Conditions: Breast Cancer Female
Keywords: Breast Cancer Breast Carcinoma Breast Tumours Early Breast Cancer
Allocation: NA | Intervention model: SINGLE_GROUP
Enrollment: 36
Healthy volunteers: no

Brief summary:
The purpose of this study is to evaluate the effect of short-term treatment with darolutamide on breast cancer cells (i.e., how the treatment may change the genes or proteins in breast cancer cells) and to evaluate its safety and the way it is tolerated by subjects.

The intent is to study these changes in order to have a better understanding of the potential use of darolutamide for women with EBC, know which patients are likely or unlikely to respond to this treatment, and determine how darolutamide may be combined with other anti-cancer drugs.

Detailed description:
This current study will enroll EBC subjects with differing breast cancer (BC) subtypes, with the intent of characterizing the molecular alterations in BC tissue before and after short-term exposure to the anti-androgen darolutamide. Studying the biological mechanisms in which darolutamide targets the androgen receptor (AR) in BC will be crucial in understanding its' potential role, as well as provide the foundation for further development of darolutamide in this disease.

TRIO030 will evaluate the following objectives: Primary -To identify the molecular alterations that occur in human BC tissue, following short-term exposure to darolutamide in female subjects with EBC; Secondary - To evaluate the safety and tolerability of short-term exposure to darolutamide in female subjects with EBC.

Design: This will be a multi-center, open-label, tissue-acquisition study involving up to 60 subjects from approximately 20 sites in North America and Europe, and will enroll EBC subjects of differing subtypes (i.e., triple negative \[TNBC\], ER+/HER2-, and HER2+), with the intent of characterizing the molecular alterations in BC tissue before and after short-term exposure to the anti-androgen darolutamide.

Such information may suggest that darolutamide be combined with other modalities; for example, if treatment with the drug is discovered to affect multiple signaling pathways. Molecular profiling of tumor samples before and after darolutamide treatment may permit the identification of patients likely or unlikely to respond to the agent based on the biological and molecular characteristics of their tumors.

Duration: It is recommended that the surgery date is defined prior to starting protocol treatment and then the start date of darolutamide (Day 1) will be derived as minus 14 to 21 days from the scheduled surgery date. Treatment will be taken until the day prior to surgery or when the subject is ordered to stop all oral intake (i.e., "nil per os" (npo)), which ever occurs first. If for some reason surgery takes place more than 21 days after treatment start, it is acceptable that the subject receives darolutamide for more than 21 days and up to a maximum of 35 days; subject should continue protocol treatment until the day prior to surgery, or npo is ordered. In these cases it is strongly recommended to have surgery performed as soon as possible after 21 days of treatment are completed.

After surgery, an End of Study (EoS) visit will occur 30 days (+/- 3 days) after the subject's last intake of darolutamide. In case of ongoing protocol treatment-related AEs/SAEs at the time of the EoS visit, monitoring of these events will continue as clinically indicated until (a) the events have resolved or (b) the events have reached a status which, in the Investigator's opinion, is unlikely to resolve due to the nature of the condition and/or the subject's underlying disease.

Total Number of Sites: A total of 14 sites were opened worldwide, with 5 sites located within the U.S, 6 in Canada, and 3 in Germany.

Sample Size/Patient Population: Up to 60 subjects will be enrolled in the study. To be able to assess the molecular alterations after darolutamide, exposure in different BC subtypes, subjects being either triple-negative, or ER+/HER2 negative, or HER2 positive, will be enrolled in the study. Up to 20 subjects with each of these subtypes will be included (with an acceptable minimum of 8 evaluable patients in each cohort).

Dosage Regimen: Darolutamide will be given at a dose of 600 mg (2 x 300 mg tablets) twice daily (b.i.d) to a daily dose of 1200 mg.

The first day of darolutamide administration in the study is considered Day 1. The last darolutamide intake will be on the day prior to breast cancer surgery, when the pre-surgery visit should occur.

darolutamide should be taken approximately at the same time each day twice a day. It is recommended that darolutamide be taken with food; recommendation is to take it with breakfast and dinner each day.

Supportive measures and dose modifications: A subject who experiences a treatment-related grade 3 or higher adverse event (AE) must be withdrawn from protocol treatment and should proceed to surgery and undergo an EoS visit. In the case of a grade 1-2 treatment-related AE, the Investigator should contact the Medical Monitor if a dose reduction or treatment hold is required.

Washout pre-trial: Prior treatment of ovarian hormone replacement therapy should be stopped at least 28 days prior to registration. Use of other investigational drugs should be stopped within 28 days of enrollment. No major surgery within 28 days before enrollment (Major surgery is defined as requiring a general anesthesia or respiratory assistance; involving openings into the great cavities of the body, organs removed, or normal anatomy altered; implying risks of severe hemorrhage; implying risk for life of the patient or severe disability).

Concomitant Medication: Subjects will be instructed to consult with the Investigator before taking any medications (including over-the-counter medications). Supportive medications may be provided prophylactically or therapeutically per Investigator discretion.

Any concomitant treatment NOT listed below is considered permitted in the study and may be prescribed as clinically appropriate during the study:

* Investigational agents other than the protocol treatment.
* Any additional standard or investigational anticancer agents, such as chemotherapy, immunotherapy, targeted/biologic therapy, endocrine therapy, etc., even if utilized as treatment of non-cancer indications. darolutamide must be permanently discontinued upon initiation of a non-protocol standard or investigational antineoplastic therapy prior to surgery.

Rescue Medication and Risk Management: No specific pre-medication is required. Based on the current available data , there are no special warnings and precautions associated with the use of darolutamide.

Premature Withdrawal / Discontinuation Criteria: The patient may withdraw from the study at any time without prejudicing future medical treatment. In any case, the withdrawal should be clearly documented in the subject's clinical records.

Should a patient decide to withdraw consent, all efforts will be made to complete and report the observations as thoroughly as possible. A complete final evaluation at the time of the patient's withdrawal should be made with an explanation of why the patient is withdrawing from the study. After complete withdrawal of consent, no further study procedure is performed and no further data will be collected.

Discontinuation: The Investigator will also discontinue protocol treatment if any of the following conditions is met:

* Intercurrent illness or a change in subject´s condition or unacceptable toxicity that warrants protocol treatment discontinuation according to Investigator's judgment
* Any event, condition, criterion which would warrant discontinuation of darolutamide (i.e., severe toxicities), including grade ≥ 3 protocol-treatment related AE
* Any event, condition, reason which would warrant darolutamide to be held for more than the maximum acceptable delay of 7 consecutive days
* Subject receives non-protocol anti-cancer therapy at any time during the protocol treatment
* Subject's decision to withdraw protocol treatment
* Lost to follow up
* Death
* Pregnancy
* Investigator's decision
* Discontinuation of the study by the sponsor
* Patient is enrolled but does not receive protocol treatment for any reason

Arms:
arm 1: Oral 300 mg darolutamide tablet; dose of 600 mg (2 x 300 mg tablets) b.i.d.

Interventions: intervention 1: darolutamide
Intervention descriptions: intervention 1: Oral 300 mg tablets; 600 mg (2 x 300 mg tablets) taken twice per day, to a daily dose of 1200 mg.

Protocol document:
Prot_000.pdf: 
 
 
 
 
 
Clinical Trial Protocol 
 
 
 
 
Study Title: A Presurgical Tissue-Acquisition Study to Evaluate Molecular Alterations in 
Human Breast Cancer Tissue Following Short-Term Exposure to the Androgen Receptor 
Antagonist Darolutamide (ODM-201) 
 
 
Study Number:  TRIO030 
 
 
Protocol Version, Date:  3.0, 16 February 2018 
 
 
ClinicalTrials.gov Identifier:  NCT03004534 
 


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TRIO030 Protocol v. 3.0 – Rationale for Amendment
TRIO030 protocol was amended due to the following reasons:

Enrollment difficulties faced by participating sites in some of the cohorts defined in the study

Updated data from the most current Investigator´s Brochure

Administrative changes, clarification and other minor edits
TRIO030 protocol was amended as follows:

Sections 1.3.1, 3.1, 4.1, 4.2, 5.3.2.1, 5.3.3, 5.3.4, 5.4.1, 5.4.2, 5.5.2, 6.3, 6.4.3, 6.4.5, 6.5.7.1, 
8.6, and Appendix 2: Wording modified to allow patients candidates for neoadjuvant therapy 
to participate on the trial, as long as patients and Investigators are willing to start such 
therapy once treatment with darolutamide is completed and to undergo a biopsy prior to 
starting neoadjuvant therapy. 

Sections 1.2 and 5.5.1: Sections have been updated as per safety data from Amendment 1 to 
Investigator’s Brochure version 3.0, dated 22-Dec-2017.

Sections 3.2, 6.3, and 6.5.7.1: Allowance of 2 cores for screening samples if 3 are not feasible.

Sections 3.1, 3.2, 6.1.2.2: Allowance of a minimum of 8 evaluable patients per cohort.

Sections 3.1, 6.3, and 6.4.1: Wording added to clarify that patients enrolled but never treated 
will be discontinued from study.

Section 3.3: Potential reasons for the premature termination of the study by the sponsor 
were added. 

Sections 4.1 and 4.2: Criterion modified to allow patients with T1 ≥ 1.0 cm.

Section 4.2: Modifications done on exclusion criteria related to previous or concurrent anti-
cancer treatments. 

Section 5.5.3: Conditions required to consider vasectomised male partner as a highly 
effective contraceptive method were added as a footnote.

Sections 6.3 and 6.4.3: Wording added to allow Pre-surgery visit to take place the same day 
as BC surgery/pre-NAST biopsy.

Section 6.4.3: Section was modified to request that laboratory safety tests are available and 
reviewed in the Pre-surgery visit before surgery is performed.

Section 8.5: Wording modified to perform interim review of data at given time points 
depending on the recruitment, for example once 20, around 35 and once all patients have 
been enrolled and undergone the EoS as per protocol.

Term darolutamide utilized throughout the protocol instead of ODM-201 for consistency with 
Investigator’s Brochure.

TRIO Project Team updated as per latest changes in its composition.


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
Term “patient” utilized throughout the protocol to refer to women participating in the study, 
instead of other terms (e.g. “subject”).

Abbreviation change (HR+ to substitute ER+ where applicable).

Acronym “NAST” was utilized throughout the protocol to refer to Neoadjuvant Systemic 
Therapy.

“Pre-NAST biopsy” utilized throughout the protocol to refer to biopsy to be performed prior 
to start of neoadjuvant treatment in patients who are candidates for such therapy.

Figure 1: Surgery visit name changed to “BC sugery/pre-NAST biopsy”.

Table 2: Schedule of Procedures: footnotes 1 and 12 revised to clarify the timelines for tumor 
samples collection at screening.


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TABLE OF CONTENTS
LIST OF ABBREVIATIONS ..........................................................................................................................................9
PROTOCOL SYNOPSIS.............................................................................................................................................11
1.
BACKGROUND AND RATIONALE....................................................................................................................17
1.1
DISEASE OVERVIEW ....................................................................................................................................17
1.2
INVESTIGATIONAL MEDICINAL PRODUCT OVERVIEW............................................................................................17
1.3
RATIONALE ...............................................................................................................................................18
1.3.1
Rationale for Conducting the Study....................................................................................................18
1.3.2
Rationale for Regimen and Dose Selection..........................................................................................20
2.
OBJECTIVES ...................................................................................................................................................20
2.1
PRIMARY OBJECTIVE....................................................................................................................................20
2.2
SECONDARY OBJECTIVES ..............................................................................................................................20
3.
STUDY DESIGN...............................................................................................................................................20
3.1
OVERALL STUDY DESIGN AND PLAN: DESCRIPTION..............................................................................................20
3.2
NUMBER OF PATIENTS AND INVESTIGATIONAL SITES............................................................................................22
3.3
STUDY END...............................................................................................................................................22
4.
PATIENT SELECTION.......................................................................................................................................22
4.1
INCLUSION CRITERIA....................................................................................................................................23
4.2
EXCLUSION CRITERIA ...................................................................................................................................24
5.
TREATMENT ..................................................................................................................................................25
5.1
PROTOCOL TREATMENT: DAROLUTAMIDE.........................................................................................................25
5.2
PROTOCOL TREATMENT MANAGEMENT ...........................................................................................................25
5.2.1
Packaging and Labeling .....................................................................................................................25
5.2.2
Storage and Accountability................................................................................................................25
5.2.3
Drug Return/Destruction....................................................................................................................25
5.3
PROTOCOL TREATMENT ADMINISTRATION ........................................................................................................25
5.3.1
Pre-medication and Special Precautions.............................................................................................25
5.3.2
Standard Treatment Schedule............................................................................................................26
5.3.3
Protocol Treatment Discontinuation...................................................................................................27
5.3.4
Surgery and (Neo) Adjuvant Systemic Therapy ...................................................................................28
5.4
ADVERSE EVENTS MANAGEMENT AND TREATMENT SCHEDULE ADJUSTMENTS ...........................................................28
5.4.1
General Rules ....................................................................................................................................28
5.4.2
Guidance for Darolutamide Dose Modifications..................................................................................28
5.5
CONCOMITANT MEDICATIONS AND PROCEDURES ...............................................................................................29
5.5.1
Precautions – Drug Interactions .........................................................................................................29
5.5.2
Prohibited Treatments and/or Procedures..........................................................................................30
5.5.3
Contraception....................................................................................................................................30
6.
SCHEDULE OF VISITS AND PROCEDURES........................................................................................................31
6.1
PATIENT INCLUSION ....................................................................................................................................31
6.1.1
Informed Consent ..............................................................................................................................31
6.1.2
Registration and Enrollment ..............................................................................................................32
6.2
STUDY PARTICIPATION DISCONTINUATION ........................................................................................................33
6.2.1
Discontinuation from Protocol Treatment ..........................................................................................33
6.2.2
Discontinuation from Study Participation ...........................................................................................33


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6.3
SCHEDULE OF PROCEDURES...........................................................................................................................34
6.4
STUDY VISITS AND ASSESSMENTS/INVESTIGATIONS .............................................................................................38
6.4.1
Screening Period................................................................................................................................38
6.4.2
Treatment Period Visits......................................................................................................................38
6.4.3
Pre-surgery Visit ................................................................................................................................39
6.4.4
BC Surgery/Pre-NAST biopsy ..............................................................................................................40
6.4.5
End of Study Visit...............................................................................................................................40
6.4.6
Unscheduled Visit Procedures ............................................................................................................40
6.5
DESCRIPTION OF ASSESSMENTS/INVESTIGATIONS ...............................................................................................40
6.5.1
Demographics and Medical History....................................................................................................40
6.5.2
Physical Examination and Vital Signs..................................................................................................41
6.5.3
ECOG Performance Status..................................................................................................................41
6.5.4
Adverse Events and Concomitant Treatments Assessment..................................................................41
6.5.5
Laboratory Assessments ....................................................................................................................41
6.5.6
Assessment of Efficacy.......................................................................................................................41
6.5.7
Tumor Tissue Collection and Molecular Evaluation.............................................................................41
6.5.8
Other Procedures/Assessments..........................................................................................................43
7.
SAFETY MONITORING....................................................................................................................................43
7.1
DEFINITIONS .............................................................................................................................................43
7.1.1
Period of Observation ........................................................................................................................43
7.1.2
Adverse Event....................................................................................................................................43
7.1.3
Serious Adverse Events.......................................................................................................................44
7.1.4
Unexpected Adverse Event.................................................................................................................45
7.2
PERFORMING ADVERSE EVENT ASSESSMENT .....................................................................................................45
7.2.1
Collection of Adverse Event Information.............................................................................................45
7.2.2
Assessment of Causality.....................................................................................................................45
7.3
REPORTING OF SERIOUS ADVERSE EVENTS AND PREGNANCIES ...............................................................................45
7.3.1
Reporting of Serious Adverse Events and Pregnancies from the sites to TRIO ......................................45
7.3.2
SAE information Provided by TRIO to the Manufacturer......................................................................46
7.4
MANAGEMENT OF SPECIFIC CASES..................................................................................................................46
7.4.1
Medication Errors..............................................................................................................................46
7.4.2
Pregnancy .........................................................................................................................................46
7.5
REPORTING TO ETHICS COMMITTEES, REGULATORY AUTHORITIES AND OTHER INVESTIGATORS.......................................47
8.
STATISTICAL CONSIDERATIONS .....................................................................................................................47
8.1
STUDY POPULATIONS ..................................................................................................................................47
8.2
DEMOGRAPHICS AND BASELINE CHARACTERISTICS ..............................................................................................47
8.3
PROTOCOL TREATMENT ...............................................................................................................................47
8.4
SAFETY EVALUATION ...................................................................................................................................47
8.5
SAMPLE SIZE .............................................................................................................................................48
8.6
MOLECULAR ANALYSES................................................................................................................................48
9.
ADMINISTRATIVE, ETHICAL AND REGULATORY STANDARDS.........................................................................48
9.1
STEERING COMMITTEE.................................................................................................................................48
9.2
ETHICAL CONDUCT OF THE STUDY...................................................................................................................49
9.3
INSTITUTIONAL REVIEW BOARD/INDEPENDENT ETHICS COMMITTEE (IRB/IEC)..........................................................49
9.4
COMPLIANCE WITH THE PROTOCOL AND PROTOCOL AMENDMENTS ........................................................................49
9.5
MONITORING, AUDITING AND INSPECTING .......................................................................................................50
9.6
RECORDING, PROCESSING AND RETENTION OF DATA...........................................................................................50
9.7
DATA PROTECTION .....................................................................................................................................50


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9.8
CONFIDENTIALITY AND DATA PROTECTION........................................................................................................51
9.9
INSURANCE OF LIABILITIES.............................................................................................................................51
9.10
USE OF INFORMATION AND PUBLICATION .........................................................................................................51
10.
REFERENCES..............................................................................................................................................51
11.
APPENDICES ..............................................................................................................................................53
APPENDIX 1: EASTERN COOPERATIVE ONCOLOGY GROUP PERFORMANCE STATUS SCALE....................................53
APPENDIX 2: PATIENT DIARY TEMPLATE................................................................................................................54
LIST OF FIGURES
Figure 1: Study Schema ........................................................................................................................... 22
LIST OF TABLES
Table 1: Dose Modifications*...................................................................................................................28
Table 2: Schedule of Procedures..............................................................................................................35
Table 3: Reporting Period of AEs/SAEs.....................................................................................................43
Table 4: SAE and Pregnancies Reporting Timelines..................................................................................46


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INVESTIGATORS
Approximately 20 sites in North America and Europe will participate in the study. The list of Investigators 
participating in the study will be maintained in TRIO’s Clinical Trial Management System.
STUDY STEERING COMMITTEE
List of members available in the Steering Committee Charter


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TRIO PROJECT TEAM
TRIO – 11 rue Watt 75013 PARIS – FRANCE
Karen AFENJAR
Global Project Coordinator
+ 33 1 58 10 09 79
Stéphanie CARREZ
Senior Study Manager
+ 33 1 58 10 08 87
Véronique MOINEAU
Director, Monitoring Resources
+ 33 1 58 10 09 03
Matthieu RUPIN
Director, Strategy and Clinical 
Collaborations
+ 33 1 58 10 08 89
TRIO – Suite 1100, 9925 – 109th Street EDMONTON, AB T5K2J8 - CANADA
Launa ASPESLET
Chief Executive Officer
+ 1 780 702 2260
Melissa BURTON
Senior Data Manager
+ 1 780 702 0228
Yan YANG
Senior Safety Manager
+ 1 780 701 0393
Tamara IRVIN
Project Manager, Regulatory 
Affairs
+ 1 780 702 8398
Helena FUNG
Senior Biostatistician
+ 1 780 702 0246
TRIO – Luis A. de Herrera 1248/1806, 11300 MONTEVIDEO - URUGUAY
Carlos MEYER M.D.
Clinical Research Physician
TRIO030 Lead Medical Monitor
+ 598 2622 2120 ext. 3


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LIST OF ABBREVIATIONS
AE
Adverse Event
AI
Aromatase Inhibitors
ALT
Alanine Aminotransferase / GPT
ANC
Absolute Neutrophil Count
AR
Androgen Receptor
AST
Aspartate Transaminase / GOT
AUC
Area under the curve
BC
Breast Cancer
BCRP
Breast Cancer Resistance Protein
b.i.d.
Twice a Day
CIOMS
Council for International Organizations of Medical Sciences
Cmax
Maximum Serum Concentration
CRPC
Castration-Resistant Prostate Cancer
CSR
Clinical Study Report
CYP
Cytochrome P
EBC
Early Breast Cancer
ECOG
Eastern Cooperative Oncology Group
eCRF
Electronic Case Report Form
EoS
End of Study
ER
Estrogen Receptor
FDA
Food and Drug Administration
g/dL
Grams per deciliter
GCP
Good Clinical Practices
HER2
Human Epidermal Growth Factor Receptor 2
HIPPA
Health Insurance Portability and Accountability Act
HIV
Human Immunodeficiency Virus
HR
Hormone Receptor
IB
Investigator’s Brochure
ICH
International Council for Harmonization
IEC
Independent Ethics Committee
IMP
Investigational Medicinal Product
IRB
Institutional Review Board
ITT
Intention-To-Treat
mCRPC
Metastatic Castration-Resistant Prostate Cancer
MedDRA
Medical Dictionary for Regulatory Activities
mg
Milligram
NAST
Neoadjuvant Systemic Therapy
NCI CTCAE
National Cancer Institute Common Terminology Criteria for Adverse Events
npo
nil per os, nothing by mouth
NYHA
New York Heart Association
P-gp
P-Glycoprotein
PgR
Progesterone Receptor
PICF
Patient Informed Consent Form
PS
Performance status
PSA
Prostate-Specific Antigen
RDC
Remote Data Capture
SAE
Serious Adverse Event
SERM
Selective Estrogen Receptor Modulators
SLNB
Sentinel Lymph Node Biopsy
SoC
Standard of Care
SSC
Study Steering Committee
SUSAR
Suspected Unexpected Serious Adverse Reaction
TNBC
Triple Negative Breast Cancer
TORL
Translational Oncology Research Laboratories
TRIO
Translational Research In Oncology
UCLA
University of California, Los Angeles
ULN
Upper Limit of Normal
US
United States
WBC
White Blood Cells


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WoCBP
Women of Childbearing Potential


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PROTOCOL SYNOPSIS
Protocol Title
A Presurgical Tissue-Acquisition Study to Evaluate Molecular Alterations in Human Breast 
Cancer Tissue Following Short-Term Exposure to the Androgen Receptor Antagonist
Darolutamide (ODM-201)
Protocol #
TRIO030
Study Duration
Enrollment is expected to run over a 12-month period approximately. The overall study is 
expected to last around 14 months (from First Patient In until Last Patient Last Visit).
Sponsor/Study 
Chair
This is a TRIO-sponsored study which will be chaired by Dr. Dennis Slamon (Professor of 
Medicine, Chief, Division of Hematology-Oncology, UCLA's Department of Medicine, CA, 
USA). The study is financially supported by Bayer.
Participating 
Investigators/Sites
Approximately 20 sites in North America and Europe will participate in the study. 
Target Population
Women with early-stage breast cancer (BC) candidates for surgery as primary treatment 
modality or candidates for neoadjuvant treatment who accept to undergo a second biopsy 
before starting such therapy.
Background and 
Rationale
Despite progresses made in diagnostic techniques and treatments for BC, there is still a 
significant number of patients undergoing treatment for early BC (EBC) who will relapse or 
patients with advanced BC that do not respond or progress to their treatments. The medical 
need for new and potentially more active and better tolerated agents in BC remains very 
high.
Endocrine treatment is one of the strategies with a major therapeutic value in patients with 
estrogen-receptor (ER) positive tumors. Although, the role of ER and progesterone receptor 
(PgR) is well established in BC as predictive and prognostic factors, little is known about the 
clinical significance of the androgen receptor (AR). Preclinical studies show that the 
androgen signaling pathway plays a critical role in the development of normal and malignant 
breast tissue. AR is expressed in approximately 80% and 60% of primary and metastatic BC, 
respectively. Its expression varies across the different subtypes, approximately 84-95% in 
ER+ tumors, 50-63% in ER-/HER2+ cases and 10-53% in TNBC. 
In ER+ BC it has been suggested that acquired resistance to antiestrogen therapies could 
result from adaptation from estrogen dependence to androgen dependence. Recently, 
clinical reports of AR antagonism have been promising, especially in TNBC. Bicalutamide and 
enzalutamide have been tested in phase II trials in TNBC and have shown promising clinical 
activity. 
These developments in combination with an improved understanding of AR signaling in 
different BC subtypes led to the renewed interest in targeting AR. However, the value of AR
antagonists in the management of BC, the molecular alterations that occur after exposure to 
these agents and whether there are biomarkers useful to predict their effectiveness, still 
need to be fully elucidated. Increased understanding of AR blockade and its potential value 
in different BC subtypes and clinical testing of newer and more potent AR antagonists like 
darolutamide, is thus timely and necessary. 
The current study will enroll EBC female patients with differing BC subtypes, with the intent 
of characterizing the molecular alterations in BC tissue before and after a short-term 
exposure to darolutamide. Darolutamide is a new AR antagonist, that thus far has been 
found to be very well-tolerated.
Studying the biological mechanisms in which darolutamide targets the AR in BC will be 
crucial in understanding its potential role in this disease, as well as providing the foundation 
for the rational development of darolutamide in the disease. Molecular profiling of tumor 
samples before and after darolutamide treatment may permit the identification of patients 
likely or unlikely to respond to the agent based on the biological and molecular 
characteristics of their tumors. 
Since it is essential to evaluate the molecular effects of darolutamide in tumors not exposed 
to chemotherapy or radiation, patients with untreated primary breast tumors amenable to a 


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pre-treatment biopsy followed by a post-darolutamide sample at definitive surgery or 
prior to starting neoadjuvant systemic therapy (NAST) represent an ideal population to 
study. Although no direct benefit from the treatment with darolutamide is expected in the 
participating patients, the good tolerability and favourable safety profile of darolutamide at 
the selected dose, the proposed brief duration of treatment, and the fact that patients 
enrolled in the study will not be required to undergo any delay in definitive surgery/start of 
neoadjuvant therapy, should ensure the safety and ethical set-up of the trial.
Objectives 
Primary Objective:
To identify the molecular alterations that occur in human BC tissue, following short-term 
exposure to darolutamide in female patients with EBC.
Secondary Objectives:
To evaluate the safety and tolerability of short-term exposure to darolutamide in female 
patients with EBC.
Study Design
This is a multi-center, open-label, tissue-acquisition study involving up to 60 patients with 
early-stage invasive BC. 
Patients with (a) suspicion of invasive BC based on clinical and/or radiological findings, or 
(b) a confirmation of invasive BC based on previous cytology/core/incisional biopsy will be 
invited to participate in the study: (a) In cases without pathological confirmation of BC 
diagnosis, signature of patient informed consent form (PICF) before BC confirmation 
biopsy is preferred to allow the collection of study-specific biopsies to be conducted.  
(b) For patients who have already had a confirmatory biopsy, they may also participate in 
the study if willing to undergo a subsequent study-specific biopsy procedure according to 
the protocol or if their diagnostic biopsy has been done according to the protocol 
requirements (in terms of number of cores/tissue, fresh frozen collection, etc). 
At time of PICF signature, patients will be registered in the study. Primary tumor samples will 
be collected by core needle biopsy or incisional biopsy (excisional biopsy will not be 
allowed). A minimum of three cores of tissue using a 14-gauge needle is recommended (or 
equivalent amount with an incisional biopsy). Two of the cores will be snap-frozen and the 
third one should be prepared in paraffin according to site’s practice. In cases, it is not
possible to provide enough material, collection of 2 cores (or equivalent amount with 
incisional biopsy) will be acceptable: one core will be snap-frozen and the other one will be 
prepared in paraffin.
To be able to assess the molecular alterations after darolutamide exposure in different BC 
subtypes, patients being either triple-negative, or HR+/HER2 negative, or HER2 positive will 
be enrolled in the study (up to 20 patients with each of these subtypes, with an acceptable 
minimum of 8 evaluable patients in each cohort if enrollment of recommended 20 patients 
is not feasible for any reason).
Eligible patients will be enrolled and receive darolutamide at a dose of 600 mg (2 × 300 mg 
tablets) b.i.d. to a daily dose of 1,200 mg. Protocol treatment should start within 5 days of 
enrollment, and within 42 days of the study-specific tumor sample collection. Minimum 
duration of treatment with darolutamide is 14 days with a recommended maximum 
duration of 21 days. It is recommended that the BC surgery/pre-NAST biopsy date is defined 
prior to starting protocol treatment. The start date of darolutamide (Day 1) will be derived 
as minus 14 to 21 days from the scheduled BC surgery/pre-NAST biopsy date. If for some 
reason BC surgery/pre-NAST biopsy takes place more than 21 days after treatment start (e.g. 
scheduling issues, delays, etc.), it is acceptable that patient receives darolutamide for more 
than 21 days and up to a maximum of 35 days. Patient should continue protocol treatment 
until the day prior to BC surgery/pre-NAST biopsy or “nil per os” (npo) is ordered. In these 
cases it is strongly recommended to have BC surgery/pre-NAST biopsy performed as soon as 
possible after 21 days of treatment are completed. Also, in any case that treatment is 
extended beyond 21 days, TRIO Medical Monitor should be contacted to discuss 
continuation of treatment and the need for assessments.
At the time of BC surgery (if NAST is not indicated), or before NAST starts if indicated, BC 
tissue will be collected: two cores (or an equivalent amount of tissue), one fresh-frozen and 


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one paraffin-embedded. 
End of Study (EoS) visit will occur 30 days (+/- 3 days) after the patient's last intake of 
darolutamide.
Eligibility Criteria
Inclusion Criteria
1.
Signed and dated PICF obtained prior to initiation of any study-specific procedure and 
treatment.
2.
Female ≥ 18 years old.
3.
Histologically proven invasive breast carcinoma (through either a core needle biopsy or 
an incisional biopsy) for which surgery is indicated as the primary treatment modality. 
Patients for which NAST is indicated are also eligible provided they are willing to 
undergo a biopsy after completing treatment with darolutamide and prior to NAST 
start.
4.
Known ER, PgR and HER2 statuses.
5.
Tumor must be confined to either the breast or to the breast and ipsilateral axilla (Note: 
patients with multifocal/multicentric tumors are eligible). Patient must have (according 
to TNM 7th edition rules): 
T1 with T ≥1.0 cm, T2 or T3 by at least one radiographic or clinical measurement
Either clinically positive (N1 only) or clinically negative axillary nodes (N0)
M0
6.
Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1.
7.
Adequate organ function within 28 days prior to enrollment, as defined by the 
following criteria:
Hematology: Hemoglobin ≥ 9.0 g/dL, Absolute neutrophil count (ANC) ≥ 1.5 × 109/L, 
Platelet count ≥ 100 × 109/L
Liver function: Alanine aminotransferase (ALT) and aspartate transaminase (AST) ≤ 
2.5 × upper limit of normal (ULN), Total bilirubin ≤ 1.5 × ULN (or 3 times ULN for 
patients with documented Gilbert’s syndrome or for whom indirect bilirubin 
concentrations suggest an extra-hepatic source of elevation).
Renal function: Creatinine ≤ 2.0 × ULN 
8.
No more than 42 days should elapse from the day the study-specific tumor sample is 
taken at initial diagnosis (or subsequent procedure) to the day of the first intake of 
darolutamide.
9.
Women of childbearing potential (WoCBP) must agree to use acceptable non-hormonal 
contraceptive methods of birth control from the day of the screening pregnancy test 
and up to 3 months after the last intake of darolutamide. 
10. For WoCBP negative serum pregnancy test within 7 days of enrollment. 
11. Willingness and ability to comply with scheduled visits, treatment plans, laboratory 
tests, and biopsies as detailed in the protocol.
Exclusion Criteria
1.
Any T0, Tis, T1 < 1.0 cm, T4; or N2-3; or M1 BC.
2.
Bilateral invasive BC.
3.
Patient that underwent excisional biopsy of the primary tumor.
4.
Medical indication or patient desire to undergo BC surgery or start NAST prior to 
completing at least 14 days of treatment with darolutamide, and/or refusal of patient 
to undergo corresponding biopsy in case NAST is planned.
5.
Prior or concurrent systemic anticancer therapy for BC treatment (immunotherapy, 
biologic/targeted therapy, chemotherapy, investigational agents).
6.
Prior or concurrent ipsilateral radiation therapy for invasive or noninvasive BC.
7.
Prior or concurrent treatment or preventative use of any hormonal agent such as 
aromatase inhibitors (AI), fulvestrant, raloxifene, tamoxifen or other selective estrogen 
receptor modulators (SERM), or with any other hormonal agent used for the treatment 
or prevention of BC or for any other indication (e.g. osteoporosis).
8.
Concurrent use of ovarian hormone replacement therapy. Prior treatment should be 
stopped at least 28 days prior to registration.
9.
Prior or concurrent treatment with AR antagonists or CYP17 enzyme inhibitor.
10. Use of other investigational drug within 28 days of enrollment.
11. Major surgery within 28 days before enrollment.


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12. Any concurrent or previous malignancy within 5 years prior to enrollment except for 
basal or squamous skin cancer, or carcinoma in situ of the cervix, or other non-
invasive/in-situ neoplasm, all of which must have been adequately and radically 
treated. A patient with previous history of invasive malignancy (other than adequately 
and radically treated basal or squamous skin cancer) is eligible provided that she has 
been disease free for more than 5 years.
13. Severe or uncontrolled concurrent disease, infection or comorbidity.
14. Known active viral hepatitis, human immunodeficiency virus (HIV) or chronic liver 
disease.
15. Other serious illness or medical condition within 6 months before enrollment, including 
any of the following: Concurrent congestive heart failure New York Heart Association 
(NYHA) Class III or IV, severe/unstable angina pectoris, myocardial infarction, 
uncontrolled hypertension,  coronary/peripheral artery bypass graft, high-risk  
uncontrolled arrhythmias, stroke.
16. Any contraindication to oral agents or gastrointestinal disorder or procedure which 
expects to interfere significantly with absorption of protocol treatment.
17. History or current evidence of any condition, therapy, or laboratory abnormality that 
might confound the results of the trial, interfere with the patient’s participation for the 
full duration of the trial, or is not in the best interest of the patient to participate, in the 
opinion of the treating Investigator.
18. Known psychiatric or substance abuse disorders that would interfere with cooperation 
with the requirements of the trial.
19. Known allergy to darolutamide or any of the excipients.
20. Pregnant or lactating patients.
Protocol Treatment
Darolutamide will be given at a dose of 600 mg (2 × 300 mg tablets) b.i.d. corresponding to a 
daily dose of 1,200 mg.
The following timelines will be considered for darolutamide administration:

Darolutamide shall be administered starting within 5 days of enrollment.

No more than 42 days should elapse from the day the study-specific tumor sample 
is taken at initial diagnosis (or in a subsequent procedure) to the day of the first 
intake of darolutamide.

Minimum duration of treatment will be 14 days prior to BC surgery or to the pre-
NAST biopsy (if NAST is indicated).

A maximum duration of 21 days of treatment is recommended. . It is recommended 
that the date of BC surgery/pre-NAST biopsy is defined prior to starting protocol 
treatment. The start date of darolutamide (Day 1) will be derived as minus 14 to 21 
days from  the scheduled BC surgery/pre-NAST biopsy date.

Treatment will be given until the day prior to the BC surgery, or the day before pre-
NAST biopsy or when npo is ordered, whichever occurs first. 

If for some reason BC surgery/pre-NAST biopsy takes place more than 21 days after 
treatment start, it is acceptable that patient receives darolutamide for more than 
21 days and up to a maxiumum of 35 days; patient should continue protocol 
treatment until the day prior to BC surgery/pre-NAST biopsy or npo is ordered. In 
these cases it is strongly recommended to have BC surgery/pre-NAST biopsy done 
as soon as possible after 21 days of treatment are completed. Also, in any case 
that treatment is extended beyond 21 days, TRIO Medical Monitor should be 
contacted to discuss continuation of treatment and the need for assessments.


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Visits and 
Assessments
Activity
Screening
Treatment 
Period
Pre-
surgery
BC 
surgery/pre-
NAST biopsy
End of 
Study
Informed 
consent/Registration 
and Enrollment
X
Demographics, medical, 
surgical, disease history
X
Complete physical 
examination 
X
X
Symptom-directed 
physical examination
X
X
ECOG PS
X
X
X
X
Weight, height, blood 
pressure
X
Serum pregnancy test
X
Hematology, Blood 
chemistry
X
X
X
Treatment compliance 
assessment
X
X
AEs assessments
X
X
X
X
Concomitant 
medication
X
X
X
X
Tumor sample
X
X
Statistical Methods 
Up to 60 patients (a minimum of 8 evaluable patients and up to 20 patients in each breast 
cancer subtype: triple-negative, HR+/HER2 negative, HER2 positive) will be required for the 
performance of the molecular studies and possible hypothesis generation. 
A patient will be considered evaluable if the following criteria are met:
Minimum duration of treatment with darolutamide is of 14 days, with a minimum of 10 
consecutive days of treatment prior to the definitive BC surgery/pre-NAST biopsy. 
Mandatory tumor tissue is collected at screening and at BC surgery/pre-NAST biopsy
according to the protocol and submitted to the central lab.
Adequacy for molecular assessment of the tumor tissue collected before and after the 
protocol treatment initiation. The adequacy will be evaluated by the central lab.
Assessment of patient evaluability, safety and distribution of patients with respect to 
hormone receptor status and HER2 status will be performed by the Study Steering 
Committee (SSC) at given time points depending on the recruitment, primarily planned once 
20, around 35 and once all patients have been enrolled and undergone the EoS per protocol. 
This will allow for the SSC to ascertain that the appropriate number of evaluable patients is 
being accrued, that more or less patients may be required, or in the extreme case, that 
accrual should close if the patient evaluability rate is very low and/or safety concerns arise.
The SSC will define the evaluability of patients that discontinue treatment more than 3 days 
before surgery: molecular analyses may be taken into consideration for the decision about 
the evaluability of such patients.
Study Populations
Intention-to-treat (ITT) Population: the ITT population includes all patients who are 
enrolled in the study regardless of whether they have been treated or not.
Safety Population: the Safety population includes all patients who have taken at least one 


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tablet of darolutamide.
Evaluable population: includes all patients who are considered evaluable per protocol.


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1. BACKGROUND AND RATIONALE
1.1
Disease Overview
Breast cancer (BC) is the second most common cancer in the world and by far, the most frequent 
cancer among women.1 In the United States (US), approximately 246,000 new BC cases and 40,000 
deaths due to this disease are expected to occur in women during 2016. This represents 29% of all 
cancers diagnosed annually in women and 14% of deaths due to cancer annually in women.2
BC is a highly heterogeneous disease with marked differences regarding molecular, biological and 
clinical features. Molecular subtypes have been described with corresponding differences in 
prognosis and response to treatments. The current classification is based on the expression of 
estrogen and progesterone receptors (ER and PgR respectively) and HER2, and BC are broadly 
divided into three subtypes: HR+/HER2 negative, HER2+ and triple negative BC (TNBC). The 
characterization of which subtype of BC a patient has, allows to adapt the different treatment 
modalities due to their predictive value and has also prognostic implications. 
Despite progress made in diagnostic techniques and treatments for BC, there is still a significant 
number of patients undergoing treatment for early BC (EBC) who will relapse or patients with 
advanced BC whose tumor will not respond to standard treatments or progress after them. The 
medical need for new and potentially more active and better tolerated agents in BC remains very 
high. 
1.2
Investigational Medicinal Product Overview
Darolutamide (ODM-201) is a new AR antagonist that is being jointly developed by Bayer AG and 
Orion Corporation for the treatment of castration-resistant prostate cancer (CRPC). As of the version 
date of this protocol, it has not been marketed or authorized to be marketed in any country.
Darolutamide potently inhibits testosterone binding to AR and retains antagonistic properties in cells 
expressing increased AR levels better than bicalutamide. Darolutamide diminishes AR-signaling by 
inhibiting testosterone mediated nuclear localization of AR more efficiently than bicalutamide. It
shows antiandrogenic and antitumor activity in in vitro and in vivo models of CRPC and in clinical 
studies in patients with metastatic CRPC (mCRPC).
Darolutamide is a 1:1 mixture of 2 pharmacologically active diastereomers: ORM-16497 and ORM-
16555 which are able to interconvert. Steady-state plasma concentrations of darolutamide are
reached by 7 days of repeated dosing. The mean elimination half-life is in the range of 11.5-16.1 
hours.
Data from an in vivo mass balance study in rats suggests that about two-thirds of oral darolutamide
dose is excreted in feces and the remaining amount in urine. After a single dose, excretion is almost 
complete by 48 hours post-dose.
In general, the potential for darolutamide to inhibit CYP reaction pathways in vitro in human liver 
microsomes is low. The CYP450 induction potential of darolutamide was evaluated in vitro in 
primary cultures of human hepatocytes and a risk of weak to moderate CYP3A4 induction by 
darolutamide was observed.
In vivo data from study 17726 indicates that co-administration of rifampicin -a strong CYP3A4 
inducer- (600 mg, fasted state) and a single dose of darolutamide (600 mg; 2 tablets of 300 mg) 
together with food, resulted in a 3-and 2-fold decrease of AUC (0-72) and Cmax of darolutamide to 
28% and 48% respectively. Therefore, repeated co-administration of strong or moderate CYP3A4 
inducers (e.g. carbamazepine, phenobarbital, St. John's Wort) with darolutamide is expected to 
reduce darolutamide plasma concentrations. The concomitant intake of these substances during 
study treatment should hence be avoided.


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In vitro data indicate that darolutamide is a substrate of P-glycoprotein (P-gp) and Breast cancer 
resistance protein (BCRP). Therefore administration of strong inhibitors of P-gp (e.g., verapamil, 
dronedarone) and BCRP (e.g., pantoprazole, eltrombopaq) may increase the plasma concentrations 
of Darolutamide and should be used with caution. However, in vivo data from Study 17726 showed 
that co-administration of itraconazole –a strong CYP3A4 and P-gp inhibitor- (200 mg) and a single 
dose of darolutamide (600 mg; 2 tablets of 300 mg) together with food, resulted in a 1.7-fold 
increase in AUC (0-72) and a 1.4-fold increase of C max of darolutamide. This indicates that co-
administration of darolutamide with a strong CYP3A4 or P-gp inhibitor does not result in a clinically 
relevant increase of darolutamide (ODM-201) plasma concentrations.
Plasma concentration of drugs that are sensitive P-gp or BCRP substrates might be increased by 
darolutamide. Medicinal products that are sensitive substrates for P-gp (e.g., digoxin) or BCRP (e.g.,
methotrexate) should be used with caution when co-administered with darolutamide. In vivo data 
shows that the administration of 600 mg of darolutamide b.i.d. over 4 days prior to administration 
of a single dose of 5 mg rosuvastatin, a BCRP substrate, together with food resulted in a 5.2-fold 
increase in mean exposure (AUC) of rosuvastatin and a 4.9-fold increase in maximum serum 
concentration (Cmax). This indicates that co-administration of darolutamide can increase the plasma 
concentrations of other concomitant BCRP substrates (e.g. methotrexate, sulfasalazine, fluvastatin, 
atorvastatin).
As of 04 January 2017, darolutamide has been given to 173 patients with CRPC. Until now, 
darolutamide has been studied in 6 clinical studies. The majority of the phase I/II studies were 
conducted in mCRPC patients. One phase I study was conducted in healthy male volunteers with a 
single administration of darolutamide. Non-metastatic CRPC patients are being investigated in an
ongoing phase III study (ARAMIS trial).
Antitumor activity of darolutamide was evaluated in 3 studies in mCRPC. They have shown that 
darolutamide has antitumor activity at all dose levels as evaluated by Prostate Specific Antigen 
(PSA), circulating tumor cells and soft and bone lesion imaging. PSA decline ≥ 50% was observed at 
12 weeks in 74% of chemotherapy-naïve patients treated with darolutamide.
Darolutamide has thus far been found to be well-tolerated and to have negligible blood-brain barrier 
penetration. The integrated safety data from clinical trials showed 93.9% of patients developed at 
least 1 adverse event (AE) and grade 3 or higher AEs were reported in 28% of patients. The most 
common AEs have been fatigue in 26.8% of patients and back pain in 22% of patients. The most 
common AEs considered related to protocol treatment by the Investigator have been fatigue (9.8%) 
and hot flush (4.3%). The majority of AEs were classified as grade 1 (59.7% of the events) and grade 
2 (31.2% of the events). Of the serious adverse events (SAEs) reported in clinical trials (27.4 %) none 
of them was considered related to the protocol treatment by the Investigator or by the Sponsor.
In summary, the results indicate that darolutamide exhibits a good safety and tolerability profile and 
promising antitumor activity in men with CRPC.
Additional information on the clinical efficacy, safety, pharmacology of darolutamide is provided in 
the darolutamide Investigator brochure (IB).  
1.3
Rationale
1.3.1
Rationale for Conducting the Study
The androgen receptor (AR) is a member of the nuclear steroid hormone receptor family, which also 
includes ER and PgR. These are critical components of signaling pathways and regulate gene 
expression. AR expression in BC has long been noted and is the most common nuclear hormone 
receptor. AR is expressed in approximately 80% and 60% of primary and metastatic BC, respectively.3
Its expression varies across the different subtypes, approximately 84-95% in ER+ tumors, 50-63% in 
ER-/HER2+ cases and 10-53% in TNBC.3


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The role of AR in BC still needs to be fully elucidated, differently from the well-known ER and PgR. 
Preclinical studies show that the androgen signaling pathway plays a critical role in the development 
of normal and malignant breast tissue. Epidemiologic studies suggest that increased levels of 
circulating androgens are associated with an increased risk for BC, primarily ER/PgR-positive BC.4
AR expression in ER+ BC associates with improved outcomes, but AR seems to be oncogenic in the 
tamoxifen-resistant setting.3 Approximately 30 to 50% of all ER+ BC tumors display de novo
resistance to standard endocrine therapies and most metastatic ER+ tumors do eventually acquire 
resistance over time.5 AR positivity increases resistance to tamoxifen in BC models in vitro and in 
vivo.6 Thus, it is possible that acquired resistance to antiestrogen therapies could result from 
adaptation from estrogen dependence to androgen dependence.7  
In the case of TNBC, signaling through the AR may play a role in pathogenesis. Studies which have 
focused on AR expression in TNBC have provided conflicting results in regards to the prognostic 
value of AR expression.8 While some studies suggest that AR expression is a favorable prognostic 
factor in patients with TNBC, others suggest that it predicts reduced survival in early TNBC.8, 9
Irrespective of this, AR might represent a new target in TNBC, which has so far a worse prognosis
and very limited standard therapeutic options, none of them targeted.
As a result of these findings, AR has emerged as a potential useful marker for the further 
subclassification of BC and as a potential therapeutic target.
Modulation of AR signaling, either inhibitory or stimulatory, has previously been explored in BC with 
somewhat contrasting results. Androgen therapy has historically been used but largely abandoned in 
the 1970s due to its toxicity and the emergence of ER-directed therapies.
More recently, clinical reports of AR antagonism have been promising, especially in TNBC. 
Bicalutamide has been assessed in a phase II, single-arm trial in AR+, ER- advanced BC, and showed 
clinical benefit rate of 19% with a good safety profile.10 Some newer drugs like enzalutamide, have 
also been tested in TNBC. A phase II study in AR+ TNBC proved that this therapy was well-tolerated 
and showed promising clinical activity in selected patients.11
These developments in combination with an improved understanding of AR signaling in different BC
subtypes led to the renewed interest in targeting AR. However, the value of AR antagonists in the 
management of BC, the molecular alterations that occur after exposure to these agents and whether 
there are biomarkers useful to predict their effectiveness, still need to be fully elucidated. Increased 
understanding of AR blockade and its potential value in different BC subtypes and clinical testing of 
newer and more potent AR antagonists like darolutamide, is thus timely and necessary.
Translational Oncology Research Laboratories (TORL, at University of California Los Angeles, US) has 
conducted cell viability studies in 49 BC cell lines exposed to different concentrations of 
darolutamide. The studies have shown that cell lines belonging to each of the three BC subtypes 
(HR+, HER2+, TN) responded different to darolutamide. Responses are mainly seen in cell lines 
expressing AR but also in some lines that lack AR expression, suggesting that AR expression may not 
be the only biomarker predicting response to darolutamide (unpublished).
The current study will enroll EBC female patients with differing BC subtypes, with the intent of 
characterizing the molecular alterations in BC tissue before and after short-term exposure to the 
antiandrogen darolutamide. Studying the biological mechanisms in which darolutamide targets the 
AR in BC will be crucial in understanding its potential role in this disease, as well as providing the 
foundation for the rational development of darolutamide in the disease. Such information may 
suggest that darolutamide be combined with other modalities; for example, if treatment with the 
drug is discovered to affect multiple signaling pathways. Molecular profiling of tumor samples before 
and after darolutamide treatment may permit the identification of patients likely or unlikely to 
respond to the agent based on the biological and molecular characteristics of their tumors. Since it is 
essential to evaluate the molecular effects of darolutamide in tumors not previously exposed to 
chemotherapy or radiation, patients with untreated primary breast tumors amenable to a pre-


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treatment biopsy followed by a post-darolutamide sample at definitive surgery or prior to starting 
neoadjuvant systemic therapy (NAST) represent an ideal population to study. Male patients will not 
be enrolled due to pathological and biological differences in male breast cancer.12
Although no direct benefit from the treatment with darolutamide is expected in the participating 
patients, the good tolerability and favourable safety profile of darolutamide at the selected dose, the 
proposed brief duration of treatment, and the fact that patients enrolled in the study will not be 
required to undergo any delay in definitive surgery or start of NAST if indicated, should ensure the 
safety and ethical set-up of the trial.
1.3.2
Rationale for Regimen and Dose Selection
In TRIO030, darolutamide will be given at a dose of 600 mg (2 × 300 mg tablets) b.i.d. to a daily dose 
of 1,200 mg. Darolutamide should be taken with food since exposure and Cmax are reduced in fasting 
conditions. 
Darolutamide has been studied in doses ranging from 100 mg to 900 mg b.i.d. in a phase I dose 
escalation and dose finding study. No dose-limiting toxicities were observed in this study and the 
maximum tolerated dose was not reached. Systemic exposure to darolutamide increased dose-
proportionally in the dosing range from 100 to 700 mg b.i.d. At 900 mg b.i.d. dose there was no 
further increase in Cmax or AUC observed when compared to 700 mg b.i.d. Based on the results of 
this phase I study, the recommended dose was 600 mg b.i.d. for further development. At the dose of 
600 mg b.i.d. most AEs were grade 1-2 and assessed by the Investigators as not related to protocol 
treatment. 
Therefore, the dose selected in the study is considered as safe and is the dose that is being used in 
current darolutamide trials in patients with other primary cancer sites.
2. OBJECTIVES
2.1
Primary Objective

To identify the molecular alterations that occur in human BC tissue, following short-term 
exposure to darolutamide in female patients with EBC.
2.2
Secondary Objectives

To evaluate the safety and tolerability of short-term exposure to darolutamide in female 
patients with EBC.
3. STUDY DESIGN
3.1
Overall Study Design and Plan: Description
This is a multi-center, open-label, tissue-acquisition study involving up to 60 patients with early-
stage invasive BC. 
Female patients with (a) suspicion of invasive BC based on clinical and/or radiological findings, or (b) 
those who have a confirmation of invasive BC based on previous cytology/core/incisional biopsy will 
be invited to participate in the study:
(a) In cases without pathological confirmation of BC diagnosis, signature of patient informed 
consent form (PICF) before BC confirmation biopsy is preferred to allow the collection of study-
specific biopsies to be conducted. 


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(b) For patients who have already had a confirmatory biopsy, they may also participate in the 
study if willing to undergo a subsequent study-specific biopsy procedure according to the 
protocol or if their diagnostic biopsy has been done according to the protocol requirements (in 
terms of number of cores/tissue, fresh frozen collection, etc.; refer to section 6.5.7.1).
At time of PICF signature, patients will be registered in the study. Primary tumor samples will be 
collected by core needle biopsy or incisional biopsy (excisional biopsy will not be allowed). A 
minimum of three cores of tissue using a 14-gauge needle is recommended (or equivalent amount 
with an incisional biopsy). Two of the cores will be snap-frozen and the third one should be prepared 
in paraffin according to site’s practice. In cases that it is not possible to provide enough material, 
collection of 2 cores (or equivalent amount with incisional biopsy) will be acceptable; one core will 
be snap-frozen and the other one will be prepared in paraffin. 
After all screening procedures and confirmation of eligibility, patients will be enrolled and will begin 
intake of darolutamide. To be able to assess the molecular alterations after darolutamide exposure 
in different BC subtypes, female patients being either triple-negative, or HR+/HER2 negative, or 
HER2 positive will be enrolled in the study. Up to 20 patients with each of these subtypes will be 
included (refer to section 6.1.2.2 for further details on the enrollment according to BC subtype). 
However, a minimum of 8 evaluable patients in each cohort is considered acceptable if enrollment of 
20 patients is not feasible within a cohort for any reason.
Eligible patients will be enrolled and receive darolutamide at a dose of 600 mg (2 × 300 mg tablets) 
b.i.d. corresponding to a daily dose of 1,200 mg. Protocol treatment should start within 5 days of 
enrollment and within 42 days of the study-specific tumor sample collection. Patients enrolled, but 
not treated, will be discontinued from study and managed as per the site’s standard of care (SoC).
Minimum duration of treatment with darolutamide is 14 days with a recommended maximum 
duration up to 21 days. It is recommended that the BC surgery/pre-NAST biopsy date is defined prior 
to starting protocol treatment. The start date of darolutamide (Day 1) will be derived as minus 14 to 
21 days from the scheduled BC surgery/pre-NAST biopsy date. Treatment will be taken until the day 
prior to the BC surgery (if NAST is not indicated) or the day before NAST starts if indicated; or when 
the patient is ordered to stop all oral intake, whichever occurs first (refer to section 5.3.2.1). If for 
some reason surgery takes place more than 21 days after treatment start (e.g. surgery scheduling 
issues, surgery delays, etc.) it is acceptable that patient receives darolutamide for more than 21 days 
and up to a maximum of 35 days. Patient should continue protocol treatment until the day prior to 
BC surgery/pre-NAST biopsy or “nil per os” (npo) is ordered. In these cases it is strongly 
recommended to have BC surgery/pre-NAST biopsy performed as soon as possible after 21 days of 
treatment are completed.
At the time of BC surgery (if NAST is not indicated), or before NAST starts if indicated, BC tissue will 
be collected: two cores (or an equivalent amount of tissue), one fresh-frozen and one paraffin-
embedded.
End of Study (EoS) visit will occur 30 days (+/- 3 days) after the patient's last intake of darolutamide.
In case protocol treatment-related AEs/SAEs ongoing at the time of the EoS visit, monitoring of these 
events will continue as clinically indicated until (a) the events have resolved or (b) the events have 
reached a status which, in the Investigator’s opinion, is unlikely to resolve due to the nature of the 
condition and/or the patient’s underlying disease. 
Treatment for BC including NAST (if applicable), definitive surgery, and adjuvant therapy will be at 
the Investigator’s discretion.


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Figure 1: Study Schema 
BC= Breast cancer
* In cases with absence of a pathological confirmation of BC diagnosis, PICF signature prior to the biopsy for BC
confirmation is preferred. For patients who already had such confirmatory biopsy, they may also participate in the study if 
willing to undergo a subsequent study-specific biopsy procedure according to the protocol or if their diagnostic biopsy has 
been done according to the protocol requirements (in terms of number of cores/tissue, fresh frozen collection, etc.).
3.2
Number of Patients and Investigational Sites
Up to 60 patients will be enrolled in the study, from approximately 20 sites in North America and 
Europe, with an acceptable minimum of 8 evaluable patients in each if enrollment of 20 patients is 
not feasible within a cohort for any reason. The list of Investigators participating in the study will be 
maintained in TRIO’s Clinical Trial Management System. 
3.3
Study End 
The study will end once the last patient undergoes the EoS visit and all data required for the analyses
have been observed, collected and cleaned. The trial may be prematurely terminated at any time by 
Ethics Committees (EC)/Institutional Review Boards (IRB), competent authorities, or the sponsor. 
Potential reasons for premature termination of the study by the sponsor are (not exhaustive):

Regulatory authority decision

Change in opinion of the EC/IRB

Drug safety concerns

Manufacturer’s decision to discontinue development of darolutamide

Inability to run the study because of operational issues (e.g., low recruitment)
If the study is prematurely terminated or discontinued, TRIO will promptly notify the Investigators 
and all other relevant parties.
4. PATIENT SELECTION
Patients can be enrolled in the study only if the eligibility criteria are met. This study can fulfill its 
objectives only if appropriate patients are enrolled. Patients who fail to meet all inclusion criteria or 
fulfill at least one exclusion criteria cannot be enrolled in the study. The sponsor will not grant any 
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If a patient whom does not meet the eligibility criteria is incorrectly enrolled, the Investigator should 
immediately inform the Medical Monitor. The decision about the continuation or discontinuation of 
this patient on protocol treatment will be based on medical judgment, treatment benefit and safety 
risks for her.
4.1
Inclusion Criteria
1.
Signed and dated PICF obtained prior to initiation of any study-specific procedure and 
treatment.
2.
Female ≥ 18 years old.
3.
Histologically proven invasive breast carcinoma (through either a core needle biopsy or an 
incisional biopsy) for which surgery is indicated as the primary treatment modality. Patients 
for which NAST is indicated are also eligible provided they are willing to undergo a biopsy (as 
per requirements in section 6.5.7) after completing treatment with darolutamide and prior to 
NAST start.
4.
Known ER, PgR and HER2 statuses.
5.
Tumor must be confined to either the breast or to the breast and ipsilateral axilla (Note: 
patients with multifocal/multicentric tumors are eligible). Patient must have (according to 
TNM 7th edition rules): 

T1 with T ≥1.0 cm, T2 or T3 by at least one radiographic or clinical measurement

Either clinically positive (N1 only) or clinically negative axillary nodes (N0)

M0
6.
Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1.
7.
Adequate organ function within 28 days prior to enrollment, as defined by the following 
criteria:

Hematology: 
- Hemoglobin ≥ 9.0 g/dL,
- ANC ≥ 1.5 × 109/L, 
- Platelet count ≥ 100 × 109/L

Liver function: 
- ALT and AST ≤ 2.5 × ULN, 
- Total bilirubin ≤ 1.5 × ULN (or 3 times ULN for patients with documented 
Gilbert’s syndrome or for whom indirect bilirubin concentrations suggest an extra-
hepatic source of elevation)

Renal function: Creatinine ≤ 2.0 × ULN
8.
No more than 42 days should elapse from the day study-specific tumor sample is taken at 
initial diagnosis (or subsequent procedure) to the day of the first intake of darolutamide.
9.
WoCBP* must agree to use acceptable non-hormonal contraceptive methods of birth control 
from the day of the screening pregnancy test and up to 3 months after the last intake of 
darolutamide. 
10. For WoCBP* negative serum pregnancy test within 7 days of enrollment.
11. Willingness and ability to comply with scheduled visits, treatment plans, laboratory tests, and 
biopsies as detailed in the protocol.


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* Note: Women of childbearing potential (WoCBP) are any women between menarche and menopause who 
have not been permanently sterilized, capable of procreation. Permanent sterilization includes 
hysterectomy and/or bilateral oophorectomy and/or bilateral salpingectomy but excludes bilateral tubal 
ligation/occlusion. Postmenopause is defined as:
-
Bilateral oophorectomy
-
Age ≥ 60
-
Age < 60 and amenorrheic for ≥12 months in the absence of an alternative medical cause and FSH and 
estradiol in postmenopausal ranges. 
4.2
Exclusion Criteria
1.
Any T0, Tis, T1 < 1.0 cm, T4; or N2-3; or M1 BC.
2.
Bilateral invasive BC.
3.
Patient that underwent excisional biopsy of the primary tumor.
4.
Medical indication or patient desire to undergo BC surgery or start NAST prior to completing 
at least 14 days of treatment with darolutamide, and/or refusal of patient to undergo 
corresponding biopsy in case NAST is planned. 
5.
Prior or concurrent systemic anticancer therapy for BC treatment (immunotherapy, 
biologic/targeted therapy, chemotherapy, investigational agents).
6.
Prior or concurrent ipsilateral radiation therapy for invasive or noninvasive BC.
7.
Prior or concurrent treatment or preventative use of any hormonal agent such as aromatase 
inhibitors (AI), fulvestrant, raloxifene, tamoxifen or other SERM, or with any other hormonal 
agent used for the treatment or prevention of BC or for any other indication (e.g. 
osteoporosis).
8.
Concurrent use of ovarian hormone replacement therapy. Prior treatment should be stopped 
at least 28 days prior to registration.
9.
Prior or concurrent treatment with AR antagonists or CYP17 enzyme inhibitor.
10. Use of other investigational drug within 28 days of enrollment.
11. Major surgery* within 28 days before enrollment.
12. Any concurrent or previous malignancy within 5 years prior to enrollment except for basal or 
squamous skin cancer, or carcinoma in situ of the cervix, or other non-invasive/in-situ 
neoplasm, all of which must have been adequately and radically treated. A patient with 
previous history of invasive malignancy (other than adequately and radically treated basal or 
squamous skin cancer) is eligible provided that she has been disease free for more than 5 
years.
13. Severe or uncontrolled concurrent disease, infection or comorbidity.
14. Known active viral hepatitis, HIV or chronic liver disease.
15. Other serious illness or medical condition within 6 months before enrollment, including any of 
the following: Concurrent congestive heart failure NYHA Class III or IV, severe/unstable angina 
pectoris, myocardial infarction, uncontrolled hypertension, coronary/peripheral artery bypass 
graft, high-risk uncontrolled arrhythmias, stroke.
16. Any contraindication to oral agents or gastrointestinal disorder or procedure which expects to 
interfere significantly with absorption of protocol treatment.
17. History or current evidence of any condition, therapy, or laboratory abnormality that might 
confound the results of the trial, interfere with the patient’s participation for the full duration 


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of the trial, or is not in the best interest of the patient to participate, in the opinion of the 
treating Investigator.
18. Known psychiatric or substance abuse disorders that would interfere with cooperation with 
the requirements of the trial.
19. Known allergy to darolutamide or any of the excipients.
20. Pregnant or lactating patients.
* Note: Major surgery defined as requiring a general anesthesia or respiratory assistance; involving 
openings into the great cavities of the body, organs removed, or normal anatomy altered; implying  risks  of 
severe hemorrhage; implying risk for life of the patient or severe disability.
5. TREATMENT
5.1
Protocol Treatment: Darolutamide
Darolutamide is the only protocol treatment in the trial and is considered an Investigational 
Medicinal Product (IMP). Darolutamide will be supplied to the investigational sites as 300 mg tablets, 
free of charge.
For the purpose of this trial, the Single Reference Document to be used for darolutamide is the IB.
5.2
Protocol Treatment Management
5.2.1
Packaging and Labeling
Darolutamide is provided as coated immediate-release 0.3 g (300 mg) tablets, in a high-density 
polyethylene (HDPE) white opaque plastic bottle containing 140 tablets.
Clinical supplies will be affixed with a clinical label in accordance with applicable regulatory 
requirements, and include a statement regarding its limitation to investigational use only. 
5.2.2
Storage and Accountability
Study drugs must be stored according to labeled requirements, at room temperature not exceeding 
30°C in a secure, limited-access location. Receipt and dispensing of trial medication must be 
recorded by an authorized person at the site. Clinical supplies may not be used for any purpose 
other than that stated in the protocol.
The Investigator/designee is responsible for keeping accurate records of the clinical supplies 
received from the Sponsor or designee, the amount administered or dispensed to and returned by 
the patients, and the amount remaining at the conclusion of the trial.
TRIO will provide appropriate documents that must be completed for drug accountability and 
destruction/return.
5.2.3
Drug Return/Destruction
Patients must return all unused medication and empty containers to the Investigator/designee. 
Unused/expired darolutamide should be destroyed locally (where requirements permit) once 
accountability is performed and drug supplies are reconciled. Procedures for proper disposal of 
darolutamide should be applied according to standards established at each site.
5.3
Protocol Treatment Administration
5.3.1
Pre-medication and Special Precautions


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No specific pre-medication is required. Based on the available data on latest vesion of IB, there are 
no special warnings and precautions for use of darolutamide.
5.3.2
Standard Treatment Schedule
5.3.2.1 Dosage and Administration
Darolutamide will be given at a dose of 600 mg (2 × 300 mg tablets) b.i.d. to a daily dose of 
1,200 mg.
The following timelines will be considered for darolutamide administration (refer to Figure 1: Study 
Schema):

Darolutamide shall be administered starting within 5 days of enrollment.

No more than 42 days should elapse from the day the study-specific tumor sample is taken 
at initial diagnosis (or in a subsequent procedure) to the day of the first intake of 
darolutamide (Note: for patients who already had their initial diagnostic biopsy prior to 
registration and this procedure has been done according to the protocol requirements, the 
42- day window should be counted from the date of such biopsy).

Minimum duration of treatment will be 14 days prior to BC surgery or to the pre-NAST 
biopsy (if NAST is indicated) (Note: a day of treatment is considered as any day where the 
patient takes any darolutamide intake)

A maximum duration of 21 days of treatment is recommended. It is recommended that the 
date of BC surgery/pre-NAST biopsy is defined prior to starting protocol treatment. The start 
date of darolutamide (Day 1) will be derived as minus 14 to 21 days from  the scheduled BC 
surgery/pre-NAST biopsy date.

Treatment will be given until the day prior to the BC surgery, or the day before pre-NAST 
biopsy or when npo is ordered, whichever occurs first. 

If for some reason BC surgery/pre-NAST biopsy takes place more than 21 days after 
treatment start, it is acceptable that patient receives darolutamide for more than 21 days 
and up to a maximum of 35 days; patient should continue protocol treatment until the day 
prior to BC surgery/pre-NAST biopsy or npo is ordered. In these cases it is strongly 
recommended to have BC surgery/pre-NAST biopsy done as soon as possible after 21 days of 
treatment are completed. Also, in any case that treatment is extended beyond 21 days, TRIO 
Medical Monitor should be contacted to discuss continuation of treatment and the need for 
assessments.

The first day of darolutamide administration in the study is considered Day 1. On or prior to 
Day 1 the patient will be provided with a bottle with 140 tablets containing 300 mg of 
darolutamide each (enough for 35 days of treatment at a daily dose of 1,200 mg). Patient 
will be instructed that the last protocol treatment intake should be the one in the morning 
of the Pre-surgery visit. The last darolutamide intake will be on the day prior to BC surgery, 
or the day before pre-NAST biopsy when the Pre-surgery visit should occur. Exceptions to 
this rule are when patient is ordered to stop oral intake before the day prior to BC 
surgery/pre-NAST biopsy, when BC surgery/pre-NAST biopsy is planned for a Monday or the 
day following a holiday, or in case that performing Pre-surgery visit on the same day of BC 
surgery/pre-NAST biopsy is required by site due to logistical reasons or patient’s/physician’s 
convenience (refer to section 6.4.3 for guidance). 
Darolutamide should be taken approximately at the same time each day, twice a day. It is 
recommended that darolutamide be taken with food; recommendation is to take it with breakfast 
and dinner each day. Patients will ingest darolutamide whole tablets with water. No tablet should be 
ingested if it is broken, cracked, or otherwise not intact.


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If the patient misses an intake or vomits following darolutamide administration, the missed/vomited
dose should be skipped and the patient should take the next intake when scheduled. No “make-up 
dose” or increased dosing should occur.
If a patient accidentally or intentionally takes a higher daily dose than the one recommended per
protocol and up to 1,800 mg (i.e. up to 6 × 300 mg tablets), protocol treatment should continue as 
scheduled without any dose reduction or skip. For example, if by mistake the patient takes 4 tablets 
in the first daily intake she can still take 2 tablets in the second daily intake since a total daily dose of 
1,800 mg is not exceeded. The Medical Monitor should be contacted for any dose taken that 
exceeds 1,800 mg daily.
5.3.2.2 Treatment Compliance
The administration of the protocol treatment should be recorded in the appropriate sections of the 
electronic case report form (eCRF).
Additionally, treatment compliance will be assessed at each visit that may occur during treatment 
phase (see section 6.4.2) and at Pre-surgery visit, and it should be properly documented in source 
documents. Patients will complete a diary (Appendix 2) to document their daily intakes. They will be 
instructed to return all unused drugs (partially used and empty containers) and their diary at each 
visit during the treatment period (if any) and at Pre-surgery visit. Site staff will perform 
accountability of the returned drug and will assess compliance of the patient. Site staff must ensure 
that the patient clearly understands the directions for self-medication and follows the schedule
adequately.
5.3.3
Protocol Treatment Discontinuation
Patients are treated until the day before BC surgery/pre-NAST biopsy (if NAST will be given), or when 
patient is ordered to stop oral intake, whichever occurs first. 
The Investigator will also discontinue protocol treatment if any of the following conditions is met:

Intercurrent illness or a change in patient´s condition or unacceptable toxicity that warrants 
protocol treatment discontinuation according to Investigator’s judgment

Any event, condition, criterion which would warrant discontinuation of darolutamide as per 
section 5.4, including Grade 3 or higher protocol-treatment related AE

Any event, condition, reason which would warrant darolutamide to be held for more than 
the maximum acceptable delay of 7 consecutive days

Patient receives non-protocol anticancer therapy at any time during the protocol treatment

Patient’s decision to withdraw protocol treatment

Lost to follow up

Death

Pregnancy 

Investigator’s decision

Discontinuation of the study by the sponsor (refer to section 3.3)
The reason and date of protocol treatment discontinuation for all patients will be documented in the 
eCRF (e.g. AE, lost to follow-up, etc.). The Investigator will attempt to complete all discharge 
procedures at the time a patient is discontinued from the study including an assessment of the AEs, 
collect all study-related materials from the patient (i.e. patient diary), and will complete the EoS
form. Patients are free to discontinue their participation in the study at any time, without prejudice 
to further treatment. 


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5.3.4
Surgery and (Neo) Adjuvant Systemic Therapy
BC surgery, NAST and post surgical BC treatments are left at the discretion of the Investigator and 
are not considered protocol treatments.
5.4
Adverse Events Management and Treatment Schedule Adjustments 
5.4.1
General Rules
Regular assessment and monitoring of AEs is required from PICF signature, throughout protocol
treatment period and at least up until surgery or BC pre-NAST biopsy if patient will receive NAST.
Toxicity will be assessed utilizing the National Cancer Institute Common Terminology Criteria for AEs
(NCI CTCAE) v4.03. For toxicities not specifically listed in the NCI CTCAE, the following grading will 
apply for assessing severity:

Grade 1: Mild

Grade 2: Moderate 

Grade 3: Severe

Grade 4: Life-threatening 

Grade 5: Death related to AE
Patients will be instructed to notify their physician immediately for any and all AEs.
Doses of protocol treatment may be modified (delayed or reduced) in case of clinically significant 
toxicities that are considered by the Investigator to be related to protocol treatment, according to 
the guidance in section 5.4.2. Assessment of causality (chronology, confounding factors, 
concomitant medications, diagnostic tests, and previous experience with the protocol treatment)
should be conducted by the Investigator prior to dose modification and/or delay whenever possible.
All dose modifications should be based on the AE requiring the greatest modification and should be 
properly documented in source documents and the eCRF.
5.4.2
Guidance for Darolutamide Dose Modifications
A patient who experiences a treatment-related Grade 3 or higher AE must be withdrawn from 
protocol treatment and should proceed to surgery/pre-NAST biopsy, and undergo EoS visit.
In the case of a grade 1-2 treatment-related AE, the Investigator should contact the Medical Monitor 
in case it is considered that a dose reduction or treatment hold is required due to the related AE.
Refer to Table 1: Dose Modifications.
Table 1: Dose Modifications*
Severity grade of 
treatment-related 
AE
Dose modifications
Protocol treatment withdrawal
Grade 1-2
No dose modification required. 
Intake per regular schedule.
Medical Monitor must be contacted in 
case the Investigator considers that a 
dose reduction or treatment hold is 
required due to a grade 1-2 treatment-
related AE. After agreement with the 
Medical Monitor, treatment may be 
Refer to Dose modifications on the left.


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held for a maximum of 7 consecutive
days and/or dose reduced to 300 mg 
b.i.d.a Protocol treatment must be 
discontinued if further dose reduction 
is required or treatment is held for 
more than 7 consectutive days. 
Grade 3 or higher
No dose reduction or delay is 
permitted.
For any grade 3 AE or higher
treatment-related AE, the patient must 
be withdrawn from treatment and 
should undergo BC surgery/pre-NAST 
biopsy. 
* Excludes non-clinically significant and asymptomatic laboratory abnormalities.
a In case of dose reduction, when AE returns to baseline or is resolved, dose escalation to 600 mg b.i.d. 
may be considered at the discretion of the Investigator.
5.5
Concomitant Medications and Procedures
Concomitant medications and procedures to be documented in patients’ medical sources and 
collected in eCRF include any medication or therapeutic procedure (e.g. prescription drugs, over-the-
counter drugs, herbal/homeopathic remedies, nutritional supplements, blood transfusions, etc.) 
used by a patient from 7 days prior to enrollment until the EoS visit.
Patients will be instructed to consult with the Investigator before taking any medications (including 
over-the-counter medications).  
Supportive medications may be provided prophylactically or therapeutically per Investigator
discretion.
Any concomitant treatment not listed in the section 5.5.2 is considered permitted in the study and 
may be prescribed as clinically appropriate during the study.
Concomitant therapies will be assessed at all visits and must be recorded on the appropriate eCRF
according to the rules in section 6.5.4.
5.5.1
Precautions – Drug Interactions

Drugs metabolized via CYP3A4 enzyme: Darolutamide may decrease the plasma 
concentrations of concomitantly given drugs which are mainly metabolized via CYP3A4 
enzyme (e.g., simvastatin, verapamil, dronedarone). Although the clinical significance of this 
potential interaction is not fully known, the concomitant intake of these substances during 
study treatment should be avoided. In case patients must continue receiving them, they
should be monitored for signs of decreased efficacy of these concomitant medications.
In vivo data from study 17726 indicates that co-administration of rifampicin -a strong 
CYP3A4 inducer- resulted in a significant decrease of darolutamide plasma concentrations.
Therefore, repeated co-administration of strong or moderate CYP3A4 inducers (e.g. 
carbamazepine, phenobarbital, St. John's Wort) with darolutamide is expected to reduce 
darolutamide plasma concentrations. Thus, the concomitant intake of these substances 
during study treatment should also be avoided.

P-gp and BCRP inhibitors/substrates: In vitro data indicate that darolutamide is a substrate 
of P-gp and BCRP. Therefore administration of strong inhibitors of P-gp (e.g. verapamil,


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dronedarone) and BCRP (e.g., pantoprazole, eltrombopaq) may increase the plasma 
concentrations of darolutamide and should be used with caution. Despite this fact, in vivo
data from study 17726 showed that co-administration of darolutamide with itraconazole -a 
strong CYP3A4 and P-gp inhibitor- does not result in a clinically relevant increase of 
darolutamide plasma concentrations. In any case, caution is recommended.
Plasma concentration of drugs that are sensitive P-gp or BCRP substrates might be increased
by darolutamide. Therefore sensitive substrates for P-gp (e.g., digoxin) should be used with 
caution when co-administered with darolutamide. Co-administration of darolutamide can 
also increase the plasma concentrations of other concomitant BCRP substrates (e.g. 
methotrexate, sulfasalazine, fluvastatin, atorvastatin). Therefore, the patients should be 
closely monitored for signs and symptoms of increased exposure to such BCRP substrates.
Dose modification of BCRP substrates should be considered based on the prescriber
information or such compounds should be avoided.
Refer to the section “Drug Development and Drug Interactions: Table of Substrates, Inhibitors and 
Inducers” of the FDA website for a list of potential interactions (as of the date of this protocol
available at: 
http://www.fda.gov/Drugs/DevelopmentApprovalProcess/DevelopmentResources/DrugInteractions
Labeling/ucm093664.htm
5.5.2
Prohibited Treatments and/or Procedures
Patients are prohibited from receiving the following therapies from the time of PICF signature until 
BC surgery/pre-NAST biopsy:

Investigational agents other than the protocol treatment.

Any additional standard or investigational anticancer agents, such as chemotherapy, 
immunotherapy, targeted/biologic therapy, endocrine therapy, etc., even if utilized as 
treatment of non-cancer indications. Darolutamide must be permanently discontinued upon 
initiation of a non-protocol standard or investigational antineoplastic therapy prior to BC 
surgery/pre-NAST biopsy.  
5.5.3
Contraception
WoCBP must agree to use acceptable non-hormonal contraceptive methods of birth control from 
the day of the screening pregnancy test up to 3 months after the last intake of protocol treatment. 
Refer to section 4.1 for the definition of WoCBP.
Acceptable forms of contraception are:
(1) pracZce absZnence† from heterosexual acZvity; 
OR 
(2) use (or have their partner use) acceptable contracepZon during heterosexual acZvity‡:  

Single method (one of the following is acceptable):
o
intrauterine device (IUD)
o
vasectomy of a female patient’s male partner¥

Combination method (requires use of two of the following):
o
diaphragm with spermicide (cannot be used in conjunction with cervical 
cap/spermicide)
o
cervical cap with spermicide (nulliparous women only)  


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o
contraceptive sponge (nulliparous women only) 
o
male condom or female condom (cannot be used together)
†AbsZnence (relaZve to heterosexual acZvity) can be used as the sole method of contracepZon if it is 
consistently employed as the patient’s preferred and usual lifestyle and if considered acceptable by local 
regulatory agencies and Ethics Committees. Periodic abstinence (e.g., calendar, ovulation, sympto-thermal, 
post-ovulation methods, etc.) and withdrawal are not acceptable methods of contraception. 
‡ If a contraceptive method listed in this section is restricted by local regulations/guidelines, then it does not 
qualify as an acceptable method of contraception for patients participating at sites in this country/region.
¥ Vasectomised partner is a highly effective birth control method provided that partner is the sole sexual 
partner of the WoCBP trial participant and that the vasectomised partner has received medical assessment of 
the surgical success.
6. SCHEDULE OF VISITS AND PROCEDURES
6.1
Patient Inclusion
6.1.1
Informed Consent
A written, dated and signed and freely given informed consent must be obtained prior to any study-
specific procedure.
Prior to any study-specific screening evaluation, the patient will be informed of the nature of the 
study and will be given pertinent information as to the intended purpose, possible benefits, and 
possible adverse experiences. The procedures and possible hazards to which the patient will be 
exposed will be explained.
An approved PICF will then be read and signed by the patient and, when required, a witness, and the 
Investigator or a person designated by the Investigator, as per local regulations. The patient will be 
provided with a copy of the signed PICF.
Patients with a suspicion of invasive BC based on clinical and/or radiological findings or those who 
have confirmation of invasive BC based on previous cytology/core/incisional biopsy deemed 
adequate candidate by the Investigator will be invited to participate in the study. In cases without 
pathological confirmation of BC diagnosis, signature of PICF before BC confirmation biopsy is an 
option to allow the collection of study-specific biopsies to be conducted at the same time as the 
diagnostic biopsy. For patients who have already had a confirmatory biopsy, they may participate in 
the study if their diagnostic biopsy has been done according to the protocol requirements. If their 
diagnostic biopsy has not been done according to the protocol requirement, they may also 
participate in the study if willing to undergo a subsequent study-specific biopsy procedure according 
to the protocol.
The patient may withdraw from the study at anytime without prejudicing future medical treatment. 
In any case, withdrawal should be documented on patient´s clinical records.
If a potential patient is illiterate or visually impaired, the Investigator must provide an impartial 
witness to read the PICF to the patient and must allow for questions. Thereafter, the patient and the 
witness must sign the PICF to attest that informed consent was freely given and understood. 
The PICF will include a statement by which the patient allows the Sponsor’s duly authorized 
personnel, the EC/IRB, and the regulatory authorities to have direct access to her data that will be 
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If important new information becomes available that may be relevant to the patient’s consent and 
willingness to continue participation in the study, the PICF will be revised and submitted to IRB/IEC
and regulatory authorities (if applicable) for approval/favorable opinion. The new information will be 
then discussed with the patient in a timely manner and if she agrees to continue participation in the 
study, the revised PICF will be signed and dated and the patient will receive a copy.
In line with each local applicable regulations, the source should also support the documentation of 
the consent process, for each patient.
6.1.2
Registration and Enrollment 
6.1.2.1 Registration and Enrollment Procedures
At time of PICF signature patients will be registered in the study and then will undergo screening 
procedures. After all screening procedures and confirmation of eligibility, patients will be enrolled.
The following steps for registering and enrolling a patient are chronologically performed:
1.
The patient must provide written informed consent prior to performing any study-specific 
procedure not considered standard practice by the institution.
2.
The site will ask TRIO for patient’s registration immediately after signature of PICF. A unique 
registration number will be assigned to each patient in the study.
3.
The site will proceed to perform the protocol-required biopsy and all screening procedures 
according to section 6.3. Upon completion of screening procedures, the Investigator or 
his/her delegate will check if the patient meets all eligibility criteria: 
Patients who do not meet all eligibility criteria: sites will have the screen failure form 
completed in Remote Data Capture (RDC) with corresponding screen failure reason 
captured.
Patients who meet all eligibility criteria: sites will enter the screening data in RDC 
which will be reviewed centrally by TRIO. After the review of the eligibility criteria, 
TRIO may address, if necessary, some discrepancies that will have to be answered by 
the site. Once the eligibility central review is completed and discrepancies have been 
resolved (if applicable) TRIO will provide the confirmation that the patient is eligible 
and the site will be able to move forward with the patient’s enrollment in RDC. The 
site will receive a confirmation of enrollment. If the patient is deemed not eligible 
following the central eligibility review, the site will be asked to report the screen 
failure reason in RDC.
6.1.2.2 Enrollment According to BC Subtype
To be able to assess the molecular alterations after darolutamide exposure in different BC subtypes,
patients being either TNBC, or HR+/HER2-, or HER2+ will be enrolled in the study. Up to 20 patients 
with each of these subtypes will be included, with an acceptable minimum of 8 evaluable patients in 
each cohort if enrollment of recommended 20 patients is not feasible within a cohort for any reason.
Subtypes are defined based on local assessment as:

TNBC: ER, PgR and HER2 negative

HR+/HER2-: ER and/or PgR positive and HER2 negative

HER2+: HER2 positive, irrespective of ER/PgR status
ER and PgR positivity is defined according to the American Society of Clinical Oncology/College of 
American Pathologists Guideline Recommendations, as cases with ≥ 1% of tumor cell nuclei that are 
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HER2 positivity is defined as IHC 3+ and/or ISH amplification. HER2 negativity is defined as IHC 0-1 
and/or ISH non-amplification. In case HER2 status is Equivocal, patients will not be assigned to any 
cohort until central review is done and HER2 status is defined as either positive or negative.
Although patient inclusion in each of these three cohorts will be based on local testing, the central 
laboratory (TORL) will also test for ER, PgR and HER2. Assessment of distribution of patients with 
respect to centrally-assessed ER/PgR/HER2 will be done on an ongoing basis by the central 
laboratory and will also be performed by the Study Steering Committee (SSC) once the minimum 
requirement of patients in a cohort have undergone the EoS per protocol. This will allow the SSC to 
ascertain the rates of discordance between local and central testings and determine if more or less 
patients may be required in one or more cohort(s).
6.2
Study Participation Discontinuation
The Investigator has the right to discontinue a patient from protocol treatment or withdraw a 
patient from the study at any time. In addition, patients have the right to voluntarily discontinue 
protocol treatment or withdraw from the study at any time for any reason. 
6.2.1
Discontinuation from Protocol Treatment
Refer to section 5.3.3.
6.2.2
Discontinuation from Study Participation
The primary reason for study participation discontinuation should be documented on the eCRF. 
Reasons for patient´s discontinuation from the study include (but may not be limited to):

Patient is enrolled but does not receive protocol treatment for any reason

Patient completes the study 

Death

Consent withdrawal from entire study participation

Patient non-compliance

Lost to follow-up
Should a patient be enrolled, but has never started protocol treatment, she will be discontinued 
from the trial, and managed according to the site’s SoC.
Should a patient decide to withdraw consent, all efforts will be made to complete and report the 
observations as thoroughly as possible. The Investigator should schedule a visit with the patient, or 
contact the patient by telephone to establish as completely as possible the reason for the 
withdrawal. A complete final evaluation at the time of the patient’s withdrawal should be made with 
an explanation of why the patient is withdrawing from the study. After complete withdrawal of 
consent, no further study procedure is performed and no further data will be collected.
If a patient withdraws consent to the use of study-specific tumor samples:

The Investigator has the responsibility to notify TRIO immediately.

TRIO will ensure that the laboratory(ies) holding the samples is/are informed about the 
withdrawn consent immediately and that samples are either returned to the study site, or 
disposed of/destroyed by the central laboratory as per site’s request/local requirements, 
and the action documented by the central laboratory(ies). 
In the event that analysis/research has already been performed, TRIO or its representatives, in strict 
compliance with the applicable local regulations, should retain the results and associated data for 
regulatory reasons but these will not be used in any subsequent analyses.In the case a patient does 


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not show up for study visits, site staff should make reasonable contact attempts before declaring the 
patient as lost to follow-up. These attempts need to be documented in the medical records. 
6.3
Schedule of Procedures
All study visits and procedures are detailed in the table presented in this section. The detailed 
description of the study visits and assessments is in sections 6.4 and 6.5.


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Table 2: Schedule of Procedures
Protocol Activities
Screening1
Treatment Period2
Pre-surgery3
BC surgery/pre-
NAST biopsy 
End of Study4
Informed consent/Registration and Enrollment5
X
Demographics, medical, surgical, disease history
X
Complete physical examination6
X
X
Symptom-directed physical examination
X
X
ECOG PS
X
X
X
X
Weight, height, blood pressure7
X
Serum pregnancy test
X8
Hematology, Blood chemistry9
X
X
X
AEs assessments
X
X
X
X
Concomitant medication and therapeutic 
procedures10
X
X
X
X
Treatment compliance assessment11
X
X
Tumor sample12
X
X
Note: All assessments should be performed within 3 days of the scheduled visit, unless otherwise specified. 
Footnotes:
1: Screening procedures (except PICF signature and screening tumor samples – see footnote 12) to be done within 28 days of enrollment, unless otherwise 
specified
2: Protocol treatment should begin within 5 days of enrollment and no more than 42 days after breast tissue collection. Treatment should last at least 14 
days, and no more than 35 days (recommended maximum duration 21 days). The need for a clinic visit during this period will depend on the duration of 
protocol treatment. If duration of treatment with ODM-201 is 14 to 21 days only the Pre-surgery visit needs to be performed after Day 1, unless otherwise 
clinically indicated. If ODM-201 is taken/expected to be taken for > 21 days (due to planned date of surgery/pre-NAST biopsy, delay in surgery/pre-NAST 
biopsy, etc.) a visit is required as close as possible to Day 21 +/- 3 days (except in case the pre-surgery visit is planned within Day 21 + / - 3 days).
3: The Pre-surgery visit should be performed the day before BC surgery or the day before BC pre-NAST biopsy is performed. The only exceptions should be: 


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(a) If for some reason the patient is ordered to stop all oral intakes earlier than the day prior to BC surgery/pre-NAST biopsy, the Pre-surgery visit should be 
scheduled on the day patient is ordered to stop oral intake; (b) In case the BC surgery/pre-NAST biopsy is planned for a Monday or the day following a 
holiday; in such cases the Pre-surgery visit should be done on a working day as close as possible to the BC surgery/pre-NAST biopsy (e.g. on a Friday if surgery 
is scheduled on Monday); or (c) In case that performing Pre-surgery visit on the same day of BC surgery/pre-NAST biopsy is required by site due to logistical 
reasons or patient’s/physician’s convenience. Refer to section 6.4.3 for further guidance about the Pre-surgery visit planning and procedures.
4: End of Study visit must be scheduled 30 days (+/- 3 days) after the last intake of darolutamide, even for patients who did not complete the minimum 14 
days of protocol treatment. The EoS will be the last study visit for patients. However, for patients experiencing protocol treatment-related AEs/SAEs that are 
ongoing at the time of the EoS visit, monitoring of these events will continue as clinically indicated until (a) the events have resolved or (b) the events have 
reached a status which, in the Investigator’s opinion, is unlikely to resolve due to the nature of the condition and/or the patient’s underlying disease. 
Frequency of visits will be according to Investigator’s judgment. 
5: There is no window to obtain the signed and dated PICF. Study-specific screening assessments not considered standard practice by the institution must 
start only once the PICF has been obtained. Registration takes place as soon as the patient signs the PICF. After screening procedures and if patient meets all 
eligibility criteria, enrollment will be done according to the process described in section 6.1.2.
6: Complete physical examination will include a complete assessment of body systems (i.e., general appearance, head, eyes, ears, nose, mouth, skin, heart, 
lungs, lymph nodes, gastrointestinal, genitourinary, skeletal, and neurologic). A complete examination of the breast and regional lymph nodes will also be 
done. 
7: Weight, height and blood pressure must be assessed and collected during screening. Afterwards they will be assessed (as well as other vital signs) 
according to the SoC.
8: Within 7 days of enrollment, only in WoCBP. 
9: Hematology: Hemoglobin, WBC, ANC, platelets. Blood chemistry: AST, ALT, total bilirubin, creatinine.
10: Concomitant medication and procedures include any medication or therapeutic procedure used by the patient within 7 days prior to enrollment until the 
EoS visit. Refer to section 6.5.4 for the reporting rules in the eCRF.
11: All unused drugs (partially used and empty containers) and the patient diary will be handed over to Investigators at each treatment visit (if any) and at the 
Pre-surgery visit.
12: Mandatory tissue collection:
-
Pre-treatment: A breast tumor sample must be collected at the time of the initial diagnosis (or in a subsequent procedure) either by core needle or 
incisional biopsy. Excisional biopsy will not be allowed. A minimum of 3 core biopsies using a 14-gauge needle (or the equivalent with an incisional 
biopsy) is recommended (2 fresh-frozen + 1 paraffin-embedded). In cases it is not possible to provide enough material, collection of 2 cores (or 
equivalent amount with incisional biopsy) will be acceptable: one core will be snap-frozen and the other one will be prepared in paraffin. For patients 
who already had such confirmatory biopsy prior to registration, there is no need to repeat it provided that their diagnostic biopsy has been done 
according to the protocol requirements (in terms of number of cores/tissue, fresh frozen collection, etc.). The screening tumor samples must be 
collected before start of treatment with darolutamide at the latest.


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- BC surgery/pre-NAST biopsy: Two cores (or equivalent amount) of breast tumor tissue must be collected at the time of the patient’s surgery or before 
starting NAST if indicated (1 fresh-frozen + 1 paraffin-embedded).  


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6.4
Study Visits and Assessments/Investigations
During all visits below, assessments should be done according to the Schedule of Procedures (section 
6.3).
6.4.1
Screening Period
The screening period starts with the registration (at time of PICF signature) and ends when the 
patient is enrolled (following eligibility central review and confirmation by TRIO) or screen failed. 
When scheduling the screening procedures it should be taken into account that treatment with 
darolutamide should start within 42 days of the study-specific tumor sample collection.
During the screening period the following assessments will be done:

Demographics, medical, surgical, disease history

Complete physical examination (including weight, height and blood pressure measurements)

ECOG PS

Serum pregnancy test: only in WoCBP, within 7 days of enrollment (refer to section 4.1 for 
the definition of WoCBP).

Laboratory assessments:
o Hematology: Hemoglobin, WBC, ANC and platelets
o Blood chemistry: AST, ALT, total bilirubin and creatinine

Adverse events assessment. For information on the assessment and collection of AEs refer 
to section 7 and to section 6.3

Concomitant medications

Tumor biopsy (refer to section 6.5.7)
Once all screening assessments have been conducted, the site will request enrollment of patients 
who meet all eligibility criteria through completion of all screening data in RDC (see section 6.1.2.1), 
and TRIO will review patient´s eligibility. No enrollment must take place before formal confirmation 
of eligibility is provided by TRIO to the site.
Once the patient is enrolled, she will be instructed to begin protocol treatment with darolutamide. If 
for some reason the patient does not start their study treatment, she will be discontinued from 
study, and managed as per site’s SoC. 
If for some reason laboratory tests are repeated between these tests done at screening and 
treatment start, the most recent measurement obtained prior to treatment start must be within the 
required values according to the inclusion criteria, to begin protocol treatment.
6.4.2
Treatment Period Visits
Refer to section 5.3.2.1 for timelines regarding protocol treatment. 
The need for a clinic visit during this period will depend on the duration of protocol treatment.
If duration of treatment with ODM-201 is 14 to 21 days, only the Pre-surgery visit needs to be 
performed after Day 1, unless otherwise clinically indicated.


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If ODM-201 is taken/expected to be taken for > 21 days (due to planned date of BC surgery/pre-
NAST biopsy, delay in BC surgery/pre-NAST biopsy, etc.) a visit is required as close as possible to Day 
21 +/- 3 days (except in case the pre-surgery visit is planned within Day 21 + / - 3 days).
During the treatment period visit(s) (if any) the following will be assessed:

Symptom-directed physical examination 

ECOG PS

AEs 

Concomitant medication
6.4.3
Pre-surgery Visit
As a general rule, the Pre-surgery visit will be performed the day before BC surgery or the day before 
pre-NAST biopsy if NAST is indicated.* If after the Pre-surgery visit takes place, the BC surgery/pre-
NAST biopsy planned for the following day is delayed for any reason, the Investigator should contact 
the Medical Monitor to discuss about potential continuation of darolutamide after the Pre-surgery
visit.
During this visit the following assessments will be done:

Complete physical examination 

ECOG PS

Safety laboratory assessments:
o Hematology: Hemoglobin, WBC, ANC and platelets
o Blood chemistry: AST, ALT, total bilirubin and creatinine

AEs

Concomitant medication
Other pre-surgical assessments should be done according to the standard of care (SoC) at the site.
The only exceptions to scheduling the Pre-surgery visit on the day prior to BC surgery/pre-NAST 
biopsy will be in the following cases:
a.
If the patient is ordered to stop all oral intakes earlier than the day prior to BC 
surgery/pre-NAST biopsy. In such case the Pre-surgery visit should be scheduled on 
the day patient is ordered to stop oral intake and all test/assessments should be 
done at that point.
b.
In case the BC surgery/pre-NAST biopsy is planned for a Monday or the day 
following a holiday and the Pre-surgery visit cannot be scheduled the day prior to BC 
sugery/pre-NAST biopsy. In such case the following will be done:
The Pre-surgery visit should be done on a working day as close as possible to 
the BC surgery/pre-NAST biopsy (e.g. on a Friday if surgery is scheduled on 
Monday).
The patient will be instructed to continue with darolutamide intakes until 
the day prior to BC surgery/pre-NAST biopsy or when she is instructed to 
stop all oral intakes, whichever occurs first.


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c.
In case that performing Pre-surgery visit on the same day of BC surgery/pre-NAST 
biopsy is required by site due to logistical reasons or patient’s/physician’s 
convenience. In such case, it is acceptable that Pre-surgery visit is performed on the 
same day that BC surgery/pre-NAST biopsy takes place, but before any of those 
procedures are performed. The results from the laboratory safety tests must be
available and reviewed by the Investigator before the BC surgery takes place.
*Note: Should they coincide, it is acceptable for the Pre-surgery visit to take place on the day of a 
treatment period visit (if any). In such cases, assessments should be done according to the Pre-
surgery visit requirements.
6.4.4
BC Surgery/Pre-NAST biopsy
Surgery will be done according to site standards. In patients not receving NAST, BC tissue will be 
collected at time of surgery according to section 6.5.7. In patients receiving NAST, a breast tumor 
sample will be collected during the pre-NAST biopsy (refer to section 6.5.7) after completing 
darolutamide and before NAST start. As much as possible, sites should avoid scheduling BC 
surgery/pre-NAST biopsy on Mondays or on the day following a holiday, so that Pre-surgery visit may 
take place the day prior to BC surgery/pre-NAST biopsy according to section 6.4.3.
In the event that a sentinel lymph node biopsy (SLNB) is performed, periareolar injection is to be 
conducted.  The dye/colloid should not be injected into the tumor itself as this may alter the tissue 
and affect the molecular analyses to be performed on tumor tissue sample.  
6.4.5
End of Study Visit
EoS visit must be scheduled 30 days (+/- 3 days) after the last intake of darolutamide, even for 
patients who did not complete the minimum 14 days of protocol treatment.
The primary purpose of this visit is to follow-up any AEs/laboratory abnormality ongoing at the time 
of protocol treatment discontinuation and to assess any new AEs/abnormality that may have 
occurred since protocol treatment discontinuation. Symptom-directed physical examination, ECOG 
PS assessment and hematology/blood chemistry should be done at this visit. All AEs will be collected 
(even if due to neoadjuvant treatment, surgery or adjuvant treatment, regardless of the relationship 
to darolutamide), as well as relevant concomitant treatment according to rules in section 6.5.4.
EoS will be the last study visit with the exception of patients with protocol treatment-related 
AEs/SAEs that are ongoing at the time of the EoS visit. In these cases, monitoring of these events will 
continue as clinically indicated until (a) the events have resolved or (b) the events have reached a 
status which, in the Investigator’s opinion, is unlikely to resolve due to the nature of the condition 
and/or the patient’s underlying disease. Frequency of visits will be according to Investigator´s
judgment.
6.4.6
Unscheduled Visit Procedures
Unscheduled visits may be performed any time to assess or follow AEs, to perform any clinically-
indicated investigation, or at the request of the patient or Investigator.
6.5
Description of Assessments/Investigations
6.5.1
Demographics and Medical History
The Investigator will collect and report in the eCRF demographics and complete history of malignant 
and clinically-significant non-malignant diseases including known hypersensitivity reactions as well 
as clinically-significant symptoms.


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6.5.2
Physical Examination and Vital Signs
At screening and at the Pre-surgery visit, physical examinations will include a complete assessment 
of body systems (i.e., general appearance, head, eyes, ears, nose, mouth, skin, heart, lungs, lymph 
nodes, gastrointestinal, genitourinary, skeletal, and neurologic) per SoC at the study site or as 
clinically indicated by symptoms. As well, a complete examination of the breast and regional lymph 
nodes will be done. Weight, height and blood pressure will also be measured at screening, and 
therafter according to SoC.
During protocol treatment and EoS visits, a symptom-directed physical examination will be done.
6.5.3
ECOG Performance Status
Assessment of ECOG PS (Appendix 1) is required to assess patient´s functional status for study 
eligibility purposes and will be performed throughout the study according to the section 6.3.
6.5.4
Adverse Events and Concomitant Treatments Assessment

Adverse events assessment: For information on the assessment and collection of AEs refer 
to section 7 and to section 6.3.

Concomitant Treatments: Concomitant treatment includes any medication or therapeutic 
procedure (e.g. prescription drugs, over-the-counter drugs, herbal/homeopathic remedies, 
nutritional supplements, blood transfusions, etc.) used by a patient from 7 days prior to 
enrollment until EoS. The concomitant treatments to be collected and reported in the eCRF
are:
o
Any treatment used by the patient within 7 days prior to enrollment
o
After enrollment, only if: 

Medication is given to treat an AE

Medication is listed in sections 5.5.1 or 5.5.2
6.5.5
Laboratory Assessments
At screening:

Hematology: Hemoglobin, WBC, ANC and platelets

Blood chemistry: AST, ALT, total bilirubin and creatinine

Serum pregnancy test: within 7 days of enrollment, only in WoCBP
At the Pre-surgery and EoS visits:

Hematology: Hemoglobin, WBC, ANC and platelets

Blood chemistry: AST, ALT, total bilirubin and creatinine
6.5.6
Assessment of Efficacy
No efficacy assessments are planned in this study.  With the exception of the examination of breast 
and regional lymph nodes according to section 6.5.2, disease assessments will be conducted 
according to the SoC at the study site.  
6.5.7
Tumor Tissue Collection and Molecular Evaluation
All tumor samples will be submitted to the study Central Laboratory and will be stored for up to 20 
years, unless otherwise required per local regulations, Investigator, IRB/IEC and/or the patient.


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Full details on the analyses to be performed on the biologic samples and procedures for sample 
collection, labeling, storage and shipment/return of biological samples will be described in separate 
document(s) (e.g. Laboratory Manual, Samples Plan).
6.5.7.1 Breast Tumor Tissue Collection
Collection of the following samples is mandatory in the study:

Pre-treatment: A breast tumor sample must be collected at the time of the initial diagnosis 
(or in a subsequent procedure) either by core needle or incisional biopsy. Excisional biopsy 
will not be allowed. 
A minimum of 3 core biopsies using a 14-gauge needle (or the equivalent with an incisional 
biopsy) is recommended to perform the protocol-defined molecular analyses. Once the core 
biopsies have been removed, two of the cores (or equivalent amount of tumor) must be 
immediately snap-frozen using the procedure outlined in the study laboratory manual, then 
stored in liquid nitrogen or at -70 to -80 degrees Celsius at the site. The third sample should 
be prepared in paraffin according to site’s practice. In cases it is not possible to provide 
enough material, collection of 2 cores (or equivalent amount with incisional biopsy) will be 
acceptable: one core will be snap-frozen and the other one will be prepared in paraffin.
In patients with multifocal or multicentric tumors, efforts will be made to obtain samples 
from each discrete lesion.
For patients who already had the confirmatory biopsy prior to registration, there is no need 
to repeat it provided that their diagnostic biopsy has been done according to the protocol 
requirements (in terms of number of cores/tissue, fresh frozen collection, etc.).

BC surgery/Pre-NAST biopsy: An amount of BC tissue equivalent to two cores (or 2 cores of 
tissue) must be collected at the time of the patient’s surgery (if no NAST indicated) or at 
time of the pre-NAST biopsy (if NAST is indicated). The pre-NAST biopsy should be done 
before NAST start. Half of the tissue will be prepared as fresh-frozen, the other half in 
paraffin.  
6.5.7.2 Molecular Evaluation
The primary endpoint of the study is to explore and define the molecular alterations that may occur 
following darolutamide administration through comparisons of BC tissue before and after short-term 
exposure to darolutamide.
At least, the following markers will be assessed on the collected samples. In all experiments, 
appropriate positive and negative controls will be included.

AR

ER

PgR

HER2 

ki67

Cytokeratins 5 and 6
Tissue microarrays will be developed from all tumor specimens. Other in situ assays for activated 
downstream signaling molecules will be performed using tissue sections and/or tissue microarrays. 
Promising candidate genes obtained from the microarray analysis will be validated using the frozen


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tissue arrays generated above and using appropriate probes (i.e. antibodies, if available, or nucleic 
acid probes).
Additional assays in example but not limited to Reverse Phase Protein Analysis (RPPA), sequenciation 
of the RNA, immunohistochemistry (IHC) etc. may be performed based on the molecular analysis 
finding and/or in the future as new technology and data become available.
6.5.8
Other Procedures/Assessments
In addition to the protocol-required assessments, the Investigator may perform any clinically 
indicated test and procedure according to SoC (e.g. coagulation test prior to surgery, etc.) although 
these will not be collected in the eCRF.
7. SAFETY MONITORING
7.1
Definitions
7.1.1
Period of Observation
At each visit and any contact during the study and follow-up period, the Investigator or designee will 
inquire about the occurrence of AEs (and corresponding medications administered) and will 
document them in the eCRF.
For the purposes of this study, the period of observation for SAEs and AEs extends from the time the 
patient signs the PICF until the EOS, except for patients that are experiencing AEs or SAEs related to 
darolutamide ongoing at the time of EoS visit. In such cases, these events will be monitored beyond 
the EoS visit as clinically indicated until (a) the events have resolved or (b) the events have reached a 
status which, in the Investigator’s opinion, is unlikely to resolve due to the nature of the condition 
and/or the patient’s underlying disease. 
The Investigator or designee will report the AEs/SAEs in the eCRF (and, if applicable, to TRIO Safety 
Department) when required as per the table below:
Table 3: Reporting Period of AEs/SAEs
Study period
AEs (non-serious)
SAEs
From PICF to 1st intake
Only if related to study 
participation
All (regardless of relationship)
From 1st intake until 
EoS visit
All (regardless of relationship)
All (regardless of relationship)
After the EoS visit
Only if related to protocol
treatment
Only if related to protocol 
treatment
7.1.2
Adverse Event
An AE is defined as any untoward medical occurrence, including an exacerbation of a pre-existing
condition, in a patient in a clinical investigation who received a pharmaceutical product. The event 
does not necessarily have to have a causal relationship with this treatment.
Worsening of pre-existing conditions: any pre-existing condition present at baseline, which remains 
unchanged during the trial, does not need to be recorded as an AE.


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Any worsening of any pre-existing baseline condition should be reported as an AE in the eCRF. 
Examples of worsening of a preexisting condition that should be recorded as an AE are given below:   

Worsening of condition meets the criteria for an AE or SAE

Action is taken with the investigational drug (e.g. dose is reduced or treatment is 
discontinued)

Treatment is required (concomitant medication is added or changed)

The Investigator believes a patient has shown a clear deterioration from baseline symptoms
Expected fluctuations or expected deterioration of the BC (symptoms of disease progression) should
not be recorded as an AE.
Changes in vital signs, physical examination and laboratory test results: these will only be recorded
as an AE in the eCRF if they are judged clinically relevant by the Investigator or led to protocol 
treatment modifications (dose delay or dose reduction).
Surgical procedures: these are not AEs per se; they are therapeutic measures for conditions that 
require surgery. The condition for which the surgery is required may be an AE. Planned surgical 
measures permitted by the clinical study protocol and the condition(s) leading to these measures are 
not AEs.
Worsening of a sign or symptom of the condition under treatment: these situations (i.e. BC) should 
not be recorded as AEs.
7.1.3
Serious Adverse Events
A SAE is one that at any dose:

Results in death

Is life-threatening1

Requires inpatient hospitalization or prolongation of existing hospitalization

Results in persistent or significant disability or incapacity2

Is a congenital anomaly or birth defect

Is an important medical event3
1 “Life-threatening” means that the patient was at immediate risk of death at the time of the SAE; it does not 
refer to a SAE that hypothetically might have caused death if it were more severe.
2 “Persistent or significant disability or incapacity” means that there is a substantial disruption of a person’s 
ability to carry out normal life functions.
3 Medical and scientific judgment should be exercised in deciding whether expedited reporting is appropriate 
in situations where none of the outcomes listed above occurred. Important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize the patient or may 
require intervention to prevent one of the other outcomes listed in the definition above should also usually 
be considered serious. Examples of such 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, the development of drug dependency or drug abuse, etc.
7.1.3.1 Disease-related Events
An event that is part of the natural course of the disease under study (i.e. breast cancer) does not
need to be reported as an SAE.


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However, if the Investigator considers that there was a causal relationship between treatment with
investigational product or protocol design/procedures and a disease progression, then this must be
reported as an SAE. Any new primary cancer must be reported as an SAE.
7.1.3.2 Planned or Administrative Hospitalization
Patients may be hospitalized for administrative or social reasons during the trial (e.g. long distance
from home to site, etc). These and other hospitalizations planned up to enrollment do not need to
be reported as an SAE but should be properly documented in the source data.
7.1.4
Unexpected Adverse Event
The Darolutamide IB provides an overview of the AEs reported across each clinical trial. 
Any SAE assessed as related to darolutamide but not presented in the IB section 8.6 Undesirable 
effects will be documented by TRIO as a suspected unexpected serious adverse reaction (SUSAR) and 
will be submitted according to applicable local regulations.
7.2
Performing Adverse Event Assessment
7.2.1
Collection of Adverse Event Information
The following information will be collected in the eCRF: Description of event (a single medical 
condition and not a collection of signs/symptoms), start/stop dates, worst grade experienced 
(severity/intensity), seriousness, action taken on protocol treatment and relationship to protocol
treatment.
Intensity of AE: The intensity of AEs will be classified and recorded according to the NCI CTCAE
version 4.03 in the eCRF, or according to the grading defined in section 5.4.1 if the AE is not 
specifically listed in NCI CTCAE.
All adverse clinical experiences, whether observed by the Investigator or reported by the patient, 
must be recorded, with details about the duration and intensity of each episode, the action taken 
with respect to the test drug (refer to section 5.4.2 for recommendations on dose modifications), 
and the patient’s outcome.  The Investigator must evaluate each adverse experience for its 
relationship to the test drug and for its seriousness.
The Investigator must appraise all abnormal laboratory results for their clinical significance. If any 
abnormal laboratory result is considered clinically significant, the Investigator must provide details 
about the action taken with respect to the protocol treatments and about the patient’s outcome.
7.2.2
Assessment of Causality
Medical judgment should be used to determine the relationship, considering all relevant factors, 
including pattern of reaction, temporal relationship, de-challenge or re-challenge, confounding 
factors such as concomitant medication, concomitant diseases and relevant history. Assessment of 
causal relationship must be recorded for each adverse event.
Relationship to study drugs will be reported as either “Yes” or “No”.

Yes: there is a reasonable causal relationship between the IMP administered and the AE.

No: there is no reasonable causal relationship between the IMP administered and the AE.
7.3
Reporting of Serious Adverse Events and Pregnancies
7.3.1
Reporting of Serious Adverse Events and Pregnancies from the sites to TRIO


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SAEs and pregnancies will be reported within 24 hours of awareness by completing and emailing or 
faxing the SAE Reporting Form to TRIO Drug Safety Unit.
TRIO Drug Safety Unit contact information:

Email: safety@trioncology.org

Fax number: + 1 780-702-2273
Table 4: SAE and Pregnancies Reporting Timelines
Initial Reports
Follow-up Information
Type of Event
Time Frame
Documents
Time Frame
Documents
All SAEs
Within 24 h of 
awareness
SAE reporting 
form
Within 24 h of 
awareness
SAE reporting 
form
Pregnancy
Within 24 h of 
awareness
Pregnancy
reporting form
Within 24 h of 
awareness
Pregnancy
reporting form
7.3.2
SAE information Provided by TRIO to the Manufacturer
TRIO will report all SAEs and pregnancies as well as follow up information for these events to Bayer
by email or other secure method within 24 hours of awareness.
7.4
Management of Specific Cases
7.4.1
Medication Errors
In the event of a medication error, the patient should be observed closely for signs of toxicity. 
Medication errors include the accidental or intentional use of darolutamide in a daily dose greater 
than the protocol-specified daily dose. Appropriate supportive treatment should be provided if 
clinically indicated. In case of toxicity derived from the medication error, dose modification 
recommendations from section 5.4.2 should be followed. 
The highest darolutamide dose administered in clinical trials is 1,800 mg daily, with no maximum 
tolerated dose achieved and no dose-limiting toxicity experienced by patients. 
If an AE is associated with (“results from”) a medication error, the corresponding AE should be 
reported as SAE, even if no other seriousness criteria are met.
In cases of a medication error without any associated AE, the medication error should be reported as 
a non-serious AE, using terminology like “accidental or intentional overdose without adverse effect”, 
“medication error without adverse effect” or similar.
7.4.2
Pregnancy
Should a pregnancy occur, the Investigator must discontinue treatment immediately, capture any 
drug exposure during pregnancy, report the pregnancy through the Pregnancy Reporting Form and 
follow the pregnancy until outcome is known. Information related to outcome of the pregnancy, 
delivery, postpartum recovery and the clinical condition of the offspring during the neonatal 
period must also be collected.
Pregnancy is not automatically considered as a SAE, however, it must be reported following SAE 
timeline as detailed in section 7.3.


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A SAE report must be forwarded to the TRIO if the outcome of the pregnancy results in an 
abortion/miscarriage or the occurrence of any other SAE(s).
7.5
Reporting to Ethics Committees, Regulatory Authorities and other 
Investigators
Prompt notification of SAEs by the Investigator to TRIO is essential so that legal obligations and 
ethical responsibilities towards the safety of patients are met.
TRIO will comply with local regulatory requirements relating to safety reporting to the regulatory 
authorities, IRB/IEC and Investigators.
TRIO will report SAEs and related safety information to the FDA as per the definitions and 
reccomendations in 21 CFR 312 and per the FDA guidance for Industry and Investigators (available at
http://www.fda.gov/downloads/Drugs/.../Guidances/UCM227351.pdf).
Investigator safety reports are prepared for SUSARs according to local regulatory requirements and 
are forwarded to Investigators as necessary.
An Investigator who receives an Investigator safety report describing a SAE(s) or other specific safety 
information (e.g., summary or listing of SAEs) from TRIO will file it with the IB and will notify the 
IRB/IEC, if appropriate according to local requirements.
8. STATISTICAL CONSIDERATIONS
8.1
Study Populations

Intention-to-treat (ITT) Population: The ITT population includes all patients who are enrolled 
in the study regardless of whether they have been treated or not.

Safety Population: The Safety population includes all patients who have taken at least one 
tablet of darolutamide.

Evaluable population: Includes all patients who satisfied all criteria in section 8.5.
8.2
Demographics and Baseline Characteristics
Demographics and baseline characteristics data will be listed and summarized by each BC subtype 
using the ITT population.
8.3
Protocol Treatment
Duration of treatment, duration of exposure, cumulative dose and relative dose intensity will be
summarized by each BC subtype using the Safety population. The patients with dose modifications 
will also be presented, along with reasons for the dose change.
8.4
Safety Evaluation
The assessment of safety will be made on the Safety population and will be based mainly on the 
frequency of AEs. The latest version of the Medical Dictionary for Regulatory Activities (MedDRA) 
will be used to code all AEs to a preferred term and system organ class. AEs will be graded using the
NCI CTCAE v4.03 or according to the applicable grading if the AE is not specifically listed in NCI 
CTCAE. Patient’s incidence of AEs will be tabulated by system organ class, preferred term and 
toxicity grade for each BC subtype. AEs leading to death or drug discontinuation, drug related and
SAEs will also be summarized.
Detailed listings for all AEs will also be provided.


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Laboratory parameters: these will be summarized at baseline, along visits and at the EoS visit by
each BC subtype, graded according to the NCI CTCAE v4.03. Tables of shifts in toxicity will also be
provided.
Other Safety Data: ECOG will be summarized at baseline, along visits and at the EoS visit by BC 
subtype. Tables of shifts in toxicity will also be provided when applicable.
8.5
Sample Size 
The primary objective of this trial is to evaluate molecular alterations occurring in BC tissue following 
short-term darolutamide administration. 
Up to 60 patients (minimum of 8 but up to 20 patients in each BC subtype: triple-negative, 
HR+/HER2 negative, HER2 positive) will be required for the performance of the molecular studies 
and possible hypothesis generation. 
A patient will be considered evaluable if the following criteria are met:

Minimum duration of treatment with darolutamide is of 14 days, with a minimum of 10 
consecutive days of treatment prior to the definitive BC surgery/pre-NAST biopsy. 

Mandatory tumor tissue is collected at screening and at BC surgery/pre-NAST biopsy 
according to the protocol and submitted to the central laboratory.

Adequacy for molecular assessment of the tumor tissue collected before and after the 
protocol treatment initiation. The adequacy will be evaluated by the central laboratory.
Assessment of patient evaluability, safety and distribution of patients with respect to hormone 
receptor status and HER2 status will be performed by the SSC at given time points depending on the 
recruitment, for example once 20, around 35 and once all patients have been enrolled and 
undergone the EoS per protocol. This will allow SSC to ascertain that the appropriate number of 
evaluable patients is being accrued, that more or less patients may be required, or in the extreme 
case, that accrual should close if the patient evaluability rate is very low and/or safety concerns 
arise.
The SSC will define the evaluability of patients that discontinue treatment more than 3 days before 
BC surgery/pre-NAST biopsy; molecular analyses may be taken into consideration for the decision 
about the evaluability of such patients. 
8.6
Molecular Analyses
The molecular analyses will be performed on the evaluable population. Descriptive statistics will be 
used to compare the pre-and post molecular findings.
A correlation analysis between the molecular alterations from the baseline tumor samples and the 
sample collected following administration of darolutamide, during the definitive BC surgery/pre-
NAST biopsy will be conducted.
9. ADMINISTRATIVE, ETHICAL AND REGULATORY STANDARDS
9.1
Steering Committee
The SSC of the trial will be composed of the study Chair(s) and representatives from TRIO. The SSC 
will include also a number of study principal Investigators. Other members may be added after 
consultation with the SSC members. The SSC will be constituted prior to the enrollment of the first 
patient. 


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The SSC will have the sole responsibility for the scientific conduct and integrity of the trial. 
Responsibilities include development and approval of the protocol document, monitoring of accrual, 
compliance and safety during the conduct of the trial. The SSC will be solely responsible for the 
analysis, interpretation and public disclosure of the results of the trial in accordance with the 
statistical plan. The SSC should endeavor to ensure that the study is conducted at all times to the 
standards set out in Guidelines for Good Clinical Practice (GCP). In all of the deliberations of the SSC, 
the rights, safety and well being of the study participants are the most important considerations.
9.2
Ethical Conduct of the Study
This clinical study was designed and shall be implemented and reported in accordance with the
clinical trial protocol, the ICH Harmonized Tripartite Guidelines for Good Clinical Practice, applicable 
local regulations (including European Directives 2001/20/EC and 2005/28/EC and the US Code of 
Federal Regulations Title 21), the Council for International Organizations of Medical Sciences 
(CIOMS), and the ethical principles laid down in the Declaration of Helsinki. This study will be 
conducted under ethical, scientific and medical standards that protect the rights and welfare of 
participants. 
9.3
Institutional Review Board/Independent Ethics Committee (IRB/IEC) 
Each participating institution must provide for the review and approval of this protocol and the 
associated informed consent documents and recruitment material by an appropriate IEC or IRB. The 
IEC/IRB decision concerning the conduct of the study will be made in writing and kept with the 
Investigator study file. A copy of this decision will also be provided to TRIO.
Particular attention is drawn to the Food and Drug Administration (FDA’s requirements for IRBs 
under 21 CFR Part 56). By signing the "Statement of Investigator" form (Form FDA 1572), the 
Investigator provides TRIO with the necessary assurance that an IRB is responsible for the 
initial/continuing review and subsequent approval of the proposed clinical study in accordance with 
these regulations when applicable.
The Investigator will agree to make required progress reports to the IEC/IRB, as well as report any 
SAEs, life-threatening problems or deaths if applicable per local regulations. The IEC/IRB will be 
informed of SAEs in other clinical studies conducted with the study drug by the Investigator or the
Sponsor according to the local regulations. The IEC/IRB must be informed by the Investigator of the 
termination of the study.
9.4
Compliance with the Protocol and Protocol Amendments
Investigators ascertain they will apply due diligence to avoid protocol deviations. Under no 
circumstances should the Investigator contact TRIO or its agents monitoring the trial, if any, to 
request approval of a protocol deviation, as no such authorized deviations are permitted. 
If the Investigator feels a protocol deviation would improve the conduct of the study this must be 
considered a protocol amendment, and unless such an amendment is agreed upon by TRIO and 
approved by the IRB/IEC it cannot be implemented. All significant protocol deviations will be 
recorded and reported in the Clinical Study Report (CSR).
Any modifications to the protocol which may impact on the conduct of the study, potential benefit 
of the patient or may affect patient safety, including changes of study objectives, study design, 
patient population, sample sizes, study procedures, or significant administrative aspects will require 
a formal amendment to the protocol. Such amendment will be agreed upon by TRIO, and approved 
by the Ethics Committee/IRB and Health Authorities where required, prior to implementation. Only 
amendments that are required to eliminate an immediate hazard to patients for patient safety can


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be implemented prior to IRB/IEC approval. Notwithstanding the need for approval of formal protocol 
amendments, the Investigator is expected to take any immediate action required for the safety of 
any patient included in this study, even if this action represents a deviation from the protocol. In 
such cases, TRIO should be notified of this action and the IRB/IEC at the study site should be 
informed.
9.5
Monitoring, Auditing and Inspecting
TRIO or its representatives must be allowed to visit all study site locations periodically to assess the 
data, quality, and study integrity. On site, they will notably review study records and directly 
compare them with source documents, review regulatory documents, discuss the conduct of the 
study with the Investigator, verify study drug accountability, and verify that the facilities remain 
acceptable.
9.6
Recording, Processing and Retention of Data
The Investigator is responsible for the preparation and maintenance of adequate case histories 
designed to record all observations and other relevant data. All patient data reported on the eCRF
must be derived from source documents and as such be consistent with the source documents, or 
the discrepancies must be explained.
Data will be entered and collected via an electronic data capture system using eCRFs. RDC is a 
validated system designed for entry of data in a way that is attributable, secure, and accurate, in 
compliance with U.S. FDA regulations for electronic records (21 CFR Part 11). Study site staff will 
receive training and have access to a manual for appropriate eCRF completion. All eCRFs should be 
completed by designated, trained site staff. eCRFs should be reviewed and electronically signed and 
dated by the Investigator or authorized designee. Only identified and trained users may view the 
data as their actions become part of the audit trial.
While study sites will be responsible for data entry, TRIO will be responsible for data management of 
this study, including quality checks.
The eCRF must be completed within 7 calendar days of data availability. All requested information 
must be entered on the eCRF. If an item is not available or is not applicable, it must be documented 
as such. The completed eCRF must be reviewed and approved by the Investigator or authorized 
designee. In the event of discrepant data, TRIO will request data clarification from the sites. The sites 
will resolve the discrepancy electronically in the RDC system. eCRF and data clarification 
documentation will be maintained in the RDC system’s audit trail.
At the end of the study, the Investigator will receive patient data, for their site, in a readable format 
on CD, DVD, or other similar storage format that must be kept with the study records. 
Acknowledgement of receipt of the storage disc or similar is required.
The Investigator should retain the study documentation according to ICH-GCP, local regulations or as 
indicated in the contractual agreement whichever is longer.
9.7
Data Protection
Patient confidentiality is strictly held in trust by the participating Investigators, their staff, TRIO or 
affiliates. This confidentiality is extended to cover testing of biological samples and genetic tests in 
addition to the clinical information relating to participating patients.
Patient and Investigator personal data which may be included in the study database shall be treated 
in compliance with all applicable laws and regulations. 


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When archiving or processing personal data, TRIO shall take all appropriate measures to safeguard 
and prevent access to this data by any unauthorized third party.
The study protocol, documentation, data and all other information generated will be held in strict 
confidence. No information concerning the study or the data will be released to any unauthorized 
third party without prior written approval by TRIO.
The study monitor or TRIO’s other authorized representatives will verify documents and records 
required to be maintained by the Investigator, including but not limited to, medical records (office, 
clinic or hospital) and pharmacy records for the patients in this study. The clinical study site will 
permit access to such records.
9.8
Confidentiality and Data Protection
All information concerning the study supplied by TRIO, in connection with this study and/or by any 
other party collaborating with TRIO, and not previously published, is considered confidential and 
proprietary information. This information includes but is not limited to the IB, clinical protocol, 
workbooks (if applicable), eCRFs, assay methods, TRIO technical methodology, and other technical 
and scientific data. This confidential information shall remain the sole property of TRIO and shall not 
be disclosed to others without prior written consent from TRIO.  Information shall not be used 
except in the performance of this study.
9.9
Insurance of Liabilities
If required, the Investigator may forward to the IEC/IRB a copy of the insurance document required
by TRIO, in order to cover his/her liabilities, and those of any other participating parties.
9.10 Use of Information and Publication
All information concerning the study drug or in connection with this study, supplied by TRIO and/or 
by any other party collaborating with TRIO within this study, and not previously published, is 
considered confidential and proprietary information. This information includes, but is not limited to, 
the IB, clinical protocol, workbooks (if applicable), eCRFs, assay methods, TRIO technical 
methodology, and basic scientific data. This confidential information shall remain the sole property 
of TRIO, the respective collaborating party, and shall not be disclosed to others without prior written 
consent from TRIO.  Information shall not be used except in the performance of this study.
To allow for the use of the information derived from this clinical study and to ensure compliance to 
current regulations, the Investigator is obliged to provide TRIO with complete test results and all 
data developed in this study. No publication, abstract or presentation of the study will be made 
without the approval of the SSC. The SSC will review the manuscript to prevent forfeiture of patent 
rights to data not in the public domain. Prior to publication, the authorship list will be agreed upon 
by the SSC. For the purpose of analyses, the names on the author list will be given according to the 
participation in the concept of the study design as well as accrual input (number of eligible patients 
accrued) by the Investigators at each center. The maximum number of authors will be determined 
by the publication policy established by the targeted journal. Abstracts and publications will be 
submitted to the authors and to the SSC at least 30 days prior to the expected date of submission to 
the intended publisher.
10.REFERENCES
1. Globocan 2012 [Internet][cited 2016 May 5] Available from: http://globocan.iarc.fr/Default.aspx
2. Siegel RL, Miller KD, Jemal A: Cancer statistics, 2016. CA Cancer J Clin 66:7–30, 2016


TRIO030 Protocol 
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3. Chia K, O’Brien M, Brown M, et al: Targeting the androgen receptor in breast cancer. Curr Oncol Rep 17:4, 
2015
4. Rampurwala M, Wisinski KB, O’Regan R: Role of the androgen receptor in triple-negative breast cancer. Clin 
Adv Hematol Oncol HO 14:186–193, 2016
5. Bergh J, Jonsson P-E, Lidbrink EK, et al: FACT: An Open-Label Randomized Phase III Study of Fulvestrant and 
Anastrozole in Combination Compared With Anastrozole Alone As First-Line Therapy for Patients With 
Receptor-Positive Postmenopausal Breast Cancer. J Clin Oncol 30:1919–1925, 2012
6. De Amicis F, Thirugnansampanthan J, Cui Y, et al: Androgen receptor overexpression induces tamoxifen 
resistance in human breast cancer cells. Breast Cancer Res Treat 121:1–11, 2010
7. Cochrane DR, Bernales S, Jacobsen BM, et al: Role of the androgen receptor in breast cancer and preclinical 
analysis of enzalutamide. Breast Cancer Res 16:R7, 2014
8. He J, Peng R, Yuan Z, et al: Prognostic value of androgen receptor expression in operable triple-negative 
breast cancer: a retrospective analysis based on a tissue microarray. Med Oncol 29:406–410, 2012
9. Choi JE, Kang SH, Lee SJ, et al: Androgen receptor expression predicts decreased survival in early stage triple-
negative breast cancer. Ann Surg Oncol 22:82–89, 2015
10. Gucalp A, Tolaney S, Isakoff SJ, et al: Phase II trial of bicalutamide in patients with androgen receptor-
positive, estrogen receptor-negative metastatic Breast Cancer. Clin Cancer Res Off J Am Assoc Cancer Res 
19:5505–5512, 2013
11. Traina TA: Results from a phase 2 study of enzalutamide (ENZA), an androgen receptor (AR) inhibitor, in 
advanced AR+ triple-negative breast cancer (TNBC). [Internet]. J Clin Oncol [cited 2016 Jun 16] Available from: 
http://meetinglibrary.asco.org/content/150040-156
12. Vermeulen M, Slaets L, Cardoso F, et al: Pathologic prognostic factors of male breast cancer: results of the 
EORTC 10085/TBCRC/BIG/NABG International Male Breast Cancer Program [Internet], in 2016[cited 2016 Nov 
22] Available from: http://www.ecco-org.eu/Events/EBCC10/Abstract-search?abstractid=24211
13. Hammond MEH, Hayes DF, Dowsett M, et al: American Society of Clinical Oncology/College of American 
Pathologists Guideline Recommendations for Immunohistochemical Testing of Estrogen and Progesterone 
Receptors in Breast Cancer. J Clin Oncol 28:2784–2795, 2010


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11. APPENDICES
Appendix 1: Eastern Cooperative Oncology Group Performance Status Scale
Grade
Description
0
Fully active, able to carry on all pre-disease performance without restriction.
1
Restricted in physically strenuous activity but ambulatory and able to carry
out work of a light and sedentary nature, e.g., light housework, office work.
2
Ambulatory and capable of all self-care, but unable to carry out any work
activities. Up and about more than 50% of waking hours.
3
Capable of limited self-care, confined to bed or chair more than 50% of
waking hours.
4
Completely disabled. Cannot carry on any self-care. Totally confined to bed
or chair.
5
Dead


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Appendix 2: Patient Diary Template
See next page. The remainder of this page is intentinonally left blank.


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Site Staff Instructions
Record the Patient ID.
Minimum duration of treatment with ODM-201 is 14 days, with a recommended maximum 
duration of 21 days. Treatment must not last more than 35 days.
Record in the below table, the calendar dates (dd/mmm/yyyy, Morning or Evening) 
corresponding to the initial treatment period with ODM-201 1200 mg per day (Sequence 1).
For grade 1-2 treatment-related AEs only: should any treatment interruption / dose 
modification take place (to be agreed on with TRIO Medical Monitor), please record in the 
subsequent row (Sequence 2) the treatment start / stop dates and treatment dose to be taken 
by the patient. Note: for grade ≥ 3 treatment-related AEs, treatment should be permanently 
discontinued.
Depending on the day / time when the patient is provided with the ODM-201 bottle, treatment 
may start with either a Morning or an Evening intake.
The last ODM-201 intake will be on the day prior to breast cancer surgery or before starting 
neoadjuvant treatment if indicated, in the morning of the Pre-surgery visit (except if the patient 
is ordered to stop oral intake earlier – please refer to protocol section 6.4.3 “Pre-surgery visit” 
for guidance).
Please refer to protocol sections 5.3 and 5.4.2 for more details.
Date of ODM-201 bottle dispensation:  _ _ / _ _ _ / _ _ _ _
(dd/mmm/yyyy)
Seq.
Treatment Period
Daily Dose
Study 
Treatment 
container
1
From:
Morning
_ _ / _ _ _ / _ _ _ _
Evening
(dd/mmm/yyyy)
To:
Morning
_ _ / _ _ _ / _ _ _ _
Evening
(dd/mmm/yyyy)
1200 mg
2 tablets twice a day 
(with food, at breakfast and dinner)
Bottle
(containing 
140 tablets of 
ODM-201 300 mg)
2
From:
Morning
_ _ / _ _ _ / _ _ _ _
Evening
(dd/mmm/yyyy)
To:
Morning
_ _ / _ _ _ / _ _ _ _
Evening
(dd/mmm/yyyy)
1200 mg
2 tablets twice a day
(with food, at breakfast and dinner)
Or
600 mg
1 tablet twice a day
(with food, at breakfast and dinner)
Date of Pre-surgery visit:  _ _ / _ _ _ / _ _ _ _
Date of surgery:  _ _ / _ _ _ / _ _ _ 
_
(dd/mmm/yyyy)
(dd/mmm/yyyy)


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Patient Instructions:

Swallow the ODM-201 whole tablets with water and do not chew them prior to swallowing. 

Take the tablets together with food if possible at approximately the same time twice a day 
(for example, with breakfast and dinner each day). 

Your study doctor may decide to reduce or hold your study treatment. Please follow his/her 
instructions carefully.

If you miss an intake or vomit following ODM-201 intake, you should skip the missed/vomited 
intake and you should take the next one as scheduled. You should not make it up.

If you inadvertently take more than the required number of tablets at a given intake, please 
take your next intake as scheduled. Please report this on your patient diary and inform your 
study doctor immediately.

If a tablet is broken or damaged, the tablet should not be used. Inform your study doctor or 
nurse.

If you lose a tablet accidentally (e.g. tablet drops in the sink, falls on the ground etc), please 
report it on your patient diary (in the section “Other comments”) and inform your study 
doctor or nurse.

Keep the tablets in the bottle provided and do not transfer them to any other container.

Record the number of tablets taken at each intake in the diary section below. 

To prevent any dosage error, you should complete your diary prior to taking the study 
treatment to ensure you will take the correct number of tablets. 

ALWAYS bring back the bottle and your completed diary to the clinic at each visit (treatment 
visit if any and Pre-surgery visit).

In case of any question or problem please contact : 

Name of study site staff: _______________________________

Phone #:_____________________________________________


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Patient Diary
DAY
DATE
(dd/mmm/yyyy)
INTAKE
TIME
Hour/Min 
(24:00 clock)
Number of tablets 
TAKEN
Report in the cells below the 
number of tablets taken (e.g. 
2, 1 or 0) at each intake.
COMMENTS
Please use this section to 
write down any side effects 
you are experiencing on a 
given day, explanation for any 
extra tablet taken or missed 
tablet, etc.
Example:
Day X
Example:
15/DEC/2016
Morning intake
8:00
2
Evening intake
19:00
1
Only 1 tab taken, the other one was 
missed in error
Day 1
Morning intake 
Evening intake 
Day 2
Morning intake 
Evening intake 
Day 3
Morning intake 
Evening intake 
Day 4
Morning intake 
Evening intake 
Day 5
Morning intake 
Evening intake 
Day 6
Morning intake 
Evening intake 
Day 7
Morning intake 
Evening intake 
Day 8
Morning intake 
Evening intake 
Day 9
Morning intake 
Evening intake 
Day 10
Morning intake 
Evening intake 
Day 11
Morning intake 
Evening intake 
Day 12
Morning intake 
Evening intake 


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DAY
DATE
(dd/mmm/yyyy)
INTAKE
TIME
Hour/Min 
(24:00 clock)
Number of tablets 
TAKEN
Report in the cells below the 
number of tablets taken (e.g. 
2, 1 or 0) at each intake.
COMMENTS
Please use this section to 
write down any side effects 
you are experiencing on a 
given day, explanation for any 
extra tablet taken or missed 
tablet, etc.
Day 13
Morning intake 
Evening intake 
Day 14
Morning intake 
Evening intake 
Day 15
Morning intake 
Evening intake 
Day 16
Morning intake 
Evening intake 
Day 17
Morning intake 
Evening intake 
Day 18
Morning intake 
Evening intake 
Day 19
Morning intake 
Evening intake 
Day 20
Morning intake 
Evening intake 
Day 21
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 


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DAY
DATE
(dd/mmm/yyyy)
INTAKE
TIME
Hour/Min 
(24:00 clock)
Number of tablets 
TAKEN
Report in the cells below the 
number of tablets taken (e.g. 
2, 1 or 0) at each intake.
COMMENTS
Please use this section to 
write down any side effects 
you are experiencing on a 
given day, explanation for any 
extra tablet taken or missed 
tablet, etc.
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Day __
Morning intake 
Evening intake 
Other comments (e.g.: lost or damaged tablets): 
_________________________________________________________________________
_________________________________________________________________________
______


Signature Manifest
Document Number: TRIO030-MLD-0001
Revision: 03
Title: Clinical Trial Protocol
All dates and times are in Mountain Time.
Clinical Trial Protocol
Approval
Name/Signature
Title
Date
Meaning/Reason
Rodrigo Fresco (RFRESCO)
15 Feb 2018, 12:01:16 PM
Approved
Launa Aspeslet (LASPESLE)
15 Feb 2018, 12:21:43 PM
Approved
Valerie Bee (VBEE)
15 Feb 2018, 12:55:27 PM
Approved
Naouel Belhani (NBELHANI)
16 Feb 2018, 08:53:38 AM
Approved
Carlos Meyer (CMEYER)
20 Feb 2018, 04:30:20 AM
Approved
Set Effective Date
Name/Signature
Title
Date
Meaning/Reason
Carlos Meyer (CMEYER)
20 Feb 2018, 04:39:20 AM
Approved
Reapproval: TRIO030-MLD-0001
Reapproval
Name/Signature
Title
Date
Meaning/Reason
Carlos Meyer (CMEYER)
21 Feb 2018, 04:12:41 AM
Approved
Reapproval: TRIO030-MLD-0001 rev 03
Reapproval
Name/Signature
Title
Date
Meaning/Reason
Carlos Meyer (CMEYER)
26 Feb 2018, 09:41:49 AM
Approved

SAP_001.pdf: 
 
 
 
 
 
Statistical Analysis Plan 
 
 
 
 
Study Title: A Presurgical Tissue-Acquisition Study to Evaluate Molecular Alterations in 
Human Breast Cancer Tissue Following Short-Term Exposure to the Androgen Receptor 
Antagonist Darolutamide (ODM-201) 
 
 
Study Number:  TRIO030 
 
 
Version, Date:  3.0, 30 September 2019 
 
 
ClinicalTrials.gov Identifier:  NCT03004534 
 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 2 of 27
Revision History
The subjective version of the SAP 3.0 is updated to align with the latest protocol revision, and 
update the sample size as a result of the enrollment closure dated 14 February 2019. Briefly, 
only seven patients in the triple negative subgroup have been enrolled; one less than planned
as per protocol.
Table 1 – Revision History
Section #
Type of Change 
Change
3.5.1
Content
Removed 
formula 
‘c’ 
for 
Duration of Treatment (days)
and 
Treatment 
Exposure 
(days)
4.1
Content
Added additional description 
for Laboratory Values and 
ECOG Performance Status
to include total number of 
patients 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 3 of 27
Table of Content
1.
Introduction ........................................................................................................................6
2.
Study Design and Objectives..............................................................................................6
2.1.
Study Design ................................................................................................................6
2.2.
Study Objectives...........................................................................................................6
2.2.1.
Primary Objective ..................................................................................................6
2.2.2.
Secondary Objective..............................................................................................6
2.3.
Sample Size..................................................................................................................6
2.4.
Timing of Analyses........................................................................................................7
3.
Statistical Methods .............................................................................................................7
3.1.
Analysis Population.......................................................................................................7
3.1.1.
Intention-to-Treat Population..................................................................................7
3.1.2.
Safety Population...................................................................................................7
3.1.3.
Evaluable Population .............................................................................................7
3.2.
Data Handling...............................................................................................................8
3.2.1.
Baseline Assessment/Measurement......................................................................8
3.2.2.
Date of First Dose..................................................................................................8
3.2.3.
Date of Last Dose..................................................................................................8
3.2.4.
Duration.................................................................................................................8
3.2.5.
Missing Data..........................................................................................................8
3.2.6.
Partial Dates ..........................................................................................................8
3.3.
Statistical Analysis ........................................................................................................9
3.3.1.
Patient Disposition.................................................................................................9
3.3.2.
Demographics and Disease Characteristics.........................................................10
3.4.
Efficacy Analyses........................................................................................................10
3.5.
Safety Analyses..........................................................................................................10
3.5.1.
Extent of Exposure...............................................................................................10
3.5.2.
Adverse Events (AEs)..........................................................................................11
3.5.3.
Laboratory Values................................................................................................11
3.5.4.
ECOG Performance Status..................................................................................12
3.5.5.
Concomitant Medications.....................................................................................12


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 4 of 27
3.5.6.
Death...................................................................................................................12
4.
List of Planned Statistical Outputs....................................................................................12
4.1.
Planned Tables...........................................................................................................12
4.2.
Planned Listings..........................................................................................................14
4.3.
Planned Graphs..........................................................................................................16
5.
Appendix..........................................................................................................................16
5.1.
Protocol Synopsis – Protocol v3.0...............................................................................16
5.2.
Protocol Synopsis – Protocol v5.0 (Germany).............................................................22


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 5 of 27
List of Abbreviations 
ADSL
Subject-Level Analysis Dataset
ADaM
Analysis Data Model
AE
Adverse Events
CM
Concomitant Medications
CTCAE
Common Terminology Criteria for Adverse Events
EBC
Early Breast Cancer
EoS
End of Study
ITT
Intent-to-treat or Intention-to-treat
ITTFL
Intent-to-treat Population Flag
MedDRA
Medical Dictionary for Regulatory Activities
NCI
National Cancer Institute
PT
Preferred Term
Q1
First Quartile
Q3
Third Quartile
RDI
Relative Dose Intensity
SAE
Serious Adverse Event
SAFFL
Safety Population Flag
SAP
Statistical Analysis Plan
SAS®
Statistical Analysis System Software
SOC
System Organ Class
SSC
Study Steering Committee
TEAE
Treatment Emergent Adverse Event
TRIO
Translational Research in Oncology
WHODD
World Health Organization Drug Dictionary


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 6 of 27
1. Introduction
This Statistical Analysis Plan (SAP) describes the detailed statistical methodology for executing 
the statistical analyses of the Secondary Objective according to TRIO030 protocol version 3.0
and version 5.0 (Germany).
Statistical analysis will be performed by TRIO using SAS® software Version 9.4 or higher.
2. Study Design and Objectives
2.1.Study Design
The protocol synopsis is provided in Appendix 5.1 and Appendix 5.2. Refer to the protocol for 
additional details.
2.2.Study Objectives
2.2.1. Primary Objective
The primary objective for this study is to identify the molecular alterations that occur in human 
Breast Cancer (BC) tissue, following short-term exposure to darolutamide (ODM-201) in female 
subjects with Early Breast Cancer (EBC).
2.2.2. Secondary Objective
To evaluate safety and tolerability following short-term exposure to darolutamide (ODM-201) in 
female patients with EBC.
2.3.Sample Size
Up to 60 patients (a minimum of eight evaluable patients, and up to 20 patients in each BC 
Subtype: triple-negative, HR+/HER2 negative, HER2 positive) will be required for the 
performance of the molecular trials and possible hypothesis generation.
At the time of enrollment closure on 14 February 2019, 36 patients were enrolled with the 
following characteristics per local or central assessment: seven triple-negative, 19 HR+/HER2 
negative, and 10 HER2 positive. Although the triple-negative subgroup did not reach the 
minimum number of eight planned patients as per protocol, it was deemed acceptable for the 
following reasons:

The primary objective of the study is to identify the molecular alterations that occur in
human BC tissue, following short-term exposure to darolutamide in female patients with
EBC. There are no efficacy objectives in this trial.

The number of patients per cohort was not defined after a formal sample size 
calculation, but was a number considered appropriate for the performance of the 
molecular studies according to the recommendations by the central laboratory 
responsible for the molecular analyses.

The central laboratory now considers that seven patients in the triple-negative cohort is 
sufficient to perform the evaluation of the molecular alterations, and to eventually 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 7 of 27
generate the relevant hypothesis in this subgroup. Seven patients is then deemed 
sufficient to adequately characterize the study objectives in this tissue-acquisition 
window of opportunity study.

The Sponsor, along with the Study Chair, have determined that trial objectives have 
been reached in terms of the sample banking needed for biomarker analyses.

In terms of statistical analysis (specifically the safety profile), there is no comparison
between subgroups, so this change will have no effect to the final analysis.
2.4.Timing of Analyses
Safety outputs will be generated for data review performed by the Study Steering Committee 
(SSC) once 20, around 35, and once all patients have been enrolled and undergone the End of 
Study (EoS) as per protocol.
A Final Safety Analysis will be performed when all patients have completed the trial.
3. Statistical Methods
3.1.Analysis Population
3.1.1. Intention-to-Treat Population
The Intention-to-Treat (ITT) Population includes all patients who are enrolled in the trial, 
regardless of whether they actually received investigational medication. This population will be 
identified using the Intent-to-Treat Population Flag (ITTFL) as defined in Subject-Level Analysis 
Dataset (ADSL) according to Analysis Data Model (ADaM).
3.1.2. Safety Population
The Safety Population includes all patients who receive at least one tablet of darolutamide, 
and will be used for the analysis of safety data of the trial. This population set will be identified 
using the Safety Population Flag (SAFFL) as defined in ADSL according to ADaM.
3.1.3. Evaluable Population
The Evaluable Population includes all patients who satisfied the following criteria:

Minimum duration of treatment with darolutamide is 14 days, with a minimum of 10 
consecutive days of treatment prior to the definitive surgery. 

Mandatory tumor tissue is collected at screening and at surgery according to the 
protocol and submitted to the central laboratory. 

Adequacy for molecular assessment of the tumor tissue collected before and after the 
protocol treatment initiation. The adequacy will be evaluated by the central laboratory. 
The evaluable population will be used for the performance of molecular trials and possible 
hypothesis generation. More information regarding Molecular Analyses will be discussed in a 
separate document.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 8 of 27
3.2.Data Handling
3.2.1. Baseline Assessment/Measurement
The last available assessment/measurement of a particular data point on or prior to the latter of 
the date of enrollment and the date of first dose of investigational treatment. If the patient has no 
value as defined above, the baseline value will be missing.
3.2.2. Date of First Dose
The date of first dose (of investigational drug) is derived as the first date when a non-zero and 
non-missing dose of investigational drug is administered. This is also referred to as the start of 
the treatment.
3.2.3. Date of Last Dose
The date of last dose (of investigational drug) is derived as the last date when a non-zero and 
non-missing dose of investigational drug is administered.
3.2.4. Duration
Duration, except for duration of treatment, is calculated as:

Duration (days): (End Date – Start Date + 1).

Duration (weeks): (End Date – Start Date + 1) / 7

Duration (months): (End Date – Start Date + 1) / 30.4375

Duration (years): (End Date – Start Date + 1) / 365.25
3.2.5. Missing Data
All analyses and descriptive summaries will be based on the observed data. Unless otherwise 
specified, missing data will not be imputed or “carried forward.”
3.2.6. Partial Dates
Only partial or missing dates for Adverse Events (AE) and Concomitant Medications (CM) will 
be imputed according to the following:
3.2.6.1. Imputation Rules for Partial or Missing Stop Dates:
If the month and year are present, impute the last day of that month.
If only the year is present, impute December 31 of that year.
If the stop date is entirely missing, assume the event or medication is ongoing.
If a partial or complete stop date is present and the ‘ongoing’ box is checked, then it will be 
assumed that the AE or CM stopped and the stop date will be imputed if partial.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 9 of 27
3.2.6.2. Imputation Rules for Partial or Missing Start Dates
Start Date
Stop Date
Complete: 
yyyymmdd
Partial: yyyymm
Partial: yyyy
Missing
< 1st
dose
≥ 1st
dose
< 1st
dose 
yyyymm
≥ 1st
dose 
yyyymm
< 1st
dose 
yyyy
≥ 1st
dose 
yyyy
Partial: 
yyyymm
= 
1st
dose 
yyyymm
2
1
2
1
n/a
1
1
≠ 
1st
dose 
yyyymm
2
2
2
2
2
Partial: 
yyyy
= 
1st
dose 
yyyy
3
1
3
1
n/a
1
1
≠ 
1st
dose 
yyyy
3
3
3
3
3
Missing
4
1
4
1
4
1
1

1 = Impute the date of first dose

2 = Impute the first of the month

3 = Impute January 1 of the year

4 = Impute January 1 of the stop year
3.3.Statistical Analysis
Categorical variables will be summarized in frequency tables, with the counts and percentage of 
patients in each category. Percentages given in the summary tables will be rounded and thus 
may not always add up to exactly 100 percent. For continuous variables, summary statistics will 
include number of patients, mean, standard deviation, first quartile (Q1), median, third quartile 
(Q3), and minimum and maximum values (range).
EBC Subtype determined by the central assessment will be used for statistical analyses.
3.3.1. Patient Disposition
The patient disposition will be tabulated using the ITT. Frequency count and respective 
percentages of patients who were in each population (i.e. ITT, Safety, and Evaluable), and 
discontinued from treatment and/or trial will be summarized by EBC Subtype. The reason for 
discontinuation (from treatment and/or from trial participation where applicable) will be 
summarized considering the categories specified in the case report forms. Listings of disposition 
information and analysis population will be provided as well.
Protocol deviations, including eligibility deviations of the inclusion/exclusion criteria, will be 
summarized and listed for ITT, by EBC Subtype.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 10 of 27
3.3.2. Demographics and Disease Characteristics
The following patient demographics and disease characteristics will be summarized and listed 
for the ITT and Safety Population by EBC Subtype:
Patient Demographics:

Age, the integer part of [(Date of enrollment – Date of birth) / 365.25)]
o
Note: Since only the year of birth is collected (YYYY), we assume the full date to 
be 31-Dec-YYYY

Race

Ethnicity

Menopausal Status

Weight at Baseline

Height at Baseline

ECOG Status at Baseline
Disease Characteristics:

Time from Initial Diagnosis to Enrollment

Primary Tumor Location

Staging at Initial Diagnosis

Histopathological Type at Initial Diagnosis

Histopathological Grade at Initial Diagnosis

Local Estrogen Receptor Status

Local Progesterone Receptor Status 

Local HER2 Status
Medical History:
The medical history is coded by the most current version of Medical Dictionary for Regulatory 
Activities (MedDRA), and will be summarized for ITT by System Organ Class (SOC) and 
Preferred Term (PT) by EBC Subtype.
3.4.Efficacy Analyses
Not applicable.
3.5.Safety Analyses
Safety analyses will be performed on the safety population.
3.5.1. Extent of Exposure
Duration of Treatment, Treatment Exposure, Cumulative Dose and Relative Dose Intensity 
(RDI) will be summarized by EBC Subtype for the safety population.

Duration of Treatment (days) = sum of duration of each administration period
o
Duration of each administration period = [(End Date - Start Date) + 1]

Treatment Exposure (days) = [(Date of last dose - date of first dose) + 1]


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 11 of 27

Cumulative Dose (mg) = sum of [duration of each administration period × Actual Daily 
Dose] 

Actual Dose Intensity (mg/day) = (Cumulative Dose) ÷ (Treatment Exposure)

Planned Dose Intensity (mg/day) = 1200 mg

RDI (%) = (Actual Dose Intensity ÷ Planned Dose Intensity) × 100 
The RDI will be additionally presented categorized (i.e., number and percentage of patients with 
RDI of < 60%, 60 - < 80%, 80 - < 90%, 90 - < 110%, ≥ 110%).
The number and percentage of patients with dose modifications, along with their respective 
reasons will be summarized.
The compliance is assessed for the entire treatment period and will be calculated as:
Compliance (%) =
Number of Tabs Taken
∑[Duration of each administration period × (Actual Daily Dose ÷ 300)] × 100
3.5.2. Adverse Events (AEs)
AE information will be collected from the time the patient signs the informed consent and 
continues throughout the trial until EoS. After EoS, AE/SAE information will be collected only if 
related to the study treatment.
The reported AE term will be coded using the current version of the MedDRA. The severity of 
AE will be presented as reported by the Investigator based on National Cancer Institute (NCI) 
Common Terminology Criteria for Adverse Events (CTCAE) grades version 4.03.
Treatment-Emergent Adverse Events (TEAEs) is defined as AEs that start on or after the first 
dose of investigational treatment or AEs that start before the first dose of treatment but 
worsened after investigational treatment is initiated.
Incidence of TEAE will be tabulated by SOC, PT for:

All TEAEs

TEAEs by relationship to investigational treatment and maximum severity grade

TEAEs with action taken dose reduced/drug interrupted

TEAEs with action taken drug withdrawn

Serious Adverse Events (SAE) (not Serious TEAEs)

AEs leading to deaths (not Fatal TEAEs) – An AE is considered as leading to death if it 
reported as Grade 5.
In the event a patient experiences repeat episodes of the same AE according to PT, the event 
with the highest severity grade will be used for purposes of incidence tabulations.
Detailed listings for all AEs will be provided, including listing with AEs per cohort. A flag will be 
used to identify which AE is considered as treatment emergent.
3.5.3. Laboratory Values
Lab values will be converted to standard units and categorized according to NCI-CTCAE 
version 4.03. Laboratory results not corresponding to NCI-CTCAE terms will not be graded.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 12 of 27
Hematology and serum blood chemistry will be summarized in descriptive statistics by 
calculating the mean, standard deviation, median and range for the following:

Baseline value

Minimum post baseline value

Maximum post baseline value
Shift tables in laboratory toxicity grades (comparing baseline grade with worst post-baseline 
grade) will be analyzed using standard shift tables, presenting number and proportion of 
patients and their maximum grade shift. 
3.5.4. ECOG Performance Status
Tabulation of ECOG Performance Status results will be done for the following:

Baseline value

Minimum post baseline value

Maximum post baseline value
A shift table will be provided as well from baseline status to the worst status of post-baseline 
time points.
3.5.5. Concomitant Medications
Prior and concomitant medications are coded to the current version of World Health 
Organization Drug Dictionary (WHODD). The Preferred Name will be tabulated and listed by 
patient.
3.5.6. Death
All deaths will be reported in a patient listing, which will include date of death, date of last dose, 
the primary cause of death, and the number of days between the date of last dose of 
investigational drug and death (if applicable).
4. List of Planned Statistical Outputs
4.1.Planned Tables
Title
Population
Description
Patient Disposition 
ITT
Tabulates the disposition of all patients by EBC 
Subtype, including the number of patients in each 
population set as well as the number of patients 
discontinued from investigational treatment, and 
the number of patients discontinued from trial. 
The reason for drug discontinuation and trial
discontinuation will also be summarized.
Protocol Deviations
ITT
Tabulate the number of patients who reported 
protocol deviations by deviation category for each 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 13 of 27
Title
Population
Description
EBC Subtype.
Eligibility Deviations
ITT
Tabulates the number of patients who violates 
inclusion/exclusion criteria by criterion for each 
EBC Subtype.
Patient Demographics 
ITT
Safety
Tabulates summary statistics of the 
demographics (age, race, ethnicity, menopausal 
status, weight, height and ECOG status at 
baseline). 
Disease 
Characteristics 
ITT
Tabulates summary statistics for time from initial 
diagnosis to enrollment, primary tumor location, 
staging at initial diagnosis, histopathological type 
at initial diagnosis, histopathological grade at 
initial diagnosis, hormonal receptor status, and 
HER2 status. 
Medical History
ITT
Summarizes medical history by SOC and PT for 
each EBC Subtype.
Study Drug Exposure 
Safety
Summarizes the duration of treatment, 
cumulative dose, and RDI by EBC Subtype. The 
number of dose modifications will also be 
summarized.
Compliance
Safety
Compliance assessment will be summarized.
TEAEs Overview
Safety
Tabulates the number of patients with TEAEs, 
Related TEAEs, Grade 3 / 4 TEAEs, TEAEs with 
action taken as dose reduced or drug interrupted, 
AEs leading to drug withdrawn, SAE, Fatal AE.
TEAEs
Safety
Summary of number of patients and events by 
SOC and PT for each EBC Subtype.
TEAEs per Maximum 
Grade
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
Related TEAEs
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
Related TEAEs per 
Maximum Grade
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
Grade 3 / 4 TEAEs
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
TEAEs with Action 
Taken as Dose 
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype. 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 14 of 27
Title
Population
Description
Reduced
TEAEs with Action 
Taken as Drug 
Interrupted
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
TEAEs Leading to 
Treatment 
Discontinuation
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
SAE
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
AEs Leading to Death
Safety
Summary by SOC and PT of number of patients 
and events for each EBC Subtype.
Laboratory Values
Safety
Summary of each laboratory parameter at 
baseline, minimum post baseline value, and 
maximum post baseline value for each EBC 
Subtype and the total number of patients. In 
addition, shift tables will be presented.
ECOG Performance 
Status 
Safety
Summary of each laboratory parameter at 
baseline, minimum post baseline value, and 
maximum post baseline value for each EBC 
Subtype and the total number of patients. In 
addition, shift tables will be presented.
Concomitant 
Medications 
Safety
Tabulate the frequency and percentage of 
patients according to Preferred Name. Summary 
will be presented separately for prior medications 
and concomitant medications.
4.2.Planned Listings
Title
Population
Description
Patient Accountability
ITT
Listing of all patients and with population 
indicators, EBC Subtype. For patients who 
discontinued from investigational treatment 
and/or trial, include the reason of respective 
discontinuations.
Demographics
ITT
Listing of demographics.
Disease 
Characteristics
ITT
Listing of disease characteristics.
Local Hormonal 
ITT
Listing of local hormonal receptors and HER2 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 15 of 27
Title
Population
Description
Receptors and HER2 
Status
status.
Deaths
ITT
Listing of patients who died, including primary 
cause of death, date of death, date of last dose, 
and the number of days from date of last dose to 
death (if applicable).
Study Drug Exposure
Safety
Listing of patients for extent of exposure.
Study Drug Exposure 
Summary
Safety
Listing of patients for summary of exposure (e.g. 
duration of treatment, cumulative dose, RDI).
AEs
Safety
Listing of all AE for each patient; the listing 
includes the verbatim term, PT, SOC, 
seriousness, worst grade, start date, outcome, 
stop date, relationship to study drug, action 
taken for study drug, and treatment emergent 
flag. 
Related TEAEs
Safety
Listing of TEAEs in this category using the same 
structure as the AEs listing with the omission of 
treatment emergent flag.
Grade 3 or 4 TEAE
Safety
Listing of TEAEs in this category using the same 
structure as the AEs listing with the omission of 
treatment emergent flag.
TEAEs with Action 
Taken for Dose 
Reduced
Safety
Listing of TEAEs in this category using the same 
structure as the AEs listing with the omission of 
treatment emergent flag.
TEAEs with Action 
Taken for Drug 
Interrupted
Safety
Listing of TEAEs in this category using the same 
structure as the AEs listing with the omission of 
treatment emergent flag. 
TEAEs Leading to 
Study Treatment 
Discontinuation
Safety
Listing of TEAEs in this category using the same 
structure as the AEs listing 
SAEs
Safety
Listing of AEs in this category using the same 
structure as the AEs listing. 
Related SAEs
Safety
Listing of AEs in this category using the same 
structure as the AEs listing.
AEs Leading to Death
Safety
Listing of AEs in this category using the same 
structure as the AEs listing.
Laboratory Values
Safety
Listing of all laboratory values and their assigned 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 16 of 27
Title
Population
Description
toxicity grade (where applicable).
ECOG Performance 
Status
Safety
Listing of ECOG for each reported assessment.
Prior and Concomitant 
Medications
Safety
Listing of all prior and concomitant medications; 
the listing includes verbatim product name, 
generic name, start date, end date (or ongoing) 
and indication.
4.3.Planned Graphs
Not applicable.
5. Appendix
5.1.Protocol Synopsis – Protocol v3.0
Protocol Title
A Presurgical Tissue-Acquisition Study to Evaluate Molecular Alterations in Human Breast 
Cancer Tissue Following Short-Term Exposure to the Androgen Receptor Antagonist 
Darolutamide (ODM-201)
Protocol #
TRIO030
Study Duration
Enrollment is expected to run over a 12-month period approximately. The overall study is
expected to last around 14 months (from First Patient In until Last Patient Last Visit).
Sponsor/Study
Chair
This is a TRIO-sponsored study which will be chaired by Dr. Dennis Slamon (Professor of
Medicine, Chief, Division of Hematology-Oncology, UCLA's Department of Medicine, CA,
USA). The study is financiallysupported by Bayer.
Participating
Investigators/Sites
Approximately20 sites in North America and Europe will participate in the study.
Target Population
Women with early-stage breast cancer (BC) candidates for surgery as primary treatment
modality or candidates for neoadjuvant treatment who accept to undergo a second biopsy
before starting such therapy
Backgroundand
Rationale
Despite progresses made in diagnostic techniques and treatments for BC, there is still a 
significant number of patients undergoing treatment for early BC (EBC) who will relapse or 
patients with advanced BC that do not respond or progress to their treatments. The 
medical need for new and potentially more active and better tolerated agents in BC 
remains very high.
Endocrine treatment is one of the strategies with a major therapeutic value in patients 
with estrogen-receptor (ER) positive tumors. Although, the role of ER and progesterone 
receptor (PgR) is well established in BC as predictive and Despite progresses made in 
diagnostic techniques and treatments for BC, there is still a significant number of patients 
undergoing treatment for early BC (EBC) who will relapse or patients with advanced BC 
that do not respond or progress to their treatments. The medical need for new and 
potentially more active and better tolerated agents in BC remains very high. Endocrine 
treatment is one of the strategies with a major therapeutic value in patients with
estrogen-receptor (ER) positive tumors. Although, the role of ER and progesterone 
receptor (PgR) is well established in BC as predictive and prognostic factors, little is known 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 17 of 27
about the clinical significance of the androgen receptor (AR). Preclinical studies show that 
the androgen signaling pathway plays a critical role in the development of normal and 
malignant breast tissue. AR is expressed in approximately 80% and 60% of primary and 
metastatic BC, respectively. Its expression varies across the different subtypes, 
approximately 84-95% in ER+ tumors, 50-63% in ER-/HER2+ cases and 10-53% in TNBC. 
prognostic factors, little is known about the clinical significance of the androgen receptor 
(AR). Preclinical studies show that the androgen signaling pathway plays a critical role in 
the development of normal and malignant breast tissue. AR is expressed in approximately 
80% and 60% of primary and metastatic BC, respectively. Its expression varies across the 
different subtypes, approximately 84-95% in ER+ tumors, 50-63% in ER-/HER2+ cases and 
10-53% in TNBC.
In ER+ BC it has been suggested that acquired resistance to antiestrogen therapies could
result from adaptation from estrogen dependence to androgen dependence. Recently,
clinical reports of AR antagonism have been promising, especially in TNBC. Bicalutamide 
and enzalutamide have been tested in phase II trials in TNBC and have shown promising 
clinical activity.
These developments in combination with an improved understanding of AR signaling in
different BC subtypes led to the renewed interest in targeting AR. However, the value of 
AR antagonists in the management of BC, the molecular alterations that occur after 
exposure to these agents and whether there are biomarkers useful to predict their 
effectiveness, still need to be fully elucidated. Increased understanding of AR blockade 
and its potential value in different BC subtypes and clinical testing of newer and more 
potent AR antagonists like darolutamide, is thus timely and necessary.
The current study will enroll EBC female patients with differing BC subtypes, with the 
intent of characterizing the molecular alterations in BC tissue before and after a short-
term exposure to darolutamide. Darolutamide is a new AR antagonist, that thus far has 
been found to be very well-tolerated.
Studying the biological mechanisms in which darolutamide targets the AR in BC will be
crucial in understanding its potential role in this disease, as well as providing the 
foundation for the rational development of darolutamide in the disease. Molecular 
profiling of tumor samples before and after darolutamide treatment may permit the 
identification of patients likely or unlikely to respond to the agent based on the biological 
and molecular characteristics of their tumors. Since it is essential to evaluate the 
molecular effects of darolutamide in tumors not exposed to chemotherapy or radiation, 
patients with untreated primary breast tumors amenable to a pre-treatment biopsy 
followed by a post-darolutamide sample at definitive surgery or prior to starting 
neoadjuvant systemic therapy (NAST) represent an ideal population to study. Although no 
direct benefit from the treatment with darolutamide is expected in the participating 
patients, the good tolerability and favourable safety profile of darolutamide at the 
selected dose, the proposed brief duration of treatment, and the fact that patients 
enrolled in the study will not be required to undergo any delay in definitive surgery/start 
of neoadjuvant therapy, should ensure the safety and ethical set-up of the trial.
Objectives
Primary Objective:

To identify the molecular alterations that occur in human BC tissue, following short-
term exposure to ODM-201in female subjects with EBC.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 18 of 27
Secondary Objectives:

To evaluate the safety and tolerability of short-term exposure to darolutamide in
female subjects with EBC.
Study Design
This is a multi-center, open-label, tissue-acquisition study involving up to 60 patients with 
early-stage invasive BC.
Patients with (a) suspicion of invasive BC based on clinical and/or radiological findings, or 
(b) a confirmation of invasive BC based on previous cytology/core/incisional biopsy will be 
invited to participate in the study: (a) In cases without pathological confirmation of BC 
diagnosis, signature of patient informed consent form (PICF) before BC confirmation 
biopsy is preferred to allow the collection of study-specific biopsies to be conducted. (b) 
For patients who have already had a confirmatory biopsy, they may also participate in the 
study if willing to undergo a subsequent study-specific biopsy procedure according to the 
protocol or if their diagnostic biopsy has been done according to the protocol 
requirements (in terms of number of cores/tissue, fresh frozen collection, etc). 
At time of PICF signature, patients will be registered in the study. Primary tumor samples 
will be collected by core needle biopsy or incisional biopsy (excisional biopsy will not be 
allowed). A minimum of three cores of tissue using a 14-gauge needle is recommended (or 
equivalent amount with an incisional biopsy). Two of the cores will be snap-frozen and the 
third one should be prepared in paraffin according to site’s practice. In cases, it is not 
possible to provide enough material, collection of 2 cores (or equivalent amount with 
incisional biopsy) will be acceptable: one core will be snap-frozen and the other one will 
be prepared in paraffin.
To be able to assess the molecular alterations after darolutamide exposure in different BC 
subtypes, patients being either triple-negative, or HR+/HER2 negative, or HER2 positive 
will be enrolled in the study (up to 20 patients with each of these subtypes, with an 
acceptable minimum of 8 evaluable patients in each cohort if enrollment of recommended 
20 patients is not feasible for any reason).
Eligible patients will be enrolled and receive darolutamide at a dose of 600 mg (2 × 300 mg 
tablets) b.i.d. to a daily dose of 1,200 mg. Protocol treatment should start within 5 days of 
enrollment, and within 42 days of the study-specific tumor sample collection. Minimum 
duration of treatment with darolutamide is 14 days with a recommended maximum 
duration of 21 days. It is recommended that the BC surgery/pre-NAST biopsy date is 
defined prior to starting protocol treatment. The start date of darolutamide (Day 1) will be 
derived as minus 14 to 21 days from the scheduled BC surgery/pre-NAST biopsy date. If 
for some reason BC surgery/pre-NAST biopsy takes place more than 21 days after 
treatment start (e.g. scheduling issues, delays, etc.), it is acceptable that patient receives 
darolutamide for more than 21 days and up to a maximum of 35 days. Patient should 
continue protocol treatment until the day prior to BC surgery/pre-NAST biopsy or “nil per 
os” (npo) is ordered. In these cases it is strongly recommended to have BC surgery/pre-
NAST biopsy performed as soon as possible after 21 days of treatment are completed. 
Also, in any case that treatment is extended beyond 21 days, TRIO Medical Monitor 
should be contacted to discuss continuation of treatment and the need for assessments.
At the time of BC surgery (if NAST is not indicated), or before NAST starts if indicated, BC 
tissue will be collected: two cores (or an equivalent amount of tissue), one fresh-frozen 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 19 of 27
and one paraffin-embedded.
End of Study (EoS) visit will occur 30 days (+/- 3 days) after the patient's last intake of 
darolutamide
Eligibility Criteria
Inclusion Criteria
1.
Signed and dated PICF obtained prior to initiation of any study-specific procedure 
and treatment.
2.
Female ≥ 18 years old.
3.
Histologically proven invasive breast carcinoma (through either a core needle 
biopsy or an incisional biopsy) for which surgery is indicated as the primary 
treatment modality. Patients for which NAST is indicated are also eligible provided 
they are willing to undergo a biopsy after completing treatment with darolutamide 
and prior to NAST start.
4.
Known ER, PgR and HER2 statuses.
5.
Tumor must be confined to either the breast or to the breast and ipsilateral axilla 
(Note: patients with multifocal/multicentric tumors are eligible). Patient must have 
(according to TNM 7th edition rules):

T1 with T ≥ 1.0 cm, T2 or T3 by at least one radiographic or clinical 
measurement

Either clinically positive (N1 only) or clinically negative axillary nodes (N0)

M0
6.
Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1.
7.
Adequate organ function within 28 days prior to enrollment, as defined by the
following criteria:

Hematology: Hemoglobin ≥ 9.0 g/dL, Absolute neutrophil count (ANC) ≥ 1.5 × 
109/L, Platelet count ≥ 100 × 109/L

Liver function: Alanine aminotransferase (ALT) and aspartate transaminase 
(AST) ≤2.5 × upper limit of normal (ULN), Total bilirubin ≤ 1.5 × ULN (or  3 
times ULN for patients with documented Gilbert’s syndrome or for whom 
indirect bilirubin concentrations suggest an extra-hepatic source of elevation).

Renal function: Creatinine ≤ 2.0 × ULN
8.
No more than 42 days should elapse from the day the study-specific tumor sample 
is taken at initial diagnosis (or subsequent procedure) to the day of the first intake 
of darolutamide.
9.
Women of childbearing potential (WoCBP) must agree to use acceptable non-
hormonal contraceptive methods of birth control from the day of the screening 
pregnancy test and up to 3 months after the last intake of darolutamide.
10. For WoCBP negative serum pregnancy test within 7 days of enrollment.
11. Willingness and ability to comply with scheduled visits, treatment plans, laboratory
tests, and biopsies as detailed in the protocol.
Exclusion Criteria 
1.
Any T0, Tis, T1 < 1.0 cm, T4; or N2-3; or M1 BC.
2.
Bilateral invasive BC.
3.
Patient that underwent excisional biopsy of the primary tumor.
4.
Medical indication or patient desire to undergo BC surgery or start NAST prior to 
completing at least 14 days of treatment with darolutamide, and/or refusal of 
patient to undergo corresponding biopsy in case NAST is planned.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 20 of 27
5.
Prior or concurrent systemic anticancer therapy for BC treatment (immunotherapy, 
biologic/targeted therapy, chemotherapy, investigational agents).
6.
Prior or concurrent ipsilateral radiation therapy for invasive or noninvasive BC.
7.
Prior or concurrent treatment or preventative use of any hormonal agent such as 
aromatase inhibitors (AI), fulvestrant, raloxifene, tamoxifen or other selective 
estrogen receptor modulators (SERM), or with any other hormonal agent used for 
the treatment or prevention of BC or for any other indication (e.g. osteoporosis).
8.
Concurrent use of ovarian hormone replacement therapy. Prior treatment should 
be stopped at least 28 days prior to registration.
9.
Prior or concurrent treatment with AR antagonists or CYP17 enzyme inhibitor.
10. Use of other investigational drug within 28 days of enrollment.
11. Major surgery within 28 days before enrollment
12. basal or squamous skin cancer, or carcinoma in situ of the cervix, or other 
noninvasive/in-situ neoplasm, all of which must have been adequately and radically 
treated. A patient with previous history of invasive malignancy (other than 
adequately and radically treated basal or squamous skin cancer) is eligible provided 
that she has been disease free for more than 5 years.
13. Severe or uncontrolled concurrent disease, infection or comorbidity.
14. Known active viral hepatitis, human immunodeficiency virus (HIV) or chronic liver 
disease.
15. Other serious illness or medical condition within 6 months before enrollment, 
including any of the following: Concurrent congestive heart failure New York Heart 
Association (NYHA) Class III or IV, severe/unstable angina pectoris, myocardial 
infarction, uncontrolled hypertension, coronary/peripheral artery bypass graft, 
high-risk uncontrolled arrhythmias, stroke.
16. Any contraindication to oral agents or gastrointestinal disorder or procedure which 
expects to interfere significantly with absorption of protocol treatment.
17. History or current evidence of any condition, therapy, or laboratory abnormality 
that might confound the results of the trial, interfere with the patient’s 
participation for the full duration of the trial, or is not in the best interest of the 
patient to participate, in the opinion of the treating Investigator.
18. Known psychiatric or substance abuse disorders that would interfere with 
cooperation with the requirements of the trial.
19. Known allergy to darolutamide or any of the excipients.
20. Pregnant or lactating patient
Protocol Treatment
Darolutamide will be given at a dose of 600 mg (2 x 300 mg tablets) b.i.d. to a daily dose of 
1200 mg.
The following timelines will be considered for darolutamide administration:

Darolutamide shall be administered starting within 5 days of enrollment.

No more than 42 days should elapse from the day the study-specific tumor sample 
is taken at initial diagnosis (or in a subsequent procedure) to the day of the first 
intake of darolutamide.

Minimum duration of treatment will be 14 days prior to BC surgery or to the pre-
NAST biopsy (if NAST is indicated).

A maximum duration of 21 days of treatment is recommended. . It is 
recommended that the date of BC surgery/pre-NAST biopsy is defined prior to 
starting protocol treatment. The start date of darolutamide (Day 1) will be derived 
as minus 14 to 21 days from the scheduled BC surgery/pre-NAST biopsy date.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 21 of 27

Treatment will be given until the day prior to the BC surgery, or the day before pre-
NAST biopsy or when npo is ordered, whichever occurs first.

If for some reason BC surgery/pre-NAST biopsy takes place more than 21 days after 
treatment start, it is acceptable that patient receives darolutamide for more than 
21 days and up to a maxiumum of 35 days; patient should continue protocol 
treatment until the day prior to BC surgery/pre-NAST biopsy or npo is ordered. In 
these cases it is strongly recommended to have BC surgery/pre-NAST biopsy done 
as soon as possible after 21 days of treatment are completed. Also, in any case that 
treatment is extended beyond 21 days, TRIO Medical Monitor should be contacted 
to discuss continuation of treatment and the need for assessments
Visits and
Assessments
Activity
Screening
Treatment
Period
Pre-
Surgery
BC 
surgery/pr
e-NAST 
biopsy
End of
Study
Informed 
consent/Registration
and Enrollment
X
Demographics,
medical, 
surgical,
disease history
X
Complete
physical 
examination
X
X
Symptom-directed
physical examination
X
X
ECOG PS
X
X
X
X
Weight, height, blood 
pressure
X
Serum pregnancy test
X
Hematology,
Blood 
chemistry
X
X
X
Treatment
compliance 
assessment
X
X
AEs assessments
X
X
X
X
Concomitant
medication
X
X
X
X
Tumor sample
X
X
Statistical Methods
Up to 60 patients (a minimum of 8 evaluable patients and up to 20 patients in each breast 
cancer subtype: triple-negative, ER+/HER2 negative, HER2 positive) will be required for the 
performance of the molecular studies and possible hypothesis generation.
A patient will be considered evaluable if the following criteria are met:

Minimum duration of treatment with darolutamide is of 14 days, with a minimum of 
10 consecutive days of treatment prior to the definitive surgery.

Mandatory tumor tissue is collected at screening and at surgery according to the 
protocol and submitted to the central lab.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 22 of 27

Adequacy for molecular assessment of the tumor tissue collected before and after the 
protocol treatment initiation. The adequacy will be evaluated by the central lab.
Assessment of patient evaluability, safety and distribution of patients with respect to hormone 
receptor status and HER2 status will be performed by the Study Steering Committee (SSC) at 
given time points depending on the recruitment, primarily planned once 20, around 35 and once 
all patients have been enrolled and undergone the EoS per protocol. This will allow for the SSC 
to ascertain that the appropriate number of evaluable patients is being accrued, that more or 
less patients may be required, or in the extreme case, that accrual should close if the patient 
evaluability rate is very low and/or safety concerns arise.
The SSC will define the evaluability of patients that discontinue treatment more than 3 days 
before surgery: molecular analyses may be taken into consideration for the decision about the 
evaulabilty of such patients.
Study Populations

Intention-to-treat (ITT) Population: the ITT population includes all patients who are 
enrolled in the study regardless of whether they have been treated or not.

Safety Population: the Safety population includes all subjects who have taken at least one 
tablet of darolutamide.

Evaluable population: includes all patients who are considered evaluable per protocol.
5.2.Protocol Synopsis – Protocol v5.0 (Germany)
Protocol Title
A Presurgical Tissue-Acquisition Study to Evaluate Molecular Alterations in Human Breast 
Cancer Tissue Following Short-Term Exposure to the Androgen Receptor Antagonist 
Darolutamide (ODM-201)
Protocol #
TRIO030
Study Duration
Enrollment is expected to run over a 12-month period approximately. The overall study is 
expected to last around 14 months (from First Patient In until Last Patient Last Visit).
Sponsor/Study
Chair
This is a TRIO-sponsored study which will be chaired by Dr. Dennis Slamon (Professor of 
Medicine, Chief, Division of Hematology-Oncology, UCLA's Department of Medicine, CA, USA). 
The study is financially supported by Bayer.
Participating
Investigators/Sites
Approximately20 sites in North America and Europe will participate in the study.
Target Population
Women with early-stage breast cancer (BC) candidates for surgery as primary treatment modality 
or candidates for neoadjuvant treatment who accept to undergo a second biopsy before starting 
such therapy
Backgroundand
Rationale
Despite progresses made in diagnostic techniques and treatments for BC, there is still a 
significant number of patients undergoing treatment for early BC (EBC) who will relapse or 
patients with advanced BC that do not respond or progress to their treatments. The medical 
need for new and potentially more active and better tolerated agents in BC remains very high.
Endocrine treatment is one of the strategies with a major therapeutic value in patients with 
estrogen-receptor (ER) positive tumors. Although, the role of ER and progesterone receptor 
(PgR) is well established in BC as predictive and prognostic factors, little is known about the 
clinical significance of the androgen receptor (AR). Preclinical studies show that the androgen 
signaling pathway plays a critical role in the development of normal and malignant breast 
tissue. AR is expressed in approximately 80% and 60% of primary and metastatic BC, 
respectively. Its expression varies across the different subtypes, approximately 84-95% in ER+ 
tumors, 50-63% in ER-/HER2+ cases and 10-53% in TNBC.
In ER+ BC it has been suggested that acquired resistance to antiestrogen therapies could result 
from adaptation from estrogen dependence to androgen dependence. Recently, clinical 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 23 of 27
reports of AR antagonism have been promising, especially in TNBC. Bicalutamide and 
enzalutamide have been tested in phase II trials in TNBC and have shown promising clinical 
activity.
These developments in combination with an improved understanding of AR signaling in 
different BC subtypes led to the renewed interest in targeting AR. However, the value of AR 
antagonists in the management of BC, the molecular alterations that occur after exposure to 
these agents and whether there are biomarkers useful to predict their effectiveness, still need 
to be fully elucidated. Increased understanding of AR blockade and its potential value in 
different BC subtypes and clinical testing of newer and more potent AR antagonists like 
darolutamide, is thus timely and necessary.
The current study will enroll EBC female patients with differing BC subtypes, with the intent of 
characterizing the molecular alterations in BC tissue before and after a short-term exposure to 
darolutamide. Darolutamide is a new AR antagonist, that thus far has been found to be very 
well-tolerated.
Studying the biological mechanisms in which darolutamide targets the AR in BC will be crucial 
in understanding its potential role in this disease, as well as providing the foundation for the 
rational development of darolutamide in the disease. Molecular profiling of tumor samples 
before and after darolutamide treatment may permit the identification of patients likely or 
unlikely to respond to the agent based on the biological and molecular characteristics of their 
tumors.
Since it is essential to evaluate the molecular effects of darolutamide in tumors not exposed to 
chemotherapy or radiation, patients with untreated primary breast tumors amenable to pre-
treatment biopsy followed by a post-darolutamide sample at definitive surgery or prior to 
starting neoadjuvant systemic therapy (NAST) represent an ideal population to study. Although 
no direct benefit from the treatment with darolutamide is expected in the participating 
patients, the good tolerability and favourable safety profile of darolutamide at the selected 
dose, the proposed brief duration of treatment, and the fact that patients enrolled in the study 
will not be required to undergo any delay in definitive surgery/start of neoadjuvant therapy, 
should ensure the safety and ethical set-up of the trial
Objectives
Primary Objective:

To identify the molecular alterations that occur in human BC tissue, following short-term 
exposure to darolutamide in female patients with EBC.
Secondary Objectives:

To evaluate the safety and tolerability of short-term exposure to darolutamide in female 
patients with EBC.
Study Design
This is a multi-center, open-label, tissue-acquisition study involving up to 60 patients with early-
stage invasive BC.
Patients with (a) suspicion of invasive BC based on clinical and/or radiological findings, or (b) a 
confirmation of invasive BC based on previous cytology/core/incisional biopsy will be invited to 
participate in the study: (a) In cases without pathological confirmation of BC diagnosis, signature 
of patient informed consent form (PICF) before BC confirmation biopsy is preferred to allow the 
collection of study-specific biopsies to be conducted.
(b) For patients who have already had a confirmatory biopsy, they may also participate in the 
study if willing to undergo a subsequent study-specific biopsy procedure according to the 
protocol or if their diagnostic biopsy has been done according to the protocol requirements (in 
terms of number of cores/tissue, fresh frozen collection, etc).
At time of PICF signature, patients will be registered in the study. Primary tumor samples will be 
collected by core needle biopsy or incisional biopsy (excisional biopsy will not be allowed). A 
minimum of three cores of tissue using a 14-gauge needle is recommended (or equivalent 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 24 of 27
amount with an incisional biopsy). Two of the cores will be snap-frozen and the third one should 
be prepared in paraffin according to site’s practice. In cases, it is not possible to provide enough 
material, collection of 2 cores (or equivalent amount with incisional biopsy) will be acceptable: 
one core will be snap-frozen and the other one will be prepared in paraffin.
To be able to assess the molecular alterations after darolutamide exposure in different BC 
subtypes, patients being either triple-negative, or HR+/HER2 negative, or HER2 positive will be 
enrolled in the study (up to 20 patients with each of these subtypes, with an acceptable minimum 
of 8 evaluable patients in each cohort if enrollment of recommended 20 patients is not feasible 
for any reason).
Eligible patients will be enrolled and receive darolutamide at a dose of 600 mg (2 × 300 mg 
tablets) b.i.d. to a daily dose of 1,200 mg. Protocol treatment should start within 7 days of 
confirmation of BC diagnosis (i.e. availability of pathological report). Minimum and expected 
treatment duration should be 14 days and ODM-201 should continue until the day prior to BC 
surgery/pre-NAST biopsy or when the subject is ordered to stop all oral intake, whichever occurs 
first. In any case, surgery must be planned within 21 days of confirmation of BC diagnosis.
At the time of surgery (if NAST is not indicated), or before NAST starts if indicated, BC tissue will 
be collected: two cores (or an equivalent amount of tissue), one fresh-frozen and one paraffin-
embedded.
End of Study (EoS) visit will occur 30 days (+/- 3 days) after the patient's last intake of 
darolutamide.
Note: If the BC surgery/pre-NAST biopsy that is initially planned to take place within 21 days 
after BC diagnosis is delayed due to unforeseen circumstances (e.g. scheduling issues, delays, 
etc.) it is acceptable that the subject continues receiving ODM-201 up to a maximum of 35 days. 
However, in this case Investigator should contact the TRIO Medical Monitor to discuss the 
continuation of treatment and the need for assessments.
Eligibility Criteria
Inclusion Criteria
1.
Signed and dated PICF obtained prior to initiation of any study-specific procedure and 
treatment.
2.
Female ≥ 18 years old.
3.
Histologically proven invasive breast carcinoma (through either a core needle biopsy or 
an incisional biopsy) for which surgery is indicated as the primary treatment modality. 
Patients for which NAST is indicated are also eligible provided they are willing to undergo 
a biopsy after completing treatment with darolutamide and prior to NAST start.
4.
Known ER, PgR and HER2 statuses.
5.
Tumor must be confined to either the breast or to the breast and ipsilateral axilla (Note: 
patients with multifocal/multicentric tumors are eligible). Patient must have (according to 
TNM 7th edition rules):

T1 with T ≥ 1.0 cm, T2 or T3 by at least one radiographic or clinical measurement

Either clinically positive (N1 only) or clinically negative axillary nodes (N0)

M0
6.
Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1.
7.
Adequate organ function within 28 days prior to enrollment, as defined by the following 
criteria:

Hematology: Hemoglobin ≥ 9.0 g/dL, Absolute neutrophil count (ANC) ≥ 1.5 × 109/L, 
Platelet count ≥ 100 × 109/L

Liver function: Alanine aminotransferase (ALT) and aspartate transaminase (AST) ≤ 
2.5 × upper limit of normal (ULN), Total bilirubin ≤ 1.5 × ULN (or  3 times ULN for 
patients with documented Gilbert’s syndrome or for whom indirect bilirubin 
concentrations suggest an extra-hepatic source of elevation).

Renal function: Creatinine ≤ 2.0 × ULN
8.
No more than 7 days should elapse from the confirmation of BC diagnosis (i.e. availability 


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 25 of 27
of pathological report) to the day of the first intake of darolutamide.
9.
Women of childbearing potential (WoCBP) must agree to use acceptable non-hormonal 
contraceptive methods of birth control from the day of the screening pregnancy test and 
up to 3 months after the last intake of darolutamide.
10. For WoCBP negative serum pregnancy test within 7 days of enrollment.
11. Willingness and ability to comply with scheduled visits, treatment plans, laboratory tests, 
and biopsies as detailed in the protocol
Exclusion Criteria
1.
Any T0, Tis, T1 < 1.0 cm, T4; or N2-3; or M1 BC.
2.
Bilateral invasive BC.
3.
Patient that underwent excisional biopsy of the primary tumor.
4.
Medical indication or patient desire to undergo BC surgery or start NAST prior to 
completing at least 14 days of treatment with darolutamide, and/or refusal of patient to 
undergo corresponding biopsy in case NAST is planned.
5.
Prior or concurrent systemic anticancer therapy for BC treatment (immunotherapy, 
biologic/targeted therapy, chemotherapy, investigational agents).
6.
Prior or concurrent ipsilateral radiation therapy for invasive or noninvasive BC.
7.
Prior or concurrent treatment or preventative use of any hormonal agent such as 
aromatase inhibitors (AI), fulvestrant, raloxifene, tamoxifen or other selective estrogen 
receptor modulators (SERM), or with any other hormonal agent used for the treatment or 
prevention of BC or for any other indication (e.g. osteoporosis).
8.
Concurrent use of ovarian hormone replacement therapy. Prior treatment should be 
stopped at least 28 days prior to registration.
9.
Prior or concurrent treatment with AR antagonists or CYP17 enzyme inhibitor.
10. Use of other investigational drug within 28 days of enrollment.
11. Major surgery within 28 days before enrollment.
12. Any concurrent or previous malignancy within 5 years prior to enrollment except for 
basal or squamous skin cancer, or carcinoma in situ of the cervix, or other non-
invasive/in-situ neoplasm, all of which must have been adequately and radically treated. 
A patient with previous history of invasive malignancy (other than adequately and 
radically treated basal or squamous skin cancer) is eligible provided that she has been 
disease free for more than 5 years.
13. Severe or uncontrolled concurrent disease, infection or comorbidity.
14. Known active viral hepatitis, human immunodeficiency virus (HIV) or chronic liver disease.
15. Other serious illness or medical condition within 6 months before enrollment, including 
any of the following: Concurrent congestive heart failure New York Heart Association 
(NYHA) Class III or IV, severe/unstable angina pectoris, myocardial infarction, 
uncontrolled 
hypertension, 
coronary/peripheral 
artery 
bypass 
graft, 
high-risk 
uncontrolled arrhythmias, stroke.
16. Any contraindication to oral agents or gastrointestinal disorder or procedure which 
expects to interfere significantly with absorption of protocol treatment.
17. History or current evidence of any condition, therapy, or laboratory abnormality that 
might confound the results of the trial, interfere with the patient’s participation for the 
full duration of the trial, or is not in the best interest of the patient to participate, in the 
opinion of the treating Investigator.
18. Known psychiatric or substance abuse disorders that would interfere with cooperation 
with the requirements of the trial.
19. Known allergy to darolutamide or any of the excipients.
20. Pregnant or lactating patients
Protocol Treatment
Darolutamide will be given at a dose of 600 mg (2 × 300 mg tablets) b.i.d. corresponding to a
daily dose of 1,200 mg.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 26 of 27
The following timelines will be considered for darolutamide administration:

No more than 7 days should elapse from confirmation of BC diagnosis (i.e. availability of 
pathological report) to the day of the first intake of darolutamide.

Minimum and expected duration of treatment will be 14 days prior to BC surgery or to 
the pre-NAST biopsy (if NAST is indicated).

It is recommended that the date of BC surgery/pre-NAST biopsy is defined prior to 
starting protocol treatment. The start date of darolutamide (Day 1) will be derived as 
minus 14 days from the scheduled BC surgery/pre-NAST biopsy date. In any case, BC 
surgery/pre-NAST biopsy must be planned within 21 days of confirmation of BC 
diagnosis.

Treatment will be given until the day prior to the BC surgery, or the day before pre-NAST 
biopsy or when npo is ordered, whichever occurs first
Visits and
Assessments
Activity
Screening
Treatment
Period
Pre-
Surgery
BC 
surgery/pre-
NAST biopsy
End of
Study
Informed 
consent/Registration and 
Enrollment
X
No visits 
expected
Demographics, medical, 
surgical, disease history
X
Complete physical 
examination
X
X
Symptom-directed physical 
examination
X
ECOG PS
X
X
X
Weight, height, blood 
pressure
X
Serum pregnancy test
X
Hematology, Blood 
chemistry
X
X
X
Treatment compliance 
assessment
X
AEs assessments
X
X
X
Concomitantmedication
X
X
X
Tumor sample
X
X
Statistical Methods
Up to 60 patients (a minimum of 8 evaluable patients and up to 20 patients in each breast 
cancer subtype: triple-negative, HR+/HER2 negative, HER2 positive) will be required for the 
performance of the molecular studies and possible hypothesis generation.
A patient will be considered evaluable if the following criteria are met:

Minimum duration of treatment with darolutamide is of 14 days, with a minimum of 10 
consecutive days of treatment prior to the definitive BC surgery/pre-NAST biopsy.

Mandatory tumor tissue is collected at screening and at BC surgery/pre-NAST biopsy 
according to the protocol and submitted to the central lab.


Document prepared in accordance with:
Procedure Number:
SOP 8.1
Procedure Title:
Statistical Analysis Plan
Revision Number:
04
Originated by:
BTHIBODE
Revision:
03
Effective Date:
30 Sep 2019
Doc Number:
TRIO030-DM-0025
Title: 
Statistical Analysis Plan
Translational Research In Oncology - Confidential and Proprietary
Page 27 of 27

Adequacy for molecular assessment of the tumor tissue collected before and after the 
protocol treatment initiation. The adequacy will be evaluated by the central lab.
Assessment of patient evaluability, safety and distribution of patients with respect to hormone 
receptor status and HER2 status will be performed by the Study Steering Committee (SSC) at 
given time points depending on the recruitment, primarily planned once 20, around 35 and once 
all patients have been enrolled and undergone the EoS per protocol. This will allow for the SSC 
to ascertain that the appropriate number of evaluable patients is being accrued, that more or 
less patients may be required, or in the extreme case, that accrual should close if the patient 
evaluability rate is very low and/or safety concerns arise.
The SSC will define the evaluability of patients that discontinue treatment more than 3 days 
before surgery: molecular analyses may be taken into consideration for the decision about the 
evaluability of such patients.
Study Populations

Intention-to-treat (ITT) Population: the ITT population includes all patients who are 
enrolled in the study regardless of whether they have been treated or not.

Safety Population: the Safety population includes all patients who have taken at least 
one tablet of darolutamide.

Evaluable population: includes all patients who are considered evaluable per protocol.


Signature Manifest
Document Number: TRIO030-DM-0025
Revision: 03
Title: Statistical Analysis Plan
All dates and times are in Mountain Time.
TRIO030 SAP & Mock Shells
Approval
Name/Signature
Title
Date
Meaning/Reason
Yan Yang (YYANG)
24 Sep 2019, 02:37:36 PM
Approved
Tamara Irvin (TIRVIN)
24 Sep 2019, 02:58:11 PM
Approved
Pablo Millan (PMILLAN)
25 Sep 2019, 08:28:26 AM
Approved
Ana Popovic (APOPOVIC)
25 Sep 2019, 10:31:26 AM
Approved
Melissa Burton (MJOHNSON)
26 Sep 2019, 09:45:06 AM
Approved
Brandy Thibodeau (BTHIBODE)
26 Sep 2019, 10:17:54 AM
Approved
Stephanie Carrez (SCARREZ)
27 Sep 2019, 02:28:30 AM
Approved
Karen Afenjar (KAFENJAR)
30 Sep 2019, 07:53:27 AM
Approved
Set Effective Date
Name/Signature
Title
Date
Meaning/Reason
Brandy Thibodeau (BTHIBODE)
30 Sep 2019, 08:01:30 AM
Approved