A Phase 1 Trial Combining WEE1 Inhibitor Adavosertib (AZD1775) With Radiation Therapy for Metastatic or Inoperable and Ineligible for Definitive Chemoradiation Esophageal and Gastroesophageal Junction Cancer
Phase: PHASE1
Conditions: Clinical Stage III Esophageal Adenocarcinoma AJCC v8, Clinical Stage III Esophageal Squamous Cell Carcinoma AJCC v8, Clinical Stage III Gastroesophageal Junction Adenocarcinoma AJCC v8, Clinical Stage IV Esophageal Adenocarcinoma AJCC v8, Clinical Stage IV Esophageal Squamous Cell Carcinoma AJCC v8, Clinical Stage IV Gastroesophageal Junction Adenocarcinoma AJCC v8, Clinical Stage IVA Esophageal Adenocarcinoma AJCC v8, Clinical Stage IVA Esophageal Squamous Cell Carcinoma AJCC v8, Clinical Stage IVA Gastroesophageal Junction Adenocarcinoma AJCC v8, Clinical Stage IVB Esophageal Adenocarcinoma AJCC v8, Clinical Stage IVB Esophageal Squamous Cell Carcinoma AJCC v8, Clinical Stage IVB Gastroesophageal Junction Adenocarcinoma AJCC v8, Distal Esophagus Adenocarcinoma, Gastric Cardia Adenocarcinoma, Metastatic Esophageal Adenocarcinoma, Metastatic Esophageal Squamous Cell Carcinoma, Metastatic Gastroesophageal Junction Adenocarcinoma, Metastatic Malignant Neoplasm in the Brain, Metastatic Malignant Neoplasm in the Leptomeninges, Pathologic Stage III Esophageal Adenocarcinoma AJCC v8, Pathologic Stage III Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage III Gastroesophageal Junction Adenocarcinoma AJCC v8, Pathologic Stage IIIA Esophageal Adenocarcinoma AJCC v8, Pathologic Stage IIIA Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage IIIA Gastroesophageal Junction Adenocarcinoma AJCC v8, Pathologic Stage IIIB Esophageal Adenocarcinoma AJCC v8, Pathologic Stage IIIB Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage IIIB Gastroesophageal Junction Adenocarcinoma AJCC v8, Pathologic Stage IV Esophageal Adenocarcinoma AJCC v8, Pathologic Stage IV Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage IV Gastroesophageal Junction Adenocarcinoma AJCC v8, Pathologic Stage IVA Esophageal Adenocarcinoma AJCC v8, Pathologic Stage IVA Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage IVA Gastroesophageal Junction Adenocarcinoma AJCC v8, Pathologic Stage IVB Esophageal Adenocarcinoma AJCC v8, Pathologic Stage IVB Esophageal Squamous Cell Carcinoma AJCC v8, Pathologic Stage IVB Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage III Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage III Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage III Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIA Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIA Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIA Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIB Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIB Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage IIIB Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IV Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IV Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage IV Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IVA Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IVA Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage IVA Gastroesophageal Junction Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IVB Esophageal Adenocarcinoma AJCC v8, Postneoadjuvant Therapy Stage IVB Esophageal Squamous Cell Carcinoma AJCC v8, Postneoadjuvant Therapy Stage IVB Gastroesophageal Junction Adenocarcinoma AJCC v8, Unresectable Esophageal Adenocarcinoma, Unresectable Esophageal Carcinoma, Unresectable Esophageal Squamous Cell Carcinoma, Unresectable Gastroesophageal Junction Adenocarcinoma
Sponsor: National Cancer Institute (NCI)

NCI Protocol #:10389 
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1 
NCI Protocol #: 
10389 
 
Local Protocol #: 
TBD 
 
ClinicalTrials.gov Identifier:  NCT04460937 
 
TITLE:  A Phase 1 Trial Combining WEE1 Inhibitor Adavosertib (AZD1775) with Radiation 
Therapy for Metastatic or Inoperable and Ineligible for Definitive Chemoradiation Esophageal 
and Gastroesophageal Junction Cancer 
 
 
Corresponding Organization: LAO-OH007 / Ohio State University Comprehensive Cancer 
Center LAO 
 
Principal Investigator: 
Eric D. Miller, M.D., Ph.D. 
Ohio State University Comprehensive Cancer Center 
460 W 10th Ave 
Room A209 
Columbus, OH 43210 
614-685-4922 
614-293-4044 
Eric.Miller@osumc.edu 
 
Translational PI: 
Terence M. Williams, M.D., Ph.D. 
City of Hope Comprehensive Cancer Center 
1500 E. Duarte Rd 
Radiation Oncology 
Duarte, CA 91010 
646-456-6282 
626-218-5334 
terwilliams@coh.org 
 
 
Participating Organizations  
LAO-11030 / University Health Network Princess Margaret Cancer Center LAO 
LAO-CA043 / City of Hope Comprehensive Cancer Center LAO 
LAO-CT018 / Yale University Cancer Center LAO 
LAO-MA036 / Dana-Farber - Harvard Cancer Center LAO 
LAO-MD017 / JHU Sidney Kimmel Comprehensive Cancer Center LAO 
LAO-OH007 / Ohio State University Comprehensive Cancer Center LAO 
LAO-PA015 / University of Pittsburgh Cancer Institute LAO 
LAO-TX035 / University of Texas MD Anderson Cancer Center LAO 
LAO-NCI / National Cancer Institute LAO 

NCI Protocol #:10389 
Version Date: January 13, 2022 
 
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Gastroenterologist:  
 
Statistician: 
 
Sajid Jalil MBBS, MS, 
Lai Wei, Ph.D., M.S. 
 
OSU Comprehensive Cancer Center 
1800 Cannon Drive, 320H Lincoln Tower 
 
410 W 10th Ave 
Columbus, OH 43210 
 
Columbus, OH 43210 
614-293-6449 
 
614-293-6255 
614-293-6902 
 
614-293-8518 
Lai.Wei@osumc.edu 
      sajid.jalil@osumc.edu 
 
 
Study Coordinator:  
 
Responsible Research Nurse:  
 
OSU Multi-Center Trial Program 
OSU Multi-Center Trial Program 
 
OSU Comprehensive Cancer Center 
OSU Comprehensive Cancer Center 
 
Fax:  614-366-6652 
Fax:  614-366-6652 
 
OSUCCC-CTO-MCTP@osumc.edu 
OSUCCC-CTO-MCTP@osumc.edu 
 
 
 
Responsible Data Manager:  
OSU Multi-Center Trial Program 
Clinical Trials Office   
OSU Comprehensive Cancer CenterFax:  614-366-6652   
OSUCCC-CTO-MCTP@osumc.edu 
 
NCI-Supplied Agent:  AZD1775 (adavosertib, NSC 751084) 
 
IND #:  122697 
IND Sponsor:  DCTD, NCI 
 
 
Protocol Type / Version # / Version Date: 
Original / March 23, 2020 
 
 
 
 
Revision 01 / May 7, 2020 
 
 
 
 
Revision 02 / June 8, 2020 
 
 
 
 
Revision 03a / August 3, 2020 
 
 
 
 
Revision 4 / August 26, 2020 
 
 
 
 
Revision 5 / November 18, 2020 
 
 
 
 
Amendment 1 / August 30, 2021 
 
 
 
 
Amendment 2 / December 23, 2021 
 
 
 
 
Amendment 3 / January 13, 2022 
 
 
 

NCI Protocol #:10389 
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SCHEMA 
 
 
Dose Escalation Schedule 
Dose Level 
Dose 
AZD1775 
(mg) 
Radiation Therapy 
(Gy) 
Level -1 
150 mg QD on Days 1 and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 
Level 1* 
150 mg QD on Days 1, 3, and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 
Level 2 
200 mg QD on Days 1, 3, and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 
Level 3 
200 mg QD on Days 1-5 of weeks -
1, 1, and 3 
37.5 Gy/15 fractions 
* Starting dose 
 
 
 
Expansion Cohort 
 
 
 
 
 

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TABLE OF CONTENTS# ................................................................................................ I 
1.           OBJECTIVES……………………………………………………………………7 
1.1 
Primary Objectives...................................................................................................7 
1.2 
Secondary Objectives...............................................................................................7 
2.           BACKGROUND………………………………………………………………...7 
2.1 
Esophageal Cancer ...................................................................................................7 
2.2 
CTEP IND Agent .....................................................................................................8 
2.3 
Rationale ................................................................................................................12 
2.4 
Correlative Studies Background ............................................................................13 
3.           PATIENT SELECTION ........................................................................................16 
3.1 
Eligibility Criteria ..................................................................................................16 
3.2 
Exclusion Criteria ..................................................................................................18 
3.3 
Inclusion of Women and Minorities ......................................................................19 
4.           REGISTRATION PROCEDURES .......................................................................19 
4.1 
Investigator and Research Associate Registration with CTEP ..............................19 
4.2 
Site Registration .....................................................................................................21 
4.3 
Patient Registration ................................................................................................23 
4.4 
General Guidelines.................................................................................................25 
5.           BIOMARKER, CORRELATIVE, AND SPECIAL STUDIES ............................25 
5.1 
Summary Table for Specimen Collection ..............................................................25 
5.2 
Summary Tables for Gastroenterologist for Research Biopsies ............................26 
5.3 
Specimen Procurement Kits and Scheduling .........................................................27 
5.4 
Specimen Tracking System Instructions ................................................................27 
5.5 
Specimen Collection ..............................................................................................31 
5.6 
Shipping Specimens from Clinical Site to the ETCTN Biorepository ..................33 
5.7 
Biomarker Plan ......................................................................................................35 
5.8 
Integrated Correlative Studies................................................................................38 
5.9 
Exploratory/Ancillary Correlative Studies ............................................................38 
6.           TREATMENT PLAN ............................................................................................39 
6.1 
Agent Administration.............................................................................................39 
6.2 
Definition of Dose-Limiting Toxicity ....................................................................44 
6.3 
Dose Expansion Cohorts: .......................................................................................45 
6.4 
General Concomitant Medication and Supportive Care Guidelines ......................46 
6.5 
Duration of Therapy ...............................................................................................49 
6.6 
Duration of Follow-Up ..........................................................................................50 
7.           DOSING DELAYS/DOSE MODIFICATIONS ...................................................50 
7.1 
Dose modifications due to hematologic toxicity for adavosertib 
monotherapy ..........................................................................................................50 
7.2 
Non-hematologic toxicity dose modifications .......................................................53 
7.3 
Dose modifications to radiation therapy are defined below: .................................54 

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8.           PHARMACEUTICAL INFORMATION..............................................................55 
8.1 
AZD1775 (NSC 751084) .......................................................................................55 
9.           STATISTICAL CONSIDERATIONS ..................................................................58 
9.1 
Study Design/Endpoints.........................................................................................58 
9.2 
Sample Size/Accrual Rate ......................................................................................61 
9.3 
Analysis of Secondary Endpoints ..........................................................................62 
10.         ADVERSE EVENTS:  LIST AND REPORTING REQUIREMENTS ................62 
10.1 
Comprehensive Adverse Events and Potential Risks List (CAEPR).....................62 
10.2 
Adverse Event Characteristics ...............................................................................66 
10.3 
Expedited Adverse Event Reporting ......................................................................66 
10.4 
Routine Adverse Event Reporting .........................................................................69 
10.5 
Pregnancy ...............................................................................................................69 
10.6 
Secondary Malignancy...........................................................................................69 
10.7 
Second Malignancy ................................................................................................70 
11.         STUDY CALENDAR ...........................................................................................71 
12.         MEASUREMENT OF EFFECT ...........................................................................74 
12.1 
Antitumor Effect – Solid Tumors ..........................................................................74 
12.2 
Other Response Parameters ...................................................................................74 
13.         STUDY OVERSIGHT AND DATA REPORTING / REGULATORY 
REQUIREMENTS ................................................................74 
13.1 
Study Oversight .....................................................................................................75 
13.2 
Data Reporting .......................................................................................................75 
13.3 
CTEP Multicenter Guidelines ................................................................................77 
13.4 
Data Quality Portal ................................................................................................77 
13.5 
Collaborative Agreements Language .....................................................................78 
13.6 
Genomic Data Sharing Plan ...................................................................................80 
13.7 
Incidental/Secondary Findings Disclosure Procedure ...........................................80 
14.         REFERENCES…………………………………………………………………..81 
APPENDIX A 
PERFORMANCE STATUS CRITERIA .............................85 
APPENDIX B  
FORMULA TO ESTIMATE RENAL FUNCTION USING 
SERUM CREATININE ........................................................86 
APPENDIX C  
PATIENT MEDICATION DIARY ......................................87 
C.1 
Medication Diary for AZD1775 - Days -7 through -1 (Week -1) .........................87 
C.2 
Medication Diary for AZD1775 - Cycle 1 (Weeks 1-3) ........................................88 
APPENDIX D 
PATIENT DRUG INTERACTIONS HANDOUT AND 
WALLET CARD ..................................................................90 
APPENDIX E  
OGILVIE DYSPHAGIA SCORING ....................................93 

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APPENDIX F  
PRE-BIOPSY ASSESSMENT .............................................94 
APPENDIX G 
TISSUE BIOPSY VERIFICATION ..................................95 
APPENDIX H  
NCLN PHARMACODYNAMICS LABORATORY 
FROZEN BIOPSY COLLECTION PROCEDURE ...............1 
TABLE OF CONTENTS 
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2.0 SCOPE ......................................................................... Error! Bookmark not defined. 
3.0 ABBREVIATIONS ..................................................... Error! Bookmark not defined. 
4.0 INTRODUCTION ....................................................... Error! Bookmark not defined. 
5.0 
ROLES AND RESPONSIBILITIES ..................... Error! Bookmark not defined. 
6.0 
MATERIALS AND EQUIPMENT REQUIRED .. Error! Bookmark not defined. 
7.0 
OPERATING PROCEDURES .............................. Error! Bookmark not defined. 
8.0 
SHIP TO FNLCR FOR ANALYSIS (OPTIONAL) ............ Error! Bookmark not 
defined. 
APPENDIX 1: BATCH RECORD ................ ERROR! BOOKMARK NOT DEFINED. 
APPENDIX 2: PD SAMPLE SHIPPING MANIFEST ..... ERROR! BOOKMARK NOT 
DEFINED. 
 
 
 

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1. 
OBJECTIVES 
 
1.1 
Primary Objectives 
 
1.1.1 To identify the maximally tolerated dose of AZD1775 to be used in combination with 
radiation therapy for patients with esophageal/GEJ cancer that is metastatic or inoperable 
and not eligible for definitive chemoradiation. 
 
1.2 
Secondary Objectives 
 
1.2.1 To observe and record anti-tumor activity.  Although the clinical benefit of this drug has 
not yet been established, the intent of offering this treatment is to provide a possible 
therapeutic benefit, and thus the patient will be carefully monitored for tumor response 
and symptom relief in addition to safety and tolerability. 
 
1.2.2 To evaluate the efficacy of AZD1775 when administered in combination with radiation 
therapy by assessing changes in Ogilvie dysphagia score following treatment, time to 
second intervention for dysphagia, and overall survival. 
 
1.2.3 To identify biomarkers that are predictive for response to experimental therapy 
 
2. 
BACKGROUND 
 
2.1 
Esophageal Cancer 
 
Esophageal cancer, including gastroesophageal junction (GEJ) cancer, is the 8th most common 
cancer worldwide and the 6th leading cause of cancer-related deaths (Ferlay et al., 2015).  Globally, 
while esophageal squamous cell carcinoma remains the predominant histology in Asia, Africa, and 
South America, the incidence of esophageal adenocarcinoma has surpassed that of squamous cell 
carcinoma in North America, Australia, and Europe with predictions that by 2030 up to 1 in 100 
men will be diagnosed with esophageal adenocarcinoma during their lifetime (Arnold et al., 2017).  
In the U.S., the incidence of cancers of the esophagus and gastroesophageal junction has increased 
dramatically in recent decades, largely driven by the rising incidence of adenocarcinoma (Thrift et 
al., 2016).  An estimated 17,000 new cases were diagnosed in 2018 with nearly 16,000 deaths 
(Siegel et al., 2018).  Nearly 50-60% of patients present with inoperable disease due to tumor 
invasion of adjacent structures, evidence of distant metastasis, or because the patient is medically 
unfit for surgery (van Rossum et al., 2018).  
 
For patients with metastatic esophageal cancer or who are inoperable and not eligible for definitive 
chemoradiation, palliative radiation therapy is an established treatment for relieving dysphagia and 
improving quality of life.  However, for patients with localized disease who are not eligible for 
systemic therapy, external beam radiation therapy alone is unlikely to provide sustained 
locoregional control (Zhu et al., 2015).  Local recurrence or progression may result in dysphagia, 
gastrointestinal bleeding, and esophago-respiratory fistula, resulting in substantial morbidity with 
subsequent death.  For patients with metastatic disease, the development of more effective 
systemic therapies, including targeted agents and immunotherapy, has made durable control of the 

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primary tumor a higher priority (Bang et al., 2010; Shah et al., 2018).  Aggressive control of the 
primary tumor in patients with metastatic esophageal cancer has been associated with improved 
survival (Guttmann et al., 2017).  Strategies aimed at improving local control of the primary tumor 
in patients with metastatic disease or those who are inoperable and not eligible for definitive 
chemoradiation will certainly have an impact on quality of life and may even impact survival. 
 
2.2 
CTEP IND Agent 
 
2.2.1 AZD1775 
 
2.2.1.1 
Mechanism of Action 
 
Adavosertib, also known as AZD1775 (previously known as MK-1775), is an inhibitor of WEE1, 
a protein tyrosine kinase.  WEE1 phosphorylates and inhibits cyclin-dependent kinases 1 (CDK1) 
and 2 (CDK2), and is involved in regulation of the intra-S and G2 cell cycle checkpoints.  Proper 
functioning of these checkpoints is essential for deoxyribonucleic acid (DNA) metabolism and the 
DNA damage response (Coleman and Dunphy 1994, Parker and Piwnica-Worms 1992).  WEE1 
overexpression has been demonstrated in hepatocellular carcinoma (Masaki et al., 2003), luminal 
and HER-2 positive breast cancers (Iorns et al., 2009), colon, lung carcinoma, and seminoma 
tumor samples (Mir et al., 2010). 
 
CDK1 (also called cell division cycle 2, or CDC2) activity drives a cell from the G2 phase of the 
cell cycle into mitosis (Investigator’s Brochure 2020).  In response to DNA damage, WEE1 
inhibits CDK1 to prevent the cell from dividing until the damaged DNA is repaired (G2 checkpoint 
arrest).  Inhibition of WEE1 is expected to release a tumor cell from DNA damage-induced arrest 
at the G2/M boundary, so that un-repaired DNA damage may be taken into mitosis (M-phase).  
Since cancer cells exhibit higher levels of endogenous damage than normal cells, as well as 
exhibiting loss of one or more DNA damage response (DDR) capabilities, this is expected to 
preferentially enhance cancer cell death through mitotic catastrophe compared to normal cells.  In 
vitro experiments demonstrate that AZD1775 has synergistic cytotoxic effects when administered 
in combination with various DNA damaging agents that have divergent mechanisms of action.  
Therefore, the clinical development of AZD1775 includes its use as a chemosensitizing drug in 
combination with a cytotoxic agent (or combination of agents) for treatment of advanced solid 
tumors. 
 
CDK2 activity drives a cell into, and through the DNA synthesis (S-phase) of the cell cycle where 
the genome is duplicated in preparation for cell division (Investigator’s Brochure 2020).  An 
important aspect of CDK2 regulation of replication is the control of replication origin firing.  
WEE1, under normal circumstances, regulates CDK2 to provide sufficient time to generate the 
optimum environment for DNA synthesis, such as the generation of a sufficient pool of 
deoxynucleotide triphosphates (dNTPs), the building blocks of DNA.  Inhibition of WEE1 is 
therefore expected to cause aberrantly high CDK2 activity in S-phase cells that will have multiple 
consequences for cancer cells that already have G1/S checkpoint aberrations such as p53 mutations 
or cyclin dependent kinase inhibitor 2A (CDKN2A) deletions (Sherr 1996).  These include de-
regulated replication origin firing before sufficient dNTPs are available, resulting in a higher 
degree of replication stress (already greater in cancer cells).  The resulting increase in levels of 

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replication stress will result in replication fork stalling, the generation of unstable DNA replication 
structures and ultimately replication fork collapse where endonucleases generate DNA damage.  
Therefore, it is anticipated that AZD1775 will have independent anti-tumor activity in the absence 
of added chemotherapy, particularly in cancer cells that already have significantly higher levels of 
replication stress. 
 
Since the majority (if not all) of human cancers harbor abnormalities in their p53 G1/S checkpoint 
control, they become more dependent on S- and G2- phase checkpoints (Sherr 1996).  Thus, S- 
and G2-checkpoint abrogation caused by AZD1775 may selectively sensitize p53-deficient cells 
to anti-cancer agents (Wang et al., 2001) while single-agent activity may be seen in cancers with 
sufficiently high levels of replication stress and endogenous DNA damage. 
 
AZD1775 is a small molecule inhibitor of Wee1 kinase (IC50 = 5.18 nM), that directly inhibits 
phosphorylation of CDC2 at Tyr15 (Investigator’s Brochure, 2020).  A comprehensive review of 
AZD1775 can be found in the AZD1775 Investigator’s Brochure (2020).  AZD1775 has 
demonstrated activity in multiple cancers based on in vitro activity in NCI 60 cell lines. 
 
2.2.1.2 
Preclinical Studies 
 
Early preclinical data demonstrated enhancement of chemotherapeutic effect of AZD1775 in 
combination with carboplatin, cisplatin, and gemcitabine in p53-deficient colon and lung 
carcinoma cells (Hirai et al., 2009), and with 5-FU in p53-deficient colon and pancreatic cells, but 
not p53 wild-type colon cancer cells (Hirai et al., 2010).  The same group also demonstrated 
objective tumor regression in xenograft models using immunodeficient nude rats subcutaneously 
bearing p53 mutant colon carcinoma exposed to AZD1775 in combination with gemcitabine, in 
xenografts bearing breast carcinomas exposed to combination with capecitabine (Hirai et al., 
2009), and in p53-deficient primary pancreatic cancer xenografts, in combination with gemcitabine 
(Rajeshkumar et al., 2011).  Direct target effects were demonstrated in xenografts bearing mutant 
colon carcinoma where administration of AZD1775 (MK-1775) showed reduced pCDC2 levels in 
blood and tumor samples (Merck unpublished data). 
 
Single agent activity has been validated both in in vitro and in vivo studies (Merck unpublished 
data).  Independent evaluation of NCI-60 cell lines and hollow fiber studies support single-agent 
activity of AZD1775 in several colon cancer, melanoma, ovarian cancer, and lung cancer cell lines.  
More recent xenograft studies also support single agent activity of AZD1775 in human renal 
carcinoma cells (Merchant et. al, unpublished data), human epidermoid cells (Merck unpublished 
data), and patient-derived sarcoma samples (Kreahling et al., 2012). 
 
2.2.1.3 
Non-Clinical Pharmacokinetics 
 
The pharmacokinetics (PK) of AZD1775 were evaluated in male Sprague-Dawley rats and Beagle 
dogs following intravenous (IV) and oral administration (Investigator’s Brochure 2020).  
AZD1775 exhibited high plasma clearance in rats and dogs (57.3 and 50.8 mL/min/kg, 
respectively).  The terminal half-life (t1/2) of the compound was short in both species (1.6 and 1.1 
hours in rats and dogs, respectively).  The oral bioavailability was 59.7% in rats and 33.6% in 

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dogs.  AZD1775 (1 mcM) was moderately bound to plasma proteins from rat, dog, and human, 
with the unbound fractions being 23.2, 40.0, and 39.5%, respectively. 
 
2.2.1.4 
Metabolism 
 
The major metabolic pathway of AZD1775 in human liver preparations is oxidative metabolism 
(Investigator’s Brochure 2020).  Oxidative metabolism of AZD1775 is mediated predominantly 
by cytochrome P450 (CYP) 3A4 and Flavin Containing Monooxygenase 3 (FMO3).  AZD1775 is 
a weak reversible inhibitor of CYP2C8, CYP2C9, CYP2C19, and CYP3A4.  In addition, 
AZD1775 is a time-dependent inhibitor of CYP3A4.  
 
2.2.1.5 
Non-clinical toxicology 
 
The toxicology profile of AZD1775 in rats involved the lymphoid and hematopoietic organs and 
gastrointestinal tract, and included decreased food consumption, body weight loss, hematologic 
and serum chemistry changes, decreased urine volume and soft feces (Investigator's brochure, 
2020).  AZD1775 had no effect on CNS function in mice.  The cardiovascular effects of AZD1775 
were examined in 5 isoflurane-anesthetized and mechanically vented dogs.  At doses of 3 and 10 
mg/kg, QTc was marginally prolonged by an average of 5% and 7%, respectively.  No QTc 
changes were observed in the intact dog studies or human studies.  Evidence of reversibility of all 
toxicity changes was generally demonstrated by the end of the 2-week recovery period.  In a single 
dose oral toxicity study, severe irreversible toxicity (mortality) was seen in 1 female out of 10 
female rats after a single dose at 300 mg/kg (1800 mg/m2). 
 
2.2.1.6 
Clinical Experience 
 
As of 11 November 2019, a total of 883 patients have been exposed to AZD1775 in AZ-
sponsored or Merck-sponsored clinical studies (Investigator’s Brochure 2020).  In addition, 
approximately 897 patients have also received AZD1775 as part of externally sponsored 
scientific research. 
These patients include those who have received single doses as high as 1,300 mg of AZD1775 as 
monotherapy, 325 mg of AZD1775 in a single-dose in combination with chemotherapy, and 325 
mg twice a day (BID) in a multiple-dose regimen in combination with chemotherapy. 
 
2.2.1.7 
Clinical Efficacy 
 
Of 176 evaluable patients who received AZD1775 (either single or multiple doses) as monotherapy 
or in combination with gemcitabine, cisplatin, or carboplatin on Study PN001, a partial response 
(PR) (confirmed and unconfirmed) was observed in 17 (9.7%) patients, and stable disease (SD) 
was observed in 94 (53.4%) patients (Leijen et al., 2016a).  A phase 2 trial of AZD1775 in 
combination with carboplatin in patients with refractory or resistant TP53-mutated ovarian cancer 
(Study PN009) has also been performed, yielding a 43% overall response rate (partial response + 
complete response) (Leijen et al., 2016b).  Recently, a phase I trial was performed evaluating the 
addition of AZD1775 to gemcitabine and radiation therapy in patients with locally advanced 
pancreas cancer.  The medial overall survival for all patients was 21.7 months, which is 

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substantially higher than previous results of gemcitabine-based chemoradiation in this patient 
group (Cuneo et al., 2019). 
 
2.2.1.8 
Safety 
 
Based on the preliminary safety data available, the most frequent adverse events (AEs) observed 
were general disorders and administration site conditions (fatigue and asthenia), musculoskeletal 
and connective tissue disorders (myalgia), and gastrointestinal disorders (gastrointestinal 
hemorrhage, lower gastrointestinal hemorrhage, upper gastrointestinal hemorrhage, and 
stomatitis) (Investigator’s Brochure 2020).  The single-dose maximum tolerated dose (MTD) for 
both the gemcitabine and cisplatin combination therapies was 200 mg of AZD1775.  Dose-limiting 
toxicities (DLTs) tended to be hematological in nature in the gemcitabine group and constitutional 
in the cisplatin group.  The single-dose MTD for the combination with carboplatin was 325 mg of 
AZD1775.   
 
Hematologic DLTs have been observed in the multiple-dose AZD1775 combination treatment 
groups with gemcitabine and cisplatin (Investigator’s Brochure 2020).  An MTD of AZD1775 in 
combination with gemcitabine has been established with an interim dose of 50 mg BID on Day 1, 
25 mg BID on Day 2, and 25 mg on Day 3.  An attenuated once a day (QD) for 2 days schedule in 
combination with gemcitabine was evaluated.  The most frequent AEs included hematologic 
toxicity, nausea and vomiting, fatigue, and anorexia (Leijen et al., 2016a).  
 
The MTD for combination with cisplatin has been exceeded at the 250 mg dose level, and 
tolerability of the MTD at 200 mg BID has been confirmed (Investigator’s Brochure 2020).  DLTs 
observed in the multiple-dose carboplatin combination have been both hematological and 
constitutional in nature.  The multiple-dose MTD in combination with carboplatin is 225 mg BID.  
No MTD has been reached for AZD1775 in combination with 5-fluorouracil (5-FU).  DLTs of 
encephalopathy and hyponatremia were observed in the AZD1775 20 mg BID in combination with 
1000 mg/m2 5-FU treatment group.  For the combination of AZD1775 with topotecan plus cisplatin 
in study PN008, toxicities were generally hematological and gastrointestinal in nature.  
 
In study PN009 (Leijen et al., 2016b) and the pharmacokinetic sub-study to PN001 (Leijen et al., 
2016a) which used the pre-market formulation, (both combining 2.5-day BID dosing of AZD1775 
with carboplatin) toxicities were not qualitatively different from the ones observed in the 
carboplatin arm of PN001.  However, increased hematological toxicity was observed, which was 
attributed to a drug-drug interaction with aprepitant (which was administered as anti-nausea 
medication in these studies).  Preliminary PK analyses revealed that co-administration of 
AZD1775 and aprepitant resulted in a ~40% increase of AZD1775 exposure.  
 
2.2.1.9 
Clinical PKs 
 
Analyses of preliminary PK data from study PN001 show that the area under the plasma 
concentration-time curve from time 0 to infinity (AUC0-∞) in a single-dose regimen, the AUC 
from time 0 to trough (12 hours post dose) at steady-state for a BID dosing regimen, and the 
maximum plasma drug concentration observed (Cmax) increase approximately in a dose 

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12 
proportional manner; the time to maximum plasma drug concentration observed (Tmax) occurs at 
1.0 to 8.0 hours and the t1/2 ranged between 7.6 and 12.2 hours (Investigator’s Brochure 2020).  
 
2.2.1.10 
Potential Drug Interactions 
 
The major metabolic pathway of AZD1775 in human liver preparations is oxidative metabolism.  
Oxidative metabolism of AZD1775 is mediated predominantly by CYP3A4 and FMO3 
(Investigator’s Brochure 2020).  AZD1775 is also a weak reversible inhibitor of CYP2C8, 
CYP2C9, CYP2C19 and CYP3A4.  In addition, AZD1775 is a time-dependent inhibitor of 
CYP3A4 in in vitro human liver microsomes.  A formal clinical drug interaction study 
(D6014C00006) evaluating the effect of AZD1775 on PK of substrates for CYP3A, CYP2C19, 
and CYP1A2 has been conducted in advanced solid tumor patients.  The results of this study 
suggest that AZD1775 is a weak inhibitor of these enzymes and is unlikely to result in clinically 
significant drug interaction. 
 
2.3 
Rationale 
 
Radiation therapy induces DNA damage, resulting in activation of apoptotic pathways or inducing 
post-mitotic death due to unrepaired damage.  Following DNA damage, cells rely on cell cycle 
checkpoints to provide sufficient time for DNA repair prior to cell division.  Esophageal cancer 
cells often lack a functional G1 checkpoint due to a high frequency of TP53 mutations.  Based on 
TCGA data, up to 91% of squamous cell and 71% of adenocarcinoma esophageal cancers possess 
a TP53 mutation (The Cancer Genome Atlas Research Network, 2017), making them heavily 
dependent on the G2 checkpoint for DNA damage repair.  WEE1, a tyrosine kinase, is 
phosphorylated following DNA damage and serves as a regulator of the G2 checkpoint and, thus, 
a regulator of genomic stability by inhibitory phosphorylation of CDK1 resulting in cell cycle 
arrest and permitting DNA repair prior to proceeding with mitosis (Dominguez-Kelly et al., 2011; 
Watanabe et al., 1995).  Inhibiting WEE1 results in replication stress, loss of genomic integrity, 
nucleotide shortage, and subsequent double-strand DNA breaks (Beck et al., 2012).  AZD1775 is 
a potent and selective small molecule inhibitor of WEE1 kinase and has been shown to sensitize 
tumor cell lines to both chemotherapy and radiation (Bridges et al., 2011; Hirai et al., 2009; Kausar 
et al., 2015).  Due to the high incidence of TP53 mutations and the dependence on the WEE1/Chk1 
mediated G2-M checkpoint to repair DNA, esophageal/GEJ cancer is a logical site to consider 
the addition of a WEE1 kinase inhibitor to radiation therapy. 
 
There are several phase 1 and 2 studies which have been performed using AZD1775 both as 
monotherapy and in combination with additional chemotherapy agents (Do et al., 2015; Leijen et 
al., 2016a; Leijen et al., 2016b; Mendéz et al., 2018).  Do et al. conducted a phase 1 study of single 
agent AZD1775 for patients with refractory solid tumors (Do et al., 2015).  The MTD was 225 mg 
BID orally over 2.5 days per week for 2 weeks per 21-day cycle.  The most common toxicities 
observed were myelosuppression and diarrhea with DLTs including supraventricular 
tachyarrhythmia and myelosuppression.  Leijen et al. conducted a phase 1 study evaluating 
AZD1775 as monotherapy or in combination with gemcitabine, cisplatin, or carboplatin for 
refractory solid tumors (Leijen et al., 2016a).  The most common adverse events in the combination 
regimens consisted of fatigue, nausea/vomiting, diarrhea, and hematologic toxicity.  The rate of 
grade ≥3 gastrointestinal and hematologic toxicity was 17% and 57%, respectively.  The MTD for 

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13 
the combination regimen with carboplatin was 225 mg twice a day for 2.5 days.  Of note, the 
response rate in TP53-mutated patient was 21% vs. 12% in TP53 wild-type patients.  A subsequent 
phase 2 study from the same group in patients with TP53-mutated ovarian cancer refractory to 
first-line therapy was performed combining AZD1775 with carboplatin (Leijen et al., 2016b).  
AZD1775 was administered at a dose of 225 mg orally twice daily over 2.5 days every 21 days 
with carboplatin AUC 5.  The overall response rate was 43% with manageable toxicity with the 
most frequent grade 3 or 4 events being thrombocytopenia (48%) and neutropenia (37%).  Mendéz 
et al. performed a phase 1 study for patients with stage III/IVB head and neck squamous cell 
carcinoma (Mendéz et al., 2018).  AZD1775 was given BID over 2.5 days during the first week, 
then in combination with cisplatin/docetaxel for 3 additional weeks.  The MTD was 150 mg orally 
BID for 2.5 days with the only DLT being grade 3 diarrhea.  The most common grade ≥2 toxicities 
were diarrhea, fatigue, and neutropenia.  Eight of 10 evaluable patients had a partial response based 
on pathology and imaging with PET.  The safety of AZD1775 in combination with gemcitabine-
based chemoradiation for unresectable pancreas adenocarcinoma has been evaluated (Cuneo et al., 
2019).  In this phase 1 trial, AZD1775 was administered orally on Days 1 and 2, and on Days 8 
and 9 of a 3-week cycle, gemcitabine 1,000 mg/m2 was given on Days 1 and 8 of a 3-week 
treatment cycle, and radiation therapy was administered to a dose of 52.5 Gy in 25 fractions.  An 
AZD1775 dose of 150 mg was determined to be the optimal dose with 24% of patients 
experiencing a DLT most commonly anorexia, nausea, or fatigue.  
 
In summary, a number of phase 1 trials have been performed investigating the use of the WEE1 
inhibitor, AZD1775, either in monotherapy or in combination with multiple chemotherapy 
regimens.  Thus far, AZD1775 appears to be reasonably tolerated with the main toxicities 
including gastrointestinal, hematologic, and fatigue.  Favorable responses have been observed, in 
particular, tumors with TP53 mutations.  Combining AZD1775 with systemic therapy and 
radiation therapy has also been safely performed based on recently published data in pancreas 
cancer (Cuneo et al., 2019).  We hypothesize that inhibiting WEE1 kinase activity with AZD1775 
in combination with targeted radiation therapy will lead to improved tumor control and response 
of esophageal/GEJ cancer.  Herein, we propose a phase 1 dose escalation study to evaluate the 
safety of adding AZD1775 to radiation therapy for patients with metastatic or inoperable and 
ineligible for definitive chemoradiation esophageal/GEJ cancer. 
 
2.4 
Correlative Studies Background 
 
2.4.1 Integrated Studies 
 
2.4.1.1 
Whole exome sequencing 
 
Based on data from The Cancer Genome Atlas, esophageal cancer has an extremely high 
frequency of TP53 mutation, making these cancers highly dependent on the G2 checkpoint for 
DNA damage repair.  WEE1 is a tyrosine kinase which is phosphorylated following DNA 
damage and serves as a regulator of the G2 checkpoint by inhibitory phosphorylation of CDK1, 
resulting in cell cycle arrest and permitting DNA repair prior to proceeding with mitosis.  
However, it is currently unknown if TP53 is a potential predictive biomarker of response to 
treatment with AZD1775 combined with radiation therapy.  In addition, it is unknown if there 
are additional predictive biomarkers of response including DNA repair mutations in ATM, ATR, 

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14 
or ARX.  We hypothesize that mutations in TP53, in addition to other DNA repair mutations, will 
serve as predictive biomarkers of response to treatment with AZD1775 and radiation therapy. 
 
2.4.1.2 
ɣH2AX, pNBS1 IFA with βCATN segmentation  
 
Following double-strand DNA breaks, recruitment of MRN complex proteins serves as a sensor 
of DNA breakage and leads to phosphorylation of NBS1 (pNBS1).  pNBS1, in turn, activates 
protein kinase ATM which subsequently phosphorylates additional substrate proteins, including 
ɣH2AX which localizes at sites of DNA double-strand breaks.  Thus, both pNBS1 and ɣH2AX 
serve as markers of unrepaired double-strand DNA breaks.  We hypothesize that treatment with 
AZD1775 in the expansion cohort will result in an increase in both pNBS1 and ɣH2AX. 
The effects of AZD1775 on esophageal cancer cell cycle markers are demonstrated in Figure 1 
(Yang et al., 2020).  FLO-1 esophageal adenocarcinoma cells were treated with 0 nM or 100 nM 
of AZD1775 alone or in combination with a single exposure of 4 Gy using an RS 2000 biological 
irradiator with 165 kV X-rays.  Cells were pre-treated with the study drug for 2 hours prior to 
radiation delivery.  The cells were then lysed at 1, 4, 16, or 24 hours following radiation exposure 
and Western blotting was performed to detect alterations in cell cycle markers.  As seen in Figure 
1, AZD1775 effectively blocks WEE1 kinase activity (pWEE1 S642) leading to a decrease in 
inhibitory p-CDK1 (Y15) levels, suggesting a subsequent increase in CDK1 activity.  An increase 
in pH2A.x was also observed suggesting an increase in unrepaired double-strand DNA breaks. 
 
Figure 1:  Effects of AZD1775 on FLO-1 cell cycle markers (Yang et al., 2020). 
 
 
2.4.2 Exploratory Study 
 
2.4.2.1 
p-HH3 (S20), p-cdk1 (Y15), pWEE1(S642), and p-Chk1(S345) IHC 
 
WEE1 mediated phosphorylation and inhibition of CDK1 plays a critical role in G2/M checkpoint 
during normal cell growth and following DNA damage or replication stress.  Following DNA 
damage or replication stress, activation of CHK1 by ATR phosphorylates and stimulates WEE1 
activation to suppress CDK1 activity, which prevents entry into mitosis.  Thus, p-cdk1 and pWEE1 
serve as markers of WEE1 inhibition, pChk1 serves as a marker of DNA damage response, and 

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15 
pHH3 serves as a marker for premature mitosis.  We hypothesize that treatment with AZD1775 in 
the expansion cohort will result in a decrease in pWEE1 and p-cdk1 and an increase in pChk1 and 
p-HH3. 
 
In vivo experiments were performed where 1 to 5 million FLO-1 cells were injected in the left 
flanks of mice.  Once the tumor size reached 100-150 cm3, the mice were randomized into 4 
groups:  vehicle, AZD1775 alone, radiation alone, and AZD1775 with radiation with 8-10 mice 
per group.  In the groups that received radiation therapy, 4 Gy per day was delivered for 5 
consecutive days.  AZD1775 was given 40 mg/kg twice daily for 5 consecutive days via oral 
gavage (for those mice administered radiation, AZD1775 was given 2 hours before radiation).  
Tumor size was measured 3 times per week using a caliper and the volume calculated using the 
formula (L x W x W)/2.  A marked enhancement in growth delay in the AZD1775 + radiation 
group was observed as shown in Figure 2A, with no evidence of tumor recurrence in most mice.  
Similar results were observed in our 2nd in vivo model with 15 mice bearing OE33 cell tumors 
(Figure 2B).  To study pharmacodynamics, a group of eight mice bearing FLO-1 tumors were also 
treated with the four combinations for 3 days and the tumors were isolated, lysed, and 
immunoblotting performed.  Western blot results are shown in Figure 2C with 2 tumors per group 
represented (Yang et al., 2020).  Radiation (4 Gy) induced pWEE1 and pCDK1, suggestive of 
induction of the G2 checkpoint response.  However, a marked decrease in p-CDK1 (and pWEE1) 
was seen when comparing the radiation alone to the radiation with AZD1775, as well as reduced 
expression of cyclins A2, B1, E1, and E2.  Lastly, an increase in p-HH3 (which is a marker of 
premature mitosis) was also observed in the tumors treated with AZD1775 with or without 
radiation therapy. 
 
Figure 2:  Effects of AZD1775 on tumor growth in mice bearing FLO-1 cell tumors (A), OE33 
cell tumors (B), and cell cycle markers in excised FLO-1 cell tumors (C) (Yang et al., 2020). 

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16 
 
 
3. 
PATIENT SELECTION 
 
3.1 
Eligibility Criteria  
 
3.1.1 Patients must have histologically confirmed esophageal cancer (either squamous cell or 
adenocarcinoma), including Siewert gastroesophageal junction adenocarcinomas Types 1 
and 2, that is inoperable and not eligible for definitive chemoradiation after 
multidisciplinary review or have pathologically confirmed or imaging consistent with 
metastatic disease.  
 
3.1.2 Age ≥18 years.  Because no dosing or adverse event data are currently available on the 
use of AZD1775 in combination with radiation therapy in patients <18 years of age, 
children are excluded from this study   
 
3.1.3 ECOG performance status 0-1 (Karnofsky ≥70%, see Appendix A). 
 
3.1.4 Patients must have adequate organ and marrow function as defined below: 
 
− 
leukocytes  
 
≥3,000/mcL 
− 
absolute neutrophil count 
 
≥1,500/mcL 
− 
hemoglobin 
 
 
 
≥9 g/dL 

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− 
platelets  
 
≥100,000/mcL 
− 
total bilirubin 
 
≤1.5 × institutional upper limit of normal (ULN) 
− 
AST(SGOT)/ALT(SGPT) 
 
≤3 × institutional ULN 
− 
creatinine  
 
≤ 1.5 × institutional ULN 
OR 
− 
glomerular filtration rate (GFR) ≥60 mL /min/1.73 m2 unless data exists supporting 
safe use at lower kidney function values, no lower 
than 30 mL/min/1.73 m2 (see Appendix B) 
 
3.1.5 For patients with evidence of chronic hepatitis B virus (HBV) infection, the HBV viral 
load must be undetectable on suppressive therapy, if indicated. 
 
3.1.6 Patients with a history of hepatitis C virus (HCV) infection must have been treated and 
cured.  For patients with HCV infection who are currently on treatment, they are eligible 
if they have an undetectable HCV viral load. 
 
3.1.7 Patients with treated brain metastases are eligible if follow-up brain imaging after 
central nervous system (CNS)-directed therapy shows no evidence of progression.   
 
3.1.8 Patients with new or progressive brain metastases (active brain metastases) or 
leptomeningeal disease are eligible if the treating physician determines that immediate 
CNS specific treatment is not required and is unlikely to be required during the first cycle 
of therapy. 
 
3.1.9 Patients with a prior or concurrent malignancy whose natural history or treatment does 
not have the potential to interfere with the safety or efficacy assessment of the 
investigational regimen are eligible for this trial. 
 
3.1.10 Patients with known history or current symptoms of cardiac disease, or history of 
treatment with cardiotoxic agents, should have a clinical risk assessment of cardiac 
function using the New York Heart Association Functional Classification.  To be eligible 
for this trial, patients should be class 2B or better. 
 
3.1.11 Patients able to swallow whole capsules.  Patients with esophageal stents and/or feeding 
tubes are eligible but must be able to swallow whole capsules.  Capsules may not be 
opened or put down a feeding tube. 
 
3.1.12 Patients with a life expectancy > 3 months. 
 
3.1.13 Patients must have an electrocardiogram (ECG) within 8 weeks prior to treatment 
assignment and must have no clinically important abnormalities in rhythm, conduction 
or morphology of resting ECG 
 
3.1.14 a) Resting corrected QTc interval using the Fridericia formula (QTcF) >480 msec (as 
calculated per institutional standards) obtained from an ECG (Note:  if one ECG 
demonstrates a QTcF >480 msec, then a mean QTcF of ≤480 msec obtained from 3 

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18 
ECGs 2-5 minutes apart, is required at study entry), or b) patients with congenital long 
QT syndrome are excluded 
 
3.1.15 The effects of AZD1775 on the developing human fetus are unknown.  For this reason 
and because other therapeutic agents used in this trial are known to be teratogenic, 
women of child-bearing potential must agree to use adequate contraception (hormonal or 
barrier method of birth control; abstinence) for 2 weeks prior to study drug exposure, the 
duration of study participation, and for 1 month after completing treatment.  Women of 
child-bearing potential must have a negative serum or urine pregnancy test (minimum 
sensitivity 25 IU/L or equivalent units of HCG) within 1 week of registration.  Should a 
woman become pregnant or suspect she is pregnant while she or her partner is 
participating in this study, she should inform her treating physician immediately.  Men 
treated or enrolled on this protocol must also agree to use adequate contraception for the 
duration of study participation and for 3 months after completion of treatment.  Male 
patients should not donate sperm during exposure to study drug and for 3 months after 
study drug discontinuation. 
 
3.1.16 Ability to understand and the willingness to sign a written informed consent document.  
Participants with impaired decision-making capacity (IDMC) will not be eligible. 
 
 
3.2 
Exclusion Criteria 
 
3.2.1 Patients who have had chemotherapy or radiotherapy within 3 weeks (6 weeks for 
nitrosoureas or mitomycin C) prior to starting study therapy.  
 
3.2.2 Patients who have not recovered from adverse events due to prior anti-cancer therapy 
(i.e., have residual toxicities > Grade 1) with the exception of alopecia.   
 
3.2.3 Patients who are receiving any other investigational agents. 
 
3.2.4 History of allergic reactions attributed to compounds of similar chemical or biologic 
composition to AZD1775. 
 
3.2.5 Patients receiving any medications or substances that are strong inhibitors or inducers of 
CYP3A4 are ineligible.  Because the lists of these agents are constantly changing, it is 
important to regularly consult a frequently-updated medical reference.  As part of the 
enrollment/informed consent procedures, the patient will be counseled on the risk of 
interactions with other agents, and what to do if new medications need to be prescribed or 
if the patient is considering a new over-the-counter medicine or herbal product.   
 
3.2.6 Patients with uncontrolled intercurrent illness. 
 
3.2.7 Patients with psychiatric illness/social situations that would limit compliance with study 
requirements. 
 

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19 
3.2.8 Pregnant women are excluded from this study because AZD1775 is a WEE1 inhibiting 
agent with the potential for teratogenic or abortifacient effects.  Because there is an 
unknown but potential risk for adverse events in nursing infants secondary to treatment of 
the mother with AZD1775, breastfeeding should be discontinued if the mother is treated 
with AZD1775.  These potential risks may also apply to other agents used in this study. 
 
3.2.9 Prior thoracic or abdominal radiation therapy for cancer that would result in significant 
overlap of radiation therapy fields at the discretion of the investigator. 
 
3.2.10 Patients with congenital long QT syndrome or with a history of Torsades de pointes 
unless all risk factors contributed to Torsades have been corrected.  AZD1775 has not 
been studied in patients with ventricular arrhythmias or recent myocardial infarction. 
 
3.2.11 Eligibility of subjects receiving any medications or substances with the potential to 
affect the activity or pharmacokinetics of AZD1775 will be determined following 
review by the principal investigator. 
 
3.3 
Inclusion of Women and Minorities 
 
NIH policy requires that women and members of minority groups and their subpopulations be 
included in all NIH-supported biomedical and behavioral research projects involving NIH-
defined clinical research unless a clear and compelling rationale and justification establishes to 
the satisfaction of the funding Institute & Center (IC) Director that inclusion is inappropriate 
with respect to the health of the subjects or the purpose of the research.  Exclusion under other 
circumstances must be designated by the Director, NIH, upon the recommendation of an IC 
Director based on a compelling rationale and justification.  Cost is not an acceptable reason for 
exclusion except when the study would duplicate data from other sources.  Women of 
childbearing potential should not be routinely excluded from participation in clinical research.  
Please see http://grants.nih.gov/grants/funding/phs398/phs398.pdf. 
 
 
4. 
REGISTRATION PROCEDURES 
 
4.1 
Investigator and Research Associate Registration with CTEP 
 
Food and Drug Administration (FDA) regulations and National Cancer Institute (NCI) policy 
require all individuals contributing to NCI-sponsored trials to register and to renew their 
registration annually.  To register, all individuals must obtain a Cancer Therapy Evaluation 
Program (CTEP) Identity and Access Management (IAM) account at 
https://ctepcore.nci.nih.gov/iam.  In addition, persons with a registration type of Investigator 
(IVR), Non-Physician Investigator (NPIVR), or Associate Plus (AP) must complete their annual 
registration using CTEP’s web-based Registration and Credential Repository (RCR) at 
https://ctepcore.nci.nih.gov/rcr.    
 
RCR utilizes five person registration types. 
• IVR:  MD, DO, or international equivalent, 

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20 
• NPIVR:  advanced practice providers (e.g., NP or PA) or graduate level researchers (e.g., 
PhD), 
• AP:  clinical site staff (e.g., RN or CRA) with data entry access to CTSU applications 
such as the Roster Update Management System (RUMS), OPEN, Rave, acting as a 
primary site contact, or with consenting privileges,  
• Associate (A):  other clinical site staff involved in the conduct of NCI-sponsored trials, 
and 
• Associate Basic (AB):  individuals (e.g., pharmaceutical company employees) with 
limited access to NCI-supported systems. 
 
RCR requires the following registration documents: 
 
Documentation Required 
IVR NPIVR AP 
A 
AB 
FDA Form 1572 
✔ 
✔ 
Financial Disclosure Form 
✔ 
✔ 
✔ 
NCI Biosketch (education, training, employment, license, 
and certification) 
✔ 
✔ 
✔ 
GCP training 
✔ 
✔ 
✔ 
 
 
Agent Shipment Form (if applicable) 
✔ 
 
 
 
 
CV (optional) 
✔ 
✔ 
✔ 
 
An active CTEP-IAM user account and appropriate RCR registration is required to access all 
CTEP and Cancer Trials Support Unit (CTSU) websites and applications.  In addition, IVRs and 
NPIVRs must list all clinical practice sites and Institutional Review Boards (IRBs) covering their 
practice sites on the FDA Form 1572 in RCR to allow the following: 
 
• Addition to a site roster, 
• Assign the treating, credit, consenting, or drug shipment (IVR only) tasks in OPEN, 
• Act as the site-protocol Principal Investigator (PI) on the IRB approval, and 
• Assign the Clinical Investigator (CI) role on the Delegation of Tasks Log (DTL).  
 
In addition, all investigators acting as the Site-Protocol PI (Investigator listed on the IRB 
approval), consenting/treating/drug shipment investigator in OPEN, or as the Clinical 
Investigator (CI) on the DTL must be rostered at the enrolling site with a participating 
organization.  
 
Additional information is located on the CTEP website at 
https://ctep.cancer.gov/investigatorResources/default.htm.  For questions, please contact the 
RCR Help Desk by email at RCRHelpDesk@nih.gov. 
 
 

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21 
4.2 
Site Registration 
 
This study is supported by the NCI Cancer Trials Support Unit (CTSU). 
 
IRB Approval   
 
Sites participating with the NCI Central Institutional Review Board (NCI CIRB) must submit the 
Study Specific Worksheet for Local Context (SSW) to the CIRB using IRBManager to indicate 
their intent to open the study locally.  The NCI CIRB’s approval of the SSW is automatically 
communicated to the CTSU Regulatory Office, but sites are required to contact the CTSU 
Regulatory Office at CTSURegPref@ctsu.coccg.org to establish site preferences for applying 
NCI CIRB approvals across their Signatory Network.  Site preferences can be set at the network 
or protocol level.  Questions about establishing site preferences can be addressed to the CTSU 
Regulatory Office by email or calling 1-888-651-CTSU (2878). 
 
Sites using their local IRB or REB must submit their approval to the CTSU Regulatory Office 
using the Regulatory Submission Portal located in the Regulatory section of the CTSU website.  
Acceptable documentation of local IRB/REB approval includes: 
• Local IRB documentation, 
• IRB-signed CTSU IRB Certification Form, and/or 
• Protocol of Human Subjects Assurance Identification/IRB Certification/Declaration of 
Exemption Form. 
 
In addition, the Site-Protocol PI (i.e., the investigator on the IRB/REB approval) must meet the 
following five criteria to complete processing of the IRB/REB approval record: 
• Holds an active CTEP status, 
• Rostered at the site on the IRB/REB approval (applies to US and Canadian sites only) and 
on at least one participating roster, 
• If using NCI CIRB, rostered on the NCI CIRB Signatory record, 
• Includes the IRB number of the IRB providing approval in the Form FDA 1572 in the 
RCR profile, and 
• Holds the appropriate CTEP registration type for the protocol.  
 
Additional Requirements 
 
Additional requirements to obtain an approved site registration status include: 
• An active Federalwide Assurance (FWA) number, 
• An active roster affiliation with the Lead Protocol Organization (LPO) or a Participating 
Organization (PO), and 
• Compliance with all protocol-specific requirements (PSRs). 
 
4.2.1 Downloading Regulatory Documents 
 
Download the site registration forms from the protocol-specific page located on the CTSU 
members’ website.  Permission to view and download this protocol and its supporting documents 

NCI Protocol #:10389 
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22 
is restricted based on person and site roster assignment.  To participate, the institution and its 
associated investigators and staff must be associated with the LPO or a PO on the protocol.  One 
way to search for a protocol is listed below. 
 
• Log in to the CTSU members’ website (https://www.ctsu.org) using your CTEP-IAM 
username and password, 
• Click on Protocols in the upper left of the screen 
o Enter the protocol number in the search field at the top of the protocol tree, or 
o Click on the By Lead Organization folder to expand, then select LAO-OH007, 
and protocol number 10389, 
• Click on Documents, select Site Registration, and download and complete the forms 
provided.  (Note:  For sites under the CIRB, IRB data will load automatically to the 
CTSU.)  
 
4.2.2 Protocol Specific Requirements For NCI Protocol #10389 Site Registration 
 
• Specimen Tracking System Training Requirement: 
o All data entry users (Clinical Research Associate role) at each participating site will 
need to complete the Theradex-led training. 
o Theradex will provide a certificate of completion, which will need to be submitted to 
the CTSU through the Regulatory Submission Portal. 
o The training is a one-time only requirement per individual.  If an individual has 
previously completed the training for another ETCTN study, the training does not 
need to be completed again nor does the certificate of completion need to be 
resubmitted to the CTSU.  However, new versions of the Specimen Tracking System 
may require new training. 
o This training will need to be completed before the first patient enrollment at a given 
site. 
o Please contact STS Support at Theradex for the training 
(STS.Support@theradex.com, Theradex phone: 609-799-7580). 
 
4.2.3 Submitting Regulatory Documents 
 
Submit required forms and documents to the CTSU Regulatory Office using the Regulatory 
Submission Portal on the CTSU website.  
 
To access the Regulatory Submission Portal, log on to the CTSU members’ website go to the  
Regulatory section, and select Regulatory Submission. 
 
Institutions with patients waiting that are unable to use the Regulatory Submission Portal should 
alert the CTSU Regulatory Office immediately at 1-866-651-2878 in order to receive further 
instruction and support. 
 
Delegation of Tasks Log (DTL) 
 
Each site must complete a protocol-specific DTL using the DTL application in the Delegation 

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23 
Log section on the CTSU members’ website.  The Clinical Investigator (CI) is required to review 
and electronically sign the DTL prior to the site receiving an approved site registration status and 
enrolling patients to the study.  To maintain an approved site registration status the CI must re-
sign the DTL at least annually and when a new version of the DTL is released; and activate new 
task assignments requiring CI sign-off.  Any individual at the enrolling site on a participating 
roster may initiate the site DTL.  Once the DTL is submitted for CI approval, only the designated 
DTL Administrators or the CI may update the DTL.  Instructions on completing the DTL are 
available in the Help Topics button in the DTL application and include a Master Task List, 
which describes DTL task assignments, CI signature, and CTEP registration requirements. 
 
4.2.4 Checking Site Registration Status 
 
Site’s registration status may be verified on the CTSU website.  
 
• Click on Regulatory at the top of the screen 
• Click on Site Registration, and  
• Enter the site’s 5-character CTEP Institution Code and click on Go. 
o Additional filters are available to sort by Protocol, Registration Status, Protocol 
Status, and/or IRB Type. 
 
Note:  The status shown only reflects institutional compliance with site registration requirements 
as outlined within the protocol.  It does not reflect compliance with protocol requirements for 
individuals participating on the protocol or the enrolling investigator’s status with NCI or their 
affiliated networks. 
 
4.3 
Patient Registration 
 
4.3.1 OPEN / IWRS 
 
The Oncology Patient Enrollment Network (OPEN) is a web-based registration system available 
on a 24/7 basis.  OPEN is integrated with CTSU regulatory and roster data and with the LPOs 
registration/randomization systems or the Theradex Interactive Web Response System (IWRS) 
for retrieval of patient registration/randomization assignment.  OPEN or IWRS will populate the 
patient enrollment data in NCI’s clinical data management system, Medidata Rave. 
 
Requirements for OPEN access:   
 
• A valid CTEP-IAM account. 
• To perform enrollments or request slot reservations:  Must be on an LPO roster, ETCTN 
corresponding roster, or PO roster with the role of Registrar.  Registrars must hold a 
minimum of an Associate Plus (AP) registration type. 
• If a DTL is required for the study, the registrar must hold the OPEN Registrar task on the 
DTL for the site. 
• Have an approved site registration for the protocol prior to patient enrollment. 
 
To assign an Investigator (IVR) or Non-Physician Investigator (NPIVR) as the treating, 

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crediting, consenting, drug shipment (IVR only), or receiving investigator for a patient transfer in 
OPEN, the IVR or NPIVR must list the IRB number used on the site’s IRB approval on their 
Form FDA 1572 in RCR.  If a DTL is required for the study, the IVR or NPIVR must be 
assigned the appropriate OPEN-related tasks on the DTL. 
 
Prior to accessing OPEN, site staff should verify the following: 
 
• Patient has met all eligibility criteria within the protocol stated timeframes, and  
• All patients have signed an appropriate consent form and HIPAA authorization form (if 
applicable). 
 
Note:  The OPEN system will provide the site with a printable confirmation of registration and 
treatment information.  IWRS system also sends an email confirmation of the registration.  You 
may print this confirmation for your records.  
 
Access OPEN at https://open.ctsu.org or from the OPEN link on the CTSU members’ website.  
Further instructional information is in the OPEN section of the CTSU website at 
https://www.ctsu.org or https://open.ctsu.org.  For any additional questions, contact the CTSU 
Help Desk at 1-888-823-5923 or ctsucontact@westat.com. 
 
Patient enrollment for this study will be facilitated using the Slot Reservation System in 
conjunction with the registration system in OPEN.  Prior to discussing protocol entry with the 
patient, all site staff must use the CTSU OPEN Slot Reservation System or the IWRS Slot 
Reservation System to ensure that a slot on the protocol is available to the patient.  Once a slot 
reservation confirmation is obtained, site staff may then proceed to enroll the patient to this 
study.  
 
 
4.3.2 Special Instructions for Patient Enrollment 
 
This Study will use the ETCTN Specimen Tracking System (STS).  
 
• All biospecimens collected for this trial must be submitted using the ETCTN Specimen 
Tracking System (STS) unless otherwise noted.   
• The system is accessed through Rave user roles:  “Rave CRA” and “Rave CRA 
(Labadmin)”  for data entry at the treating institutions and “Biorepository” for users 
receiving the specimens for processing and storage at reference labs and the 
Biorepository.   
• Please refer to the Medidata Account Activation and Study Invitation Acceptance link on 
the CTSU website in the Data Management section under the Rave Home tab and then 
under Rave Resource Materials. 
• Important:  Failure to complete required fields in STS may result in a delay in 
sample processing.  Any case reimbursements associated with sample submissions will 
not be credited if samples requiring STS submission are not logged into STS. 
 
Detailed instructions on use of the STS can be found in Section 5.4. 

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4.3.3 OPEN/IWRS Questions? 
 
Further instructional information on OPEN is provided on the OPEN link of the CTSU website 
at https://www.ctsu.org or at https://open.ctsu.org.  For any additional questions contact the 
CTSU Help Desk at 1-888-823-5923 or ctsucontact@westat.com.  
 
Theradex has developed a Slot Reservations and Cohort Management User Guide, which is 
available on the Theradex website:  http://www.theradex.com/clinicalTechnologies/?National-
Cancer-Institute-NCI-11.  This link to the Theradex website is also on the CTSU website OPEN 
tab. For questions about the use of IWRS for slot reservations, contact the Theradex Helpdesk at 
609-619-7862 or Theradex main number 609-799-7580; CTMSSupport@theradex.com. 
 
4.4 
General Guidelines 
 
Following registration, patients should begin protocol treatment within 28 days.  Issues that 
would cause treatment delays should be discussed with the Principal Investigator.  If a patient 
does not receive protocol therapy following registration, the patient’s registration on the study 
may be canceled.  The Study Coordinator should be notified of cancellations as soon as possible. 
 
 
5. 
BIOMARKER, CORRELATIVE, AND SPECIAL STUDIES 
 
 
5.1 
Summary Table for Specimen Collection 
 
 
Time Point 
Specimen 
Send Specimens To: 
Archival 
 
• Archival formalin-fixed paraffin-embedded 
(FFPE) tumor tissue block (preferred)1 
(mandatory) 
 
If a block is not available, then submit: 
• 1 H&E stained slide (3-5 µm) 
• 30-50 unstained, uncharged, air-dried slides 
(10 µm) 
ETCTN Biorepository 
Baseline 
 
• 10 mL blood in EDTA tube (mandatory)3 
• 2 snap-frozen tumor tissue forceps biopsies 
(mandatory; expansion cohort only)2,4 
• 2 tumor tissue forceps biopsies in formalin 
(mandatory where clinically feasible; 
expansion cohort only)2 
ETCTN Biorepository 
 
Week -1 Day 4 or 5 (Expansion Cohort Only) 

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26 
 
• 2 
snap-frozen 
tissue 
forceps 
biopsies 
(mandatory) 2,4 
• 2 tumor tissue forceps biopsies in formalin 
(mandatory where clinically feasible)2 
ETCTN Biorepository 
 
1For archival tissue, a copy of the corresponding anatomic pathology report must be sent with 
the tissue and uploaded to Rave.  If submitting slides, then slides must be processed in order, 
and numbered sequentially (e.g., H&E stained slide is created first and labeled 1, unstained 
slides are then created and numbered 2 – 51). 
2For new biopsies, the Tissue Biopsy Verification Form (Appendix G),  a copy of the 
radiology and/or operative reports from the tissue removal procedure and the diagnostic 
anatomic pathology report must be sent with the tissue to the ETCTN Biorepository.   
3 within 28 days prior to study entry 
4 Refer to Appendix H for detailed SOP on snap frozen tumor tissue 
 
 
5.2 
Summary Tables for Gastroenterologist for Research Biopsies 
 
Biopsy #:  1 
Trial Time Point:  Baseline (Expansion cohort only) 
IR Biopsy Definition:  Research – No Clinical Impact  
Core 
Priority 
Use in the 
Trial 
Biomarker 
Name(s) 
Tumor 
Content 
Required 
Post-Biopsy Processing 
1 
Integrated 
ɣH2AX, 
pNBS1 
IFA 
with 
βCATN 
segmentation  
>50% OR 
25-50%  
Frozen 
2 
Exploratory 
p-HH3 (S20), 
p-cdk1 (Y15), 
pWEE1(S642), 
andp-
Chk1(S345) 
IHC 
Prefer 25-50% 
or >50%, but 
5-25% is 
acceptable 
Formalin 
 
Biopsy #:  2 
Trial Time Point:  Week -1 Day 4 or 5 (Expansion cohort only) 
IR Biopsy Definition:  Research – No Clinical Impact  
Core 
Priority 
Use in the 
Trial 
Biomarker 
Name(s) 
Tumor 
Content 
Required 
Post-Biopsy Processing 
1 
Integrated 
ɣH2AX, 
pNBS1 
IFA 
with 
βCATN 
segmentation  
>50% OR 
25-50%  
Frozen 

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2 
Exploratory 
p-HH3 (S20), 
p-cdk1 (Y15), 
pWEE1(S642), 
andp-
Chk1(S345) 
IHC 
Prefer 25-50% 
or >50%, but 
5-25% is 
acceptable 
Formalin 
 
Note:  Pre-biopsy assessments will be reported and tracked through a trial-specific Case Report 
Form (CRF) within the CTEP Medidata Rave system (see Appendix F).  
 
5.3 
Specimen Procurement Kits and Scheduling 
 
5.3.1 Specimen Procurement Kits  
 
Kits for the collection and shipment of specimens to the ETCTN Biorepository can be ordered 
online via the Kit Management system: 
(https://ricapps.nationwidechildrens.org/KitManagement).  
 
Users at the clinical sites will need to set up an account in the Kit Management system and select 
a specific clinical trial protocol to request a kit.  Please note that protocol may include more than 
one type of kit.  Each user may order two kits  per kit type  per day (daily max = 6 kits).  Kits are 
shipped ground, so please allow 5-7 days for receipt.  A complete list of kit contents for each kit 
type is located on the Kit Management system website. 
 
Note:  Kits or supplies are only provided for specimens shipped to the Biorepository. 
Institutional supplies must be used for all other specimen collection and processing.  
 
5.3.2 Scheduling of Specimen Collections 
 
• Tumor tissue specimens collected during biopsy procedures and fixed in formalin must 
be shipped on the same day of collection. 
• Tissue in formalin can be collected Monday through Wednesday and shipped overnight 
for arrival on Tuesday through Thursday at the ETCTN Biorepository at Nationwide 
Children’s Hospital. 
• Specimens submitted as frozen (snap-frozen tissue) can be collected on any day but must 
be stored frozen and shipped to the ETCTN Biorepository on Monday through Thursday.  
In the event that frozen specimens cannot be shipped immediately, they must be 
maintained in a -70°C to -80°C freezer. 
 
• Fresh blood specimens may be collected and shipped Monday through Friday.  
 
5.4 
Specimen Tracking System Instructions 
 
5.4.1 Specimen Tracking System Overview and Enrollment Instructions 
 
For the ETCTN STS, the following information will be requested: 

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• Protocol Number 
• Investigator Identification  
o Institution and affiliate name 
o Investigator’s name 
• Eligibility Verification:  Patients must meet all the eligibility requirements listed in 
Section 3. 
• Additional Requirements: 
o Patients must provide a signed and dated, written informed consent form. 
 
Upon enrolling a patient, IWRS will communicate with OPEN, assigning two separate and 
unique identification numbers to the patient, a Universal patient ID (UPID) and a Treatment 
patient ID.  The UPID is associated with the patient and used each and every time the patient 
engages with the portion of this or any other protocol that uses the ETCTN Specimen Tracking 
System.  The UPID contains no information or link to the treatment protocol.  IWRS will 
maintain an association between the UPID for ETCTN biobanking and molecular 
characterization and any treatment protocols the patient participates in, thereby allowing analysis 
of the molecular characterization results with the clinical data.   
 
Immediately following enrollment, the institutional anatomical pathology report for the diagnosis 
under which the patient is being enrolled must be uploaded into Rave.  The report must include 
the surgical pathology ID (SPID), collection date, block number, and the IWRS-assigned UPID 
and patient study ID for this trial.  For newly acquired biopsies, the radiology and operative 
report(s) must also be uploaded into Rave.  Important:  Remove any personally identifying 
information, including, but not limited to, the patient’s name, date of birth, initials, medical 
record number, and patient contact information from the institutional pathology report 
prior to submission. 
 
Additionally, please note that the STS software creates pop-up windows when reports are 
generated, so you will need to enable pop-ups within your web browser while using the software.   
 
For questions regarding the Specimen Tracking System, please contact STS Support at 
STS.Support@theradex.com. 
 
The Shipping List report must be included with all sample submissions. 
 
5.4.2 Specimen Labeling 
 
5.4.2.1 Blood Specimen Labels  
 
Include the following on blood specimens (including whole blood and frozen, processed blood 
products – like serum and plasma): 
• Patient Study ID 
• Universal Patient ID (UPID) 
• Specimen ID (automatically generated by Rave) 
• Time point 
• Specimen type (e.g., blood, serum) 

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• Collection date (to be added by hand) 
 
5.4.2.2 Tissue Specimen Labels  
 
Include the following on all tissue specimens or containers (e.g., formalin jar):  
• Patient Study ID 
• Universal Patient ID (UPID) 
• Specimen ID (automatically generated by Rave) 
• Time point 
• Specimen type (e.g., formalin-fixed paraffin-embedded [FFPE] Block, Formalin 
Fixed Tissue, Fresh Tissue in Media, etc.) 
• Tissue type (P for primary, M for metastatic or N for normal) 
• Surgical pathology ID (SPID) number (when applicable)   
• Block number from the corresponding pathology report (archival only) 
• Collection date (to be added by hand)  
• Slide section number (only if archival tissue is submitted as slides) (to be added by 
hand) 
 
5.4.2.3 Example of Specimen Label Generated by STS 
 
STS includes a label printing facility, accessed via the Print Label CRF in the All Specimens 
folder.  A generated PDF is emailed to the use as a result of saving that form. 
 
The following image is an example of a tissue specimen label printed on a label that is 0.5” high 
and 1.28” wide. 
 
  
 
 
The QR code in the above example is for the Specimen ID shown on the second line.   
 
Labels may be printed on a special purpose label printer, one label at a time, or on a standard 
laser printer, multiple labels per page.  Theradex recommends the use of these low temperature 
waterproof labels for standard laser printers: https://www.labtag.com/shop/product/cryo-laser-
labels-1-28-x-0-5-cl-23-colors-available/ 
 
 

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The last line item on the label includes the following data points joined together: 
1. Tissue only: Primary (P), Metastatic (M), Normal (N) tissue indicated at the 
beginning of the specimen ID; this field is blank if not relevant (e.g., for blood) 
2. Block ID or blank if not relevant 
3. SPID (Surgical Pathology ID) or blank if none 
4. An optional alpha-numeric code that is protocol specific and is only included if the 
protocol requires an additional special code classification 
 
Space is provided at the bottom of the label for the handwritten date and optional time.  
The last line on the example label is for the handwritten date and optional time. 
 
5.4.3 Overview of Process at Treating Site 
 
5.4.3.1 OPEN Registration 
 
All registrations will be performed using the Oncology Patient Enrollment Network (OPEN) 
system.  OPEN communicates automatically with the Interactive Web Response System (IWRS) 
which handles identifier assignments, any study randomization, and any prescribed slot 
assignments.  If specimen analysis is required to determine eligibility, the protocol will be setup 
with multi-step registration. 
 
Registration without eligibility specimen analysis:  
 
1. Site enters registration data into OPEN during one or more steps. 
2. IWRS receives data from OPEN, generates the Patient Study ID and the Universal Patient 
ID, both of which are sent back to OPEN. 
3. IWRS sends all applicable registration data directly to Rave at the end of the final 
registration step. 
 
Any data entry errors made during enrollment should be corrected in Rave. 
 
5.4.3.2 Rave Specimen Tracking Process Steps 
 
Step 0: Log into Rave via your CTEP-IAM account, then navigate to the appropriate participant 
 
Step 1:  Complete the Histology and Disease form (but do not upload reports until a specimen 
label can be applied to them) and the Baseline forms regarding Prior Therapies.  Enter the 
initial clinical specimen data:  
• Specimen Tracking Enrollment CRF:  Enter Time Point, Specimen Category, 
Specimen Type, Block number, Tissue type, Surgical Path ID, and number of labels 
needed (include extra labels to apply to reports to be uploaded).  CRF generates unique 
Specimen ID.  
 
Step 2:  Print labels using the Print Labels CRF located in the All Specimens folder, then  
collect specimen. 
• Label specimen containers and write collection date on each label.  After collection, store 

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labeled specimens as described in Section 5.4.2. 
• Apply an extra specimen label to each report before scanning.  Return to the Histology 
and Disease form to upload any initial Pathology, Radiology, Molecular Reports (up to 
4), Surgical (or Operative) reports and Tissue Biopsy Verification form (when 
applicable).  Return to Specimen Tracking Enrollment CRF to upload any molecular 
report (one per specimen) and/or specimen specific pathology or related report (one per 
specimen).  Uploaded reports should have protected health information (PHI) data, like 
name, date of birth, mailing address, medical record number or social security number 
(SSN), redacted.  Do not redact SPID, block number, diagnosis, or relevant dates (such as 
collection date), and include the UPID and patient study ID on each document.  
 
Step 3:  Complete specimen data entry. 
• Specimen Transmittal Form:  Enter collection date and time and other required 
specimen details. 
 
Step 4:  When ready to ship, enter shipment information. 
• Shipping Status CRF:  Enter tracking number, your contact information, recipient, 
number of sample containers and ship date once for the first specimen in a shipment. 
Copy Shipping CRF:  In the specimen folders for additional specimens (if any) that will 
be shipped with the initial specimen, please use the Copy Shipping form to derive 
common data into additional Shipping Status forms. A few unique fields will still need 
to be entered in Shipping Status. 
 
Step 5:  Print shipping list report and prepare to ship.  
• Shipping List report is available at the site level. 
• Print two copies of the shipping list, one to provide in the box, the other for your own 
records. 
• Print pathology or other required reports to include in the box.  Be sure the printed copy 
includes the specimen label. 
 
Step 6:  Send email notification.  
• For only one of the specimens in the shipment, click “Send Email Alert” checkbox on the 
Shipping Status CRF to email recipient. 
 
Step 7:  Ship the specimen(s). 
 
Step 8:  Monitor the Receiving Status form located in each specimen folder for acknowledgment 
of receipt and adequacy. 
 
5.5 
Specimen Collection  
 
5.5.1 Archival or Formalin-Fixed Paraffin-Embedded (FFPE) Tumor Specimen  
 
If previously-collected FFPE tissue will be submitted, then the following criteria must be met: 
 
• Tissue must have been collected within 24 months prior to registration  

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• FFPE tumor tissue block(s) must be submitted.  The optimal block is at least 70% tumor.  
Specimen size requirement is as follows: 
o Surface area:  25 mm2 is optimal.  Minimum is 5 mm2. 
o Volume:  1 mm3 optimal.  Minimum volume is 0.2 mm3, however the success of 
DNA extraction decreases at suboptimal tissue volume. 
 
If an existing block cannot be submitted, the following are requested, if available: 
• One (1) H&E slide (3-5 µm) 
• Thirty to fifty (30 – 50) 10 µm unstained air-dried uncharged slides  
 
Process and number slides sequentially (e.g., H&E stained slide should be created first and 
labeled with “1,” and additional unstained slides should be processed next and be labeled 2 – n). 
 
See Section 5.4.2 for labeling instructions. 
 
5.5.2 Collection of Snap-Frozen Biopsies  
 
1. Tissue should be frozen as soon as possible.  Optimally, freeze within 2 minutes from 
resection.  Refer to Appendix H for detailed SOP. 
2. Label cryovial(s) according to instructions in Section 5.4.2.  
3. Follow instructions described in Appendix H for snap-freezing tissue, including, placing 
the tissue in a pre-chilled cryovial and freeze the tube.  Keep frozen until shipment to the 
ETCTN Biorepository.  Do not ship to FNLCR. 
 
5.5.3 Formalin-Fixed Tumor Biopsies 
 
1. Label formalin-filled containers according to instructions in Section 5.4.2.  
2. Obtain 2-3 tissue forceps biopsy specimens, and place one biopsy in each cassette.  
3. Snap the cassette lids closed and place cassettes into a formalin-filled pre-labeled 
container as soon as possible after collection to prevent air drying.  Up to two cassettes 
may be placed in one formalin jar. 
4. Secure the container lids and package containers into the shipping kit according to 
instructions in Section 5.6.  Keep tissue in formalin jars at room temperature until 
shipment to the ETCTN Biorepository.  
 
5.5.4 Blood Collection  
 
5.5.4.1 Collection of Blood in EDTA Tubes for Shipping Whole Blood  
 
1. Label EDTA tubes according to the instructions in Section 5.4.2. 
2. Collect 10 mL blood in EDTA tube(s) and gently invert tube to mix. 
3. Ship on day of collection (whenever possible) according to instructions in Section 
5.6. 
4. If blood cannot be shipped on the day of collection (e.g., a late scheduled collection), 
then refrigerate until shipment.  
 

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5.6 
Shipping Specimens from Clinical Site to the ETCTN Biorepository  
 
5.6.1 General Shipping Information 
 
Core biopsies that are fixed in formalin, snap frozen core biopsies, and fresh blood should be 
shipped as one shipment in a dual chambered kit, whenever possible.  The shipping container 
sent with kit contents should be used to ship specimens to the ETCTN Biorepository.  In winter 
months, please include extra insulation, such as bubble wrap, inside the ambient side of the 
shipping container. 
 
For new biopsies, the Tissue Biopsy Verification Form (Appendix G) and a copy of the 
radiology and/or operative reports from the tissue removal procedure and the diagnostic 
anatomic pathology report must be sent with the tissues to the ETCTN Biorepository. 
  
For all archival tissue, the corresponding anatomical clinical pathology report is required both in 
the package and uploaded in the ETCTN specimen tracking system.  If this is not available at the 
time of shipment, then it must be sent to the ETCTN Biorepository as soon as possible and 
uploaded to the ETCTN specimen tracking system, or the specimen will not be processed.  The 
pathology report must state the disease diagnosis made by the reviewing pathologist. 
 
5.6.2 Specimen Shipping Instructions 
 
Tissue in formalin must be shipped on the day of collection.  Collect and ship on Monday 
through Wednesday. 
 
Frozen specimens and archival (FFPE) tissue may be shipped on Monday through Thursday.  
 
Fresh blood may be shipped on Monday through Friday.  Please select “Saturday Delivery” 
when shipping fresh blood on a Friday.  
 
5.6.2.1 
Shipping of FFPE Blocks and Glass Slides  
 
1. Before packaging blocks or slides, verify that each specimen is labeled according to 
Section 5.4.2.2. 
2. Blocks should be placed in a hard-sided container, preferably a special block holder, to 
protect the specimen.  Glass slides are to be placed in plastic slide holders.  Place tissue 
paper on top of the separated slides prior to closing the slide holder to reduce slide 
movement during shipment. 
3. Place the blocks or slides in a reinforced cardboard shipping box with appropriate 
packaging filler to minimize movement of specimens within the shipping box. 
4. Include a copy of the forms listed above and a shipping manifest from the Specimen 
Tracking System with each shipment.  
5. Please include a cold pack when shipping on hot days and extra insulation on cold days. 
6. Ship specimens to the address listed below.  FedEx Priority Overnight is strongly 
recommended to prevent delays in package receipt.  

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5.6.2.2 
Shipping Frozen and Ambient Specimens in a Dual-Chamber Kit  
 
The Dual Chambered Specimen Procurement Kit is constructed to allow the shipment of frozen 
(on dry ice) and ambient (room temperature) specimens in the same container.  Dry ice may be 
placed in either compartment of the kit but should not be put in both.  The dual chambered 
kit is only used for shipments that contain both frozen and ambient specimens.  If formalin-fixed 
tissue is shipped separately (not in the same shipment as frozen specimens), then it must be 
shipped using institutional shipping supplies.    
 
1. Before packaging specimens, verify that each specimen is labeled according to the 
instructions in 5.4.2 and that lids of all primary receptacles containing liquid are tightly 
sealed.  
2. Pre-fill one of the kit chambers about 1/3 with dry ice. 
3. Prepare the frozen specimens for shipment: 
a. Place the specimens into zip-lock bags.  
b. Place the zip-lock bags into a biohazard envelope containing absorbent material.  
Expel as much air as possible before sealing the biohazard envelope. 
c. Put each biohazard envelope into a Tyvek envelope.  Expel as much air as possible 
and then seal the Tyvek envelope. 
4. Quickly place the Tyvek envelope containing frozen specimens in the kit compartment 
that is pre-filled with dry ice.  Place the Tyvek envelope on top of the dry ice.  Cover the 
specimens with additional dry ice until the compartment is almost completely full.   
5. Place the Styrofoam lid on top to secure specimens during shipment.  Do not tape the 
inner chamber shut. 
6. Prepare the ambient specimens for shipment: 
a. Seal the lids of the formalin jars with parafilm.  Place absorbent material around 
the primary container of each liquid specimen.  Place the specimens into zip-lock 
bags. 
b. Place specimens inside the secondary pressure vessel with bubble wrap.  
c. Secure the lid on the secondary pressure vessel and set it inside the kit chamber. 
7. Insert a copy of the required forms in the kit chamber with the ambient specimens. 
8. Place the Styrofoam lid on top of the kit compartment to secure specimens during 
shipment.  Do not tape the inner chamber shut. 
9. Close the outer lid of the Specimen Procurement Kit and tape it shut with durable sealing 
tape.  Do not completely seal the container. 
10. Complete a FedEx air bill and attach to top of shipping container. 
11. Complete a dry ice label. 
12. Attach the dry ice label and an Exempt Human Specimen sticker to the side of the 
shipping container. 
13. Ship specimens via overnight courier to the address below.  FedEx Priority Overnight is 
strongly recommended to prevent delays in package receipt.  
 
5.6.3 Shipping Address 
 
Ship to the address below.  Ship fresh blood specimens the same day of specimen collection.  Do 

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not ship specimens the day before a holiday. 
 
ETCTN Biorepository 
The Research Institute at Nationwide Children's Hospital 
700 Children's Drive, WA1340 
Columbus, Ohio 43205 
PH: (614) 722-2865 
FAX: (614) 722-2897 
Email: BPCBank@nationwidechildrens.org 
 
FedEx Priority Overnight service is very strongly preferred.   
 
NOTE:  The ETCTN Biorepository FedEx Account will not be provided to submitting 
institutions.  There is no central Courier account for this study.  Sites are responsible for all costs 
for shipments to the ETCTN Biorepository. 
 
5.6.4 Contact Information for Assistance 
 
For all queries, please use the contact information below: 
 
ETCTN Biorepository  
Toll-free Phone:  (800) 347-2486 
E-mail:  BPCBank@nationwidechildrens.org 
 
 
5.7 
Biomarker Plan  
 
 

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36 
List of Biomarker Assays in Order of Priority 
Priority 
Biomarker 
Name 
Assay 
(CLIA:  
Y/N) 
Use in the Trial and 
Purpose 
Specimens Tested 
Collection Time 
Points 
Mandatory 
or 
Optional 
Assay Laboratory and Lab 
PI 
Tissue-based Biomarkers 
1 
Whole Exome 
Sequencing 
NGS 
 
CLIA:  N 
Integrated 
 
To identify additional 
predictive biomarkers 
of response to 
AZD1775 (e.g. TP53, 
DNA repair 
mutations including 
ATM, ATR, ARX or 
other mutations). 
DNA from 
Esophageal cancer 
biopsy (FFPE) 
 
Archival (FFPE) 
or baseline 
M 
NCLN Genomics Laboratory 
 
Mickey Williams, Ph.D. 
 
mickey.williams@nih.gov 
2 
ɣH2AX, 
pNBS1 IFA 
with βCATN 
segmentation  
Multiplex IF 
 
CLIA:  N 
Integrated 
 
To assess target 
engagement and 
response 
Esophageal cancer 
biopsy (Frozen 
tissue) 
For expansion 
cohort only: 
 
Baseline 
 
Week -1 Day 4 
or 5 
M 
NCLN PD Assay Laboratory 
at MD Anderson 
 
Kate Ferry-Galow, Ph.D. 
 
ferrygalowkv@mail.nih.gov 
3 
p-HH3 (S20), 
p-cdk1 (Y15), 
pWEE1(S642), 
and p-
Chk1(S345) 
IHC 
IHC 
 
CLIA:  N 
Exploratory 
 
To assess target 
engagement and 
treatment response 
Esophageal cancer 
biopsy (FFPE)  
For expansion 
cohort only: 
 
Baseline 
 
Week -1 Day 4 
or 5 
M 
City of Hope 
 
Terence Williams 
 
terwilliams@coh.org 
Blood-based Biomarkers 

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Priority 
Biomarker 
Name 
Assay 
(CLIA:  
Y/N) 
Use in the Trial and 
Purpose 
Specimens Tested 
Collection Time 
Points 
Mandatory 
or 
Optional 
Assay Laboratory and Lab 
PI 
1 
Whole Exome 
Sequencing 
NGS 
 
CLIA:  N 
Integrated 
 
To identify additional 
predictive biomarkers 
of response to 
AZD1775 (e.g. TP53, 
DNA repair 
mutations including 
ATM, ATR, ARX or 
other mutations). 
DNA from 
Peripheral blood 
Baseline (within 
28 days prior to 
study entry) 
 
M 
NCLN Genomics Laboratory 
 
Mickey Williams, Ph.D. 
 
mickey.williams@nih.gov 

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38 
 
5.8 
Integrated Correlative Studies 
 
5.8.1 Whole Exome Sequencing 
5.8.1.1 
Specimens Receipt and Processing at the ETCTN Biorepository  
 
FFPE tissue blocks will be sectioned to generate an initial hematoxylin and eosin (H&E)-stained 
slide.  All H&E stained slides will undergo a pathology QA review and annotation for 
macrodissection.  Following macrodissection, tumor tissue from unstained slides will be scraped 
for co-extraction of DNA and RNA. The nucleic acids will be analyzed to determine 
concentration and quality.  Aliquots of DNA and RNA will be shipped to the central sequencing 
laboratory for analysis. 
 
DNA will be extracted from blood collected in EDTA tubes at baseline.  Blood DNA will be 
quantitated, and then stored in a -80°C freezer until shipping for analysis. 
 
5.8.1.2 
Site Performing Correlative Study 
 
Whole exome sequencing will be performed at the NCLN Genomics Laboratory under the 
supervision of Dr. Mickey Williams (mickey.williams@nih.gov)  
 
5.8.1.3 
Contact information for notification of specimen shipment 
 
Dr. Mickey Williams  
mickey.williams@nih.gov  
5.8.2 ɣH2AX, pNBS1 IFA with βCATN segmentation 
5.8.2.1 
Specimens Receipt and Processing at the ETCTN Biorepository  
 
Upon receipt at the Biorepository, the snap-frozen tissue will be accessioned, barcoded, and 
banked in a liquid nitrogen vapor phase freezer until distribution for testing. 
 
5.8.2.2 
Site Performing Correlative Study   
 
ɣH2AX, pNBS1 IFA with βCATN segmentation will be performed at the NLCN PD Laboratory 
under the supervision of Dr. Kate Ferry-Galow (ferrygalowkv@mail.nih.gov). 
 
5.8.2.3 
Contact information for notification of specimen shipment 
 
Dr. Kate Ferry-Galow  
ferrygalowkv@mail.nih.gov 
5.9 
Exploratory/Ancillary Correlative Studies 
 
5.9.1 p-HH3 (S20), p-cdk1 (Y15), pWEE1(S642), and p-Chk1(S345) IHC 
5.9.1.1 
Specimens Receipt and Processing at the ETCTN Biorepository  
 
Tissue received in formalin will be processed and embedded in paraffin, and stored as an FFPE 

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tissue block.  Samples should be removed from formalin and placed in 70% ethanol or proceed 
to paraffin embedding 24-36 hours after collection.  FFPE blocks can be stored at room 
temperature.  FFPE tissue blocks should be sent directly from the biorepository to the Williams 
Laboratory. 
 
5.9.1.2 
Site Performing Correlative Study 
 
p-HH3 (S20), p-cdk1 (Y15), pWEE1(S642), and p-Chk1(S345) immunohistochemistry will be  
performed by the Williams Laboratory at City of Hope under the supervision of Dr. Terence 
Williams (terwilliams@coh.org). 
 
5.9.1.3 
Contact information for notification of specimen shipment 
 
Samples should be shipped to the following address for analysis: 
 
Terence Williams, M.D., Ph.D. Lab  
Lipman Graff Building B202, City of Hope 
1500 E Duarte Rd. 
Duarte, CA 91010  
Cell Phone (614) 477-6086  
Lab phones (626) 218-4357 ext. 8-4357  
Email: terwilliams@coh.org 
 
Shipments should be made Monday through Wednesday only.  
• Please notify Dr. Williams by e-mail on or before the day of submission with (1) the 
name of the contact person, (2) when to expect the sample, and (3) the overnight shipping 
carrier and tracking number of the package.  
 
6. 
TREATMENT PLAN 
 
6.1 
Agent Administration 
 
Treatment will be administered on an inpatient or outpatient basis.  Reported adverse events and 
potential risks are described in Section 10.  Appropriate dose modifications are described in 
Section 7.  No investigational or commercial agents or therapies other than those described 
below may be administered with the intent to treat the patient's malignancy. 
 
Dose Escalation Schedule 
Dose Level 
Dose 
AZD1775 
(mg) 
Radiation Therapy 
(Gy) 
Level -1 
150 mg QD on Days 1 and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 

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Level 1* 
150 mg QD on Days 1, 3, and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 
Level 2 
200 mg QD on Days 1, 3, and 5 of 
weeks -1, 1, and 3 
37.5 Gy/15 fractions 
Level 3 
200 mg QD on Days 1-5 of weeks -
1, 1, and 3 
37.5 Gy/15 fractions 
* Starting dose 
 
 
Regimen Description 
Agent 
Premedications; 
Precautions 
Dose 
Route 
Schedule 
Cycle 
Length 
AZD1775 
• 5-HT3 
antagonist, 
ondansetron 
(Zofran) 8 mg 
PO QD or 
granisetron 
(Kytril) 1 mg PO 
QD prior to each 
AZD1775 dose* 
• Dexamethasone 
4mg PO** 
*** 
PO  
2-3 hours 
before radiation 
therapy in 
weeks 1 and 3 
As 
specified in 
dose 
escalation 
table 
21 days  
(3 weeks) 
 
 
 
Radiation 
Therapy 
N/A 
37.5 Gy in 
15 
fractions 
over 3 
weeks 
N/A 
QD on 
Days 1-5, 
weeks 1-3 
*May be administered by IV as needed. 
**At a minimum, administer with AZD1775 on the first day of each 3-5 day dosing period, 
unless contraindicated or not well-tolerated. 
***Doses as appropriate for assigned dose level. 
 
All patients will be required to have a 3-week washout period prior to starting systemic therapy.  
If previously on systemic therapy, a 3-week washout period is required prior to initiating study 
therapy. 
 
The first week of treatment will consist of AZD1775 monotherapy (week -1).  AZD1775 will 
then be administered orally daily 2-3 hours prior to radiation therapy according to the dose level 
schedule during weeks 1 and 3.  Patients will receive AZD1775 concurrently with radiation.  
Radiation therapy will be delivered daily, five days per week, for a total dose of 37.5 Gy in 15 
fractions during weeks 1-3.   
 
The dose escalation component will be followed by an expansion cohort consisting of 12 patients 
to confirm safety with mandatory repeat EGD (esophagogastroduodenoscopy) on week -1 during 

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41 
AZD1775 monotherapy to verify target engagement through pharmacodynamic studies.  Of note, 
assessment of biopsy tissue for target engagement will not be conducted in real time or be used 
to inform recruitment of additional patients in the expansion cohort.  During week -1, patients 
will be treated with AZD1775 at the MTD determined during dose escalation.  On Days 4 or 5 of 
week -1, the patients will undergo repeat EGD with tumor biopsy (pharmacodynamic or PD 
biopsy) prior to receiving radiation therapy.  On week 1, patients will continue AZD1775 while 
receiving radiation therapy.  On week 2, patients will only receive radiation treatment.  On week 
3, patients will resume treatment with AZD1775 in addition to receiving radiation treatment.  
Patients on the expansion cohort will also be assessed for changes in dysphagia using the Ogilvie 
dysphagia score, a validated instrument for assessing dysphagia caused by esophageal cancer (a 
5 graded scale where 0 is normal diet with no dysphagia and 4 is inability to swallow anything) 
(see Appendix E).   
 
The patient will be requested to maintain a medication diary (Appendix C) of each dose of 
medication.  The medication diary will be returned to clinic staff at the end of each course.  Non-
compliance based on assessment of the medication diary will result in removal from the study as 
documented in 6.5. 
 
6.1.1 CTEP IND Agent 
 
For patients on QD dosing, all patients must receive a 5-HT3 antagonist, ondansetron (Zofran) 8 
mg PO QD or granisetron (Kytril) 1 mg PO QD prior to each dose of AZD1775.  If nausea and 
vomiting continue, a second dose of antiemetics can be taken 8 hours later if necessary.  In 
addition, dexamethasone 4 mg PO will be given with each AZD1775 dose, as a minimum on the 
first day of dosing AZD1775 of every 3-5 days dosing period, unless contraindicated or not well-
tolerated.  Dexamethasone or the 5-HT3 antagonist may be given by IV route as needed.  
Promethazine (Phenergan), prochlorperazine (Compazine), and benzodiazepine may still be used 
as additional adjunctive treatments during AZD1775 therapy.   
Suitable alternative medications may be used, with adequate justification, in those studies where 
the use of any of the above medications might interfere with other study procedures or are 
deemed insufficient.  
  Aprepitant and fosaprepitant are not permitted due to known drug interactions with AZD1775.    
Nausea and vomiting will be considered refractory if it does not resolve to ≤ Grade 1 with 
treatment with a combination of at least 2 of the antiemetics within 24 hours.  Patients should be 
strongly encouraged to maintain liberal oral fluid intake. 
 
6.1.2 Radiation Therapy 
Radiation therapy will be administered in 15 QD fractions 5 days per week.  The prescription 
dose per fraction is 2.5 Gy.  The total prescription dose is 37.5 Gy.  Radiation therapy is 
scheduled on trial for 5 days each week of the study, generally Monday through Friday of each 
week. 

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6.1.2.1 
Immobilization and Simulation 
 
Immobilization 
Proper immobilization is critical for this protocol.  Patient setup reproducibility must be achieved  
using appropriate clinical devices.  Patients should be positioned supine in an individualized  
immobilization device on the CT table. 
 
Simulation Imaging  
Motion assessment during simulation is not required, but may be performed on patients to account 
for the tumor and nodal excursion with respiration and the appropriate type of treatment planning 
CT required. 
 
A motion management technique-specific treatment planning CT (e.g., 4DCT, breath-hold, gating, 
Active Breathing Control (ABC) etc.) can be performed during simulation to define gross tumor 
volume (GTV), clinical target volume (CTV), and planning target volume (PTV) (see definitions 
in Section 6.1.2.2).  Slice thickness should be a maximum of 3-5 mm in slice thickness for regions 
including gross disease and 8-10 mm for other areas.  A uniform 5 mm thickness is also acceptable. 
The field of view must be large enough so that none of the patient’s anatomy along the path of the 
treatment beams is excluded.   
 
A treatment planning FDG-PET/CT scan (or FDG-PET alone) (if available for simulation) with 
the patient in the treatment position is encouraged for treatment planning, but not required. In the 
case where the CT is obtained in the treatment position as part of radiation simulation, the CT from 
this study may be used as the planning CT scan.  CT based treatment planning should include, 
superiorly, the entire thorax, and, inferiorly, below the kidneys.  In preparation for the planning 
CT scan, patients will be asked to fast for 2 hours prior to simulation and treatment to minimize 
the volume of stomach contents.   
 
Intravenous and esophageal contrast during the planning CT are optional at the discretion of the 
treating physician.  If contrast is used, the densities or RLSPs should be over-ridden or the contrast 
scan must be registered to a non-contrast scan for planning purposes. 
 
 
6.1.2.2 
Definition of Target Volumes and Margins 
 
Detailed Specifications  
Target volumes: The definitions of volumes will be in accordance with the 1999 
ICRUReport#62. 
 
GTVp_3750, IGTVp_3750:  The GTVp includes the GTV defined as the primary tumor in the 
esophagus. GTVp should be contoured on the primary CT dataset.  If 4DCT simulation is 
performed for nongated free-breathing delivery, the phases of the breath cycle should be used to 
construct the composite GTV volume (IGTVp).  Ideally, targets should be contoured on all phase 
images in order to construct IGTV.  Some commercial software like MIM may be used to 
populate the physician’s GTV contour on one-phase images to the other phase images. If such 
software is unavailable, at least 4 phase images (end of inspiration, mid-inspiration, end of 

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43 
expiration and mid-expiration) should be used to assure IGTV accuracy. Maximum Intensity 
Projected (MIP) images should not be used to generate IGTV (Chen et al., 2014).  
 
GTVn_3750, IGTVn_3750:  The GTVn is defined as any grossly involved regional lymph 
nodes, either considered suspicious on FDG-PET/CT or proven by EUS biopsy, which is 
included at the discretion of the treating radiation oncologist. GTVn should be contoured on the 
primary CT dataset.  If 4DCT simulation is performed, the phases of the breath cycle should be 
used to construct the composite GTV volume (IGTVn).  Ideally, targets should be contoured on 
all phase images in order to construct IGTVn.  Some commercial software like MIM may be 
used to populate the physician’s GTV contour on one phase images to the other phase images.  If 
such software is unavailable, at least 4 phase images (end of inspiration, mid-inspiration, end of 
expiration and mid-expiration) should be used to assure IGTVn accuracy.  Maximum Intensity 
Projected (MIP) images should not be used to generate IGTVn (Chen et al., 2014).  
 
GTV_3750, IGTV_3750:  structure combined to include GTVp_3750 and GTVn_3750 for 
GTV_3750 or IGTV_3750 which will include IGTVp_3750 plus IGTVn_3750 if 4DCT 
simulation is performed. 
 
CTV_3750:  The CTV is defined as the GTV_3750 or IGTV_3750 with a 0.5 cm uniform 
expansion and edited back as needed for structures where there is unlikely to be invasion. 
Elective nodal radiation should not be performed.  
 
PTV_3750:  Additional margin shall be added to the CTV for set up error and movement.  This 
expansion should be 0.5 to 1.0 cm and does not need to be uniform in all dimensions. 
 
 
6.1.2.3 
Definition of Critical Structures and Margins 
Contouring and dose assessment of the following normal structures is encouraged. 
 
Standard Name 
 
Description 
 
SpineCanal 
Spinal Canal 
Lungs 
Right + Left Lung  
Lung_R 
Right Lung 
Lung_L 
Left Lung 
Heart 
Heart 
Liver  
Liver 
Kidney_R 
Kidney_L 
Right Kidney 
Left Kidney 
Bowel_Small 
Small bowel 
Bowel_Large 
Large bowel 
Stomach 
Stomach 
 
 
Detailed Specifications 
SpineCanal:  Boundaries:  Cranial:  1st slice of CT; Caudal:  last slice of CT; 

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Lungs:  Boundaries:  Cranial:  From apex bilaterally; Caudal:  to bottom of L2 
Heart:  Boundaries:  Base:  Bottom of the aortic arch; Inferior:  Apical most of the 
ventricle 
Liver:  Boundaries:  Cranial:  From dome; Caudal:  Inferior tip 
Kidneys:  Entire kidneys contoured separately as Kidney_R and Kidney_L 
Bowel_Small:  Boundaries:  Level of PTV 
Bowel_Large:  Boundaries:  Level of PTV  
Stomach: Entire stomach including cardia, fundus, body, antrum, and pylorus.  
 
 
6.1.2.4 
Dose Prescription 
The PTV_3750 should receive a dose of 37.5 Gy in 15 fractions.  This table should be used as a 
guide for treatment planning purposes.   
 
Structures 
Goal 
Heart 
V42 <10 cc 
Bowel_Small 
(including duodenum) 
V39 <05 cc 
Bowel_Large 
V45 <10 cc 
Stomach 
V39 <50 cc 
Liver 
V30 <700 cc 
Kidney_R, Kidney_L 
V37.5 <15 cc 
Lungs 
V16 <950 cc 
SpineCanal 
V39 <05 cc 
Target Volume 
Goal 
PTV_3750 
D1cc < 110% 
V100% ≥ 95% 
 
6.1.2.5 
Treatment Technology 
Photon therapy is required using either 3D conformal or intensity modulated radiation therapy 
(IMRT).  IMRT may be delivered using multiple fixed fields employing dynamic multi-leaf 
collimator, helical arc therapy, or volumetric modulated arc therapy.  Proton therapy is not 
permitted. 
 
6.1.2.6 
Online treatment image guided localization and treatment 
 
Image-guided radiation should be performed daily and can be accomplished with either 2-
dimensional kilovoltage (KV) images or 3-dimensional cone beam imaging at the discretion of 
the treating radiation oncologist.   
6.2 
Definition of Dose-Limiting Toxicity 
 
Management and dose modifications associated with the above adverse events are outlined in 
Section 7. 
 
The DLT observation window will include the entire duration of AZD1775 monotherapy and 
radiation followed by a 3-week break/DLT evaluation period.  DLTs will be graded in severity 
according to CTCAE v5.0.  A DLT will be defined as an adverse event that is related (possibly, 
probably, or definitely) to administration of AZD1775 and fulfills one of the following criteria: 
 

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• grade 4 neutropenia for ≥5 days or resulting in hospitalization for treatment of 
neutropenic fever 
• grade 3 or 4 thrombocytopenia for ≥4 days 
• grade ≥3 diarrhea if it is refractory to treatment as outlined in Section 6.4.4.1, and unable 
to be corrected to Grade 2 or less within 24 hours.  Bloody or Grade 4 diarrhea will be 
dose-limiting. 
• grade 4 esophagitis 
• 
Any other grade 3 or 4 nonhematologic toxicity owing to treatment (with the exception of 
alopecia, nausea, and vomiting) requiring therapy to be held for >1 week, or recurrent or 
unacceptable grade 3 or 4 toxicity due to the treatment.  Grade 3 nausea and vomiting 
will only be considered dose-limiting if it is refractory to anti-emetic therapy and unable 
to be corrected to Grade 1 or less within 24 hours. 
• 
Rise in creatinine to Grade 3, not corrected to Grade 1 or less within 48 hours with IV 
fluids will be considered dose-limiting.  All Grade 4 rises in creatinine will be dose 
limiting. 
• Grade ≥ 3 metabolic toxicities unable to be corrected to Grade 1 or baseline within 48 
hours (hypocalcemia or hypercalcemia, hypomagnesemia or hypermagnesemia, and 
hyponatremia) will be considered dose limiting.  For hypokalemia or hyperkalemia, grade 
≥ 2 toxicities unable to be corrected to grade 1 or less within 48 hours will be considered 
dose limiting.  Grade 4 metabolic toxicities that are symptomatic will be considered dose-
limiting regardless of duration or ability to correct. 
• Failure to tolerate 90% of the dosing (both radiation therapy and AZD1775) and failure to 
remain on the study for 21 days due to toxicity will be considered a DLT. 
• Dose-limiting hematologic and non-hematologic toxicities will be defined differently and 
will be based on events that occur during study drug administration and for 3 weeks after 
the completion of radiation therapy (total of six weeks).   
 
We will employ the Bayesian optimal interval (BOIN) design to find the MTD (Liu and Yuan, 
2015; Yuan et al., 2016) (see Section 9.1).  Dose-limiting toxicity (DLT) is defined above.   
 
For the BOIN design, the target toxicity rate for the MTD is 25%.  Patients will be enrolled and 
treated in cohorts of size 2 starting at dose level 1.  The BOIN design does not require a fixed 
cohort size and allows for decision making at any time during the trial by comparing the observed 
DLT rate at the current dose with the escalation and de-escalation boundaries.  Therefore, the 
subsequent cohort size could be subject to change depending on the DLT data observed (but will 
not exceed 4).   
 
6.3 
Dose Expansion Cohorts: 
 
Once the MTD is reached, an additional 12 patients will be treated at this dose.  For the 
expansion cohort, patients will continue to be monitored for occurrence of DLT.  We will use the 
elimination boundaries in Table 2 for toxicity monitoring. If 3 of the first 5 patients or if ≥4 of 6 
patients experience DLT, the Principal Investigator will discuss with all study investigators and 
with CTEP whether further addition of patients is needed to re-assess the MTD.  Monitoring of 
all safety and toxicity data is done by the Principal Investigator and the Corresponding 
Organization on a real-time basis as data are entered into Medidata Rave using the Web 

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Reporting Module.  All participating sites are expected to notify the Principal Investigator when 
a DLT has occurred. 
 
6.4 
General Concomitant Medication and Supportive Care Guidelines 
 
Because there is a potential for interaction of AZD1775 with other concomitantly administered 
drugs, the case report form must capture the concurrent use of all other drugs, over-the-counter 
medications, or alternative therapies.  The Principal Investigator should be alerted if the patient is 
taking any agent known to affect or with the potential for drug interactions.  The study team 
should check a frequently-updated medical reference for a list of drugs to avoid or minimize use 
of.  Appendix D (Patient Drug Interactions Handout and Wallet Card) should be provided to 
patients. 
 
6.4.1 Interactions with CYP proteins 
 
AZD1775 is a weak reversible inhibitor of CYP3A4 (IC50 14 mcM).  A formal clinical drug 
interaction study (D6014C00006) evaluating the effect of AZD1775 on PK of substrates for 
CYP3A, CYP2C19, and CYP1A2 has been conducted in advanced solid tumor patients.  The 
results of this study suggest that AZD1775 is a weak inhibitor of these enzymes and is unlikely 
to result in clinically significant drug interaction.  
 
In vitro studies suggest that the major pathways of the metabolism of AZD1775 in humans 
involve CYP3A4.  Potent or moderate inhibitors or inducers of CYP3A4, sensitive CYP3A4 
substrates, and CYP3A4 substrates with a narrow therapeutic window should be avoided until 
additional data on drug-drug interaction become available. 
 
In vitro data suggests that AZD1775 may be a weak reversible inhibitor of CYP2C19 (IC50 12 
mcM).  Caution should therefore be exercised when AZD1775 is co-administered with agents 
that are sensitive substrates of CYP2C19, or substrates of this enzyme with a narrow therapeutic 
range. 
 
6.4.2 Interactions with protein transporters  
 
Transporter studies (in vitro) have shown that AZD1775 is both a substrate and inhibitor (IC50 20 
mcM) of P-glycoprotein (P-gp).  Maximum impact of these finding is likely to occur for drugs 
administered orally at the same time as AZD1775.  Caution should therefore be exercised when 
agents that are inhibitors or substrates of P-gp are administered concomitantly with AZD1775. 
 
Recent in vitro transporter studies have shown AZD1775 to be an inhibitor of breast cancer 
resistance protein (BCRP) (IC50 5.1 mcM).  This finding is particularly relevant for drugs 
administered orally where exposure is normally limited by BCRP-mediated efflux, in particular 
some statins such as rosuvastatin.  Other drugs where the disposition is mediated via BCRP 
should be administered with caution, dose modification should be considered, or the drug should 
be substituted by an alternative drug.   
 
AZD1775 has been shown to be an inhibitor of multidrug and toxin extrusion protein 1 

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(MATE1) (IC50 0.132 mcM) and multidrug and toxin extrusion protein 2 (MATE2K) 
transporters (IC50 0.307 mcM).  A drug interaction with substrates of either transporter cannot be 
ruled out, the most important substrate known to date being metformin. 
 
6.4.3 Drug-drug interactions 
 
An exploratory assessment of the effect of aprepitant on AZD1775 exposure in oncology patients 
suggests that there is a drug interaction between AZD1775 and aprepitant, as exposure to 
AZD1775 increased by ~40% when aprepitant was co-administered with AZD1775.  The 
observed increase in AZD1775 exposure is likely the result of CYP3A4 inhibition by aprepitant.  
This increase in exposure is statistically significant.  At the selected MTDs, this increase may 
also be of clinical importance.  Therefore, it is recommended that AZD1775 should not be co-
administered with aprepitant (or a prodrug of aprepitant) until further evaluation. 
 
The administration of AZD1775 with substances known to prolong the QTc interval is not 
recommended.  Special note: the potential risk of concomitant use of AZD1775 with ondansetron 
(known to prolong the QTc interval) should be taken into account. 
 
Potent or moderate inhibitors or inducers of CYP3A4, sensitive CYP3A4 substrates, and 
CYP3A4 substrates with a narrow therapeutic window should be avoided until additional data on 
drug-drug interactions (DDI) becomes available.  The use of sensitive substrates of CYP3A4, 
such as atorvastatin, simvastatin and lovastatin, is prohibited in this study.  As grapefruit and 
Seville oranges are known to contain moderate inhibitors of CYP3A4, these fruits or their 
products (including marmalade, juice, etc.) should be avoided while taking AZD1775. 
 
6.4.4 AE Management guidelines 
 
6.4.4.1 
Diarrhea 
 
If diarrhea develops and does not have an identifiable cause other than study drug administration 
or radiation therapy, anti-diarrheals such as Lomotil (diphenoxylate HCl 2.5 mg + atropine 
sulfate 0.025 mg/tablet) dosed according to package insert or loperamide 4 mg po after the first 
unformed stool with 2 mg po with every 2 hours as long as unformed stools continue (4 mg 
every 4 hours while asleep).  No more than 16 mg of loperamide should be taken during a 24-
hour period.  This regimen can be repeated for each diarrheal episode.  Diarrhea will be 
considered refractory if it does not resolve within 24 hours ≤ to Grade 2 with the above 
medications.  If the patient develops blood or mucus in the stool, dehydration, or hemodynamic 
instability, or fever along with the diarrhea, anti-diarrheals will be discontinued and the patient 
will be treated with IV fluids and antibiotics as medically indicated. 
 
6.4.5 Neutropenia 
 
To reduce the risk of severe myelosuppression events, a complete blood count (CBC) should be 
performed weekly.  Febrile neutropenia is a life-threatening complication requiring 
hospitalization and urgent broad spectrum antibiotics, as well as an aggressive search for the 
source and microbial cause of the episode.  Growth factors to prevent neutropenia will not be 

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administered prophylactically.  If necessary, they may be administered according to accepted 
American Society of Clinical Oncology (ASCO) guidelines to allow re-treatment. 
 
6.4.6 Anemia 
 
Symptomatic anemia should be treated with red blood cell transfusion and is recommended if the 
hemoglobin falls below 8 g/dL.  The initiation of erythropoietic therapy for the management of 
chemotherapy-induced anemia follows the American Society of Hematology/ASCO clinical 
practice guidelines (http://www.asco.org). 
 
6.4.7 Thrombocytopenia 
 
Thrombocytopenia will be treated conservatively.  In the absence of bleeding, or a necessary 
invasive procedure, platelet transfusions should be given for a platelet count ≤10,000/mm3.  If 
invasive procedure(s) is (are) planned, or the patient develops bleeding, platelet transfusions 
should be administered in accordance with the standard of practice, usually maintaining a platelet 
count above 50,000/mm3. 
 
6.4.8 Cardiac disorders 
 
AZD1775 partially reversibly inhibited the hERG current with an IC50 value of 6.9 mcM in 
whole-cell voltage clamp studies.  Tachycardia, and QTc prolongation have been observed in 
clinical studies with AZD1775, either as monotherapy or in combination with cytotoxic 
chemotherapy. 
 
Attention should be paid to any detected increases in QTc interval (See Section 7.2).   
 
6.4.9 Gastrointestinal hemorrhage 
 
Gastrointestinal hemorrhage is a potential risk for which a causal association with AZD1775 has 
not yet been established.  As of 11 November 2019, AstraZeneca has received five SAEs of 
gastrointestinal hemorrhagic events originating from AstraZeneca sponsored studies in 
AZD1775 monotherapy (Investigator’s Brochure 2020).   
 
Hematemesis and/or melaena are clear indicators of gastrointestinal hemorrhage.  Localization 
and control of the bleeding should be the primary method of medical intervention.  When 
required, transfusions may be indicated.  Where investigations for sub-clinical or occult 
gastrointestinal hemorrhage prove negative, other causes of refractory anemia should be 
considered and investigated. 
 
6.4.10 Infections and Infestations (Sepsis) 
 
Sepsis has been added as an important potential risk for AZD1775.  As part of the normal sepsis 
surveillance of oncology patients on known myelosuppressive treatments, Investigators should 
also be aware of the possibility of sepsis arising in association with the use of AZD1775 and be 
prepared to reduce dosing of AZD1775 or withdraw it where a causal relationship is suspected. 

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49 
 
Sepsis can be so severe that it could lead to death.  Patient discussions and patient's decisions must be 
documented about: 1) whether the physician has determined that therapy is beneficial and, if so, 2) 
whether or not the patient has chosen to continue the treatment. 
 
6.4.11 Nausea and vomiting (Emesis) 
 
All patients on QD dosing will receive anti-emetics as described in Section 6.1.1.  
 
6.5 
Duration of Therapy 
 
In the absence of treatment delays due to adverse event(s), treatment may continue for one cycle 
or until one of the following criteria applies: 
 
• 
Disease progression 
 
• 
Intercurrent illness that prevents further administration of treatment 
 
• 
Unacceptable adverse event(s) 
 
• 
Patient decides to withdraw from the study 
 
• 
General or specific changes in the patient's condition render the patient unacceptable for 
further treatment in the judgment of the investigator 
 
• 
Clinical progression 
 
• 
Patient non-compliance 
 
• 
Pregnancy 
 
o All women of child bearing potential should be instructed to contact the investigator 
immediately if they suspect they might be pregnant (e.g., missed or late menstrual 
period) at any time during study participation. 
 
o The investigator must immediately notify CTEP in the event of a confirmed 
pregnancy in a patient participating in the study. 
 
• 
Termination of the study by sponsor 
 
• 
The drug manufacturer can no longer provide the study agent 
 
The reason(s) for protocol therapy discontinuation, the reason(s) for study removal, and the 
corresponding dates must be documented in the Case Report Form (CRF). 
 

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6.6 
Duration of Follow-Up 
 
Patients will be examined 3 weeks after completing treatment, then followed every 3 months by 
a doctor visit for the first 2 years after completion of therapy, after removal from study, or until 
death, whichever occurs first.  Patients will then be followed every 6 months for 3 years by 
doctor visit.  Patients removed from study for unacceptable adverse event(s) will be followed 
until resolution or stabilization of the adverse event. 
 
 
7. 
DOSING DELAYS/DOSE MODIFICATIONS 
 
 
Dose Level 
AZD1775 Dose 
-1 
150 mg QD on Days 1 and 5 of weeks -1, 1, 
and 3 
1* 
150 mg QD on Days 1, 3, and 5 of weeks -1, 1, 
and 3 
2 
200 mg QD on Days 1, 3, and 5 of weeks -1, 1, 
and 3 
3 
200 mg QD on Days 1-5 of weeks -1, 1, and 3 
* Starting dose 
 
Patients will be allowed up to two dose reductions. 
 
Dose adjustments will be based on the organ system exhibiting the greatest degree of toxicity.  
Dose reductions or holds and initiation of supportive care are allowed as clinically indicated by 
the treating physician. 
If further dose modification is considered necessary, a discussion must take place with the 
Medical Monitor and the Medical Monitor needs to provide approval for the patient to continue.  
Any patient requiring a toxicity-related dose delay of more than 21 days from the intended day of 
the next scheduled dose must be discontinued from the study unless there is approval from the 
Medical Monitor for the patient to continue. 
7.1 
Dose modifications due to hematologic toxicity for adavosertib monotherapy 
 
Complete blood counts (CBC) will be obtained for all patients at the beginning of each 
treatment week and reviewed prior to dose administration.  If hematologic toxicity occurs, 
treatment should be held, and ANC and platelets should be monitored at least weekly.  Patients 
should be managed as medically indicated.  Treatment should not be resumed until recovery to 
Grade 1 (ANC ≥1.5 x 109/L or 1,500/ mcL; platelets ≥75,000/ mcL). 
 
 

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51 
Table 1. Neutrophil and Platelet Blood Counts and Study Drug Action  
Neutrophil count 
Action 
Adavosertib 
 
2nd 
event 
Action 
Adavosertib 
 
3rd 
event 
Action 
Adavosertib 
 
Grade 2  
<1.5-1.0 x109/L 
Investigator judgement to continue adavosertib or interrupt until Grade 1 
(≥1.5-1.0 x 109/L) 
 
 
Grade 3 
<1.0-0.5 x109/L 
Hold 
Resume at same 
dose 
 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
2 
Grade 3 
<1.0-0.5 x109/L 
with documented 
infection and/or 
fever 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
2 
 
Hold 
Contact 
Medical 
Monitor 
Grade 4 
<0.5 x109/L 
 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
2 
 
Discontinue 
and follow for 
disease 
progression 
Grade 4  
Febrile 
neutropenia or 
Grade 4 Infection 
with neutropenia 
Discontinue and 
follow for 
disease 
progression 
 
 
 
 
Platelet count 
Action 
Adavosertib 
 
2nd 
event 
Action 
Adavosertib 
 
3rd 
event 
Action 
Adavosertib 
 
Grade 2  
<75,000-50,000/ 
μL 
Hold 
Resume at same 
dose 
 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
1 

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52 
Table 1. Neutrophil and Platelet Blood Counts and Study Drug Action  
Grade 3 
<50,000- 25,000/ 
μL 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
2 
 
Hold 
Resume at DL-
2 
Grade 4 
 <25,000/ μL 
without any 
evidence of 
bleeding 
Hold 
Resume at DL-
1 
 
Hold 
Resume at DL-
2 
 
Discontinue 
and follow for 
disease 
progression 
Thrombocytopenic 
haemorrhage 
(gross occult 
bleeding) 
associated with 
platelet count 
<50,000/ μL 
Discontinue and 
follow for 
disease 
progression 
 
 
 
 
DL-1 = One dose level lower; DL-2 = Two dose levels lower 
 
Please consider using G-CSF in the event of severe neutropenia or febrile neutropenia according 
to institutional standards.  
 
No more than two dose reductions will be allowed for any patient.  Patients requiring further 
dose reduction due to toxicity must discontinue study treatment.  Dose re-escalation is not 
allowed.  If the patient has concurrent neutropenia and thrombocytopenia, please follow the most 
conservative guidance in the table below and discuss with Medical Monitor as needed. 
 
If hematological parameters do not recover within 21 days, the patient should be removed from 
the study treatment.   
 

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53 
Neutropenia, infection, febrile neutropenia dose modifications and management 
Any day 
 
Grade 3 neutropenic fever (ANC <1000/µL + 
Temperature ≥101°F [38.5°C]) or neutropenic 
infection 
Documented infection with Grade 3 
neutropenia (ANC <1000/µL)  
Grade 4 neutropenia 
(ANC <500/μL >7 days 
Grade 4 thrombocytopenia (platelet count 
<25,000/μL >7 days) 
 
Hold dose until recovery.   
Then, upon resuming dosing, reduce 
AZD1775a.  
 
Grade 4 febrile neutropenia or Grade 4 
infection with neutropenia (both defined as 
septic shock) 
Thrombocytopenic haemorrhage (gross occult 
bleeding) associated with a platelet count 
<50,000/μL 
 
Discontinue treatment and follow for disease 
progression. 
 
a 
No more than two dose reduction will be allowed for any patient.  Patients requiring an 
additional dose modification to AZD1775 due to toxicity will discontinue study treatment. 
 
 
7.2 
Non-hematologic toxicity dose modifications 
 
Substantial acute toxicities should be managed as medically indicated and with temporary 
suspension of investigational product, as appropriate.  Dose reductions or holds and initiation of 
supportive care are allowed as clinically indicated by the treating physician.  Dose reductions of 
AZD1775 should be considered if the toxicity is considered to be related to AZD1775, i.e. in 
monotherapy studies or in combination studies if relationship cannot be wholly attributed to the 
combination agent (each combination agent should be considered on an individual basis).  Dose 
re-escalation is not permitted.  In general, if a patient experiences a G1/G2 non-hematological 
toxicity, no dose modification is required.  If a patient experiences a G3 or G4 toxicity which is 
not attributable to the disease or disease-related processes under investigation, dosing will be 
interrupted and/or the dose reduced, and supportive therapy administered as required.   Any 
patient who develops a Grade 3 or 4 non-hematologic toxicity that does not resolve to ≤ Grade 1 
within 21 days should be removed from the study treatment unless approved by the Medical 
Monitor.   
 
Table 2:  QTc interval prolongation (AZD1775 dose modifications) 
QTc Value (triplicate)a, b 
Action to be taken 
 

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54 
QTc 481-500 ms (Grade 2) c 
 
Hold treatment for up to 21 days.  Consider seeking 
cardiologist’s advice.  Once QTc interval has returned to 
Grade 1 and correction of possible electrolyte imbalance 
has been made, treatment may resume at the same dose 
level.  
QTc ≥ 501 ms or 
Shift from baseline of ≥ 
60ms (Grade 3)c 
Hold treatment for up to 21 days.  Seek cardiologist’s 
advice and correct any electrolyte imbalance. 
- 
If QTc interval recovers to ≤ 480 
ms (Grade 1) or baseline if there 
was a shift from baseline of ≥ 
60ms, then may resume treatment at 
next lower dose level, if 
cardiologist agrees. 
- 
If QTc interval doesn’t recover to ≤ 
480 ms or baseline if there was a 
shift from baseline of ≥ 60ms, then 
discontinue treatment permanently 
Torsade de pointes or 
polymorphic ventricular 
tachycardia or 
signs/symptoms of serious 
arrhythmia (Grade 4)c 
 
  
Discontinue treatment permanently.  Seek cardiologist’s 
advice.  
a Mean triplicate value 
b Based on a dedicated QT study conducted to evaluate the effect of single & multiple doses of AZD1775 225 mg on QT 
prolongation in advanced solid tumor patients, there was no significant relationship found between ΔQTcF and AZD1775 
concentration.  Model prediction of change in QTcF at the geometric mean of Cmax on Day 1 (712.8 nM) was -2.405 msec (90% 
CI: -5.939, 1.129).  Similarly, model prediction of QTcF at the geometric mean Cmax on Day 3 (1462 nM) was -0.7568 msec 
(90% CI: -5.148, 3.634).  The upper limit of the predicted 90% CIs for ∆QTcF were well below 10 msec, supporting the 
conclusion that AZD1775 does not have a clinically relevant impact on cardiac repolarization.   
c Per CTCAE V 5.0 
ECG should be assessed if electrolyte abnormalities are detected or suspected (e.g. in relation to gastrointestinal 
toxicity).  If a prolongation of the QTc interval is detected this must be closely monitored as clinically indicated. 
 
7.3 
Dose modifications to radiation therapy are defined below: 
 
▪ Radiation therapy will typically continue as prescribed without modifications while drug-
related adverse events are being managed. 
 
▪ Radiation therapy will be interrupted for grade >3 non-hematologic toxicity at least 
possibly related to radiation therapy.  If the patient develops > grade 3 toxicity at least 
possibly related to radiation therapy, radiation therapy and AZD1775 should be withheld.  
Interruption of therapy may continue until the toxicity has regressed to grade ≤2 to allow 
resumption of therapy; however, every effort should be made to limit treatment 

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55 
interruptions to 1-2 weeks.  
 
▪ Management of radiation-related toxicity is left to the discretion of the treating radiation 
oncologist. 
 
▪ Missed radiation treatments should ideally be made up if reasonable to do so, such that 
the total dose delivered is 37.5 Gy in 15 fractions.  Ultimate discretion to complete the 
radiation course is left to the treating physician team. 
 
▪ In the unlikely event that radiation must be permanently discontinued, patients will be 
taken off-study. 
 
Patients experiencing grade >2 toxicity during treatment are required to visit the study doctor 
after completion of treatment every other week until toxicity resolves to grade ≤2. 
Definition of Complete Course: 
 
A cycle is defined as 21 days.  If a patient receives 90% of scheduled treatments of AZD1775 
and radiation therapy and remains in the study until D21, the patient will be considered to have 
completed the therapy.  Patients who do not complete the therapy for reasons other than toxicity 
will be replaced.  Patients who do not complete the therapy due to toxicity will not be replaced.  
All patients should have completed therapy (unless removed from the study due to toxicity) to be 
evaluable. 
 
 
8. 
PHARMACEUTICAL INFORMATION 
 
A list of the adverse events and potential risks associated with the investigational agent 
administered in this study can be found in Section 10.1. 
 
8.1 
AZD1775 (NSC 751084) 
 
Chemical Name: 2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-6-[4-(4-methylpiperazin-1-
yl)anilino]pyrazolo[3,4-d]pyrimidin-3-one hemihydrate 
 
Other Names: MK-1775, adavosertib 
 
Classification: inhibitor of WEE1 kinase 
 
CAS: 1277170-60-1 
 
Molecular Formula:  C27H32N8O2·0.5H2O  
M.W.: 500.6 
 
Mode of Action: AZD1775 inhibits WEE1 which phosphorylates and inhibits cyclin-dependent 
kinases 1 (CDK1) and 2 (CDK2), and is involved in regulation of the intra-S and G2 cell cycle 
checkpoints.  In in vitro and in vivo preclinical models, AZD1775 selectively enhanced 
chemotherapy induced death of cells deficient in p53 signaling. 

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56 
 
Description: AZD1775 is a crystalline, hemihydrate form of the drug substance. 
 
How Supplied: AZD1775 is supplied by AstraZeneca and distributed by the Pharmaceutical 
Management Branch, CTEP/DCTD/NCI as capsules available in 25 mg (yellow color, size 2 gelatin 
capsule) and 100 mg (orange color, size 2 gelatin capsule) strengths.  The dry-filled capsules consist 
of a roller-compacted granule of drug substance, lactose monohydrate, microcrystalline cellulose, 
croscarmellose sodium, and magnesium stearate.  Each high density polypropylene (HDPE) bottle 
contains 20 capsules. 
 
Storage: Store at 2 to 30˚C (36 to 86˚F).  Do not freeze. 
If a storage temperature excursion is identified, promptly return AZD1775 to between 2-30°C and 
quarantine the supplies.  Provide a detailed report of the excursion (including documentation of 
temperature monitoring and duration of the excursion) to PMBAfterHours@mail.nih.gov for 
determination of suitability. 
 
Stability:  Shelf-life stability studies of AZD1775 capsules are ongoing. 
 
Route of Administration: Oral administration.  Capsules should not be opened. 
 
Potential Drug Interactions: AZD1775 is primarily metabolized by CYP3A4 and is a weak, time-
dependent inhibitor of CYP3A4. Avoid concomitant CYP3A4 moderate or strong 
inhibitors/inducers, and sensitive substrates with a narrow therapeutic index.  AZD1775 is also a 
weak inhibitor of CYP2C19.  Caution should be exercised with concomitant administration of 
sensitive substrates or substrates with a narrow therapeutic index. 
 
In vitro transporter studies have shown that AZD1775 was an inhibitor of OATP1B1, OATP1B3, 
MATE1, MATE2K, P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), and a 
substrate for P-gp and BCRP.  The PK parameters of AZD1775 could be altered if AZD1775 is 
coadministered with P-gp and BCRP inhibitors/inducers, and there is potential for drug-drug 
interactions when coadministered with OATP1B1, OATP1B3, MATE1, MATE2K, P-gp and 
BCRP substrates.  This finding is particularly relevant for drugs administered orally where 
exposure is normally limited by BCRP-mediated efflux, in particular some statins.  Modelling has 
predicted a substantial increase in the exposure of atorvastatin when coadministered with 
AZD1775 and the use of atorvastatin is therefore prohibited. 
 
 
Availability 
 
AZD1775 is an investigational agent supplied to investigators by the Division of Cancer 
Treatment and Diagnosis (DCTD), NCI.  AZD1775 is provided to the NCI under a Collaborative 
Agreement between the Pharmaceutical Collaborator and the DCTD, NCI (see Section 13.5). 
 
8.1.1 Agent Ordering and Agent Accountability 
 

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57 
8.1.1.1 
NCI-supplied agents may be requested by eligible participating Investigators (or their 
authorized designee) at each participating institution.  The CTEP-assigned protocol 
number must be used for ordering all CTEP-supplied investigational agents.  The 
eligible participating investigators at each participating institution must be registered 
with CTEP, DCTD through an annual submission of FDA Form 1572 (Statement of 
Investigator), NCI Biosketch, Agent Shipment Form, and Financial Disclosure Form 
(FDF).  If there are several participating investigators at one institution, CTEP-
supplied investigational agents for the study should be ordered under the name of one 
lead participating investigator at that institution. 
 
In general, sites may order initial agent supplies when a subject is being screened for 
enrollment onto the study. 
Submit agent requests through the PMB Online Agent Order Processing (OAOP) 
application.  Access to OAOP requires the establishment of a CTEP Identity and 
Access Management (IAM) account and the maintenance of an “active” account 
status, a “current” password, and active person registration status.  For questions 
about drug orders, transfers, returns, or accountability, call or email PMB any time.  
Refer to the PMB’s website for specific policies and guidelines related to agent 
management.   
 
8.1.1.2 
Agent Inventory Records – The investigator, or a responsible party designated by the 
investigator, must maintain a careful record of the receipt, dispensing and final 
disposition of all agents received from the PMB using the appropriate NCI 
Investigational Agent (Drug) Accountability Record (DARF) available on the CTEP 
forms page.  Store and maintain separate NCI Investigational Agent Accountability 
Records for each agent, strength, formulation and ordering investigator on this 
protocol. 
 
8.1.2 Investigator Brochure Availability 
 
The current versions of the IBs for the agents will be accessible to site investigators and research 
staff through the PMB OAOP application.  Access to OAOP requires the establishment of a 
CTEP IAM account and the maintenance of an “active” account status, a “current” password and 
active person registration status.  Questions about IB access may be directed to the PMB IB 
Coordinator via email. 
 
8.1.3 Useful Links and Contacts 
• CTEP Forms, Templates, Documents:  http://ctep.cancer.gov/forms/ 
• NCI CTEP Investigator Registration:  RCRHelpDesk@nih.gov 
• PMB policies and guidelines: 
http://ctep.cancer.gov/branches/pmb/agent_management.htm  
• 
PMB Online Agent Order Processing (OAOP) application:  
https://ctepcore.nci.nih.gov/OAOP 
• CTEP Identity and Access Management (IAM) account: https://ctepcore.nci.nih.gov/iam/  
• CTEP IAM account help:  ctepreghelp@ctep.nci.nih.gov  
• IB Coordinator:  IBCoordinator@mail.nih.gov  

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58 
• PMB email:  PMBAfterHours@mail.nih.gov 
• PMB phone and hours of service: (240) 276-6575 Monday through Friday between 8:30 
am and 4:30 pm (ET) 
 
9. 
STATISTICAL CONSIDERATIONS 
 
9.1 
Study Design/Endpoints 
 
This is a phase 1 study to evaluate the safety of adding a WEE1 kinase inhibitor, AZD1775, to 
palliative radiation therapy followed by a 12-patient expansion cohort to verify safety and target 
engagement at the MTD.  The phase 1 portion of the study will employ a BOIN design where the 
target toxicity rate for the MTD is 25% with 75% dose-elimination cut-off and the maximum sample 
size is 18 DLT evaluable patients.  Patients will be enrolled and treated in cohorts of size 2 starting 
at dose level 1 with the study design described as follows:  
 
1. The first cohort of 2 patients is treated at dose level 1. 
2. A dose escalation/de-escalation will be conducted according to the rule displayed in Table 3 
below to assign a dose to the next cohort of patients (subsequent cohort size may be subject 
to change, but will not exceed 4).  First find the column for the total number of patients 
treated at the current dose in Table 3 below: 
a. If the cumulated total # of DLTs at the current dose is less than or equal to n1 in the 
column of the corresponding total number of patients treated at the current dose, there 
will be dose escalation.  
b. If the cumulated total # of DLTs at the current dose is greater than or equal to n2 in 
the column of the corresponding total number of patients treated at the current dose, 
there will be dose de-escalation.  
c. If the cumulated total number of DLTs at the current dose is greater than or equal to 
n3, the current and higher doses will be eliminated.  “Eliminate” means that the 
current and higher doses will be eliminated from the trial to prevent treating any 
future patients at these doses because they are overly toxic.  
d. When a dose is eliminated, there will automatically be de-escalation from the dose to 
the next lower level.  When the lowest dose is eliminated, the trial will be stopped for 
safety.  In this case, no dose should be selected as the MTD.  
e. If none of the actions (i.e., escalation, de-escalation or elimination) is triggered, new 
patients will be treated at the current dose.  
f. If the current dose is the lowest dose (dose level -1) and the rule indicates dose de-
escalation, new patients will be treated at the lowest dose unless the number of DLTs 
reaches the elimination boundary, at which point the trial will be terminated for 
safety.  
g. If the current dose is the highest dose and the rule indicates dose escalation, new 
patients will be treated at the highest dose.  
 
3. Repeat step 2 until the maximum sample size of 18 DLT-evaluable patients is reached or stop 
the trial if the number of patients treated at the current dose reaches 10. 
 

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59 
After the study is completed, the MTD will be selected based on isotonic regression, as specified.  
Specifically, the MTD will be selected as the dose for which the isotonic estimate of the toxicity 
rate is closest to the target toxicity rate (i.e. 25%).  If there are ties, the higher dose level will be 
selected when the isotonic estimate is lower than the target toxicity rate; and the lower dose level 
will be selected when the isotonic estimate is greater than the target toxicity rate.  Of note, the 
protocol team can choose, for safety reasons and clinical judgment, to not escalate doses when 
permitted by design and select a dose lower than the MTD as the recommended phase 2 dose. 
 
 
Table 3:  Dose escalation/de-escalation rule for the BOIN design. 
Actions 
The number of patients treated at the current dose 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
Escalate if # 
of DLT ≤  
0 
0 
0 
0 
0 
1 
1 
1 
1 
1 
De-escalate 
if # of DLT 
≥ 
1 
1 
1 
2 
2 
2 
3 
3 
3 
3 
Eliminate if 
# of DLT ≥ 
NA 
NA 
2 
2 
2 
2 
3 
3 
3 
4 
 
 
 

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60 
 
BOIN Design Flowchart 
 
 
 
 
Once we determine the MTD, an additional 12 patients (expansion cohort) will be enrolled for 
additional experience with safety and efficacy.  We will use the elimination boundaries in Table 
3 for toxicity monitoring. 
 
Frequency and severity of adverse events and tolerability of the regimen will be collected and 
summarized with descriptive statistics.  The maximum grade for each type of toxicity will be 
recorded for each patient, and frequency tables will be reviewed to determine toxicity patterns.  
All patients who have received at least one dose of the therapeutic agents will be evaluable for 
toxicity and tolerability. 
 
Operating characteristics 
 
Table 4 shows the operating characteristics of the trial design based on 10,000 simulations of the 
trial using the BOIN Design Desktop Program (Venier et al., 2018).  The operating 
characteristics show that the design selects the true MTD, if any, with high probability and 
allocates more patients to the dose levels with the DLT rate closest to the target of 0.25. 
 

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61 
Table 4. Operating Characteristics of the BOIN design. 
 
 
Dose Level 
Number of 
Patients 
% Early 
Stopping 
 
-1 
1 
2 
3 
 
Scenario 1 
True DLT Rate 
0.25 
0.42 
0.48 
0.53 
 
11.15 
46.6 
Selection % 
40.7 
9.8 
2.4 
0.5 
 
# Pts Treated 
6.6 
3.3 
1.0 
0.3 
 
Scenario 2 
True DLT Rate 
0.01 
0.10 
0.25 
0.40 
 
17.28 
0.2 
Selection % 
8.9 
40.7 
37.9 
12.3 
 
# Pts Treated 
3.0 
5.6 
5.6 
3.1 
 
Scenario 3 
True DLT Rate 
0.02 
0.06 
0.10 
0.25 
 
17.23 
0.4 
Selection % 
3.0 
9.2 
41.3 
46.1 
 
# Pts Treated 
2.6 
3.3 
5.2 
6.2 
 
Scenario 4 
True DLT Rate 
0.05 
0.07 
0.10 
0.12 
 
16.63 
2.1 
Selection % 
3.7 
7.5 
14.3 
72.4 
 
# Pts Treated 
2.9 
3.2 
3.5 
7.1 
 
 
 
9.2 
Sample Size/Accrual Rate 
 
The maximum number of patients included in this study will be 18 for the phase 1 portion 
and 12 for the expansion cohort with an additional three patients recruited for ~10% attrition.  
Planned accrual is one patient per month.   
 
 
 
 
 
 

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62 
PLANNED ENROLLMENT REPORT 
 
DOMESTIC PLANNED ENROLLMENT REPORT (TREATMENT) 
Racial Categories 
Ethnic Categories 
Total 
Not Hispanic or Latino 
Hispanic or Latino 
Female 
Male 
Female 
Male 
American Indian/ 
Alaska Native 
0 
0 
0 
0 
0 
Asian 
1 
2 
0 
0 
3 
Native Hawaiian or 
Other Pacific 
Islander 
1 
2 
0 
0 
3 
Black or African 
American 
2 
2 
0 
0 
4 
White 
4 
19 
0 
0 
23 
More Than One 
Race 
0 
0 
0 
0 
0 
Total 
8 
25 
0 
0 
33 
 
PHS 398 / PHS 2590 (Rev. 03/20 Approved Through 2/28/2023)  
OMB No. 0925-0001/0002 
 
 
9.3 
Analysis of Secondary Endpoints 
 
Symptom relief rate will be calculated with 95% binomial confidence intervals (assuming the 
number of patients is binomially distributed).  Time to second intervention for dysphagia will be 
defined as the time from initiation of therapy to the time of second intervention for dysphagia.  
Overall survival will be defined as the duration from patient enrollment to death due to any 
cause.  Patients last known to be alive will censored at the date of last contact.  Survival will 
initially be analyzed using Kaplan-Meier methods, resulting in median survival times with 95% 
CI.  Ogilvie dysphagia scores will be summarized at baseline and post-treatment, and the 
improvement will be compared using paired t-test or Wilcoxon signed-rank test between baseline 
and post-treatment.  Biomarkers will be described graphically or summary measures (e.g. mean 
and standard errors, or median and range) and compared between responders and non-responders 
using a two sample t-test or Wilcoxon test if the data is not normally distributed. 
 
10. ADVERSE EVENTS:  LIST AND REPORTING REQUIREMENTS  
 
Adverse event (AE) monitoring and reporting is a routine part of every clinical trial.  The 
following list of AEs (Section 10.1) and the characteristics of an observed AE (Sections 10.2 and 
10.3) will determine whether the event requires expedited reporting via the CTEP Adverse Event 
Reporting System (CTEP-AERS) in addition to routine reporting. 
 
10.1 
Comprehensive Adverse Events and Potential Risks List (CAEPR) 
 
 
10.1.1 CAEPR for CTEP IND Agent:  AZD1775 (adavosertib, NSC 751084) 
 
The Comprehensive Adverse Events and Potential Risks list (CAEPR) provides a single list of 
reported and/or potential adverse events (AE) associated with an agent using a uniform 
presentation of events by body system. In addition to the comprehensive list, a subset, the 

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63 
Specific Protocol Exceptions to Expedited Reporting (SPEER), appears in a separate column and 
is identified with bold and italicized text. This subset of AEs (SPEER) is a list of events that are 
protocol specific exceptions to expedited reporting to NCI (except as noted below).  Refer to the 
'CTEP, NCI Guidelines: Adverse Event Reporting Requirements' 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/docs/aeguidelines.pdf for 
further clarification. Frequency is provided based on 323 patients. Below is the CAEPR for 
AZD1775 (adavosertib). 
NOTE: Report AEs on the SPEER ONLY IF they exceed the grade noted in parentheses 
next to the AE in the SPEER.  If this CAEPR is part of a combination protocol using multiple 
investigational agents and has an AE listed on different SPEERs, use the lower of the grades 
to determine if expedited reporting is required. 
 
 Version 2.7, April 27, 20201 
 
 Adverse Events with Possible  
 Relationship to AZD1775 (adavosertib) 
 (CTCAE 5.0 Term) 
[n= 323] 
 
 
 Specific Protocol 
Exceptions to Expedited 
Reporting  
Likely (>20%) 
Less Likely (<=20%) 
Rare but Serious (<3%)  
 
BLOOD AND LYMPHATIC SYSTEM DISORDERS 
  
 
Anemia 
 
 Anemia (Gr 3) 
 
 
Febrile neutropenia 
  
CARDIAC DISORDERS 
  
 
 
Atrial fibrillation 
  
 
 
Supraventricular tachycardia   
GASTROINTESTINAL DISORDERS 
  
 
Abdominal pain 
 
 Abdominal pain (Gr 2) 
 
Constipation 
 
 Constipation (Gr 2) 
Diarrhea 
 
 
 Diarrhea (Gr 3) 
 
Dyspepsia 
 
  
 
 
Gastrointestinal hemorrhage2   
 
Mucositis oral 
 
 Mucositis oral (Gr 2) 
Nausea 
 
 
 Nausea (Gr 3) 
Vomiting 
 
 
 Vomiting (Gr 3) 
GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS 
  
 
Edema limbs 
 
 Edema limbs (Gr 2) 
Fatigue 
 
 
 Fatigue (Gr 3) 
 
Fever 
 
 Fever (Gr 2) 
HEPATOBILIARY DISORDERS 
  
 
 
Hepatobiliary disorders - 
Other (hepatitis) 
  
INFECTIONS AND INFESTATIONS 
  
 
Infection3 
 
 Infection3 (Gr 3) 
INVESTIGATIONS 
  
 
Alanine aminotransferase 
increased 
 
 Alanine aminotransferase 
increased (Gr 3) 
 
 
Electrocardiogram QT 
corrected interval prolonged 
  
 
Lymphocyte count decreased 
 
  
 
Neutrophil count decreased 
 
 Neutrophil count decreased 
(Gr 4) 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
64 
 
 Adverse Events with Possible  
 Relationship to AZD1775 (adavosertib) 
 (CTCAE 5.0 Term) 
[n= 323] 
 
 
 Specific Protocol 
Exceptions to Expedited 
Reporting  
Likely (>20%) 
Less Likely (<=20%) 
Rare but Serious (<3%)  
 
 
Platelet count decreased 
 
 Platelet count decreased (Gr 4) 
 
Weight loss 
 
  
 
White blood cell decreased 
 
 White blood cell decreased (Gr 
4) 
METABOLISM AND NUTRITION DISORDERS 
  
 
Anorexia 
 
 Anorexia (Gr 2) 
 
Dehydration 
 
  
 
Hypokalemia 
 
 Hypokalemia (Gr 2) 
 
Hypomagnesemia 
 
 Hypomagnesemia (Gr 2) 
MUSCULOSKELETAL AND CONNECTIVE TISSUE DISORDERS 
  
 
Back pain 
 
 Back pain (Gr 2) 
 
Muscle cramp 
 
  
 
Myalgia 
 
 Myalgia (Gr 2) 
NERVOUS SYSTEM DISORDERS 
  
` 
Dizziness 
 
 Dizziness (Gr 2) 
 
Headache 
 
 Headache (Gr 2) 
 
 
Intracranial hemorrhage 
  
PSYCHIATRIC DISORDERS 
  
 
Insomnia 
 
  
RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS 
  
 
Cough 
 
 Cough (Gr 2) 
 
Dyspnea 
 
 Dyspnea (Gr 2) 
 
 
Hypoxia 
  
SKIN AND SUBCUTANEOUS TISSUE DISORDERS 
  
 
Rash4 
 
 Rash4 (Gr 2) 
VASCULAR DISORDERS 
  
 
 
Phlebitis 
  
 
 
1This table will be updated as the toxicity profile of the agent is revised.  Updates will be distributed to all 
Principal Investigators at the time of revision.  The current version can be obtained by contacting 
PIO@CTEP.NCI.NIH.GOV.  Your name, the name of the investigator, the protocol and the agent should 
be included in the e-mail. 
 
2Gastrointestinal hemorrhage includes Anal hemorrhage, Cecal hemorrhage, Colonic hemorrhage, 
Duodenal hemorrhage, Esophageal hemorrhage, Esophageal varices hemorrhage, Gastric hemorrhage, 
Hemorrhoidal hemorrhage, Ileal hemorrhage, Intra-abdominal hemorrhage, Jejunal hemorrhage, Lower 
gastrointestinal hemorrhage, Oral hemorrhage, Pancreatic hemorrhage, Rectal hemorrhage, 
Retroperitoneal hemorrhage, and Upper gastrointestinal hemorrhage under the GASTROINTESTINAL 
DISORDERS SOC. 
 
3Infection includes all 75 sites of infection under the INFECTIONS AND INFESTATIONS SOC. 
 
4Rash may include rash, erythema, eczema, and rash maculo-papular. 
 
5Peripheral neuropathy includes both peripheral motor neuropathy and peripheral sensory neuropathy. 
 
6Acute kidney injury includes renal impairment and acute renal insufficiency. 

NCI Protocol #: 10389 
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65 
 
Adverse events reported on AZD1775 (adavosertib) trials, but for which there is insufficient 
evidence to suggest that there was a reasonable possibility that AZD1775 (adavosertib) caused 
the adverse event: 
 
BLOOD AND LYMPHATIC SYSTEM DISORDERS - Blood and lymphatic system disorders - Other 
(pancytopenia); Blood and lymphatic system disorders - Other (thrombocytosis); Blood and lymphatic 
system disorders - Other (right leg deep vein thrombosis); Leukocytosis 
Cardiac disorders - Cardiac disorders - Other (cardiomegaly); Chest pain - cardiac; Myocardial 
infarction; Palpitations; Sinus bradycardia; Sinus tachycardia 
EAR AND LABYRINTH DISORDERS - Ear pain; Hearing impaired; Tinnitus 
EYE DISORDERS - Blurred vision; Cataract; Eye disorders - Other (eye swelling); Eye pain; Keratitis; 
Photophobia; Scleral disorder; Vision decreased; Watering eyes 
GASTROINTESTINAL DISORDERS - Abdominal distension; Anal pain; Ascites; Belching; Bloating; 
Cheilitis; Colitis; Colonic obstruction; Dry mouth; Duodenal ulcer; Dysphagia; Enterocolitis; Flatulence; 
Gastric ulcer; Gastritis; Hemorrhoids; Oral pain; Rectal pain; Small intestinal obstruction 
GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS - Chills; Death NOS; Edema 
trunk; Flu like symptoms; Gait disturbance; General disorders and administration site conditions - Other 
(catheter site pain); Infusion site extravasation; Malaise; Non-cardiac chest pain; Pain 
IMMUNE SYSTEM DISORDERS - Allergic reaction; Anaphylaxis; Cytokine release syndrome 
INJURY, POISONING AND PROCEDURAL COMPLICATIONS - Fall; Injury, poisoning and procedural 
complications - Other (excoriation); Injury, poisoning and procedural complications - Other (ligament 
sprain) 
INVESTIGATIONS - Alkaline phosphatase increased; Aspartate aminotransferase increased; Blood 
bilirubin increased; Creatinine increased; GGT increased; Investigations - Other (blood urea increased); 
Lymphocyte count increased 
METABOLISM AND NUTRITION DISORDERS - Alkalosis; Hypercalcemia; Hyperglycemia; 
Hyperkalemia; Hyperuricemia; Hypoalbuminemia; Hypocalcemia; Hyponatremia; Hypophosphatemia; 
Tumor lysis syndrome 
MUSCULOSKELETAL AND CONNECTIVE TISSUE DISORDERS - Arthralgia; Arthritis; Bone pain; 
Flank pain; Generalized muscle weakness; Muscle weakness lower limb; Musculoskeletal and connective 
tissue disorder - Other (groin pain); Neck pain; Pain in extremity 
NEOPLASMS BENIGN, MALIGNANT AND UNSPECIFIED (INCL CYSTS AND POLYPS) - Neoplasms 
benign, malignant and unspecified (incl cysts and polyps) - Other (carcinoid tumor); Tumor pain 
NERVOUS SYSTEM DISORDERS - Central nervous system necrosis; Cognitive disturbance; Depressed 
level of consciousness; Dysesthesia; Dysgeusia; Encephalopathy; Lethargy; Nervous system disorders - 
Other (hemiparesis); Paresthesia; Peripheral neuropathy5; Presyncope; Somnolence; Syncope 
PSYCHIATRIC DISORDERS - Agitation; Anxiety; Confusion; Depression 
RENAL AND URINARY DISORDERS - Acute kidney injury6; Hematuria; Urinary frequency; Urinary 
incontinence; Urinary retention; Urinary tract pain 
REPRODUCTIVE SYSTEM AND BREAST DISORDERS - Genital edema; Reproductive system and 
breast disorders - Other (female genital tract fistula) 
RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS - Allergic rhinitis; Apnea; 
Bronchopulmonary hemorrhage; Epistaxis; Hiccups; Nasal congestion; Pleural effusion; Pneumonitis; 
Pulmonary hypertension; Respiratory, thoracic and mediastinal disorders - Other (diaphragmalgia); Voice 
alteration; Wheezing 
SKIN AND SUBCUTANEOUS TISSUE DISORDERS - Alopecia; Bullous dermatitis; Dry skin; 
Hyperhidrosis; Pain of skin; Palmar-plantar erythrodysesthesia syndrome; Pruritus; Purpura; Rash 
acneiform; Skin ulceration; Urticaria 
VASCULAR DISORDERS - Flushing; Hematoma; Hot flashes; Hypertension; Hypotension; 
Thromboembolic event 
 
 
 

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66 
Note: AZD1775 (adavosertib) in combination with other agents could cause an exacerbation of any 
adverse event currently known to be caused by the other agent, or the combination may result in events 
never previously associated with either agent. 
 
10.2 
Adverse Event Characteristics  
 
• CTCAE term (AE description) and grade:  The descriptions and grading scales found 
in the revised NCI Common Terminology Criteria for Adverse Events (CTCAE) version 
5.0 will be utilized for AE reporting.  All appropriate treatment areas should have access 
to a copy of the CTCAE version 5.0.  A copy of the CTCAE version 5.0 can be 
downloaded from the CTEP website 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm. 
 
• For expedited reporting purposes only:   
- 
AEs for the agent that are bold and italicized in the CAEPR (i.e., those listed in the 
SPEER column, Section 10.1) should be reported through CTEP-AERS only if the 
grade is above the grade provided in the SPEER. 
 
• Attribution of the AE: 
- 
Definite – The AE is clearly related to the study treatment. 
- 
Probable – The AE is likely related to the study treatment. 
- 
Possible – The AE may be related to the study treatment. 
- 
Unlikely – The AE is doubtfully related to the study treatment. 
- 
Unrelated – The AE is clearly NOT related to the study treatment. 
 
10.3 
Expedited Adverse Event Reporting 
 
10.3.1 Rave-CTEP-AERS Integration 
 
The Rave Cancer Therapy Evaluation Program Adverse Event Reporting System (CTEP-
AERS) integration enables evaluation of post-baseline AEs entered in Rave to determine 
whether they require expedited reporting, and facilitates entry in CTEP-AERS for those 
AEs requiring expedited reporting.    
 
All AEs that occur after baseline are collected in Medidata Rave using the Adverse Event 
form, which is available for entry at each treatment or reporting period, and used to 
collect AEs that start during the period or persist from the previous reporting period.  The 
CRA will enter AEs that occur prior to the start of treatment on a baseline form that is not 
included in the Rave-CTEP-AERS integration.  AEs that occur prior to enrollment must 
begin and end on the baseline Adverse Event form and should not be included on the 
standard Adverse Events form that is available at treatment unless there has been an 
increase in grade. 
 
Prior to sending AEs through the rules evaluation process, site staff should verify the 
following on the Adverse Event form in Rave: 
 

NCI Protocol #: 10389 
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67 
• The reporting period (course/cycle) is correct, and 
• AEs are recorded and complete (no missing fields) and the form is query-free  
 
The CRA reports AEs in Rave at the time the Investigator learns of the event.  If the CRA 
modifies an AE, it must be re-submitted for rules evaluation. 
 
Upon completion of AE entry in Medidata Rave, the CRA submits the AE for rules 
evaluation by completing the Expedited Reporting Evaluation form.  Both NCI and 
protocol-specific reporting rules evaluate the AEs submitted for expedited reporting.  A 
report is initiated in CTEP-AERS using information entered in Medidata Rave for AEs 
that meet reporting requirements.  The CRA completes the report by accessing CTEP-
AERS via a direct link on the Medidata Rave Expedited Reporting Evaluation form. 
 
In the rare occurrence that Internet connectivity is lost, a 24-hour notification is to be 
made to CTEP by telephone at 301-897-7497.  Once internet connectivity is restored, the 
24-hour notification that was phoned in must be entered immediately into CTEP-AERS 
using the direct link from Medidata Rave. 
 
Additional information about the CTEP-AERS integration is available on the CTSU 
website: 
 
• Study specific documents:  Protocols > Documents > Education and Promotion, 
and   
• Expedited Safety Reporting Rules Evaluation user guide:  Resources > CTSU 
Operations Information > User Guides & Help Topics. 
 
NCI requirements for SAE reporting are available on the CTEP website:  
 
• NCI Guidelines for Investigators:  Adverse Event Reporting Requirements is 
available at 
https://ctep.cancer.gov/protocolDevelopment/electronic_applications/docs/aeguid
elines.pdf. 
 
10.3.2 Distribution of Adverse Event Reports 
 
CTEP-AERS is programmed for automatic electronic distribution of reports to the following 
individuals:  Principal Investigator and Adverse Event Coordinator(s) (if applicable) of the 
Corresponding Organization or Lead Organization, the local treating physician, and the Reporter 
and Submitter.  CTEP-AERS provides a copy feature for other e-mail recipients.  
 
10.3.3 Expedited Reporting Guidelines 
 
Use the NCI protocol number and the protocol-specific patient ID assigned during trial 
registration on all reports. 
 
Note:  A death on study requires both routine and expedited reporting, regardless of 

NCI Protocol #: 10389 
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68 
causality as long as the death occurred within 30 days after the last administration of the 
investigational agent.  Attribution to treatment or other cause must be provided. 
 
Death due to progressive disease should be reported as Grade 5 “Disease progression” in the 
system organ class (SOC) “General disorders and administration site conditions.”  Evidence that 
the death was a manifestation of underlying disease (e.g., radiological changes suggesting tumor 
growth or progression: clinical deterioration associated with a disease process) should be 
submitted.  
 
 
Phase 1 and Early Phase 2 Studies:  Expedited Reporting Requirements for Adverse 
Events that Occur on Studies under an IND/IDE within 30 Days of the Last Administration 
of the Investigational Agent/Intervention 1, 2 
FDA REPORTING REQUIREMENTS FOR SERIOUS ADVERSE EVENTS (21 CFR Part 312) 
NOTE:  Investigators MUST immediately report to the sponsor (NCI) ANY Serious Adverse Events, whether or not 
they are considered related to the investigational agent(s)/intervention (21 CFR 312.64) 
An adverse event is considered serious if it results in ANY of the following outcomes:   
1) 
Death 
2) 
A life-threatening adverse event  
3) 
An adverse event that results in inpatient hospitalization or prolongation of existing hospitalization for ≥ 24 
hours  
4) 
A persistent or significant incapacity or substantial disruption of the ability to conduct normal life functions  
5) 
A congenital anomaly/birth defect.  
6) 
Important Medical Events (IME) that may not result in death, be life threatening, or require hospitalization 
may be considered serious when, based upon medical judgment, they may jeopardize the patient or 
subject and may require medical or surgical intervention to prevent one of the outcomes listed in this 
definition. (FDA, 21 CFR 312.32; ICH E2A and ICH E6). 
 
ALL SERIOUS adverse events that meet the above criteria MUST be immediately reported to the NCI via electronic 
submission within the timeframes detailed in the table below. 
Hospitalization 
Grade 1 and Grade 2 Timeframes 
Grade 3-5 
Timeframes 
Resulting in 
Hospitalization  
≥ 24 hrs 
10 Calendar Days 
24-Hour 5 Calendar 
Days 
Not resulting in 
Hospitalization  
≥ 24 hrs 
Not required 
NOTE:  Protocol specific exceptions to expedited reporting of serious adverse events are found in the Specific 
Protocol Exceptions to Expedited Reporting (SPEER) portion of the CAEPR. 
Expedited AE reporting timelines are defined as: 
o 
“24-Hour; 5 Calendar Days” - The AE must initially be submitted electronically within 24 hours of learning 
of the AE, followed by a complete expedited report within 5 calendar days of the initial 24-hour report. 
o 
“10 Calendar Days” - A complete expedited report on the AE must be submitted electronically within 10 
calendar days of learning of the AE. 
1Serious adverse events that occur more than 30 days after the last administration of investigational 
agent/intervention and have an attribution of possible, probable, or definite require reporting as follows:  
Expedited 24-hour notification followed by complete report within 5 calendar days for: 
• 
All Grade 3, 4, and Grade 5 AEs 
Expedited 10 calendar day reports for: 
• 
Grade 2 AEs resulting in hospitalization or prolongation of hospitalization 

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69 
2For studies using PET or SPECT IND agents, the AE reporting period is limited to 10 radioactive half-lives, 
rounded UP to the nearest whole day, after the agent/intervention was last administered.  Footnote “1” above 
applies after this reporting period. 
Effective Date:  May 5, 2011 
 
 
10.4 
Routine Adverse Event Reporting 
 
All Adverse Events must be reported in routine study data submissions.  AEs reported 
expeditiously through CTEP-AERS must also be reported in routine study data 
submissions. 
 
Adverse event data collection and reporting, which are required as part of every clinical trial, are 
done to ensure the safety of patients enrolled in the studies as well as those who will enroll in 
future studies using similar agents.  AEs are reported in a routine manner at scheduled times 
during the trial using Medidata Rave.  For this trial the Adverse Event CRF is used for routine 
AE reporting in Rave. 
 
10.5 
Pregnancy 
 
Although not an adverse event in and of itself, pregnancy as well as its outcome must be 
documented via CTEP-AERS.  In addition, the Pregnancy Information Form included within 
the NCI Guidelines for Adverse Event Reporting Requirements must be completed and 
submitted to CTEP.  Any pregnancy occurring in a patient or patient’s partner from the time of 
consent to 90 days after the last dose of study drug must be reported and then followed for 
outcome.  Newborn infants should be followed until 30 days old.  Please see the “NCI 
Guidelines for Investigators: Adverse Event Reporting Requirements for DCTD (CTEP and CIP) 
and DCP INDs and IDEs” (at http://ctep.cancer.gov/protocolDevelopment/adverse_effects.htm) 
for more details on how to report pregnancy and its outcome to CTEP.  
 
10.6 
Secondary Malignancy 
 
A secondary malignancy is a cancer caused by treatment for a previous malignancy (e.g., 
treatment with investigational agent/intervention, radiation or chemotherapy). A secondary 
malignancy is not considered a metastasis of the initial neoplasm. 
 
CTEP requires all secondary malignancies that occur following treatment with an agent under an 
NCI IND/IDE be reported expeditiously via CTEP-AERS.  Three options are available to 
describe the event:  
 
• Leukemia secondary to oncology chemotherapy (e.g., acute myelocytic leukemia [AML])  
• Myelodysplastic syndrome (MDS)  
• Treatment-related secondary malignancy  
 
Any malignancy possibly related to cancer treatment (including AML/MDS) should also be 
reported via the routine reporting mechanisms outlined in each protocol.  

NCI Protocol #: 10389 
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70 
 
10.7 
Second Malignancy 
 
A second malignancy is one unrelated to the treatment of a prior malignancy (and is NOT a 
metastasis from the initial malignancy).  Second malignancies require ONLY routine AE 
reporting unless otherwise specified.

NCI Protocol #: 10389 
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71 
 
11. STUDY CALENDAR 
 
Baseline evaluations are to be conducted within 2 weeks prior to start of protocol therapy.  Scans must be done <4 weeks prior to the 
start of therapy.  In the event that the patient’s condition is deteriorating, laboratory evaluations should be repeated within 48 hours 
prior to initiation of the next cycle of therapy. 
 
 
Pre-Treatment 
During Treatment 
Follow-Up 
 
≤12 
months 
prior to 
study 
entry 
≤28 
days 
prior to 
study 
entry 
≤14 days 
prior to 
study 
entry 
Wk 
-1 
 
Wk 
1 
Wk 
2 
Wk 
3 
Wk 
6 
Q3 months (+/- 1 
month) for first 2 years 
Q6 months (+/- 1 
month) for years 2-5 
AZD1775a 
 
 
 
X 
X 
 
X 
 
 
 
Informed consent 
 
X 
 
 
 
 
 
 
 
 
History and physical exam 
 
 
X 
 
X 
X 
X 
X 
X 
X 
Vital signs 
 
 
X 
 
X 
X 
X 
X 
X 
X 
Weight 
 
 
X 
 
X 
X 
X 
X 
X 
X 
Performance statusc 
 
 
X 
 
X 
X 
X 
X 
X 
X 
CBC w/diff, plts 
 
 
X 
 
X 
X 
X 
X 
X 
X 
Comprehensive Chemistry 
Paneld 
 
 
X 
 
X 
X 
X 
X 
X 
X 
EKG 
 
X 
As clinically indicated 
 
 
Pregnancy teste 
 
X 
 
Xf 
 
 
 
 
 
 
Adverse event evaluationg 
 
 
 
X--------------------------------------------------------X 
 
 
CT chest, abdomen, and pelvis 
or CT chest, MRI abdomen 
and pelvis, or whole-body 
PET-CT 
 
X 
 
 
 
 
 
 
 
 
EGD with biopsy 
X 
 
Xb 
Xb,h 
 
 
 
 
 
 

NCI Protocol #: 10389 
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72 
 
Pre-Treatment 
During Treatment 
Follow-Up 
 
≤12 
months 
prior to 
study 
entry 
≤28 
days 
prior to 
study 
entry 
≤14 days 
prior to 
study 
entry 
Wk 
-1 
 
Wk 
1 
Wk 
2 
Wk 
3 
Wk 
6 
Q3 months (+/- 1 
month) for first 2 years 
Q6 months (+/- 1 
month) for years 2-5 
Radiation therapyi 
 
 
 
 
X 
X 
X 
 
 
 
Ogilvie dysphagia score 
(expansion cohort)j 
 
 
X 
 
 
 
 
X 
X 
X 
Whole Exome Sequencing 
(Formalin Fixed Biopsy) 
 
Xk 
 
 
 
 
 
 
 
 
Whole Exome Sequencing 
(blood in EDTA) 
 
X 
 
 
 
 
 
 
 
 
ɣH2AX, pNBS1 IFA with 
βCATN segmentation (Snap 
Frozen Biopsy) 
 
 
Xb 
Xb,h 
 
 
 
 
 
 
p-HH3 (S20), p-cdk1 (Y15), 
pWEE1(S642), and p-
Chk1(S345) IHC (Formalin 
Fixed Biopsy) 
 
 
Xb 
Xb,h 
 
 
 
 
 
 

NCI Protocol #: 10389 
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73 
 
Pre-Treatment 
During Treatment 
Follow-Up 
 
≤12 
months 
prior to 
study 
entry 
≤28 
days 
prior to 
study 
entry 
≤14 days 
prior to 
study 
entry 
Wk 
-1 
 
Wk 
1 
Wk 
2 
Wk 
3 
Wk 
6 
Q3 months (+/- 1 
month) for first 2 years 
Q6 months (+/- 1 
month) for years 2-5 
 
a: 
Dosing days as assigned.  Dose should be taken 2-3 hours prior to radiation therapy in weeks 1 and 3. 
b: 
Expansion cohort only. 
c: 
Note:  Performance status evaluations are based on a 4 week cycle.  At minimum, performance status should be evaluated at the beginning of every cycle. 
d: 
Albumin, alkaline phosphatase, total bilirubin, bicarbonate, BUN, calcium, chloride, creatinine, glucose, LDH, phosphorus (as clinically indicated), potassium, 
total protein, SGOT [AST], SGPT [ALT], sodium. 
e: 
Pregnancy test for women of childbearing potential. 
f: 
To be performed on Day 1. 
g: 
Patients with toxicity during treatment must visit the study doctor after completion of treatment every other week until toxicity resolves to grade ≤2. 
h: 
To be performed on Day 4 or 5 
i: 
Delivered five days per week, for a total dose of 37.5 Gy in 15 fractions. 
j: 
See Appendix E for Ogilvie dysphagia scoring  
k: 
If archival (FFPE) tissue is not available, a baseline biopsy is mandatory. 
 
 
 

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74 
 
12. MEASUREMENT OF EFFECT 
 
Although the clinical benefit of this drug has not yet been established, the intent of offering this 
treatment is to provide a possible therapeutic benefit, and thus the patient will be carefully 
monitored for tumor response and symptom relief in addition to safety and tolerability.   
 
12.1 
Antitumor Effect – Solid Tumors 
 
12.1.1 Definitions 
 
Evaluable for Toxicity.  All patients will be evaluable for toxicity from the time of their first 
treatment with AZD1775. 
 
Evaluable for Objective Response.  Only those patients who have measurable disease present at 
baseline, have received at least one cycle of therapy, and have had their disease re-evaluated will 
be considered evaluable for response.  These patients will have their response classified 
according to the definitions stated below.  (Note:  Patients who exhibit objective disease 
progression prior to the end of cycle 1 will also be considered evaluable.) 
 
12.1.2 Disease Parameters 
 
Patient dysphagia will be evaluated using the Ogilvie dysphagia score, comparing pre-treatment 
to post-treatment scores (Appendix E). 
 
12.1.3 Overall Survival 
 
Overall survival will be defined as the duration from patient enrollment to death due to any 
cause.  Patients last known to be alive will censored at the date of last contact.  Survival will 
initially be analyzed using Kaplan-Meier methods, resulting in median survival times with 95% 
CI. 
 
 
12.2 
Other Response Parameters 
 
12.2.1 Dysphagia 
 
Time to second intervention for dysphagia will be defined as the time from initiation of therapy 
to the time of second intervention for dysphagia.   
 
 
13. STUDY OVERSIGHT AND DATA REPORTING / REGULATORY 
REQUIREMENTS 
 
Adverse event lists, guidelines, and instructions for AE reporting can be found in Section 10 
(Adverse Events: List and Reporting Requirements). 

NCI Protocol #: 10389 
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75 
 
13.1 
Study Oversight 
 
This protocol is monitored at several levels, as described in this section.  The Protocol Principal 
Investigator is responsible for monitoring the conduct and progress of the clinical trial, including 
the ongoing review of accrual, patient-specific clinical and laboratory data, and routine and 
serious adverse events; reporting of expedited adverse events; and accumulation of reported 
adverse events from other trials testing the same drug(s).  The Protocol Principal Investigator and 
statistician have access to the data at all times through the CTMS web-based reporting portal.   
 
For the Phase 1 portion of this study, all decisions regarding dose escalation/expansion/de-
escalation require sign-off by the Protocol Principal Investigator through the CTMS/IWRS.  In 
addition, for the Phase 1 portion, the Protocol Principal Investigator will have at least monthly, 
or more frequently, conference calls with the Study Investigators and the CTEP Medical 
Officer(s) to review accrual, progress, and adverse events and unanticipated problems.  
 
All Study Investigators at participating sites who register/enroll patients on a given protocol are 
responsible for timely submission of data via Medidata Rave and timely reporting of adverse 
events for that particular study.  This includes timely review of data collected on the electronic 
CRFs submitted via Medidata Rave. 
 
All studies are also reviewed in accordance with the enrolling institution’s data safety monitoring 
plan. 
 
Mandatory safety and trial review teleconferences will be scheduled and moderated by the Multi-
Center Trial Program (MCTP). All sites involved in the study are expected to have a 
representative present for every call to review and discuss patients on study and other applicable 
agenda items. Meeting minutes will be used to document each teleconference. The minutes will 
be stored in the MCTP protocol files. Teleconferences must minimally be held monthly and may 
be held more frequently, as needed. For studies closed to accrual with patients expected to 
remain on long-term treatment and/or follow-up, teleconferences may be extended to occur every 
two months or quarterly. Decreasing frequency of teleconferences requires OSU PI and MCTP 
approval. 
 
13.2 
Data Reporting 
 
Medidata Rave is a clinical data management system being used for data collection for this 
trial/study.  Access to the trial in Rave is controlled through the CTEP-IAM system and role 
assignments.   
 
Requirements to access Rave via iMedidata: 
 
• 
A valid account, and  
• 
Assigned a Rave role on the relevant LPO or PO roster at the enrolling site of:  Rave 
CRA, Rave Read Only, Rave CRA (LabAdmin), Rave SLA, or Rave Investigator.   
• 
Rave role requirements: 

NCI Protocol #: 10389 
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76 
o Rave CRA or Rave CRA (Lab Admin) role, must have a minimum of an 
Associate Plus (AP) registration type, 
o Rave Investigator role, must be registered as an Non-Physician Investigator 
(NPIVR) or Investigator (IVR), and 
o Rave Read Only role, must have at a minimum an Associates (A) registration 
type. 
• 
Refer to https://ctep.cancer.gov/investigatorResources/default.htm for registration types 
and documentation required. 
 
If the study has a DTL, individuals requiring write access to Rave must also be assigned the 
appropriate Rave tasks on the DTL. 
 
Upon initial site registration approval for the study in Regulatory Support System (RSS), all 
persons with Rave roles assigned on the appropriate roster will be sent a study invitation e-mail 
from iMedidata.  To accept the invitation, site staff must log in to the Select Login 
(https://login.imedidata.com/selectlogin) using their CTEP-IAM username and password, and 
click on the accept link in the upper right-corner of the iMedidata page.  Site staff will not be 
able to access the study in Rave until all required Medidata and study specific trainings are 
completed.  Trainings will be in the form of electronic learnings (eLearnings), and can be 
accessed by clicking on the link in the upper right pane of the iMedidata screen.  If an eLearning 
is required and has not yet been taken, the link to the eLearning will appear under the study name 
in iMedidata instead of the Rave EDC link; once the successful completion of the eLearning has 
been recorded, access to the study in Rave will be granted, and a Rave EDC link will display 
under the study name.  
 
Site staff that have not previously activated their iMedidata/Rave account at the time of initial 
site registration approval for the study in RSS will receive a separate invitation from iMedidata 
to activate their account.  Account activation instructions are located on the CTSU website in the 
Data Management section under the Rave resource materials (Medidata Account Activation and 
Study Invitation Acceptance).  Additional information on iMedidata/Rave is available on the 
CTSU members’ website in the Data Management > Rave section at www.ctsu.org/RAVE/ or by 
contacting the CTSU Help Desk at 1-888-823-5923 or by e-mail at ctsucontact@westat.com.   
 
13.2.1 Method 
 
This study will be monitored by the Clinical Trials Monitoring Service (CTMS).  Data will be 
submitted to CTMS at least once every two weeks via Medidata Rave (or other modality if 
approved by CTEP).  Information on CTMS reporting is available at 
http://www.theradex.com/clinicalTechnologies/?National-Cancer-Institute-NCI-11.  On-site 
audits will be conducted three times annually (one annual site visit and two data audits).  For 
CTMS monitored studies, after users have activated their accounts, please contact the Theradex 
Help Desk at (609) 619-7862 or by email at CTMSSupport@theradex.com for additional support 
with Rave and completion of CRFs. 
 
13.2.2 Responsibility for Data Submission 
 

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77 
For ETCTN trials, it is the responsibility of the PI(s) at the site to ensure that all investigators at 
the ETCTN Sites understand the procedures for data submission for each ETCTN protocol and 
that protocol specified data are submitted accurately and in a timely manner to the CTMS via the 
electronic data capture system, Medidata Rave.   
 
Data are to be submitted via Medidata Rave to CTMS on a real-time basis, but no less than once 
every 2 weeks.  The timeliness of data submissions and timeliness in resolving data queries will 
be tracked by CTMS.  Metrics for timeliness will be followed and assessed on a quarterly basis.  
For the purpose of Institutional Performance Monitoring, data will be considered delinquent if it 
is greater than 4 weeks past due. 
 
Data from Medidata Rave and CTEP-AERS is reviewed by the CTMS on an ongoing basis as 
data is received.  Queries will be issued by CTMS directly within Rave.  The queries will appear 
on the Task Summary Tab within Rave for the CRA at the ETCTN to resolve.  Monthly web-
based reports are posted for review by the Drug Monitors in the IDB, CTEP.  Onsite audits will 
be conducted by the CTMS to ensure compliance with regulatory requirements, GCP, and NCI 
policies and procedures with the overarching goal of ensuring the integrity of data generated 
from NCI-sponsored clinical trials, as described in the ETCTN Program Guidelines, which may 
be found on the CTEP 
(http://ctep.cancer.gov/protocolDevelopment/electronic_applications/adverse_events.htm)  and 
CTSU websites. 
 
CTMS will utilize a core set of eCRFs that are Cancer Data Standards Registry and Repository 
(caDSR) compliant (http://cbiit.nci.nih.gov/ncip/biomedical-informatics-
resources/interoperability-and-semantics/metadata-and-models).  Customized eCRFs will be 
included when appropriate to meet unique study requirements.  The PI is encouraged to review 
the eCRFs, working closely with CTMS to ensure prospectively that all required items are 
appropriately captured in the eCRFs prior to study activation.  CTMS will prepare the eCRFs 
with built-in edit checks to the extent possible to promote data integrity. 
 
CDUS data submissions for ETCTN trials activated after March 1, 2014, will be carried out by 
the CTMS contractor, Theradex.  CDUS submissions are performed by Theradex on a monthly 
basis.  The trial’s lead institution is responsible for timely submission to CTMS via Rave, as 
above. 
 
Further information on data submission procedures can be found in the ETCTN Program 
Guidelines 
(http://ctep.cancer.gov/protocolDevelopment/electronic_applications/adverse_events.htm). 
 
13.3 
CTEP Multicenter Guidelines 
 
N/A 
 
13.4 
Data Quality Portal 
 
The Data Quality Portal (DQP) provides a central location for site staff to manage 

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78 
unanswered queries and form delinquencies, monitor data quality and timeliness, 
generate reports, and review metrics.  
 
The DQP is located on the CTSU members’ website under Data Management.  The Rave 
Home section displays a table providing summary counts of Total Delinquencies and 
Total Queries.  DQP Queries, DQP Delinquent Forms, and the DQP Reports modules are 
available to access details and reports of unanswered queries, delinquent forms, and 
timeliness reports.  Review the DQP modules on a regular basis to manage specified 
queries and delinquent forms. 
 
The DQP is accessible by site staff that are rostered to a site and have access to the CTSU 
website.  Staff that have Rave study access can access the Rave study data using a direct 
link on the DQP. 
 
To learn more about DQP use and access, click on the Help icon displayed on the Rave 
Home, DQP Queries, and DQP Delinquent Forms modules. 
 
Note:  Some Rave protocols may not have delinquent form details or reports specified on 
the DQP.  A protocol must have the Calendar functionality implemented in Rave by the 
Lead Protocol Organization (LPO) for delinquent form details and reports to be available 
on the DQP.  Site staff should contact the LPO Data Manager for their protocol regarding 
questions about Rave Calendaring functionality.  
 
 
13.5 
Collaborative Agreements Language 
 
The agent(s) supplied by CTEP, DCTD, NCI used in this protocol is/are provided to the NCI 
under a Collaborative Agreement (CRADA, CTA, CSA) between the Pharmaceutical 
Company(ies) (hereinafter referred to as “Collaborator(s)”) and the NCI Division of Cancer 
Treatment and Diagnosis.  Therefore, the following obligations/guidelines, in addition to the 
provisions in the “Intellectual Property Option to Collaborator”  
(http://ctep.cancer.gov/industryCollaborations2/intellectual_property.htm) contained within the 
terms of award, apply to the use of the Agent(s) in this study: 
 
1. Agent(s) may not be used for any purpose outside the scope of this protocol, nor can 
Agent(s) be transferred or licensed to any party not participating in the clinical study.  
Collaborator(s) data for Agent(s) are confidential and proprietary to Collaborator(s) and shall 
be maintained as such by the investigators.  The protocol documents for studies utilizing 
Agents contain confidential information and should not be shared or distributed without the 
permission of the NCI.  If a copy of this protocol is requested by a patient or patient’s family 
member participating on the study, the individual should sign a confidentiality agreement.  A 
suitable model agreement can be downloaded from: http://ctep.cancer.gov. 
 
2. For a clinical protocol where there is an investigational Agent used in combination with 
(an)other Agent(s), each the subject of different Collaborative Agreements, the access to and 
use of data by each Collaborator shall be as follows (data pertaining to such combination use 

NCI Protocol #: 10389 
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79 
shall hereinafter be referred to as "Multi-Party Data”): 
 
a. NCI will provide all Collaborators with prior written notice regarding the existence and 
nature of any agreements governing their collaboration with NCI, the design of the 
proposed combination protocol, and the existence of any obligations that would tend to 
restrict NCI's participation in the proposed combination protocol. 
 
b. Each Collaborator shall agree to permit use of the Multi-Party Data from the clinical trial 
by any other Collaborator solely to the extent necessary to allow said other Collaborator 
to develop, obtain regulatory approval or commercialize its own Agent. 
 
c. Any Collaborator having the right to use the Multi-Party Data from these trials must 
agree in writing prior to the commencement of the trials that it will use the Multi-Party 
Data solely for development, regulatory approval, and commercialization of its own 
Agent. 
 
3. Clinical Trial Data and Results and Raw Data developed under a Collaborative Agreement 
will be made available to Collaborator(s), the NCI, and the FDA, as appropriate and unless 
additional disclosure is required by law or court order as described in the IP Option to 
Collaborator (http://ctep.cancer.gov/industryCollaborations2/intellectual_property.htm).  
Additionally, all Clinical Data and Results and Raw Data will be collected, used and 
disclosed consistent with all applicable federal statutes and regulations for the protection of 
human subjects, including, if applicable, the Standards for Privacy of Individually 
Identifiable Health Information set forth in 45 C.F.R. Part 164. 
 
4. When a Collaborator wishes to initiate a data request, the request should first be sent to the 
NCI, who will then notify the appropriate investigators (Group Chair for Cooperative Group 
studies, or PI for other studies) of Collaborator's wish to contact them. 
 
5. Any data provided to Collaborator(s) for Phase 3 studies must be in accordance with the 
guidelines and policies of the responsible Data Monitoring Committee (DMC), if there is a 
DMC for this clinical trial. 
 
6. Any manuscripts reporting the results of this clinical trial must be provided to CTEP by the 
Group office for Cooperative Group studies or by the principal investigator for non-
Cooperative Group studies for immediate delivery to Collaborator(s) for advisory review and 
comment prior to submission for publication.  Collaborator(s) will have 30 days from the 
date of receipt for review.  Collaborator shall have the right to request that publication be 
delayed for up to an additional 30 days in order to ensure that Collaborator’s confidential and 
proprietary data, in addition to Collaborator(s)’s intellectual property rights, are protected.  
Copies of abstracts must be provided to CTEP for forwarding to Collaborator(s) for courtesy 
review as soon as possible and preferably at least three (3) days prior to submission, but in 
any case, prior to presentation at the meeting or publication in the proceedings.  Press 
releases and other media presentations must also be forwarded to CTEP prior to release.  
Copies of any manuscript, abstract and/or press release/ media presentation should be sent to: 
 

NCI Protocol #: 10389 
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80 
Email:  ncicteppubs@mail.nih.gov 
 
The Regulatory Affairs Branch will then distribute them to Collaborator(s).  No publication, 
manuscript or other form of public disclosure shall contain any of Collaborator’s confidential/ 
proprietary information. 
 
13.6 
Genomic Data Sharing Plan 
 
The investigators and statistician and/or bioinformaticians for a study will have access to all data 
on mutations and variants stored in the Theradex Data Base.  This information will be 
sequestered from access throughout the study until it is analyzed for purposes of reporting and 
publishing of the study results.   
 
 
13.7 
Incidental/Secondary Findings Disclosure Procedure 
 
Incidental/secondary findings of germline and/or somatic mutations, confirmed by Sanger 
sequencing, will be relayed to the treating physician who is responsible for notifying the patients, 
with a genetic counselor present if that is deemed appropriate.  
 
 
 

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81 
 
14. REFERENCES 
 
Arnold, M., M. Laversanne, L.M. Brown, et al.  (2017).  Predicting the Future Burden of 
Esophageal Cancer by Histological Subtype:  International Trends in Incidence up to 2030.  Am J 
Gastroenterol.  112:1247-1255. 
 
Bang, Y.J., E. Van Cutsem, A. Feyereislova, et al.  (2010).  Trastuzumab in Combination with 
Chemotherapy Versus Chemotherapy Alone for Treatment of HER2-positive Advanced Gastric 
or Gastro-oesophageal Junction Cancer (ToGA):  A Phase 3, Open-label, Randomised 
Controlled Trial.  Lancet.  376:687-97. 
 
Beck, H., V. Nähse-Kumpf, M.S.Y. Larsen, et al.  (2012).  Cyclin-Dependent Kinase 
Suppression by WEE1 Kinase Protects the Genome through Control of Replication Initiation and 
Nucleotide Consumption.  Mol Cell Biol.  32:4226-4236. 
 
Bridges, K.A., H. Hirai, C.A. Buser, et al.  (2011).  MK-1775, a Novel Wee1 Kinase Inhibitor, 
Radiosensitizes p53-defective Human Tumor Cells.  Clin Cancer Res.  17:5638-5648. 
 
Chen, X., H. Lu, A. Tai, et al.  (2014).  Determination of Internal Target Volume for Radiation 
Treatment Planning of Esophageal Cancer by Using 4-dimensional Computed tomography 
(4DCT).  Int J Radiat Oncol Biol Phys.  90:102-9. 
 
Coleman T.R. and W.G. Dunphy.  (1994).  Cdc2 regulatory factors.  Current Opinion in Cell 
Biology.  6(6):877-882. 
 
Cueno, K.C., M.A. Morgan, V. Sahai, et al.  (2019).  Dose Escalation Trial of the Wee1 Inhibitor 
Adavosertib (AZD1775) in Combination with Gemcitabine and Radiation for Patients with 
Locally Advanced Pancreatic Cancer.  J Clin Oncol.  37:2643-2650. 
 
Do, K., D. Wilsker, J. Ji, et al.  (2015).  Phase I Study of Single-Agent AZD1775 (MK-1775), a 
Wee1 Kinase Inhibitor, in Patients with Refractory Solid Tumors.  J Clin Oncol.  33:3409-15. 
 
Dominguez-Kelly, R., Y. Martin, S. Koundrioukoff, et al.  (2011).  Wee1 Controls Genomic 
Stability during Replication by Regulating the Mus81-Eme1 endonuclease.  J Cell Biol.  
194:567-79. 
 
Ferlay, J., I. Soerjomataram, R. Dikshit, et al.  (2015).  Cancer Incidence and Mortality 
Worldwide:  Sources, Methods and Major Patterns in GLOBOCAN 2012.  Int J Cancer.  
136:E359-86. 
 
Guttmann, D.M., N. Mitra, J. Bekelman, et al.  (2017).  Improved Overall Survival with 
Aggressive Primary Tumor Radiotherapy for Patients with Metastatic Esophageal Cancer.  J 
Thorac Oncol.  12:1131-1142.  
 
Hirai, H., Y. Iwasawa, M. Okada, et al.  (2009).  Small-molecule Inhibition of Wee1 Kinase by 

NCI Protocol #: 10389 
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82 
MK-1775 Selectively Sensitizes p53-deficient tumor cells to DNA-damaging agents.  Mol 
Cancer Ther.  8:2992-3000. 
 
Hirai H., T. Arai, M. Okada, et al.  (2010).  MK-1775, a small molecule Wee1 inhibitor, 
enhances antitumor efficacy of various DNA-damaging agents, including 5-fluorouracil.  Cancer 
Biology & Therapy.  9(7):514-522. 
 
Investigator’s Brochure.  (2020).  AZD1775, Adavosertib.  AstraZeneca Pharmaceuticals, 
Wilmington, DE.  Edition 18.  January 29, 2020. 
 
Iorns E., C. J. Lord, A. Grigoriadis, et al.  (2009).  Integrated Functional, Gene Expression and 
Genomic Analysis for the Identification of Cancer Targets.  PLOS ONE.  4(4):e5120. 
 
Kausar, T., J.S. Schreiber, D. Karnak, et al.  (2015).  Sensitization of Pancreatic Cancers to 
Gemcitabine Chemoradiation by WEE1 Kinase Inhibition Depends on Homologous 
Recombination Repair.  Neoplasia. 17:757-66. 
 
Kreahling J.M., J.Y. Gemmer, D. Reed, et al.  (2012).  MK1775, a Selective Wee1 Inhibitor, 
Shows Single-Agent Antitumor Activity against Sarcoma Cells.  Molecular Cancer 
Therapeutics.  11(1):174-182. 
 
Leijen S., R.M.J.M. van Geel, A.C. Pavlick, et al.  (2016).  Phase I Study Evaluating WEE1 
Inhibitor AZD1775 As Monotherapy and in Combination With Gemcitabine, Cisplatin, or 
Carboplatin in Patients With Advanced Solid Tumors.  Journal of Clinical Oncology.  
34(36):4371-4380. 
 
Leijen S., R.M.J.M. van Geel, G.S. Sonke, et al.  (2016).  Phase II Study of WEE1 Inhibitor 
AZD1775 Plus Carboplatin in Patients With TP53-Mutated Ovarian Cancer Refractory or 
Resistant to First-Line Therapy Within 3 Months.  Journal of Clinical Oncology.  34(36):4354-
4361. 
 
Liu, S. and Y. Yuan.  (2015).  Bayesian Optimal Interval Designs for Phase I Clinical Trials.  J R 
Stat Soc C-APPL.  64:507-523. 
 
Masaki T., Y. Shiratori, W. Rengifo, et al.  (2003).  Cyclins and Cyclin-Dependent Kinases: 
Comparative: Study of Hepatocellular Carcinoma Versus Cirrhosis.  Hepatology.  37(3):534-
543. 
 
Méndez, E., C.P. Rodriguez, M.C. Kao, et al.  (2018).  A Phase I Clinical Trial of AZD1775 in 
Combination with Neoadjuvant Weekly Docetaxel and Cisplatin before Definitive Therapy in 
Head and Neck Squamous Cell Carcinoma.  Clin Cancer Res.  24:2740-2748. 
 
Mir S.E., P. C. De Witt Hamer, P. M. Krawczyk, et. al.  (2010).  In Silico Analysis of Kinase 
Expression Identifies WEE1 as a Gatekeeper against Mitotic Catastrophe in Glioblastoma.  
Cancer Cell.  18(3):244-257. 
 

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Parker L.L. and H. Piwnica-Worms.  (1992).  Inactivation of the p34CDK1-cyclin B complex by 
the human WEE1 tyrosine kinase.  Science.  57(5078):1955-7. 
 
Rajeshkumar N.V., E. De Oliveira, N. Ottenhof, et al.  (2011).  MK-1775, a Potent Wee1 
Inhibitor, Synergizes with Gemcitabine to Achieve Tumor Regressions, Selectively in p53-
Deficient Pancreatic Cancer Xenografts.  Clinical Cancer Research  17(9):2799-2806. 
 
Shah, M.A., T. Kojima, D. Hochhauser, et al.  (2019).  Efficacy and Safety of Pembrolizumab 
for Heavily Pretreated Patients with Advanced, Metastatic Adenocarcinoma or Squamous Cell 
Carcinoma of the Esophagus.  JAMA Oncol.  5:546-550. 
 
Sherr CJ.  (1996).  Cancer cell cycles.  Science.  274(5293):1672-7 
 
Siegel, R.L., K.D. Miller, and A. Jemal.  (2018).  Cancer Statistics, 2018.  CA Cancer J Clin.  
68:7-30. 
 
The Cancer Genome Atlas Research Network.  (2017).  Integrated Genomic Characterization of 
Oesophageal Carcinoma.  Nature.  541:169-175. 
 
Thrift, A.P.  (2016).  The Epidemic of Oesophageal Carcinoma: Where are We Now?  Cancer 
Epidemiol.  41:88-95. 
 
University of Michigan Cancer Center.  (2018).  Dose Escalation Trial of AZD1775 and 
Gemcitabine (+Radiation) for Unresectable Adenocarcinoma of the Pancreas.  Available from: 
https://clinicaltrials.gov/ct2/show/NCT02037230. 
 
Van Rossum, P.S.N., N.H. Mohammad, F.P. Vleggaar, and R. van Hillegersberg.  (2018).  
Treatment for Unresectable of Metastatic Oesophageal Cancer:  Current Evidence and Trends.  
Nat Rev Gastroenterol Hepatol.  15:235-249. 
 
Venier, J., R. Herrick, C. Norris, et al.  (2018).  Bayesian Optimal Interval (BOIN) Phase I 
Design (PID-862):  Version 1.0.7, Houston, Texas:  The University of Texas MD Anderson 
Cancer Center. 
 
Wang Y., Li J., Booher R.N., et al.  (2001).  Radiosensitization of p53 Mutant Cells by 
PD0166285, a Novel G2 Checkpoint Abrogator.  Cancer Res.  61(22):8211-7. 
 
Watanabe, N., M. Broome, and T. Hunter.  (1995).  Regulation of the Human WEE1Hu CDK 
Tyrosine 15-kinase during the Cell Cycle.  EMBO J.  14:1878-1891. 
 
Yang, L., C. Shen, C. Pettit, et al.  (2020).  Wee1 Kinase Inhibitor AZD1775 Effectively 
Sensitizes Esophageal Cancer to Radiotherapy.  Clin Cancer Res.  26:3740-3750. 
 
Yuan, Y., K.R. Hess, S.G. Hilsenbeck, et al.  (2016).  Bayesian Optimal Interval Design:  A 
Simple and Well-performing Design for Phase I Oncology Trials.  Clin Cancer Res.  22:4291-
4301. 

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Zhu, L.L., L. Yuan, H. Wang, et al.  (2015).  A Meta-Analysis of Concurrent 
Chemoradiotherapy for Advanced Esophageal Cancer.  PLoS One.  10:e0128616. 
 
 

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85 
APPENDIX A 
PERFORMANCE STATUS CRITERIA 
 
ECOG Performance Status Scale 
Karnofsky Performance Scale 
Grade 
Descriptions 
Percent 
Description 
0 
Normal activity.  Fully active, able 
to carry on all pre-disease 
performance without restriction. 
100 
Normal, no complaints, no evidence 
of disease. 
90 
Able to carry on normal activity; 
minor signs or symptoms of disease. 
1 
Symptoms, but ambulatory.  
Restricted in physically strenuous 
activity, but ambulatory and able 
to carry out work of a light or 
sedentary nature (e.g., light 
housework, office work). 
80 
Normal activity with effort; some 
signs or symptoms of disease. 
70 
Cares for self, unable to carry on 
normal activity or to do active work. 
2 
In bed <50% of the time.  
Ambulatory and capable of all 
self-care, but unable to carry out 
any work activities.  Up and about 
more than 50% of waking hours. 
60 
Requires occasional assistance, but 
is able to care for most of his/her 
needs. 
50 
Requires considerable assistance and 
frequent medical care. 
3 
In bed >50% of the time.  Capable 
of only limited self-care, confined 
to bed or chair more than 50% of 
waking hours. 
40 
Disabled, requires special care and 
assistance. 
30 
Severely disabled, hospitalization 
indicated.  Death not imminent. 
4 
100% bedridden.  Completely 
disabled.  Cannot carry on any 
self-care.  Totally confined to bed 
or chair. 
20 
Very sick, hospitalization indicated. 
Death not imminent. 
10 
Moribund, fatal processes 
progressing rapidly. 
5 
Dead. 
0 
Dead. 
 
 
 

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APPENDIX B 
FORMULA TO ESTIMATE RENAL FUNCTION USING SERUM 
CREATININE 
 
Formulas to estimate renal function using serum creatinine provided by the NCI’s Investigational 
Drug Steering Committee (IDSC) Pharmacological Task Force in table below. 
 
1. Estimated glomerular filtration rate (eGFR) using the Chronic Kidney Disease Epidemiology 
Collaboration (CKD-EPI) (Levey et al., 2009). 
 
Formulae: 
 
Race and 
Sex 
Serum Creatinine (SCr), 
µmol/L (mg/dL) 
Equation 
Black 
 
 
Female 
≤62 (≤0.7) 
GFR = 166 × (SCr/0.7)−0.329 × (0.993)Age 
 
>62 (>0.7) 
GFR = 166 × (SCr/0.7)−1.209 × (0.993)Age 
Male 
≤80 (≤0.9) 
GFR = 163 × (SCr/0.9)−0.411 × (0.993)Age 
 
>80 (>0.9) 
GFR = 163 × (SCr/0.9)−1.209 × (0.993)Age 
 
 
 
White or 
other 
 
 
Female 
≤62 (≤0.7) 
GFR = 144 × (SCr/0.7)−0.329 × (0.993)Age 
 
>62 (>0.7) 
GFR = 144 × (SCr/0.7)−1.209 × (0.993)Age 
Male 
≤80 (≤0.9) 
GFR = 141 × (SCr/0.9)−0.411 × (0.993)Age 
 
>80 (>0.9) 
GFR = 141 × (SCr/0.9)−1.209 × (0.993)Age 
 
SCr in mg/dL; Output is in mL/min/1.73 m2 and needs no further conversions. 
 
2. eGFR using the Modification of Diet in Renal Disease (MDRD) Study (Levey et al., 2006). 
 
175 x SCr–1.154 × age–0.203 × 0.742 (if female) × 1.212 (if black) 
Output is in mL/min/1.73 m2 and needs no further conversions. 
3. Estimated creatinine clearance (ClCr) by the Cockcroft-Gault (C-G) equation (Cockcroft and 
Gault, 1976). 
 
 
Followed by conversion to a value normalized to 1.73 m2 with the patient’s body surface area 
(BSA). 
 
References 
1. Levey, A.S., L.A. Stevens, C.H. Schmid, et al.  (2009).  A new equation to estimate 
glomerular filtration rate.  Ann Inter Med.  150:604-612. 
2. Levey, A.S., J. Coresh, T. Greene, et al.  (2006).  Using standardized serum creatinine 
values in the modification of diet in renal disease study equation for estimating 
glomerular filtration rate.  Ann Intern Med.  145:247-254. 
3. Cockcroft, D.W. and M.H. Gault.  (1976).  Prediction of creatinine clearance from serum 
creatinine.  Nephron.  16:31-41. 
 

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87 
APPENDIX C 
PATIENT MEDICATION DIARY 
 
C.1 
Medication Diary for AZD1775 - Days -7 through -1 (Week -1)  
 
 
 
 
PATIENT’S MEDICATION DIARY – AZD1775 
 
     Today’s date _______________________________ 
Agent ___________AZD1775___________ 
     Patient Name_________________________ (initials acceptable) 
Patient Study ID _____________________ 
INSTRUCTIONS TO THE PATIENT: 
1. 
Complete one form for each month. 
2. 
Your doctor will tell you what days you should take AZD1775.  You will take ___25 mg capsules and ___ 100 mg capsules on 
the designated treatment days.  You should swallow the capsules whole.  Do not chew or open capsules. 
3.     If you vomit shortly after you take your medications:  the dose should only be replaced if all of the intact capsules can be seen  
and counted.  Should you miss a scheduled dose for whatever reason (e.g., as a result of forgetting to take the tablets or  
vomiting), you can take the scheduled dose up to a maximum of 2 hours after that scheduled dose time.  If greater than 2 hours  
after the scheduled dose time, do not take the missed dose.  Take your assigned dose at the next scheduled time. 
4.     Record the date, the number of capsules of each size you took, and when you took them.   
5. 
If you have any comments or notice any side effects, please record them in the Comments column.   
6.     Notify your doctor at the first sign of poorly formed or loose stools, or an increased frequency of bowel movements.  
Loperamide (Imodium) should be kept on hand and should be taken as recommended by your doctor. 
7.     Please return the forms to your physician when you go for your next appointment. 
 
Day 
 
Date 
What time was 
dose taken? 
# of capsules taken 
Comments 
25 mg 
100 mg 
1 
 
 
 
 
 
2 
 
 
 
 
 
3 
 
 
 
 
 
4 
 
 
 
 
 
5 
 
 
 
 
 
6 
 
 
 
 
 
7 
 
 
 
 
 
Physician’s Office will complete this section: 
1. 
Date patient started protocol treatment____________________________________________________________________ 
2. 
Date patient was removed from study_____________________________________________________________________ 
3. 
Patient’s planned total daily dose________________________________________________________________________ 
4. 
Total number of capsules taken this month (each size)___________________________________________________________ 
5. 
Physician/Nurse/Data Manager’s Signature ________________________________________________________________ 
 
    Patient’s 
signature:____________________________________________________________________________________ 
 
 
 
 
CTEP-assigned Protocol #__10389____ 
 
Local Protocol # ___________________ 

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88 
C.2 
Medication Diary for AZD1775 - Cycle 1 (Weeks 1-3) 
 
 
 
 
PATIENT’S MEDICATION DIARY – AZD1775 
 
     Today’s date _______________________________ 
Agent ___________AZD1775___________ 
     Patient Name_________________________ (initials acceptable) 
Patient Study ID _____________________ 
INSTRUCTIONS TO THE PATIENT: 
1. 
Complete one form for each month. 
2. 
Your doctor will tell you what days you should take AZD1775.  You will take your dose of AZD1775 on designated days 2-3 
hours prior to radiation therapy.  You will take ___25 mg capsules and ___ 100 mg capsules on the designated treatment days.  
You should swallow the capsules whole.  Do not chew or open capsules. 
3. 
If you vomit shortly after you take your medications:  the dose should only be replaced if all of the intact capsules can be seen  
and counted.  Should you miss a scheduled dose for whatever reason (e.g., as a result of forgetting to take the tablets or  
vomiting), you can take the scheduled dose up to a maximum of 2 hours after that scheduled dose time.  If greater than 2 hours  
after the scheduled dose time, do not take the missed dose.  Take your assigned dose at the next scheduled time. 
4.    Shaded rows indicate the days that you WILL NOT take AZD1775. 
5.     Record the date, the number of capsules of each size you took, and when you took them.   
6. 
If you have any comments or notice any side effects, please record them in the Comments column.   
7.     Notify your doctor at the first sign of poorly formed or loose stools, or an increased frequency of bowel movements.  
Loperamide (Imodium) should be kept on hand and should be taken as recommended by your doctor. 
8.     Please return the forms to your physician when you go for your next appointment. 
 
Day 
 
Date 
What time was 
dose taken? 
# of capsules taken 
Comments 
25 mg 
100 mg 
1 
 
 
 
 
 
2 
 
 
 
 
 
3 
 
 
 
 
 
4 
 
 
 
 
 
5 
 
 
 
 
 
6 
 
 
 
 
 
7 
 
 
 
 
 
8 
 
X 
X 
X 
X 
9 
 
X 
X 
X 
X 
10 
 
X 
X 
X 
X 
11 
 
X 
X 
X 
X 
12 
 
X 
X 
X 
X 
13 
 
X 
X 
X 
X 
14 
 
X 
X 
X 
X 
15 
 
 
 
 
 
16 
 
 
 
 
 
17 
 
 
 
 
 
18 
 
 
 
 
 
19 
 
 
 
 
 
20 
 
 
 
 
 
21 
 
 
 
 
 
Physician’s Office will complete this section: 
1. 
Date patient started protocol treatment____________________________________________________________________ 
 
CTEP-assigned Protocol #__10389____ 
 
Local Protocol # ___________________ 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
89 
2. 
Date patient was removed from study_____________________________________________________________________ 
3. 
Patient’s planned total daily dose________________________________________________________________________ 
4. 
Total number of capsules taken this month (each size)___________________________________________________________ 
5. 
Physician/Nurse/Data Manager’s Signature ________________________________________________________________ 
 
    Patient’s 
signature:____________________________________________________________________________________ 
 
 
 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
90 
APPENDIX D 
PATIENT DRUG INTERACTIONS HANDOUT AND WALLET 
CARD 
 
Information for Patients, Their Caregivers and Non-Study Healthcare Team on Possible 
Interactions with Other Drugs and Herbal Supplements  
 
Patient 
Name: 
      
Diagnosis:  
      
Trial #: 
10389 
Study 
Doctor: 
      
Study Doctor 
Phone #:  
      
Study 
Drug(s): 
AZD1775 
 
Please show this paper to all your healthcare providers (doctors, physician assistants, nurse 
practitioners, pharmacists), and tell them you are taking part in a clinical trial sponsored by the 
National Cancer Institute.  
 
AZD1775 (adavosertib) interacts with certain specific enzymes in the liver and certain transport 
proteins that help move drugs in and out of cells. 
 
• The enzymes in question are CYP 3A4 and 2C19.  AZD1775 (adavosertib) is metabolized 
by CYP3A4 and may be affected by other drugs that inhibit or induce these enzymes.  
AZD1775 (adavosertib) is an inhibitor of CYP 3A4 and 2C19 and may affect the 
metabolism of other drugs. 
• The proteins in question are OATP1B1, OATP1B3, MATE1, MATE2K, P-gp, and BCRP.  
AZD1775 (adavosertib) is a substrate of P-gp and BCRP and may be affected by other 
drugs that inhibit or induce these transporters.  AZD1775 (adavosertib) is an inhibitor of 
OATP1B1, OATP1B3, MATE1, MATE2K, P-gp, and BCRP and may affect transport of 
other drugs in and out of cells. 
 
 
To the patient: Take this paper with you to your medical appointments and keep the 
attached information card in your wallet.   
 
AZD1775 (adavosertib) may interact with other drugs which can cause side effects.  For this 
reason, it is very important to tell your study doctors of any medicines you are taking before you 
enroll onto this clinical trial.  It is also very important to tell your doctors if you stop taking any 
regular medicines, or if you start taking a new medicine while you take part in this study.  When 
you talk about your current medications with your doctors, include medicine you buy without a 
prescription (over-the-counter remedy), or herbal supplements such as St. John’s Wort.  It is 
helpful to bring your medication bottles or an updated medication list with you. 
 
Many health care providers can write prescriptions.  You must tell all of your health care 
providers (doctors, physician assistants, nurse practitioners, or pharmacists) you are taking part 
in a clinical trial.  
 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
91 
These are the things that you and they need to know:  
 
AZD1775 (adavosertib) must be used very carefully with other medicines that need certain liver 
enzymes or transport proteins to be effective or to be cleared from your system.  Before you 
enroll onto the clinical trial, your study doctor will work with your regular health care providers 
to review any medicines and herbal supplements that are considered “strong inducers/inhibitors 
or substrates of CYP 3A4, 2C19, OATP1B1, OATP1B3, MATE1, MATE2K, P-gp, and 
BCRP.” 
 
•  Please be very careful!  Over-the-counter drugs (including herbal supplements) may 
contain ingredients that could interact with your study drug.  Speak to your doctors or 
pharmacist to determine if there could be any side effects.  
•  Your regular health care provider should check a frequently updated medical reference or 
call your study doctor before prescribing any new medicine or discontinuing any 
medicine.  Your study doctor’s name is  
 
_____________________________________ and he or she can be contacted at  
 
_____________________________________.                                                                                                         May 2015 
 
(Next page: Patient Drug Interaction Wallet Card) 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
92 
PATIENT DRUG INTERACTION WALLET CARD 
 
EMERGENCY INFORMATION 
 
DRUG INTERACTIONS  
Show this card to all of your 
healthcare providers. Keep 
it with you in case you go to 
the emergency room. 
Tell your doctors before you 
start or stop any medicines. 
Check with your doctor or 
pharmacist if you need to use 
an over-the-counter medicine 
or herbal supplement! 
Carry this card with you at all times   
AZD1775 interacts with certain specific enzymes in the liver or 
other tissues like the gut and certain transport proteins that help 
move drugs in and out of cells and must be used very carefully with 
other medicines. 
Patient Name:       
Use caution and avoid the 
following drugs if possible:  
 
 
 
Your healthcare providers should be aware of any medicines that 
are strong inducers/inhibitors or substrates of CYP3A4, CYP2C19, 
OATP1B1, OATP1B3, MATE1, MATE2K, P-gp, and BCRP. 
Diagnosis:       
Study Doctor:       
Study Doctor Phone #:       
NCI Trial #: 10389 
Before prescribing new medicines, your health care provider 
should check a frequently-updated medical reference for a list of 
drugs to avoid or contact your study doctor. 
Study Drug(S):  AZD1775 
(adavosertib) 
 
             Version 05/2015 
 
For more information: 1-800-4-
CANCER 
For more information: 1-800-4-
CANCER 
For more information: 1-800-4-
CANCER 
For more information: 1-800-4-
CANCER 
cancer.gov | clinicaltrials.gov 
cancer.gov | clinicaltrials.gov 
cancer.gov | clinicaltrials.gov 
cancer.gov | clinicaltrials.gov 
 
Fold at dotted lines: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
93 
APPENDIX E 
OGILVIE DYSPHAGIA SCORING 
 
Ogilvie’s dysphagia score classification is as follows: 
 
Grade 
Description 
0 
Able to eat the ordinary diet 
1 
Able to eat solid food 
2 
Able to eat semisolids 
3 
Able to take liquids 
4 
Able to eat nothing 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
94 
APPENDIX F 
PRE-BIOPSY ASSESSMENT 
 
A pre-biopsy lesion assessment can increase trial safety and efficiency.  By agreement between 
all investigators, an attempt at biopsy will be made if the clinical trial team determines that a 
biopsy poses minimal relative risk, provides potential clinical gain to the participant, and will 
likely yield sufficient tissue for analysis.   
 
Pre-biopsy assessments will be reported and tracked through a trial-specific CRF within the 
CTEP Medidata Rave system.  Additional information can be found in the Investigational 
Radiology SOP available at:   
https://ctep.cancer.gov/initiativesPrograms/docs/ETCTN_IR_Research_Biopsy_SOP.pdf.  
 
Individual Patient Pre-Biopsy Assessment.  Gastroenterologist co-investigators are encouraged 
to apply this pre-biopsy scoring and correlation system to assist in the determination of biopsy 
appropriateness. 
 
• Gastroenterologist co-investigators assign a subjective score of 1-3 based on likelihood of 
success due to lesion characteristics.  
 
1.  Biopsy should not be done  
A. Due to safety concerns 
B. Due to lack of suitable lesion for biopsy 
 
2.  Uncertainty about success  
A. Due to access path to lesion 
B. Due to lesion characteristics  
 
3.  Likely successful  
 
• Lesion characteristics to be considered 
• Size (small) (<2 cm) 
• Location/path to lesion 
• Morphologic features (necrosis, sub-solid, sclerosis, ill-defined/infiltrative) 
• PET (+/-), avidity  
• Organ/site (sclerotic bone is low yield; fine needle aspiration to be used) 

NCI Protocol #: 10389 
Version Date:  January 13, 2022 
 
95 
 
APPENDIX G 
TISSUE BIOPSY VERIFICATION 
 
A copy of the diagnostic pathology report must be shipped with all tissue specimens sent to the 
ETCTN Biorepository. 
 
If the corresponding pathology report is not available for the biopsy, then a copy of the 
radiology report or operative report from the biopsy procedure and the diagnostic 
pathology report must be sent to the ETCTN Biorepository. A completed copy of this 
appendix (i.e., Tissue Biopsy Verification) must also be submitted to the ETCTN 
Biorepository. 
 
Note: If this information is not provided with the biopsy specimen, then it will not be accepted 
by the ETCTN Biorepository. 
 
 
 
Please have the Clinician* responsible for signing out this patient’s case complete the following: 
 
ETCTN Universal Patient ID:  
 
 
ETCTN Patient Study ID:   
 
 
Date of Procedure (mm/dd/yyyy):   
 
 
Tissue Type (circle one): 
Primary 
 
 
Time point (circle one):  Baseline 
Week -1 Day 4 or 5 
 
 
 
Site Tissue Taken From:   
 
 
Diagnosis:   
 
 
I agree that this tissue may be released for research purposes only and that the release of this 
tissue will not have any impact on the patient’s care. 
 
 
 
Clinician Signature 
Date 
 
 
 
Clinician Printed Name 
 
*Note: For the purposes of this form, Clinician could include the Nurse Practitioner, Registered Nurse, Pathologist, 
Radiologist, Interventional Radiologist, Surgeon, Oncologist, Internist, or other medical professional responsible for 
the patient’s care.  
 
Version: 1 
Effective Date: 9/2019 

 
 
APPENDIX H 
NCLN PHARMACODYNAMICS LABORATORY FROZEN BIOPSY 
COLLECTION PROCEDURE 
 

DCTD Standard Operating Procedures 
 
 
Date: 2019.08.02 16:29:51 -04'00' 
 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
Page 1 of 11 
Doc. #: 
SOP340507 
Revision: 
G 
Effective Date: 
6/19/2019 
 
Laboratory of Human Toxicology & Pharmacology  
Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc. 
Frederick National Laboratory for Cancer Research 
 
Corey J. Evans -S 
Technical Reviewer: 
Corey Evans 
 
Date: (Affiliate) 
 
 
Digitally signed by Corey J. Evans - 
S (Affiliate) 
Date: 2019.06.20 13:47:37 -04'00' 
Digitally signed by Jiuping Ji -S 
Jiuping Ji -S (Affiliate) (Affiliate) 
Date: 2019.07.05 10:34:23 -04'00' 
Katherine V. Ferry- 
IQC Approval: 
Katherine V. Ferry-Galow 
 
Date: galow -S (Affiliate) 
Ralph E. Parchment -S 
LHTP Approval: 
Ralph E. Parchment 
 
Date: (Affiliate) 
Digitally signed by Katherine V. 
Ferry-galow -S (Affiliate) 
Date: 2019.07.22 14:46:16 -04'00' 
Digitally signed by Ralph E. 
Parchment -S (Affiliate) 
Date: 2019.08.02 13:09:35 -04'00' 
DCTD OD Approval: 
Toby Hecht 
 
Date: Toby T. Hecht -S 
Digitally signed by Toby T. Hecht -S 
 
Change History 
 
Revision 
Approval Date 
Description 
Originator 
Approval 
G 
6/19/2019 
Minor updates to collection and shipping procedures. 
KFG 
REP 
F 
2/11/2015 
Updated contact shipping address and process for advance 
notification of shipments. 
KFG 
REP 
E 
7/3/2013 
Updated tube-type to 1.5-mL conical bottom screw cap 
tubes to allow for broader use in DCTD assays and 
minimize the need to transfer biopsies during sample 
extraction steps. Decreased maximum time from biopsy 
collection to freezing to 2 minutes. 
YAE 
REP 
D 
1/8/2013 
Update handling in surgical suite including details on 
halving of biopsy. Record total time elapsed from biopsy 
collection to freezing. 
YAE, MM 
JJ 
C 
12/29/2010 
Update sample snap freeze to dry ice/ethanol bath or liquid 
nitrogen. 
YAE 
JJ 
B 
7/24/2009 
Updated SOP format and prepared for publication to the 
DCTD Biomarkers Web site 
YAE 
JJ 
A 
10/13/2006 
Revision with New Shipping Address 
YZ 
JJ 
-- 
8/25/2006 
New Document 
YZ 
JJ 
 
Please check for revision status of the SOP at 
http://dctd.cancer.gov/ResearchResources/ResearchResources-biomarkers.htm 
and be sure to use the current version. 
NCTVL Reviewer: 
Jiuping Ji 

DCTD Standard Operating Procedures 
 
 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
Page 2 of 11 
Doc. #: 
SOP340507 
Revision: 
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Effective Date: 
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TABLE OF CONTENTS 
1.0 
PURPOSE .................................................................................................................................................... 3 
2.0 
SCOPE .......................................................................................................................................................... 3 
3.0 
ABBREVIATIONS ...................................................................................................................................... 3 
4.0 
INTRODUCTION ........................................................................................................................................ 3 
5.0 
ROLES AND RESPONSIBILITIES ............................................................................................................ 3 
6.0 
MATERIALS AND EQUIPMENT REQUIRED ........................................................................................ 4 
7.0 
OPERATING PROCEDURES .................................................................................................................... 5 
8.0 
SHIP TO FNLCR FOR ANALYSIS (OPTIONAL) .................................................................................... 7 
APPENDIX 1: BATCH RECORD ............................................................................................................................... 9 
APPENDIX 2: PD SAMPLE SHIPPING MANIFEST .............................................................................................. 11 

DCTD Standard Operating Procedures 
 
 
Title: 
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1.0 
PURPOSE 
Standardize the method for collecting and handling frozen needle tumor biopsies to enable specimen use 
for measurement of pharmacodynamic (PD) markers following treatment with anticancer agents. 
2.0 
SCOPE 
This procedure applies to all personnel involved in the collection and handling of frozen needle tumor 
biopsies for use in PD marker assays during clinical trials. The goal of this SOP and associated training is 
to ensure consistency in tumor needle biopsy collection and handling between clinical sites. 
3.0 
ABBREVIATIONS 
 
DCTD 
= Division of Cancer Treatment and Diagnosis 
ELISA 
= Enzyme-Linked ImmunoSorbent Assay 
FNLCR 
= Frederick National Laboratory for Cancer Research 
ID 
= Identification / Identifier 
IQC 
= Internal Quality Control 
LHTP 
= Laboratory of Human Toxicology and Pharmacology 
NCTVL 
= National Clinical Target Validation Laboratory 
PADIS 
= Pharmacodynamics Assay Development & Implementation Section 
PD 
= Pharmacodynamic 
SOP 
= Standard Operating Procedure 
 
4.0 
INTRODUCTION 
Specimen handling, shipping, and storage procedures (pre-analytical variables) can have a significant 
impact on the reliability of biomarker measurements in the laboratory. Following detailed steps for 
sample collection and handling procedures and recording any deviations from this procedure allows 
retrospective identification of artifactual changes in biomarker readout and increases the reliability of the 
data and validity of the analytical results. 
5.0 
ROLES AND RESPONSIBILITIES 
Laboratory Director/Supervisor      The Laboratory Director/Supervisor, directs laboratory operations, 
supervises technical personnel and reporting of findings, and is       
responsible for the proper performance of all laboratory procedures. 
Oversees the personnel who follow the SOPs in the laboratory and is 
responsible for ensuring the personnel are certified and have sufficient 
experience to handle clinical samples. 
Certified Assay Operator and/or 
A Certified Assay Operator and/or PK/PD Support Lab personnel may 
PK/PD Support Lab Personnel 
be a Laboratory Technician/Technologist, Research Associate, or 
Laboratory Scientist who has been certified through DCTD training on 
this SOP. Works under the guidance of the Laboratory 
Director/Supervisor. This person performs laboratory procedures and 
examinations in accordance with the current SOP(s), as well as any 
other procedures conducted by a laboratory, including maintaining 
equipment and records and performing quality assurance activities 
related to performance. 

DCTD Standard Operating Procedures 
 
 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
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5.1 
It is the responsibility of the Laboratory Director/Supervisor to ensure that all personnel have 
documented training and qualification on this SOP prior to the actual handling and processing of 
samples from clinical trial patients. The Laboratory Director/Supervisor is responsible for 
ensuring the Certified Assay Operator running the SOP has sufficient experience to handle and 
analyze clinical samples. 
5.2 
It is the responsibility of the Certified Assay Operator and/or PK/PD Support Lab personnel to 
confirm scheduled specimen collection time points, pre-print all labels and data collection sheets 
in advance, check documentation for accuracy, and verify that the required collection tubes, 
supplies, and equipment are available for successful collection and handling of biopsy samples. 
5.3 
The Certified Assay Operator and/or PK/PD Support Lab personnel responsible for conducting 
the specimen collection and handling procedures are to follow this SOP and complete the 
required tasks and associated documentation. The Batch Record (Appendix 1) must be completed 
in real-time for each experimental run, with each page dated and initialed, and placed with the 
clinical sample information. 
5.4 
The responsible personnel are to check the DCTD Biomarkers Web site 
(http://dctd.cancer.gov/ResearchResources/ResearchResources-biomarkers.htm) to verify that the 
latest SOP version is being followed. 
6.0 
MATERIALS AND EQUIPMENT REQUIRED 
6.1 
Stopwatch, total time in minutes and seconds required 
6.2 
1.5-mL Sarstedt o-ring screw cap, conical bottomed tubes (Sarstedt, Cat#: 72.703.416) 
6.3 
Disposable, fine-tipped tweezers (e.g., VWR, Cat#: 83009-010). Tweezer tips need to easily fit 
to bottom of a 1.5-mL Sarstedt tube 
6.4 
Printable microcentrifuge tube labels or BSI labeling system 
6.5 
81-place freezer boxes (e.g., Fisher Scientific, Cat#: 12-565-182) 
6.6 
Thermoflask cooler or polystyrene foam container 
6.7 
Ice bucket 
6.8 
Liquid nitrogen or dry ice/ethanol bath 
6.9 
Wet ice 
6.10 
-80°C freezer (or lower) 
6.11 
Biohazard specimen bags 
6.12 
Insulating Styrofoam shipping container 

DCTD Standard Operating Procedures 
 
 
Title: 
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7.0 
OPERATING PROCEDURES 
7.1 
Record the name and certification number of the Certified Assay Operator and/or PK/PD Support 
Lab personnel performing the SOP, the facility/clinic collecting the specimens, the 
Patient/Sample ID, the primary diagnosis, and the clinical protocol number in the Batch Record 
(Appendix 1). 
o The Batch Record for this SOP is sufficient for collection of a single set of biopsy 
samples collected from a single patient at a single timepoint. If collecting biopsy samples 
for more than one patient, prepare a separate Batch Record for each patient.  
7.2 
Labels 
7.2.1 
Prepare enough pre-printed specimen labels for each whole or halved biopsy sample to be 
collected and frozen as defined in the Pharmacodynamic/Correlative Study section of the 
Clinical Protocol; be sure to coordinate with the clinical center if they prepare the labels 
for sample collection. If two passes are collected from one tumor, the labels would be 
identical except that the specimen ID would be followed by a letter A/B to designate pass 
number. The specimen ID includes the CTEP protocol number followed by a unique 
patient identifier and a specimen series ID. 
NCI tumor biopsy specimen IDs for PD sampling are series 500 with consecutive 
numbers identifying the collection time point as defined in the Clinical Protocol. 
Sample pre-printed label for all frozen-tissue biopsy tubes: 
 
7.2.2 
Of the pre-printed labels prepared for each sample, one label will go on each 1.5-mL 
Sarstedt tube, one on the Batch Record (Appendix 1), and the last will be given to the 
research nurse to place into the patient record sheet. Note: be sure that no patient 
identifiable information is shown on the labels. 
7.3 
Tumor Needle Biopsy Collection and Handling 
7.3.1 
The research nurse is to notify the laboratory of scheduled PD sample collections, 
preferably giving at least 24 hours of notice. Arrive at the biopsy collection site early 
enough to allow sufficient time to set up laboratory supplies, collect relevant clinical 
information, and ensure rapid transport of specimens to the laboratory for placement at 
-80ºC (or lower) after collection. 
7.3.2 
Bring all necessary lab supplies including: disposable tweezers, a minimum of two 
1.5-mL Sarstedt tubes (one for each whole biopsy core) pre-cooled on liquid nitrogen or 
dry ice/ethanol in an insulated bucket, and one pre-printed specimen label to give to the 
research nurse for the patient record. 
Note: Pre-chill additional 1.5-mL Sarstedt tubes for specimen collection in case the 
interventional radiologist collects additional passes, or one of the other tubes is 
compromised prior to collection. 

DCTD Standard Operating Procedures 
 
 
Title: 
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7.3.3 
The total time elapsed between biopsy collection and placement into the pre-chilled tube 
is of key importance to biomarker analysis; biopsies should be frozen within 2 minutes 
of collection. The interventional radiologist will eject the biopsy onto a sterile slide (for 
optimal analyte recovery the slide should be pre-chilled). Start a stopwatch (or note the 
time) at this point (Appendix 1, Section 1) and immediately walk the slide to the sample 
preparation table. 
7.3.4 
In the Batch Record (Appendix 1, Section 1), indicate if a full or halved biopsy, as 
defined in the Pharmacodynamic/Correlative Study section of the Clinical Protocol, is 
prepared. 
7.3.4.1 
For whole biopsies: Uncap an empty, pre-chilled 1.5-mL Sarstedt tube and, 
using disposable tweezers, pick up the freshly collected needle biopsy with the 
tweezers at one end, and touch the opposite end of the biopsy to the inner 
surface of the prechilled 1.5-mL Sarstedt tube. This should attach the tissue to 
the tube, allowing it to be dropped into the tube while releasing the tissue from 
the tweezers without sticking. Dispose of the tweezers in the appropriate 
biohazardous waste container(s). 
7.3.4.2 
For halved biopsies: Use 1-2 disposable tweezers and cut/shear the biopsy in 
half cross-wise while it is on the slide (do not pull or stretch the biopsy 
longitudinally). Use the tweezers to transfer the halved biopsies to sterile pre- 
chilled tubes as indicated above. 
7.3.5 
Immediately snap freeze the biopsy by placing the tube in liquid nitrogen or a dry 
ice/ethanol bath. Note: DO NOT let the tubes tip over in the dry ice/ethanol bath. 
7.3.6 
Calculate the total time elapsed from biopsy collection to biopsy freezing and record the 
total number of minutes and seconds elapsed in the Batch Record (Appendix 1, 
Section 1). 
7.3.7 
Note the specific location of each biopsy pass collected (e.g., spleen, large left upper 
quadrant splenic mass) and a description of the appearance of the biopsy (e.g., large 
whole core or small, fragmented core) in the Batch Record (Appendix 1, Section 1). 
7.3.8 
Note the biopsy timepoint (Appendix 1, Section 1). 
7.4 
If biopsy procedure details can be obtained from the interventional radiologist or research nurse, 
record them in the Batch Record (Appendix 1, Section 2). Some information may not be 
available until a later time from the clinical staff. 
During first-in-human PD sample collection studies, information such as type of anesthesia and 
time-lag between biopsy needle withdrawal and sample freezing need to be tracked in order to 
determine the optimal sample collection procedure for the clinical community. 
7.5 
Return to the sample processing laboratory and transfer the frozen biopsy specimen(s) to -80°C 
(or lower) for storage until shipment to the PD processing laboratory. Record the date and time 
specimens are placed at -80ºC (or lower; Appendix 1, Section 3). 
7.6 
Review and finalize the Batch Record and document ANY and ALL deviations from this SOP in 
the Batch Record (Appendix 1, Section 5). 
7.7 
The Laboratory Director/Supervisor should review the Batch Record and sign to affirm the data 
contained within are correct (Appendix 1, Section 6). 

DCTD Standard Operating Procedures 
 
 
Title: 
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Effective Date: 
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8.0 
SHIP TO FNLCR FOR ANALYSIS (OPTIONAL) 
If shipping to a location other than FNLCR, use the following steps as a guide. 
8.1 
Sites are required to create a FedEx shipping label to accommodate the variable dry ice weight of 
the shipment package. Use only FedEx Priority Overnight Shipping. FNLCR PD Support will 
provide a FedEx account number to cover the cost of the shipment via 
NCI_PD_Support@mail.nih.gov. 
8.1.1 
By linking the FNLCR FedEx account number provided to the biopsy shipment, FNLCR 
PD Support can closely monitor the shipment and cover all shipping costs, and 
appropriate notification can be provided to pertinent staff of expected sample shipment 
arrival. 
8.1.2 
Please send all shipments via FedEx Priority Overnight shipping to the FNLCR PD 
Specimen Central Receiving address listed below: 
Attention: Dan Danner 
NCI-F/FNLCR 
1073 Beasley Street, Building 1073 
Fort Detrick 
Frederick, MD 21701 
Phone: (301) 846-5748 
8.2 
Once a tumor biopsy has been collected from a patient and placed at -80ºC (or lower), FNLCR 
PD Specimen Central Receiving should be notified that the specimens are ready for shipment. 
Preferably, if additional biopsies will be taken from the same patient (post-dose timepoint[s]), the 
biopsies are to be stored in a local freezer at -80ºC (or lower), and shipped together as a full set in 
one shipment. 
8.3 
Send an e-mail to FNLCR PD Specimen Central Receiving (NCI_PD_Support@mail.nih.gov) to 
advise that biopsy samples are being prepared for shipment. State “Protocol Name PD 
Specimens Ready for Shipment” in the subject line. Request a confirmation e-mail that personnel 
will be available on the expected delivery date and time. Personnel are generally available to 
receive frozen shipments Tuesday through Friday, exclusive of government holidays. If needed, 
FNLCR PD Central Receiving can be contacted directly at (240) 344-5697 (Rachel Andrews) or 
(301) 401-8070 (Amy Pantella). 
8.4 
Use the PD Sample Shipping Manifest template in Appendix 2 to generate a shipping list 
containing pertinent sample information. 
8.5 
Make a copy of the Shipping Manifest and specimen Batch Records so that one copy can be sent 
to FNLCR with the biopsy samples and one copy can be maintained at the collection site for 
internal records. 
8.6 
Day of Shipment 
8.6.1 
Just prior to shipment, place specimen tubes into a biohazard specimen bag then in an 
insulating Styrofoam shipping container. The insulating Styrofoam shipping containers 
are required to have dimensions of at least 14″×11″×9″ (length, width, height) with a 
minimum of 20 pounds of added dry ice. Sufficient dry ice is imperative to maintain the 
samples at -20°C for at least 72 hours. Expect 10 pounds of dry ice to sublimate per day 
during transit. Add additional dry ice to the required 20 pounds if shipping is anticipated 
to be longer than 24 hours. 

DCTD Standard Operating Procedures 
 
 
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Tumor Frozen Needle Biopsy Specimen Collection and Handling 
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8.6.2 
All weekly processing specimens are recommended to ship out on Mondays through 
Thursdays via FedEx Priority Overnight (excluding any day before a federal holiday). 
8.6.3 
Verify that the contents of the package match the Shipping Manifest and sign and date 
the bottom of both copies of the Shipping Manifest. Place one copy of the Shipping 
Manifest inside the shipping box along with copies of the completed Batch Records for 
all specimens. 
8.6.4 
Seal the box and print and attach the shipping address onto the outside of the shipping 
container; be sure the container is labeled as containing biohazardous specimens. 
8.6.5 
Record the shipping date, time, tracking number, and shipping information in the Batch 
Record (Appendix 1, Section 4). 
8.6.6 
E-mail FNLCR PD Specimen Central Receiving (NCI_PD_Support@mail.nih.gov) a 
shipment notification. State “Protocol Name PD Specimen Shipment” in the subject line 
and reference the tracking number in the e-mail. Please notify FNLCR PD Specimen 
Central Receiving of any issues or protocol deviations as soon as possible and provide 
written notes on the Batch Record. 
8.6.7 
Once specimens arrive at the receiving laboratory, they should be immediately placed at 
-80ºC (or lower) pending delivery to the processing laboratory for protein extraction. 

DCTD Standard Operating Procedures 
BATCH RECORD 
INITIALS:  
 
DATE:   
 
 
 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
Page 9 of 11 
Doc. #: 
SOP340507 
Revision: 
G 
Effective Date: 
6/19/2019 
 
APPENDIX 1: BATCH RECORD 
A separate Batch Record should be started for each patient sample set. 
Note: 
A pre-dose and post-dose sample from the same patient would have the 
same Patient ID, but different Specimen ID numbers. 
 
Note: 
Record times using military time (24-h designation); for example, 
specify 16:15 to indicate 4:15 PM. 
 
Certified Assay Operator:   
 
Certification Number:   
 
 Check here if PK/PD Support Lab Personnel 
 
 
Facility/Clinic Collecting Specimens:   
 
Clinical Protocol Number:   
 
Patient ID:   
 
Primary Diagnosis:   
 
 
1. 
Biopsy Collection 
 
1st Pass 
2nd Pass 
3rd Pass 
4th Pass 
Specimen ID 
Site of Biopsy (complete for all 
passes or note “same” for 
replicate cores) 
Description of Biopsy (e.g., large 
intact core or small and 
fragmented core) 
Biopsy Timepoint (Cycle, Day, 
and Hours post dose, if post 
treatment) 
Biopsy size prepared for PD or 
histological analysis: 
 Full 
 Halved 
 Full 
 Halved 
 Full 
 Halved 
 Full 
 Halved 
Required: 
Time elapsed from collection to 
placement in tube min 
sec 
min 
sec 
min 
sec 
min 
sec 
Time biopsy collected (opt) 
: 
: 
: 
: 
Time biopsy placed in tube 
(opt) 
: 
: 
: 
: 
 
 
Place 
PD Specimen Label 
Here 

DCTD Standard Operating Procedures 
BATCH RECORD 
INITIALS:  
 
DATE:   
 
 
 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
Page 10 of 11 
Doc. #: 
SOP340507 
Revision: 
G 
Effective Date: 
6/19/2019 
 
2. 
Biopsy Procedure Details 
 
Specimen ID 
Time local anesthesia administered 
: 
Dose of local anesthetic 
mg 
Name of local anesthetic used (from Research Nurse) 
Time of skin incision 
: 
Needle Type (e.g., Temno) 
Needle diameter 
gauge 
Needle Length 
cm 
Time guide needle introduced 
: 
Time guide needle placement confirmed 
: 
Time biopsy needle introduced 
: 
3. 
Biopsy Storage 
 
Date/time biopsy specimen(s) placed at 
                -80°C (or lower) 
  
/ 
/ 
 
  
: 
 
  
ºC 
4. 
Shipping to FNLCR (optional) 
 
                 Date and time samples shipped 
  
 
  
: 
 
Tracking information 
  
 
 
 
 
**Attach copy of Shipping Manifest 
5. 
Notes, including any deviations from the SOP: 
 
 
 
 
 
 
 
 
 
 
6. 
Laboratory Director/Supervisor Review of Batch Record 
                 Laboratory Director/Supervisor: 
(PRINT) 
 
   (SIGN) 
                 Date: 
  
/ 
/ 
 

 
 
DCTD Standard Operating Procedures (SOP) 
Title: 
Tumor Frozen Needle Biopsy Specimen Collection and Handling 
Page 11 of 11 
Doc. #: 
SOP340507 
Revision: 
G 
Effective Date: 
6/19/2019 
  APPENDIX 2: PD SAMPLE SHIPPING MANIFEST 
 
From: 
Phone: 
Email: 
PD Sample  
Shipping Manifest 
In 
Package 
 
Item No 
Patient/ Specimen 
ID 
Clinical 
Protocol 
 
Description 
Primary 
Diagnosis 
 
Site of Biopsy 
Time Point 
Scheduled 
Collection 
Date 
Collection 
Time 
 
Example 
1234-xx999-1023-500 
12-C-0000 
Full biopsy 
Melanoma 
Right forearm 
Pre-dose D1 
06/12/12 
08:50 
 
 
Example 
 
1234-xx999-1023-501 
 
12-C-0000 
 
Half biopsy 
 
Melanoma 
Right forearm 
(same lesion) 
 
Cycle 1, D8 
 
06/20/12 
 
16:05 
  
1 
/ 
/ 
  
2 
/ 
/ 
  
3 
/ 
/ 
  
4 
/ 
/ 
  
5 
/ 
/ 
  
6 
/ 
/ 
  
7 
/ 
/ 
  
8 
/ 
/ 
  
9 
/ 
/ 
  
10 
/ 
/ 
Verification of Contents 
Signature 
Date 
 
Contents Verified Collection Laboratory 
  
 
/ 
/ 
 
Contents Verified FNLCR PD Central Receiving 
 
/ 
/