 
number of subjects with at least 1 PCI post-baseline ECG value at that time point in that 
treatment group. A listing of all subjects with post-baseline PCI value will be provided 
including the subject number, baseline, and post-baseline PCI values. 
Table 3:  
Criteria for Potentially Clinically Important ECG Values 
Parameter 
Classification  
Criteria 
Overall evaluation 
ABNORMAL 
Overall evaluation is ABNORMAL 
Rhythm 
NON-SINUS 
RHYTHM 
Rhythm is not SINUS 
Heart rate (bpm) 
LOW and DECREASE 
≤45 and decrease of >15 from baseline value 
HIGH and INCREASE 
≥100 and increase of >15 from baseline value 
PR interval (msec) 
HIGH and INCREASE 
≥200 and increase of ≥20 from baseline value 
QRS interval (msec) 
HIGH 
≥120 
QTc interval (men) 
(msec)* 
HIGH 
>430 and increase from baseline value >30 
QTc interval (women) 
(msec)* 
HIGH 
>450 and increase from baseline value >30 
Abbreviations: bpm=beats per minute; QTc=QT interval corrected.   
*Values noted refer to both Bazett’s (QTcB) and Fridericia’s (QTcF) formula 
7.5 
Other Safety Data 
No other safety assessments/variables are planned for this study. 
8. 
PHARMACOKINETIC ANALYSIS  
8.1 
Drug Concentration 
Blood samples will be drawn from each subject during this study for the determination of 
plasma concentration of anagrelide, 3-hydroxy-anagrelide, and RL603. Serial blood 
samples will be collected on Day 1 and Day 8 for PK analysis at predose, 0.5, 1, 1.5, 2, 
2.5, 3, 4, 8, and 12 hours after administration of the investigational product. Plasma 
concentrations of anagrelide and its metabolites will be measured using a validated 
analytical method. 
Blood samples will be drawn from each subject during this study for the determination of 
plasma concentration of omeprazole. Omeprazole PK sample collections on Days 6 and 


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Day 7 will be taken at pre-dose only. Serial blood samples will be collected on treatment 
on Day 8 for PK analysis at predose, 0.5, 1, 1.5, 2, 2.5, 3, 4, 8, 12, and 24 hours after 
administration of the investigational product. Plasma concentrations of omeprazole will 
be measured using a validated analytical method. 
8.2 
Handling Below Limit of Quantitation Values 
The following procedures will be used for plasma concentrations of anagrelide and its 
metabolites and omeprazole data below the lower limit of quantification (LLOQ): 
• Samples that are below limit of quantification (BLQ) are reported as <LLOQ on 
the data listings, where LLOQ is replaced by the actual value for LLOQ for 
specific PK assay. 
• Samples that are BLQ are treated as zero in the calculation of summary statistics 
(eg, mean, SD, etc.) for the plasma concentrations at individual time points. 
Geometric mean will be set to missing where zero values exist. 
• Mean concentrations are reported as zero if all values are BLQ or zero, and no 
other descriptive statistics are reported. If the calculated mean (± SD) 
concentration is less than the LLOQ, the value will be reported as calculated. The 
mean values derived using these conventions will be used to create the mean 
plasma concentration versus time plots. 
• For calculation of area under the curve (AUC), BLQ values are set equal to zero 
in the dataset loaded into Phoenix® WinNonlin® (Certara USA, Inc, Princeton, 
NJ) for PK analysis. WinNonlin® uses the zero values that occur before the first 
time point with a concentration greater than LLOQ. All BLQ values following the 
first measurable concentration will be set to “missing” in the dataset loaded into 
WinNonlin®. 
• Missing values will not be imputed. 
8.3 
Pharmacokinetic Parameters 
Pharmacokinetic parameters will be determined on Day 1 and Day 8 for anagrelide and 
its metabolites and on Day 8 for omeprazole from the individual plasma concentration 
versus actual time data. Pharmacokinetic parameters will be estimated by 
non­compartmental analysis, as deemed appropriate, using Phoenix WinNonlin (Certara 
USA, Inc., Princeton, NJ) Version 8.0 or higher and all calculations will be based on the 
actual time since dose.  
The PK parameters for anagrelide, 3-hydroxy-anagrelide and RL603 with and without 
omeprazole administration on Day 1 and Day 8, and PK parameters for omeprazole (and 
its metabolite) will include, but may not be limited to: 


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Parameter 
Description 
AUC0-∞ 
Area under the curve extrapolated to infinity, calculated using the 
observed value of the last non-zero concentration, calculated using the 
linear up/log down trapezoidal rule (anagrelide and metabolites) 
AUC0-t 
Area under the curve from time 0 to the last time point of sample 
collection, calculated using the linear up/ log down trapezoidal rule 
(anagrelide and metabolites) 
AUC0-tau 
Area under the curve from time 0 to the end of the dosing interval 
(24 hours post dose), calculated using the linear up/log down 
trapezoidal rule (omeprazole only) 
Cmax 
Maximum concentration  
Cmin 
Minimum concentration observed over the dosing interval 
(omeprazole only) 
Ctrough 
Plasma concentration observed at the end of a dosing interval 
(collected before the next administration [predose concentration at 
Day 6, Day 7 and Day 8]) (omeprazole only) 
tmax 
Time of maximum observed concentration sampled during a dosing 
interval 
t½ 
Terminal half-life, calculated as ln(2)/λz 
CL/F 
Total body clearance, calculated as: Dose/AUC0-∞ (anagrelide only) 
and Dose/AUC0-tau (omeprazole only) 
Vz/F 
Volume of distribution associated with the terminal slope following 
extravascular administration, calculated as: 
CL/F / λz (anagrelide and omeprazole only) 
MRAUC 
Metabolite ratio (anagrelide metabolites only), calculated as: 
AUC0-∞ (metabolite) / AUC0-∞ (parent); molecular weight 
adjustment needed for metabolites. 
MRCmax 
Metabolite ratio (anagrelide metabolites only), calculated as: 
Cmax (metabolite) / Cmax (parent); molecular weight adjustment 
needed for metabolites. 
 
The primary PK parameters will include the Cmax, AUC0-t, and AUC0-∞ for anagrelide and 
its metabolites, 3-OH-anagrelide (BCH24426) (active) and RL603 (inactive). 
8.4 
Statistical Analysis of Pharmacokinetic Data 
Individual anagrelide, 3-hydroxy-anagrelide, RL603, and omeprazole concentrations will 
be listed by subject, treatment, day, and time and summarized by treatment, day, and time 
based on the PK Set. 


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The following descriptive statistics of plasma concentration summary will be provided: 
number of subjects, arithmetic mean, geometric mean, SD, arithmetic CV%, geometric 
CV%, 25th Percentile (Q1), 75th Percentile (Q3), median, minimum, and maximum.  
The mean and individual plasma anagrelide, 3­hydroxy-anagrelide, and RL603, and 
omeprazole concentrations versus time profiles by treatment will be presented in figures 
on both linear and semi-logarithmic scales. Mean (±SD) and individual anagrelide, 
3­hydroxy­anagrelide, RL603, and omeprazole plasma concentrations versus time 
profiles will be presented using nominal time based on the PK Set. 
As well as being summarized and presented graphically for all subjects combined, 
anagrelide, 3-hydroxy-anagrelide, and RL603 concentrations will also be summarized 
and presented graphically by CYP2C19 metabolizer status (EM and IM/PM). Refer to 
Section 5.2 for CYP2C19 metabolizer status criteria. 
Individual anagrelide, 3-hydroxy-anagrelide, RL603, and omeprazole PK parameters will 
be listed by subject, treatment, and day, and summarized by treatment and day based on 
the PK Set with the following descriptive statistics: n, arithmetic mean, arithmetic SD, 
95% CI of the mean, CV%, median, Q1, Q3, minimum, maximum, geometric mean, 95% 
CI of the geometric mean, and geometric CV%. 
As well as being performed for all subjects combined, anagrelide, 3-hydroxy-anagrelide, 
and RL603 parameters will also be analyzed by CYP2C19 metabolizer status (EM and 
IM/PM). 
A linear mixed model using a 2-factor analysis of variance (ANOVA) for a 
single­sequence crossover design with fixed factors for treatment and subject will be used 
to compare the log-transformed PK parameters between the 2 treatments (anagrelide with 
omeprazole versus anagrelide alone). The magnitude of the effect of omeprazole on the 
PK profile of anagrelide and its metabolites will be evaluated by the point estimate and 
ninety percent (90%) CIs for the treatment difference on the log­transformed parameters 
back-calculated to the original scale for the comparisons of Cmax, AUC0-t, and AUC0-∞. 
The geometric mean ratios of the PK parameters comparing anagrelide with omeprazole 
to anagrelide alone and the corresponding 90% CIs will be calculated for (1) all subjects, 
(2) EMs, and (3) IMs/PMs.
9.
OTHER ANALYSES
No other analyses are planned for this study.


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10. 
INTERIM ANALYSIS/ DATA MONITORING COMMITTEE  
There is no planned interim analysis or Data Monitoring Committee in this study. 
11. 
DATA HANDLING CONVENTIONS  
11.1 
General Data Reporting Conventions  
Continuous variables will be summarized using the following descriptive statistics: 
number of subjects, mean, median, standard deviation, Q1, Q3, minimum, maximum. 
Categorical and count variables will be summarized by the number of subjects and the 
percent of subjects in each category.  
See Shire TFL Standards for rules on the number of decimal places to present data and 
p­values. 
11.2 
Definition of Baseline  
Baseline for all safety analyses is defined as the last observed value for the safety 
assessment prior to taking the first dose of investigational product on Day 1 (based on 
dates or date/times).  
11.3 
Repeated or Unscheduled Assessments of Safety Parameters  
If a subject has repeated assessments before the start of investigational product, then the 
results from the final assessment made prior to the start of investigational product will be 
used as baseline. If end of study assessments are repeated or unscheduled, the last 
post­baseline assessment will be used as the end of study assessment for generating 
descriptive statistics. However, all post-baseline assessments will be used for PCI value 
determination and all assessments will be presented in the data listings. 
11.4 
Handling of Missing, Unused, and Spurious Data  
11.4.1 Missing Date Information for Prior or Concomitant Medications 
(Therapies/Procedures) 
For prior or concomitant medications, incomplete (ie, partially missing) start date and/or 
stop date will be imputed. When the start date and the stop date are both incomplete for a 
subject, impute the start date first. 


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11.4.1.1 Incomplete Start Date  
The following rules will be applied to impute the missing numerical fields. If the stop 
date is complete and the imputed start date is after the stop date, then the start date will be 
imputed using the stop date. 
11.4.1.1.1 
Missing Day and Month 
• 
If the year of the incomplete start date is the same as the year of the date of the first 
dose of investigational product, then the day and month of the date of the first dose 
of investigational product will be assigned to the missing fields. 
• 
If the year of the incomplete start date is before the year of the date of the first dose 
of investigational product, then December 31 will be assigned to the missing fields. 
• 
If the year of the incomplete start date is after the year of the date of the first dose 
of investigational product, then 01 January will be assigned to the missing fields. 
11.4.1.1.2 
Missing Month Only 
• 
The day will be treated as missing and both month and day will be replaced 
according to the above procedure. 
11.4.1.1.3 
Missing Day Only 
• 
If the month and year of the incomplete start date are the same as the month and 
year of the date of the first dose of investigational product, then the day of the date 
of the first dose of investigational product will be assigned to the missing day 
• 
If either the year is before the year of the date of the first dose of investigational 
product or if both years are the same, but the month is before the month of the date 
of the first dose of investigational product, then the last day of the month will be 
assigned to the missing day 
• 
If either the year is after the year of the date of the first dose of investigational 
product or if both years are the same, but the month is after the month of the date of 
the first dose of investigational product, then the first day of the month will be 
assigned to the missing day. 
11.4.1.2 Incomplete Stop Date  
The following rules will be applied to impute the missing numerical fields. 
11.4.1.2.1 
Missing Day and Month 
• 
If the year of the incomplete stop date is the same as the year as of the date of the 
last dose of investigational product, then the day and month of the date of the last 
dose of investigational product will be assigned to the missing fields. 
• 
If the year of the incomplete stop date is before the year of the date of the last dose 
of investigational product, then 31 December will be assigned to the missing fields. 


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• 
If the year of the incomplete stop date is after the year of the date of the last dose of 
investigational product, then 01 January will be assigned to the missing fields. 
11.4.1.2.2 
Missing Month Only 
• 
The day will be treated as missing and both month and day will be replaced 
according to the above procedure. 
11.4.1.2.3 
Missing Day Only 
• 
If the month and year of the incomplete stop date are the same as the month and 
year of the date of the last dose of investigational product, then the day of the date 
of the last dose of investigational product will be assigned to the missing day 
• 
If either the year is before the year of the date of the last dose of investigational 
product or if both years are the same, but the month is before the month of the date 
of the last dose of investigational product, then the last day of the month will be 
assigned to the missing day 
• 
If either the year is after the year of the last dose of investigational product or if 
both years are the same, but the month is after the month of the date of the last dose 
of investigational product, then the first day of the month will be assigned to the 
missing day. 
11.4.2 Missing Date Information for Adverse Events  
For AEs with partial start dates, non-missing date parts will be used to determine if the 
AE is treatment-emergent or not. If a determination cannot be made using the 
non-missing date parts as to when the AE occurred relative to study drug administration, 
eg, AE start year and month are the same as the year and month of the first dose of 
investigational product, then the AE will be classified as treatment-emergent. 
To facilitate categorization of AEs as treatment-emergent, imputation of dates can be 
used. For AEs, the default is to only impute incomplete (ie, partially missing) start dates. 
Incomplete stop dates may also be imputed when calculation of the duration of an AE is 
required per the protocol. If imputation of an incomplete stop date is required, and both 
the start date and the stop date are incomplete for a subject, impute the start date first. 
11.4.2.1 Incomplete Start Date  
Follow the same rules as in Section 11.4.1.1. 
11.4.2.2 Incomplete Stop Date  
Follow the same rules as in Section 11.4.1.2. 


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11.4.3 Missing Severity Assessment for Adverse Events  
If severity is missing for an AE starting prior to the date of the first dose of 
investigational product, then a severity of “Mild” will be assigned. If the severity is 
missing for an AE starting on or after the date of the first dose of investigational product, 
then a severity of “Severe” will be assigned. The imputed values for severity assessment 
will be used for incidence summaries, while both the actual and the imputed values will 
be used in data listings. 
11.4.4 Missing Relationship to Investigational Product for Adverse Events  
If the relationship to investigational product is missing for an AE starting on or after the 
date of the first dose of investigational product, a causality of “Related” will be assigned. 
The imputed values for relationship to investigational product will be used for incidence 
summaries, while both the actual and the imputed values will be presented in data 
listings. 
11.4.5 Character Values of Clinical Laboratory Variables 
If the reported value of a clinical laboratory variable cannot be used in a statistical 
analysis due to, for example, that a character string is reported for a numerical variable. 
The appropriately determined coded value will be used in the statistical analysis. 
However, the actual values as reported in the database will be presented in data listings. 
12. 
ANALYSIS SOFTWARE  
Statistical analyses will be performed using Version 9.4 (or newer) of SAS on a suitably 
qualified environment.  
Phoenix WinNonlin (Certara USA, Inc., Princeton, NJ) Version 8.0 (or higher) will be 
used for calculation of PK parameters.  
13. 
CHANGES TO ANALYSIS SPECIFIED IN PROTOCOL 
Not applicable. 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
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14. 
REFERENCES 
Not applicable. 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
 
 
 
            08 May 2019 
15. 
APPENDICES  
15.1 
Schedule of Activities 
Table 4: 
Schedule of Assessments 
 
Screening 
Period 
Day -28 to 
Day -2 
Treatment  
Period 
Early  
Termination a 
 
Follow Up 
Phone 
Contact 7±2 
daysb 
 
Day -1 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 6 
Day 7 
Day 8 
Day 9 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
In-Patientc 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
Inclusion/Exclusion Criteria 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Medical & Medication History 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Physical Examination  
X 
X  
X  
 
 
 
 
 
 
 
X 
X 
 
Height, weight and calculate BMId 
X 
 
 
 
 
 
 
 
 
 
 
 
Vital Signs (blood pressure and 
pulse)e 
X 
X  
X 
 
 
 
 
 
 
X 
X 
X 
 
12-lead ECG e 
X 
X 
X 
 
 
 
 
 
 
X 
X 
X 
 
Drugs of abuse and alcohol screen 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
Urine cotinine 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
HIV, HBsAg, and HCV screen 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Serum Pregnancy (all females) 
X 
X 
 
 
 
 
 
 
 
 
X 
X 
 
Biochemistry, hematology, and 
urinalysis 
X 
X 
X 
 
 
 
 
 
 
X 
X 
X 
 
TSH and T4  
X 
 
 
 
 
 
 
 
 
 
 
 
 
Omeprazole PK Sample 
Collectionf  
 
 
 
 
 
 
 
X f 
X f 
X 
X 
  
 
Anagrelide PK Sample Collection 
 
 
X 
 
 
 
 
 
 
X 
 
  
 
Administration of omeprazoleg  
 
 
 
X 
X 
X 
X 
X 
X 
Xi 
 
 
 
Administration of anagrelide h, i 
 
 
X i 
 
 
 
 
 
 
X i 
 
 
 
Concomitant Medications 
X 
X 
X 
X 
X 
X 
X 
X 
X  
X 
X 
X 
X 
Adverse events/serious adverse 
events 
X 
X 
X 
X 
X 
X 
X 
X 
X  
X 
X 
X 
X 
BMI=body mass index; ECG=electrocardiogram; HIV=human immunodeficiency virus, HBsAG=hepatitis B surface antigen, HCV=hepatitis C virus; TSH=thyroid 
stimulating hormone, T4=thyroxine, PK=pharmacokinetic 


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Statistical Analysis Plan v1.0 
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a An attempt to perform the early termination assessments and procedures will be made for any subject who withdraws or is removed from the study. 
b A follow up telephone contact (or in-person follow-up contact at the investigator’s discretion) will take place 7±2 days following the subject’s last dose of anagrelide. 
c Subjects will remain in-patient at the Clinical Research Center starting on Day -1 until discharged following the 24-hour post dose PK sample and safety assessments on the 
morning of Day 9  
d Height to be measured without shoes and weight to be taken in light clothes 
e Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs. Vital sign assessments during the screening visit only include 
blood pressure, pulse, respiratory rate and temperature.  
f Omeprazole PK sample collections on Days 6 and Day 7 are to be taken at pre-dose only. 
g Omeprazole is to be administered prior to breakfast. 
h Anagrelide should be taken by the subject at 8:00 am. If more than 1 subject is receiving anagrelide on the same day, the dosing interval between subjects may be a stagger 
of 3 minutes between dose administrations. 
i Subjects are required to fast for approximately 10 hours prior to and until 4 hours post dose (after PK and safety assessments) on Days 1 and 8. 
 
 
 
 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
 
 
 
            08 May 2019 
Table 5: 
Schedule of Assessments – Detail of Treatment Period Day 1 
 
Study 
Procedure/Relative 
to IP 
Administration 
Administration  
of  
Anagrelidea 
Anagrelide 
PK Sample 
Collection 
Physical 
Examination 
Vital Signsb 
(BP and 
pulse) 
12-lead  
ECGb 
Biochemistry, 
hematology, 
and urinalysis 
Concomitant 
Medications 
Adverse 
Events/Serious 
Adverse Events 
Pre-dosec 
 
X 
 
X 
X 
X 
X 
X 
0 
Xd 
 
 
 
 
 
X 
X 
0.5 hours 
 
X 
 
 
 
 
X 
X 
1 hour 
 
X 
 
X 
 
 
X 
X 
1.5 hours 
 
X 
 
 
 
 
X 
X 
2 hours 
 
X 
 
X 
 
 
X 
X 
2.5 hours 
 
X 
 
 
 
 
X 
X 
3 hours 
 
X 
 
 
 
 
X 
X 
4 hours 
 
X 
 
X 
 
 
X 
X 
8 hours 
 
X 
 
X 
 
 
X 
X 
12 hours 
 
X 
X 
X 
X 
X 
X 
X 
ECG=electrocardiogram; PK=pharmacokinetic; IP=investigation product 
a The subjects should be administered anagrelide starting at 8:00 am. 
b Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs.  
c Pre-dose assessments should be performed within 30 minutes prior to dose. 
d Subjects are required to fast for approximately 10 hours prior to and until 4 hours post-dose (after PK and safety assessments) on Day 1 and 8. 
 
 
 
 


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Statistical Analysis Plan v1.0 
Protocol Number: SPD422-113 
 
 
 
            08 May 2019 
Table 6: 
Schedule of Assessments – Detail of Treatment Period Day 8  
 
Study 
Procedure/Relative 
to IP 
Administration 
Administration  
of Anagrelide 
& 
Omeprazolea 
Anagrelide 
PK Sample 
Collection 
Omeprazole 
PK Sample 
Collection 
 
Physical 
Examination 
Vital Signsb 
(BP and pulse) 
12-lead  
ECGb 
Biochemistry, 
hematology, 
and urinalysis 
Concomitant 
Medications 
Adverse 
Events/Serious 
Adverse 
Events 
Pre-dosec 
 
X 
X 
 
X 
X 
X 
X 
X 
0 
Xd 
 
 
 
 
 
 
X 
X 
0.5 hours 
 
X 
X 
 
 
 
 
X 
X 
1 hour 
 
X 
X 
 
X 
 
 
X 
X 
1.5 hours 
 
X 
X 
 
 
 
 
X 
X 
2 hours 
 
X 
X 
 
X 
 
 
X 
X 
2.5 hours 
 
X 
X 
 
 
 
 
X 
X 
3 hours 
 
X 
X 
 
 
 
 
X 
X 
4 hours 
 
X 
X 
 
X 
 
 
X 
X 
8 hours 
 
X 
X 
 
X 
 
 
X 
X 
12 hours 
 
X 
X 
 
X 
X 
 
X 
X 
24 hours 
 
 
X 
X 
X 
X 
X 
X 
X 
ECG=electrocardiogram; PK=pharmacokinetic; IP=investigation product 
a The subjects should be administered anagrelide and omeprazole concurrently starting at 8:00 am. 
b Subjects should be resting in a supine position for at least 5 minutes prior to collecting vital signs and ECGs.  
c Pre-dose assessments should be performed within 30 minutes prior to dose. 
d Subjects are required to fast for approximately 10 hours prior to and until 4 hours post-dose (after PK and safety assessments) on Days 1 and Day 8.