A Phase I/II Trial of TRC102 (Methoxyamine HCl) in Combination With Temozolomide in Patients With Relapsed Solid Tumors and Lymphomas
Phase: PHASE1
Conditions: Lymphomas, Solid Tumors, NSCLC, Metastatic Colon Carcinoma, Granulosa Cell Ovarian Cancer
Sponsor: National Cancer Institute (NCI)

CTEP #9483
Clinical Center #: 13-C-0118
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TITLE:  A Phase I/II Trial of TRC102 (methoxyamine HCl) in Combination with 
Temozolomide in Patients with Relapsed Solid Tumors and Lymphomas
Abbreviated Title: Ph I/II TRC102 Temozolomide 
Coordinating Center:
Developmental Therapeutics Clinic (NCIDTC)
National Cancer Institute
10 Center Drive, Bethesda, MD 20892
Principal Investigator:
A. P. Chen, MD A-E
31 Center Drive
Bldg 31 Room 3A44
Bethesda, MD 20892 
Phone: 240-781-3320; Fax: 240-541-4515 
chenali@mail.nih.gov
Referral Contact:
Jennifer Zlott, RN, DCTD, NCI A,B,E
Bldg 10/Rm 8D53 
Phone: (240) 760-6046 
Fax: (301) 451-5625 
zlottjh@mail.nih.gov
 
Roles: Aobtains information by intervening or interacting with living individuals for research 
purposes; Bobtains identifiable private information about living individuals; Cobtains the 
voluntary informed consent of individuals to be subjects; Dmakes decisions about subject 
eligibility; Estudies, interprets, or analyzes identifiable private information or data/specimens 
for research purposes
NCI Supplied Agent:    
TRC102 - NSC 3801
Commercial Agent:
Temozolomide – NSC 362856
IND Number:                 
118052
NCT Number:
NCT01851369
Protocol Version Date:  
July 6, 2022

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PRÉCIS
Background:
Base excision repair (BER) of DNA repair pathway has been implicated in resistance to 
both alkylating and antimetabolite chemotherapy. 
TRC102 (methoxyamine HCl) acts through a novel mechanism to inhibit BER and has 
demonstrated the ability to potentiate the activity of the alkylating agent temozolomide 
(TMZ), in vitro and in vivo.  We hypothesize that TRC102 can be safely co-administered 
with TMZ and would potentiate DNA damage caused by TMZ, resulting in antitumor 
responses.
Based on responses measured during the Phase I portion of the trial, we will further 
explore the efficacy of this combination in patients with metastatic colon carcinoma, non-
small cell lung cancer (NSCLC), and granulosa cell ovarian cancer 
Primary Objective:
To establish the safety, tolerability, and maximum tolerated dose (MTD) of oral TRC102 
in combination with oral TMZ in patients with refractory solid tumors 
Evaluate the pharmacokinetic (PK) profile of oral TRC102 when administered in 
combination with TMZ.
To explore the response rate of this combination in patients with colon cancer, NSCLC, 
and granulosa cell ovarian cancer
Secondary Objective:
To explore the progression free survival rate of this combination in patients with colon 
cancer, NSCLC, and granulosa cell ovarian cancer
Exploratory Objectives:
Investigate tumor genomic and transcriptomic alterations potentially associated with 
sensitivity and/or the development of resistance to TRC102 and temozolomide.
Determine the effects of the study treatment on the level of histone γH2AX in circulating 
tumor cells (CTCs) and tumor and correlate the γH2AX response in tumor and CTCs
Determine the effects of the study treatment on the levels of cleaved caspase 3, epithelial-
mesenchymal transition, and APE in tumor and CTCs     
Determine and characterize the effects of study treatment on erythrocytes
Characterize the clinical presentation of hemolysis observed in earlier study subjects and 
explore the possible mechanisms 
Eligibility:
Phase I: histologically confirmed solid tumors that have progressed on standard therapy 
known to prolong survival or for which no standard treatment options exist
Phase II:  histologically confirmed adenocarcinoma of the colon post at least two lines of 
therapy, NSCLC post at least two lines of therapy, or granulosa cell ovarian cancer post at 
least one line of therapy
No major surgery, radiation, or chemotherapy within 4 weeks prior to entering the study
Adequate organ function

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Healthy adult volunteers ≥ 18 years of age will be consented to donate research blood 
Please note: As of Amendment R (5/12/18), healthy adult volunteers will no longer be 
recruited to provide blood for this study as we will no longer perform the hemolysis 
analysis.
Study Design: 
Phase I
This is an open-label Phase I trial; traditional 3+3 design.
Oral TRC102 and oral TMZ will be administered daily, days 1-5 in 28-day cycles
Once the MTD is established, up to 15 additional patients will be enrolled at the MTD to 
further evaluate that dose for PK and PD endpoints for evidence of DNA damage and 
apoptosis.
During the escalation phase, tumor biopsies will be optional.  During the expansion phase, 
(once MTD is reached), mandatory paired tumor biopsies will be pursued in the 15 
additional patients enrolled to further evaluate PD endpoints.
Phase II
This is a 3-arm Simon 2-stage design trial evaluating independently the response rate of 
patients with colon, NSCLC, and granulosa cell ovarian cancer.   
Patients with a body surface area (BSA) of ≥1.6 m2 will receive 125 mg of TRC 102 and 
150 mg/m2 of TMZ PO qday x 5 every 28 days (DL6).  Patients with a BSA of <1.6 m2 
will receive 100 mg of TRC 102 and 150 mg/m2 of TMZ PO qday x 5 every 28 days 
(DL5).  Each cycle will be 28 days.
The accrual ceiling for the Phase II portion is 75 patients.
Mandatory paired tumor biopsies will be pursued to further evaluate PD endpoints.

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SCHEMA: PHASE I
TRC102 and TMZ will be administered daily on days 1-5
a   Biopsies will be optional during the escalation phase and mandatory in the 15 additional patients enrolled 
at the MTD during the expansion phase.  Biopsies will be collected before drug administration on study 
(baseline); on cycle 1, day 5 (±1 day in case of scheduling difficulties) at 3-4 hrs after drug; and at time of 
disease progression (optional)   
b   Blood samples for CTCs (optional) will be collected from all patients at the following time points:     
     cycle 1 prior to treatment, cycle 1 day 1, 8 hrs after drug in both the escalation and expansion phases, 
cycle 1 day 5 (±1 day in case of scheduling difficulties) in the expansion phase only: cycle 2 day 1 and 
day 1 of all subsequent cycles before drug in both the escalation and expansion phases, and at time of 
disease progression   
c   Blood samples for PK analyses (optional) will be collected in Cycle 1 only prior to drug administration 
and 1, 2, 3, 4, 8, 12, and 24 hours post-dose in both the escalation and expansion phases, and once on day 
5 prior to dose in the expansion phase only
Phase I Dose Escalation Schedule (28-day cycle)
Dose Level
TRC102 orally
TMZ orally 
Level 1
25 mg once daily D1-5
125 mg/m2 once daily D1-5
Level 2
50 mg once daily D1-5
125 mg/m2 once daily D1-5
Level 3
50 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 4
75 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 5
100 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 6
125 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 7
150 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 8*
150 mg once daily D1-5
200 mg/m2 once daily D1-5
*Patients on DL7 can escalate to DL8 from cycle 2 onwards if DL 7 is tolerated in cycle 1
Cycle 1 (duration 28 days)
Tumor Biopsya
TMZ
Day 1
CTC Samplingb 
 Day 5
TRC102
PK Samplingc

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SCHEMA: PHASE II
Patients on three separate cohorts (colon cancer, NSCLC, or granulosa cell ovarian cancer) will 
each receive one of two dose levels:

patients with a BSA ≥1.6 m2 will receive 125 mg TRC 102 and 150 mg/m2 TMZ po 
daily on days 1-5 (DL6)

patients with a BSA <1.6 m2 will receive 100 mg TRC 102 and 150 mg/m2 TMZ po 
daily on days 1-5 (DL5)
Mandatory biopsies (patients with colon and granulosa cell ovarian cancer only) will be 
collected before drug administration on study (baseline) and 3-4 hrs after drug administration on 
cycle 1, day 4 or 5, (post-dose).  One optional signal restaging follow-up biopsy may be 
performed on day 1 ( 2 days) of the cycle following any restaging at which a 10-19% increase 
in tumor volume is observed (according to RECIST criteria) if the patient has been on study for 
at least 4 cycles, or at time of disease progression.
   
Blood samples for CTCs (optional; all patients) will be collected at the following time points:  cycle 
1 prior to treatment, before treatment on cycle 1 day 4 or 5 (on the day of the post-dose biopsy), day 
1 (±1 day) of all subsequent cycles before drug, and at time of disease progression   
 
Cycle 1 (duration 28 days)
Tumor Biopsy
TMZ
Day 1
CTC Sampling 
 Day 5
TRC102

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TABLE OF CONTENTS
PRÉCIS ..........................................................................................................................................2
SCHEMA: Phase I..........................................................................................................................4
SCHEMA: Phase II ........................................................................................................................5
TABLE OF CONTENTS ...............................................................................................................6
1.
OBJECTIVES ....................................................................................................................8
1.1
Primary Objectives.....................................................................................8
1.2
Secondary Objective...................................................................................8
1.3
Exploratory Objectives:..............................................................................8
2.
BACKGROUND................................................................................................................8
2.1
Background on the Activity of TRC102 in Combination with Alkylating 
Chemotherapy ..........................................................................................10
2.2
TRC102 ....................................................................................................11
2.3
Temozolomide (TMZ)..............................................................................12
2.4
Rationale...................................................................................................14
2.5
Correlative Studies Background...............................................................15
2.6
Genetic Sequencing Research Ethics .......................................................17
3.
PATIENT SELECTION...................................................................................................18
3.1
Eligibility Criteria (Patients) ....................................................................18
3.2
Exclusion Criteria (Patients) ....................................................................19
3.3
Eligibility Criteria (Healthy volunteer blood donors) ..............................20
3.4
Screening Evaluation................................................................................20
4.
REGISTRATION PROCEDURES..................................................................................21
4.1
Patient Registration Process .....................................................................21
4.2
Investigator and Research Associate Registration with CTEP ................21
5.
TREATMENT PLAN ......................................................................................................22
5.1
Agent Administration...............................................................................24
5.2
Definition of Dose-Limiting Toxicity (Phase I only)...............................25
5.3
General Concomitant Medication and Supportive Care Guidelines ........26
5.4
Duration of Therapy .................................................................................27
5.5
Duration of Follow Up .............................................................................27
5.6
Criteria for Removal from Study..............................................................28
6.
DOSING DELAYS/DOSE MODIFICATIONS..............................................................28
6.1
Dose Reduction ........................................................................................29
7.
ADVERSE EVENTS:  LIST AND REPORTING REQUIREMENTS...........................29
7.1
Comprehensive Adverse Events and Potential Risks List(s) (CAEPRs) .29
7.2
CTEP Reporting Requirements................................................................32

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7.3
NIH Reporting Requirements...................................................................35
8.
PHARMACEUTICAL INFORMATION ........................................................................36
8.1
TRC102 (NSC 3801)................................................................................36
8.2
Temozolomide..........................................................................................37
8.3
Agent Ordering and Agent Accountability ..............................................38
9.
BIOMARKER, CORRELATIVE, AND SPECIAL STUDIES.......................................39
9.1
Pharmacokinetics (Phase I only) ..............................................................39
9.2
Pharmacodynamics...................................................................................40
9.3
Exploratory Genomic Analyses................................................................44
9.4
Sample Collection and Processing ...........................................................47
10.
STUDY CALENDAR......................................................................................................48
11.
MEASUREMENT OF EFFECT......................................................................................50
11.1
Antitumor Effect – Solid Tumors.............................................................50
12.
DATA REPORTING / REGULATORY REQUIREMENTS .........................................56
12.1
Data Reporting .........................................................................................56
12.2
CTEP Multicenter Guideline....................................................................56
12.3
Collaborative Agreements Language .......................................................56
12.4
Genomic Data Sharing Plan .....................................................................58
13.
STATISTICAL CONSIDERATIONS.............................................................................58
13.1
Study Design ............................................................................................58
13.2
Sample Size/Accrual Rate........................................................................59
14.
HUMAN SUBJECTS PROTECTIONS...........................................................................59
14.1
Justification for Subject Selection............................................................59
14.2
Evaluation of Benefits and Risks/Discomforts ........................................60
14.3
Consent and Assent Process and Documentation.....................................60
14.4
Procedure for Protecting Against or Minimizing Any Potential Risks....61
14.5
Patient Advocate.......................................................................................62
REFERENCES.............................................................................................................................63
APPENDIX A: PERFORMANCE STATUS CRITERIA...........................................................66
APPENDIX B: INFORMATION ON POSSIBLE DRUG INTERACTIONS............................67
APPENDIX C: PATIENT’S MEDICATION DIARY ................................................................68
APPENDIX D: PK/PD COLLECTION WORKSHEETS...........................................................70
APPENDIX E: PK SAMPLE HANDLING.................................................................................78
APPENDIX F: ERYTHROCYTE ANALYSIS...........................................................................79

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1. OBJECTIVES
1.1
Primary Objectives

To establish the safety, tolerability, and maximum tolerated dose of oral TRC102 
(methoxyamine HCl) in combination with oral temozolomide (TMZ) in patients with 
refractory solid tumors 

Evaluate the pharmacokinetic (PK) profile of oral TRC102 when administered in 
combination with TMZ

To explore the response rate of this combination in patients with colon cancer, 
NSCLC, and granulosa cell ovarian cancer
1.2
Secondary Objective

To explore the progression free survival rate of this combination in patients with 
colon cancer, NSCLC, and granulosa cell ovarian cancer
1.3
Exploratory Objectives:

Investigate tumor genomic and transcriptomic alterations potentially associated with 
sensitivity and/or the development of resistance to TRC102 and temozolomide.

Determine the effects of the study treatment on the level of histone γH2AX in 
circulating tumor cells (CTCs) and tumor and correlate the γH2AX response in 
tumor and CTCs

Determine the effects of the study treatment on the levels of cleaved caspase 3 and 
Ki-67 in tumor

To explore resistance mechanisms to the study drug combination 

Determine and characterize the effects of study treatment on erythrocytes.

Characterize the clinical presentation of hemolysis observed in earlier study subjects 
and explore the possible mechanisms 
2. BACKGROUND
Resistance to chemotherapy is a primary reason that patients fail treatment.  Among the various 
mechanisms by which resistance to chemotherapy can develop, the base excision repair (BER) 
pathway has been shown to promote resistance to both alkylating and antimetabolite 
chemotherapy.  TRC102 acts through a novel mechanism to inhibit BER and has demonstrated 
the ability to potentiate the activity of the alkylating agents temozolomide and carmustine, and 
the antimetabolite agents fludarabine and pemetrexed in murine models of human cancer.  
Therefore, TRC102 may be able to potentiate the activity of alkylating and antimetabolite 
chemotherapy [1].
Published studies indicate that TRC102 has the ability to interrupt the process of BER by 

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binding to apurinic/apyrimidinic (AP) sites produced during the initial step of BER [1, 2].  
TRC102-bound AP sites are not substrates for apurinic/apyrimidinic endonuclease (APE), 
which performs an essential step in BER.  TRC102-bound AP sites are, however, substrates for 
topoisomerase II (topo II), an enzyme that cleaves damaged DNA.  In vitro studies of cancer 
cells that contain high levels of topo II indicate that TRC102 effectively potentiates the activity 
of chemotherapy. 
The ability of TRC102 to interrupt the BER pathway was demonstrated in vitro [2].  TRC102 
was tested for its ability to modify DNA in a way to prevent its cleavage by APE, which 
hydrolyzes the phosphodiester backbone 5’ to the AP site generated by the action of DNA 
deglycosylase (Figure 1) [1].  An oligonucleotide containing a single uracil base was 
synthesized and then incubated with uracil deglycosylase (UDG), a DNA deglycosylase, to 
excise the uracil within the oligonucleotide and produce an AP site.  Following the reaction with 
UDG, the oligonucleotide was incubated with or without TRC102, to produce substrates 
containing an AP site or an AP site bound to TRC102, respectively.  In the absence of TRC102, 
AP sites were efficiently hydrolyzed by purified APE, demonstrating a cleavage product of 
lower molecular mass.  
Fig 1. BER pathway.
In contrast, TRC102-bound AP-sites were resistant to the hydrolytic activity of APE, indicating 
that TRC102-bound AP-sites are not substrates for AP endonuclease and providing a 
mechanism of action by which TRC102 inhibits the BER pathway (Figure 2).  As a correlative 
study, we will measure AP-site accumulation in DNA at baseline and following TRC102 plus 
TMZ treatment.  

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Fig 2.  Effect of TRC102 on the BER 
pathway.  TRC102 interrupts BER, 
because TRC102- bound AP sites are not 
substrates for APE, a critical BER 
enzyme.  TRC102-bound AP sites, 
however, are substrates for topo II-
mediated DNA cleavage.  DNA single 
and double-strand breaks produced 
through the action of topo II on TRC102-
bound DNA lead to cellular apoptosis.
2.1
Background on the Activity of TRC102 in Combination with Alkylating 
Chemotherapy 
The ability of TRC102 to potentiate chemotherapy was initially demonstrated using the 
alkylating agent temozolomide [3, 4].  Collectively, the available data indicate that treatment of 
cancer cells with temozolomide produced N7-methylguanine and N3-methyladenine DNA 
adducts that activated BER to generate AP sites within double-stranded DNA.  TRC102 bound 
covalently to AP sites to form structurally modified AP sites that were then resistant to the 
repair activity of APE.  The persistence of these lesions led to cell death through the generation 
of DNA strand breaks. 
The lethal toxicity induced by the alkylating agent temozolomide combined with TRC102 
appears to be mediated through the poisoning effect of topo II [2].  While TRC102-bound AP 
sites are refractory to the catalytic activity of APE (indicating their ability to block BER), they 
are cleaved by purified topo II or nuclear extracts from tumor cells expressing high levels of 
topo II, suggesting that TRC102-bound AP sites stimulate topo II-mediated DNA cleavage [2].  
Furthermore, cells treated with temozolomide and TRC102 demonstrate increased expression of 
topo II and increased formation of phosphorylated histone H2AX (γH2AX), a marker of DNA 
double strand breaks, that co-localizes with up-regulated topo II.  These data support the 
hypothesis that DNA double-strand breaks marked by γH2AX foci are associated with topo II in 
cells. 

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The cleavage of TRC102-bound AP sites by topo II also explains the ability of TRC102 to 
potentiate the effects of temozolomide in cancer cells (that have high levels of topo II) 
compared to normal cells 
(which have low levels of 
topo II).  For example, in 
vitro addition of TRC102 
effectively sensitized tumor 
cells, but not bone marrow 
cells, to the effects of 
temozolomide (Figure 3).
Fig 3. Levels of topo II are 
increased in human colon 
HCT116 and SW480 cancer 
cell lines, but not in bone 
marrow cells by Western blot 
(Panel A).  TRC102 (MX) 
potentiates the in vitro 
activity of temozolomide in 
inhibiting survival of SW480 
human colon cancer cells 
(Panel B) but has minimal 
effect on temozolomide activity towards human or murine bone marrow cells (Panels C and D, 
respectively) [2].
 
In murine xenograft studies, TRC102 efficiently potentiated the antitumor effect of 
temozolomide and carmustine in several colon cancer cell lines regardless of cell line genetic 
status, including O6-methylguanine DNA-methyltransferase (MGMT), mismatch repair 
(MMR), or p53 status [3].  For example, the combination of temozolomide and TRC102 caused 
regression of SW480 tumor xenografts that were resistant to treatment with temozolomide 
alone.  Importantly, treatment with temozolomide plus TRC102 did not result in higher levels of 
myelosuppression or weight loss compared to treatment with temozolomide alone [3].
      
2.2
TRC102
Clinical Experience
IV TRC102 with Oral TMZ
Initial pharmacokinetic (PK) analyses of patients receiving TRC102 as a 5-day continuous 
infusion revealed a distinct PK profile, 10-fold greater than previously estimated in dogs, where 
half-life of TRC102 was estimated to be 4.5 hours.  As a result, TRC102 administration 
adjusted from a five-day continuous infusion to a one hour intravenous infusion.  Twenty-five 
patients have enrolled, in two cohorts.  In cohort A (patients with no-CNS disease), patients 
have enrolled in dose-levels 1, 2 and 3 (TMZ 150 mg/m2/day, days 1-5 and TRC102, 15 mg/m2, 
30 mg/m2 and 60 mg/m2 respectively).  In cohort B (patients with CNS involvement), patients 

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have enrolled in dose-levels 1 and 2.  Enrollment started at a DL lower (in DL1, TMZ 100 
mg/m2/day, days 1-5 and TRC102, 15 mg/m2; for DL2 TMZ increased to 150 mg/m2/day, days 
1-5 and TRC102 remained 15 mg/m2).  The average half-life of TRC102 administered as an 
one-hour continuous infusion was 55.04 hours (range: 12.2 - 100.3 hours, n = 20), statistically 
not different from the half-life of TRC102 administered as a 5-day continuous infusion which 
was 45.1 hours (range: 32.1 - 68.8 hours, n = 6).  Pharmacodynamic (PD) results from 22 
patients showed that administration of the combination of TMZ and TRC102 resulted in 10-
40% reduction in detectable AP sites.  Comet assay results from 20 patients revealed that the 
combination of TMZ and TRC102 induced a 2 to 3-fold higher levels of DNA strand breaks 
compared to TMZ alone.  PD demonstration of TRC102’s biologic activity on patients’ 
mononuclear cells has been demonstrated even at the lowest DL.  The combination of TRC102 
and TMZ has been well-tolerated.  For the non-CNS involvement cohort, one DLT was 
observed at DL1; grade 3 psychosis in a patient with progressive disease on increasing doses of 
opioids.  A grade 3 allergic reaction classified as an idiosyncratic event resulted in further 
expansion of the cohort to 10 evaluable patients with no additional DLTs observed.  No DLTs 
have been observed at DL2.  For the CNS involved cohort, no DLTs have been observed to 
date.  Three patients had stable disease (lung, ovarian, head and neck primary) [5].
Oral TRC102 with IV Pemetrexed 
The safety, PK, and PD of oral TRC102 combined with IV pemetrexed was evaluated.  TRC102 
alone was administered on days 1- 4 of a 2-week cycle followed by TRC102 (D1-4) and 
500 mg/m2 pemetrexed (D1) every 3 weeks thereafter.  Twenty eight patients were enrolled and 
treated with a total of 93 cycles of TRC102 at 15 mg/m2 (n=4), 30 mg/m2 (n=7), 60 mg/m2 
(n=11) and 100 mg/m2 (n=6).  The MTD was exceeded at 100 mg/m2 due to grade 3 anemia in 
50% of patients.  No other DLTs were reported.  Other adverse events possibly related to 
TRC102 included grade 3/4 neutropenia without fever, grade 3 thrombocytopenia, grade 1/2 
fatigue, and grade 1/2 asthenia.  TRC102 plasma concentrations required for in vivo activity 
were achieved at all dose levels studied (Cmax > 50 ng/mL, t1/2 = 28 hr).  PD studies 
confirmed that TRC102 binds pemetrexed-induced AP sites.  One patient at 30 mg/m2 TRC102 
with oropharyngeal adenoid cystic cancer metastatic to the lung had RECIST-defined PR that 
lasted until cycle 14.  Stable disease for ≥ 3 cycles was seen in 3 patients at 30 mg/m2, 2 patients 
at 60 mg/m2, and 4 patients at 100 mg/m2, including one patient with squamous cell lung cancer 
who had SD for 9 cycles.  So this study showed that a daily oral TRC102 is well-tolerated at 
doses expected to inhibit BER and potentiate the activity of chemotherapy.  The recommended 
phase II dose is 60 mg/m2 p.o. for 4 days in combination with standard dose pemetrexed [5, 6].
2.3
Temozolomide (TMZ)
TMZ is an alkylating drug approved by the FDA (http://www.accessdata.fda.gov) for the 
treatment of adult patients with newly diagnosed glioblastoma multiforme (GBM) 
concomitantly with radiotherapy and then as maintenance treatment and refractory anaplastic 
astrocytoma patients who have experienced disease progression on a drug regimen containing 
nitrosourea and procarbazine.  TMZ is not directly active but undergoes rapid nonenzymatic 
conversion at physiologic pH to the reactive compound 5-(3-methyltriazen-1-yl)-imidazole-4-
carboxamide (MTIC).  The cytotoxicity of MTIC is thought to be primarily due to alkylation of 
DNA.  Alkylation (methylation) occurs mainly at the O6 and N7 positions of guanine.  TMZ is 

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rapidly and completely absorbed after oral administration with a peak plasma concentration 
(Cmax) achieved in a median Tmax of 1 hour.  Food reduces the rate and extent of 
temozolomide absorption.  Cytochrome P450 enzymes play only a minor role in the metabolism 
of temozolomide and MTIC.  
Safety of TMZ
The most common adverse reactions (≥10% incidence) are alopecia, fatigue, nausea, vomiting, 
headache, constipation, anorexia, convulsions, rash, hemiparesis, diarrhea, asthenia, fever, 
dizziness, coordination abnormal, viral infection, amnesia, and insomnia.  The most common 
Grade 3 to 4 hematologic laboratory abnormalities (≥10% incidence) that have developed 
during treatment with temozolomide are lymphopenia, thrombocytopenia, neutropenia, and 
leucopenia which requires monitoring absolute neutrophil count (ANC) and platelet count prior 
to dosing and throughout treatment.  Cases of myelodysplastic syndrome and secondary 
malignancies, including myeloid leukemia, have been observed.  All patients, particularly those 
receiving steroids, should be observed closely for the development of lymphopenia and 
Pneumocystis jiroveci pneumonia.  Cases of hepatic injury, including fatal hepatic failure, have 
also been observed in patients enrolled on temozolomide studies; it was noted that liver toxicity 
may occur several weeks or more after initiation of treatment or after temozolomide 
discontinuation.
Current Protocol Status
As of October 2017, we have completed accrual to the Phase 1 portion of this study (52 
patients); DL7 was established as the MTD after two DLTs occurred on DL8 (Grade 3 
hemolysis and abdominal pain).  However, because 6 patients had grade 3 anemia requiring 
transfusion in the DL7 expansion cohort, subsequent patients were treated at DL6 (125 mg TRC 
102, 150 mg/m2 TMZ) and none of the remaining Phase I patients on DL6 required 
transfusions.    
In the Phase I portion of the trial there were four confirmed PRs (one colon, one NSCLC, two 
granulosa cell ovarian cancer) and one unconfirmed PR (colon); 11 patients have had stable 
disease.  All patients had prior therapy.  The one unconfirmed PR had stroke that was not 
considered to be drug related prior to confirmation; this patient came off study at the time of 
stroke, but returned 2 years later and received an additional three cycles of TRC 102 plus 
temozolomide on Special Exception protocol E16-5001 (16-C-9950) at the Clinical Center at a 
dose of 125 mg TRC 102 and 150 mg/m2 TMZ before coming off treatment per patient 
decision. 
We began accruing to the Phase II portion of the trial in February of 2016 at DL6 and have 
accrued 26 of patients as of October 20, 2017.  Due to a recent grade 3 anemia requiring 
RBC transfusion in a patient with very low BSA (1.43 m2) receiving DL6, as of Amendment 
P (10/20/2017) we will administer a maximum of 100 mg TRC102 daily (DL5) for patient 
whose BSA is lower than 1.6.  Patients with BSA equal or higher than 1.6 will receive a 
maximum of 125mg TRC102 daily (DL6).
Of note, analysis of the pharmacokinetic data collected to date indicate that all dose levels of 
TRC102 reached Cmax >50 ng/mL required for in vivo activity in preclinical models, and that 

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combining these two agents does not alter the pharmacokinetics of either.  Due to prior 
suggestions of anemia, patients on the expansion cohort will undergo a hemolysis workup, 
which is optional and as clinically indicated.  
2.4
Rationale
TRC102 has been shown to potentiate the activity of temozolomide by preventing BER and 
allowing cleavage of TRC102 bound DNA, which will cause DNA strand breaks in cancer cells.  
We hypothesize that oral TRC102 can be safely co-administered with TMZ and would 
potentiate DNA damage caused by TMZ resulting in antitumor responses.  Based on drug 
mechanisms of action we anticipate that patient responses will be associated with an MGMT-
low/negative + DNA mismatch repair-competent phenotype, and that eventual resistance to 
study drugs will be associated with the loss of this DNA repair phenotype.  
Granulosa cell tumor of the ovary is a stromal cell neoplasm that is able to secrete sex steroids 
such as estradiol, inhibin, follicle regulatory protein, and mullerian inhibitory substance, all of 
which have been investigated as prognostic markers [7-11].  Granulosa cell tumors make up 
approximately 2-5% of ovarian cancers [12]. They are very rare and have a long natural history 
as well as tendency to recur years after their diagnosis [13].  Advanced metastatic disease is 
present in about 10% of cases.  Average age of presentation is perimenopause or 
postmenopausal women around 50 years [14, 15].  Initial treatment is similar to that for 
epithelial ovarian cancer with surgery as the backbone in early stage disease.  Radiation has also 
been used but there is not sufficient data for its use in the adjuvant setting [16]. Chemotherapy 
has been shown to be effective in series of case series both with combinations including BEP 
[17], PVB [18] as well as single agents including paclitaxel [19] and tamoxifen [20].
Lung cancer has the highest mortality rate among cancers worldwide.  Lung cancer remains the 
leading cause of cancer-related mortality in the United States although its incidence is 
decreasing.  In 2014, 224,210 new cases are projected and 146,409 persons are calculated to die 
from the disease in the United States [21].  Chemotherapy combined with surgery and 
radiotherapy are the mainstays for many patients treated with lung cancer although many feel 
that these strategies alone have reached a therapeutic plateau.  Changes in major cell-signaling 
and regulatory pathways are now thought to play a major role in tumorigenesis.  Small molecule 
targets toward specific targets (e.g., ALK, EGFR, ROSS1, ERCC1) as well as immunotherapy 
are being used and further explored alone and in combinations with traditional therapies[22]. 
Long term survival remains poor for all patients but especially for those patients who are 
diagnosed with metastatic disease and new avenues need to be found to continue to improve 
outcomes.  There is no expectation that there will be a different expectation of response in 
adenocarcinoma vs. squamous cell lung cancer; we are planning to accrue both populations.  
Colorectal cancer (CRC) is one of the most prevalent cancers and a leading cause of cancer 
mortality worldwide [21].  To better understand the biologic markers of the disease, CRC has 
undergone extensive molecular characterization, which revealed important oncogenes (e.g., 
KRAS, BRAF, PIK3CA), tumor suppressor genes (e.g., APC, TP53, PTEN) and signaling 
pathways that are critical for the development, survival, and progression these cancer cells [23]. 
These genes are involved in major signaling pathways that have been linked to cancer, 

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including the WNT/β-catenin, mitogen-activated protein kinase (MAPK), transforming growth 
factor beta (TGF-β) and phosphoinositide 3-kinase (PI3K) pathways [24]. A number of targeted 
agents against specific genomic targets have been developed.  Patients whose tumors are 
characterized based on these molecular markers still show remarkable variability in terms of 
prognosis and response to therapy [25]. Many studies have addressed sub-classification of CRC, 
focusing on epigenetic factors and gene expression profiles to help guide clinical treatment and 
practice [26].  Colorectal cancer patients with known MSI-high disease who have not been 
previously treated with immunotherapy will be excluded from the Phase II portion of this study 
based on reports that they may benefit from PD-1 blockade therapy [27].
Partial responses in granulosa cell ovarian cancer (2 patients), NSCLC (1 patient with squamous 
cell), and colorectal cancer (one almost complete response and one unconfirmed) seen in the 
Phase 1 trial warrants further investigations in terms of response and identifying predictive 
biomarkers.  There are no readily available molecular or cytological links that would predict the 
response in these distinct tumor types.
2.5
Correlative Studies Background
Tumor tissue biopsies and circulating tumor cells will be collected to measure levels of 
phosphorylated histone H2AX (γH2AX), one of the earliest markers of DNA double-strand 
breaks [28-30].  H2AX is phosphorylated at its C-terminus (serine 139 in humans) within 
minutes following DNA double-strand breaks marking the chromatin domain around the broken 
chromosomal DNA ends, thus allowing the recruitment of repair factors [28, 30-33].  Levels of 
H2AX within a cell are directly correlated to the number of DNA double-strand breaks.  Hence 
H2AX can be used as a dosimeter and biomarker for DNA double-strand break damage.  The 
Pharmacodynamic Assay Development and Implementation Section (PADIS) at the Frederick 
National Laboratory for Cancer Research (FNLCR) has validated an immunofluorescent assay 
for H2AX [34]. Biopsies will be optional in the escalation phase and mandatory for the 
expansion phase.  
In Phase I, a biopsy will be obtained at baseline (prior to drug administration), 3-4 hours post 
dose administration on day 5, and optionally at the time of disease progression.  Biopsies will 
only be obtained on patients with easily accessible, biopsiable disease in escalation phase.  All 
patients in the expansion phase should have safely biopsiable disease and be willing to undergo 
tumor biopsy.  
Biopsies are optional in the escalation phase, but with Amendment K, we revised the protocol to 
consent current patients who are responding (i.e., stable disease or partial response) to consider 
providing archival tissue, such as a diagnostic tissue block or previously collected biopsy tissue 
sent for CLIA genetic testing (biopsy core or 6 unstained and one H&E slide).  In the absence of 
this tissue, we would ask these patients to consider consenting to having a research biopsy on 
study, if the patient has biopsiable disease, with a second optional biopsy at time of disease 
progression.  While having pre-study biopsy tissue is optional for assessing drug response, 
provision of “on-study” biopsies would address whether drug resistance and eventual loss of 
response are associated with loss of the MGMT-low/negative plus DNA mismatch repair-

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competent phenotype.  No more than three on-study biopsies will be requested, and no biopsy 
will be requested unless there is a reasonable possibility that a paired sample (biopsy or archival 
tissue) is or will be available. 
In Phase II, a biopsy will be obtained at baseline (prior to drug administration) and 3-4 hours 
post dose administration on day 4 or 5.  An optional biopsy will also be collected at the time of 
disease progression or at the beginning of the cycle following any restaging at which a 10-19% 
increase in tumor volume is observed (according to RECIST criteria) if the patient has been on 
study for at least 4 cycles to study drug resistance mechanisms.  Biopsies will only be obtained 
from patients with colon cancer or granulosa cell ovarian cancer.  All patients in the expansion 
phase should have safely biopsiable disease and be willing to undergo tumor biopsy.  
Blood sampling for CTCs will be performed for all patients as described in Section 9.2.2.1.  We 
will also evaluate whether we can measure changes in the number and phenotype (epithelial-
mesenchymal transition) in CTCs and tumor tissue in patients over time to explore any 
correlation with response to treatment or disease progression.  This analysis will be performed 
with the ApoStream instrument, which uses antibody-independent CTC isolation technology 
that can isolate viable CTCs from epithelial and non-epithelial cancers.  
We additionally plan to measure cleaved caspase 3 (cCasp3) as a marker of apoptosis.  Cleaved 
caspase 3 is a key protease activated during early stages of apoptosis via cleavage of its pro-
enzyme form into a heterodimer of 17- and 12-kDa subunits.  Immunofluorescence assays for 
this marker has been developed and validated by Dr. Kinders' laboratory at FNLCR.  
FNLCR has developed an immunoassay for tumor MGMT protein expression to investigate the 
relative importance of MGMT status for this combination treatment.  Additionally, the 
laboratory of Dr. Stanton Gerson (Case Western Reserve University) has agreed to assay any 
remaining tumor tissue via IHC for the enzyme DNA-3-methyladenine glycosylase, also known as 
3-alkyladenine DNA glycosylase (AAG) or N-methylpurine DNA glycosylase (MPG), the only 
glycosylase identified to date that excises alkylation-damaged purine bases in humans, such as those 
produced by temozolomide [35, 36].  No patient data will accompany any samples sent to Dr. 
Gerson’s lab.
While treatment with the combination of TRC102 and temozolomide has been observed to lead 
to clinical responses in patients on the phase I portion of this trial, patients who respond are 
likely to eventually progress due to acquired drug resistance, making further understanding of 
the determinants and mechanisms of acquired resistance a priority.  To address this, after 
Amendment R (5/12/2018), the optional tumor biopsies collected at progression or restaging 
follow-up (i.e., after a restaging at which a 10-19% increase in tumor size is observed) will be 
sequenced with the CLIA-certified OCAv3 assay by the CLIA-certified Molecular 
Characterization (MoCha) Laboratory at the Frederick National Laboratory for Cancer 
Research.  Patients undergoing these optional biopsies will receive the Oncomine report from 
OCAv3 to share with their health care providers in order to help guide future treatment 
decisions.  Genetic information from the tumor at the time of progression will be compared with 
baseline tumor genetic information to investigate the development of resistance to the 
combination of TRC102 and temozolomide by mechanisms such as inactivation of mismatch 
repair genes or the development of increased mutational load and neoantigen production [37]. 

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MoCha will also perform exploratory whole-exome sequencing (WES) and whole transcriptome 
RNASeq gene expression assays on baseline tumor and germline tissues from patients to 
attempt to identify genomic alterations and gene expression patterns potentially associated with 
sensitivity to treatment, the results of which will not be shared with the patients.  
Peripheral blood sampling for assessment of anemia and hemolysis will be performed for all 
patients at baseline and as described in Appendix F.  In preclinical animal models, TRC102 
caused dose-dependent extravascular hemolysis, as demonstrated by development of anemia, 
elevation in serum bilirubin, reticulocytosis, and bone marrow hypercellularity reflective of 
regenerative response, and splenomegaly [Investigator Brochure].  The combination of TRC102 
and pemetrexed or TMZ did not affect hematologic changes, and there was no evidence of 
cumulative effects on hematological toxicity.  In a phase I study of TRC102 in combination 
with pemetrexed for refractory cancer treatment, anemia secondary to extravascular hemolysis 
was the only dose-limiting toxicity observed [38]. The anemia was reversible and manageable 
with standard supportive care.  The mechanisms of extravascular hemolysis associated with 
TRC102 are not well understood.  In the present trial, hemolytic anemia was the dose-limiting 
toxicity, and MTD was exceeded at dose-level 8 (TRC102 150 mg, TMZ 200 mg/m2).  Given 
negativity of direct Coombs tests in these patients who experienced hemolytic anemia, 
hemolytic anemia is considered to be non-immune mediated.  An in vitro study of 
hydroxylamines in human erythrocytes suggests impairment of RBC enzymes as the potential 
mechanism of hemolysis [39].  We hypothesize that TRC102 induces extravascular hemolysis 
by affecting erythrocyte enzymes and/or erythrocyte membrane and osmotic tolerance.  By 
identifying the underlying mechanisms and susceptible subjects with underlying RBC 
enzymopathies or RBC membrane disease, we aim to improve the clinical safety of TRC102.  
To perform this analysis, healthy adult volunteers will be accrued to provide normal research 
blood samples. Please note: As of Amendment R (5/12/18), healthy adult volunteers will no 
longer be recruited to provide blood for this study as we will no longer perform this analysis of 
the mechanisms of extravascular hemolysis.
2.6
Genetic Sequencing Research Ethics
This trial will collect identifiable genetic data from patients enrolled after Amendment R 
(5/12/2018) for exploratory studies to examine genomic alterations and gene expression patterns 
potentially associated with sensitivity and/or acquired resistance to treatment.  As patient 
clinical response data (both for this study and, potentially, after a patient is off-study) will be 
required for comparison to sequencing results, de-identifying the samples is not feasible.  
Designing the study therefore poses challenging questions about informed consent, the privacy 
of the patient and the patient’s family, the researchers’ obligation to disclose genetic 
information to the patient, and the use and storage of research data [40-42].  In the vast majority 
of cases, we do not know the medical significance of genetic variants [43, 44].  These 
challenges will continue to be evaluated to maintain the rigor and integrity of the study and the 
wellbeing of our patients.  A Certificate of Confidentiality has been obtained from the NIH to 
help protect the privacy of all study participants. 
The informed consent document for this protocol after Amendment R (5/12/2018) contains 
language informing patients about the performance of genetic studies, and patients will have the 

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option to choose whether they wish to take part in these studies.
Targeted sequencing (CLIA-certified) and whole-exome sequencing (performed for research 
purposes but non-CLIA) of tumor and blood can detect non-ambiguous germline variants, 
which may raise health and privacy implications for the patient and his or her family.  WES will 
not be validated for clinical use, and no clinical decisions can be made based on its results.  
Furthermore, no WES genetic data will be shared with the patient.
This study does not meet the criteria specified by the NIH Genomic Data Sharing (GDS) Policy 
(see Section 12.4), and therefore, a GDS plan has not been included in this protocol.
3. PATIENT SELECTION
3.1
Eligibility Criteria (Patients)
3.1.1
Phase I: histologically confirmed solid tumors that have progressed on standard therapy 
known to prolong survival or for which no standard treatment options exist. 
3.1.2
Phase II:  histologically confirmed colorectal adenocarcinoma post at least two lines of 
therapy, NSCLC post at least two lines of therapy, or granulosa cell ovarian cancer post 
at least one line of therapy.  Patients must have measurable disease.  
3.1.3
Age ≥18 years.  Because no dosing or adverse event data are currently available on the 
use of TRC102 in combination with TMZ in patients < 18 years of age, children are 
excluded from this study.  
3.1.4
Patients enrolling in the expansion cohorts must have disease amenable to biopsy and be 
willing to undergo pre-and post-treatment biopsies.
3.1.5
ECOG performance status ≤ 2 (Phase I), ≤ 1(Phase II).
3.1.6
Life expectancy of greater than 3 months
3.1.7
Patients must have normal organ and marrow function as defined below:

Absolute neutrophil count
≥1,500/mcL

Hemoglobin                            ≥10 g/dL without transfusion within 4 days prior to 
enrollment 

Platelets
≥100,000/mcL

Total bilirubin
≤1.5 X institutional ULN

AST(SGOT)/ALT(SGPT)
≤3 X institutional upper limit of normal; 5.0 x ULN     
in cases of liver metastases  

creatinine
≤1.5 X institutional ULN
OR

creatinine clearance
≥60 mL/min/1.73 m2 for patients with creatinine levels 
>1.5 mg/dL

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3.1.8
The effects of study drug on the developing human fetus are unknown.  For this reason, 
women of child-bearing potential and men must agree to use adequate contraception 
(hormonal or barrier method of birth control; abstinence) prior to study entry and for the 
duration of study participation and for at least 3 months after dosing with study drugs 
ceases.  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 prior to the study, for the duration of study participation, and 3 months 
after completion of study drug administration.
3.1.9
Patients must have completed any chemotherapy, radiation therapy, or biologic therapy 
≥ 4 weeks (or 5 half-lives, whichever is shorter) prior to entering the study (6 weeks for 
nitrosoureas or mitomycin C).  Patients must be ≥ 2 weeks since any prior 
administration of a study drug in a Phase 0 or equivalent study and ≥ 1 week from 
palliative radiation therapy.  Patients must have recovered to eligibility levels from prior 
toxicity or adverse events.  Treatment with bisphosphonates is permitted.
3.1.10 Patients must be able to swallow whole tablets or capsules; nasogastric or G-tube 
administration is not allowed.
3.1.11 Ability to understand and the willingness to sign a written informed consent document 
and to undergo tumor biopsies in the expansion phase.
3.2
Exclusion Criteria (Patients)
3.2.1
Patients who are actively receiving any other investigational agents. 
3.2.2
Patients with active brain metastases or carcinomatous meningitis are excluded from this 
clinical trial.  Patients with treated brain metastases, whose brain metastatic disease has 
remained stable for ≥ 4 weeks without requiring steroid and anti-seizure medications are 
eligible to participate.
3.2.3
Phase II only: No other prior malignancies are allowed except for the following: 

Adequately managed stage 0 (carcinoma in situ), I, or II basal cell or squamous cell 
carcinoma from which the patient is currently in complete remission.

Any other cancer from which the patient has been disease-free for three years.

Adequately managed stage I or II well differentiated thyroid or prostate cancer is 
also eligible, wherein the patient is not required to be in complete remission.
3.2.4
Phase II only: patients with colorectal cancer with known MSI-high disease who have 
not previously been treated with immunotherapy or who have refused treatment with 
immunotherapy.
3.2.5
History of allergic reactions attributed to compounds of similar chemical or biologic 

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composition to TRC102 or TMZ.
3.2.6
Uncontrolled intercurrent illness including, but not limited to, serious untreated 
infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac 
arrhythmia, or psychiatric illness/social situations that would limit compliance with 
study requirements. 
3.2.7
Pregnant women are excluded from this study because the effects of the study drugs on 
the developing fetus are unknown.  Because there is an unknown but potential risk for 
adverse events in nursing infants secondary to treatment of the mother with the study 
drugs, breastfeeding should be discontinued prior to the first dose of study drug and 
women should refrain from nursing throughout the treatment period and for 3 months 
following the last dose of study drug. 
3.2.8
HIV-positive patients on combination antiretroviral therapy are ineligible because of 
possible PK interactions with TRC102.
3.3
Eligibility Criteria (Healthy volunteer blood donors) 
Please note: As of Amendment R (5/12/18), healthy adult volunteers will no longer be 
recruited to provide blood for this study as we will no longer perform the hemolysis analysis.
3.3.1
Age >18 years; hemoglobin ≥12 g/dL; no history of bleeding problems; not taking 
aspirin or any medication that may affect erythrocyte biochemistry
3.3.2
Willingness to sign the healthy volunteer informed consent form. 
3.4
Screening Evaluation
3.4.1
Histologic confirmation: All patients are required to have histologically confirmed solid 
tumor(s) per Sections 3.1.1 or 3.1.2.  Pathology confirmation should preferably be done 
by the Laboratory of Pathology, NIH.  However, pathology reports from outside 
institutions confirming the diagnosis will also be acceptable for eligibility purposes. 
3.4.2 History and physical examination: Complete history and physical examination (including 
height, weight, vital signs, performance score, EKG) will be conducted within 8 days 
prior to enrollment. 
3.4.3 Imaging Studies (Baseline): Every participant should have an evaluation of known sites 
of disease as part of the baseline evaluation.  All patients will be required to undergo a 
CT scan of the chest/abdomen/pelvis to evaluate sites of disease within 28 days prior to 
enrollment.  In an occasional patient, MRI may be performed for a disease site instead of 
a CT scan if MRI is felt to provide more accurate measure of disease.  MRI or CT scan 
with contrast of the brain, MRI liver, MRI for other disease sites, or bone scan may be 
done as clinically indicated.

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3.4.4 Laboratory Evaluation: Baseline laboratory data are to be obtained within 8 days prior to 
enrollment:
•
Hematological Profile: CBC with differential.
•
Biochemical Profile: albumin, alkaline phosphatase, total bilirubin, BUN, calcium, 
creatinine, phosphorus, total protein, SGOT[AST], SGPT[ALT], magnesium, 
potassium, LDH, and sodium.
•
Coagulation Profile: PT, PTT, INR for patients undergoing tumor biopsy.
•
Urine and/or serum pregnancy test for female participants of childbearing potential.
•
Erythrocyte analyses per Appendix F. 
       
4. REGISTRATION PROCEDURES
4.1
Patient Registration Process
Registration and status updates (e.g., when a participant is taken off protocol 
therapy and when a participant is taken off-study) will take place per CCR SOP 
ADCR-2, CCR Participant Registration & Status Updates found here: 
https://ccrod.cancer.gov/confluence/pages/viewpage.action?pageId=73203825
Cohorts:
Cohort 1: healthy volunteers
Cohort 2: phase I solid tumor
Cohort 3: phase II CRC
Cohort 4: phase II NSCLC
Cohort 5: phase II granulosa cell
Arms:
Arm 1: TRC102 + Temozolomide
Arm 2: no treatment
Cohorts 2, 3, 4, and 5 are assigned to arm 1. Cohort 1 is assigned to arm 2.
4.2
Investigator and Research Associate Registration with CTEP
Food and Drug Administration (FDA) regulations require IND sponsors to select qualified 
investigators.  NCI policy requires all persons participating in any NCI-sponsored clinical 
trial to register and renew their registration annually.  To register, all individuals must obtain 
a CTEP Identity and Access Management (IAM) account (https://ctepcore.nci.nih.gov/iam).  
In addition, persons with a registration type of Investigator (IVR), Non-Physician 
Investigator (NPIVR), or Associate Plus (AP) (i.e., clinical site staff requiring write access to 
OPEN or RAVE or acting as a primary site contact) must complete their annual registration 
using CTEP’s web-based Registration and Credential Repository (RCR) 
(https://ctepcore.nci.nih.gov/rcr).  Documentation requirements per registration type are 
outlined in the table below.

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Documentation Required
IVR
NPIVR
AP
A
FDA Form 1572


Financial Disclosure Form



NCI Biosketch (education, training, 
employment, license, and certification)



HSP/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 CTSU (Cancer Trials Support Unit) websites and applications.  In addition, 
IVRs and NPIVRs must list all clinical practice sites and IRBs covering their practice sites 
on the FDA Form 1572 in RCR to allow the following:

Added to a site roster

Assigned the treating, credit, consenting, or drug shipment (IVR only) tasks in OPEN

Act as the site-protocol PI on the IRB approval

Assigned the Clinical Investigator (CI) role on the Delegation of Tasks Log (DTL).    
Additional information can be found 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
5. TREATMENT PLAN
Phase I: This is an open-label Phase I trial evaluating the combination of oral TRC102 with oral 
TMZ, in 28-day cycles.  The starting dose of TRC102 is based on the safety information from 
the trial of IV TRC102 combined with TMZ.  The study will follow a 3+3 design: dose will be 
escalated in cohorts of 3 patients with the individual dose of TRC102 and TMZ increased in 
successive dose levels as outlined in the dose escalation table below.  Once the MTD is 
established, up to 15 additional patients will be enrolled at the MTD (expansion phase) to 
further define the dose and evaluate PK and PD at this dose level. 
Phase II: Patients with a body surface area (BSA) of ≥1.6 m2 will be started on 125 mg TRC 
102 and 150 mg/m2 Temozolomide orally on days 1-5 of each 28-day cycle (DL6).  Patients 
with a BSA of <1.6 m2 will be started on 100 mg TRC 102 and 150 mg/m2 Temozolomide 
orally on days 1-5 of each 28-day cycle (DL5).  There will be no dose escalation for patients on 
the Phase II portion of the study; appropriate dose modifications are described in Section 6.  Up 
to 75 patients will be accrued to this part of the study.  
History and physical examination can be done up to 8 days before start of a new cycle.
Patients will be examined at baseline for the first cycle then at the start of every cycle for the 
remainder of the study (up to 3 days before start of a new cycle).  The start of the next cycle 
may be delayed for up to 1 week to accommodate scheduling conflicts. 

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Labs (CBC with differential; serum chemistries) will be performed (+/- 2 days) as follows: 
every week up to week 3 for cycle 1.  Starting with cycle 2, labs (CBC with differential; serum 
chemistries) will only be performed before the start of each cycle (laboratory evaluation may be 
performed up to 7 days prior to scheduled date to accommodate scheduling conflicts and patient 
convenience).  If clinically indicated, labs may be obtained more frequently.
Blood for erythrocyte analysis (optional) will be collected on cycle 1 days 7, 14, and 21 (+/- 2 
days) as described in Appendix F.  
CT scans will be performed at baseline, and repeat scans will be performed every 2 cycles 
(every 3 cycles for patients on study more than one year).
Phase I Dose Escalation Schedule (28-day cycle)
Dose Level
TRC 102 orally
TMZ orally 
Level -2
25 mg once daily D1-5
75 mg/m2 once daily D1-5
Level -1
25 mg once daily D1-5
100 mg/m2 once daily D1-5
Level 1
25 mg once daily D1-5
125 mg/m2 once daily D1-5
Level 2
50 mg once daily D1-5
125 mg/m2 once daily D1-5
Level 3
50 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 4
75 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 5
100 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 6
125 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 7
150 mg once daily D1-5
150 mg/m2 once daily D1-5
Level 8*
150 mg once daily D1-5
200 mg/m2 once daily D1-5
*Patients on DL7 can escalate to DL8 from cycle 2 onwards if DL 7 is tolerated in cycle 1
Tumor biopsies will be optional during the escalation phase.  For the escalation phase, 
3 patients should have completed at least one cycle of therapy prior to considering dose 
escalation in the next cohort of patients.  Dose escalation will proceed according to the dose 
escalation table.  DLT is defined in Section 5.2.  Determination of DLT will be based on 
toxicities observed in the first cycle of therapy.  Patients are considered evaluable for toxicity 
for the purpose of cohort dose escalation decisions if they either 1) experienced DLT or 2) have 
received at least 90% of the planned 28-day doses of treatment in one cycle of therapy and have 
been followed for one full cycle without DLT.  All toxicities will be reported for all patients 
who receive any amount of study drug on this study.  Evaluation of toxicity will begin with 
study drug administration on cycle 1 day 1.  
Intra-patient dose escalation is permitted if: 1) patient had ≤ Grade 1 drug-related toxicity 
except alopecia, lymphopenia; 2) Proposed higher dose should have been established as safe; 
3) Patients should not have progressive disease on study; 4) Patients on dose level 7 who 
tolerate (i.e., no grade 2 or greater drug-related adverse events) cycle 1 can escalate to dose 
level 8 from cycle 2 onwards.   

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Phase I MTD Expansion Phase 
Once the MTD is established, up to 15 additional patients will be treated and tumor biopsies 
will be obtained to assess PD effects.  Tumor biopsies will be mandatory during the expansion 
phase of the study.  Biopsies will be obtained at the time points outlined below.  Depending on 
the results obtained from the tumor biopsies in the first few patients, the timing of the biopsy 
can be adjusted, but the number of tumor biopsies and the procedure will not change.  Any 
change in the timing of the tumor biopsy will be discussed with the patient prior to obtaining 
informed consent.
For the purposes of final analysis of the expansion phase, only patients with paired tumor 
biopsies will be considered evaluable.  Patients with fewer than the 2 required biopsies will not 
be considered evaluable for the expansion phase and will need to be replaced in the accrual 
scheme of the expansion phase. 
5.1
Agent Administration
Reported adverse events and potential risks are described in Section 7.  Appropriate dose 
modifications are described in Section 6.  No investigational or commercial agents or therapies 
other than those described below may be administered with the intent to treat the patient's 
malignancy. 
TRC102 will be administered to patients as a capsule dosed according to the dose escalation 
table.  TRC102 should be administered together with TMZ capsules.  TRC102/TMZ should be 
administered either 1 hour before or 2 hours after a meal.  Patients should drink an eight ounce 
glass of water following TRC102/TMZ administration.  Antiemetic therapy may be 
administered prior to and/or following administration of TRC102/TMZ in patients who develop 
nausea and vomiting but not prophylactically.  TRC102/TMZ capsules should not be opened or 
chewed.  If capsules are accidentally opened or damaged, precautions should be taken to avoid 
inhalation or contact with the skin or mucous membranes. 
The drugs will be administered in 28-day cycles (± 1 day for scheduling).  A missed or vomited 
dose will not be replaced.  The patients will be instructed to take the next scheduled dose at the 
regularly scheduled time.  Patients will be asked to maintain a Study Medication Diary 
(Appendix C) and record each dose of medication.  Patients will be given instructions for 
completing the medication diary and will be asked to return it to the clinic staff at the end of 
each cycle.

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5.2
Definition of Dose-Limiting Toxicity (Phase I only)
For MTD determination, DLTs will be defined as toxicities occurring within the first cycle of 
treatment.  Patients who come off study for reasons other than drug-related toxicity (e.g., 
disease progression) prior to completion of more than 90% of dose of the first cycle will be 
replaced.  Dose-limiting toxicity (DLT) is defined as an adverse event that occurs during the 
first cycle and is felt to be related (possibly, probably, or definitely) to administration of study 
drugs and fulfills one of the following criteria:   
5.2.1
Hematologic Toxicity 
5.2.1.1
Grade 4 neutropenia   
5.2.1.2
Febrile neutropenia 
5.2.1.3
Neutropenic infection: Grade ≥ 3 neutropenia with grade ≥ 3 infection.
5.2.1.4
Grade ≥ 3 thrombocytopenia 
5.2.1.5
A drop in Hgb ≥3.0 g/dL over one week
5.2.1.6
Any degree of lymphopenia, or leukopenia in the absence of grade 4 neutropenia 
will not be considered dose limiting.
5.2.2
Grade ≥ 3 Non-hematologic Toxicity 
5.2.2.1
Grade ≥ 3 non-hematological toxicity (except alopecia) felt to be related to study 
medications will be considered dose-limiting with the following clarifications:
5.2.2.2
Diarrhea Grade 3 will only be considered dose-limiting if it is refractory to treatment 
as outlined in Section 5.3.2, Supportive Care Guidelines, and unable to be corrected 
to Grade 2 or less within 24 hours.  Bloody or Grade 4 diarrhea will be dose-
limiting.
5.2.2.3
Nausea and vomiting Grade 3 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 
(Section 5.3.1).
5.2.2.4
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.
5.2.2.5
Grade ≥ 3 electrolyte abnormalities will not be considered dose limiting.  Grade 4 
metabolic toxicities that are symptomatic will be considered dose-limiting regardless 
of duration or ability to correct.

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Management and dose modifications associated with the above adverse events are outlined in 
Section 6.  Dose escalation will proceed within each cohort according to the following scheme.  
Number of Patients with DLT 
at a Given Dose Level
Escalation Decision Rule
0 out of 3
Enter 3 patients at the next dose level.
≥ 2
Dose escalation will be stopped.  This dose level will be declared the 
maximally administered dose (highest dose administered).  Three (3) 
additional patients will be entered at the next lowest dose level if only 
3 patients were treated previously at that dose.
1 out of 3
Enter at least 3 more patients at this dose level.
If 0 of these 3 patients experience DLT, proceed to the next dose 
level.
If 1 or more of this group suffer DLT, then dose escalation is 
stopped, and this dose is declared the maximally administered dose.  
Three (3) additional patients will be entered at the next lowest dose 
level if only 3 patients were treated previously at that dose.
≤1 out of 6 at highest dose level 
below the maximally 
administered dose
This is generally the recommended phase 2 dose.  At least 6 patients 
must be entered at the recommended phase 2 dose.
5.3
General Concomitant Medication and Supportive Care Guidelines
All patients will be provided with the best available supportive care.  All concurrent 
medications should be documented prior to initiation of treatment, and be periodically reviewed 
with the patient.  Particular attention must be paid to medications which may cause hemolytic 
anemia.
No other approved or investigational anticancer treatment will be permitted during the study 
period, including chemotherapy, biologic response modifiers, hormone therapy, 
immunotherapy, or radiotherapy.  
5.3.1
Nausea/Vomiting
Anti-emetics will not be administered routinely prior to TMZ or TRC102.  However, if a 
patient develops nausea/vomiting, anti-emetics such as but not limited to 
prochlorperazine, metoclopramide, 5-HT3 antagonists, or aprepitant may be given.  In 
addition, if a patient develops nausea and/or vomiting that is Grade 2 or greater, anti-
emetics may be instituted prophylactically at the discretion of the investigator.  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.
5.3.2
Diarrhea
If diarrhea develops and does not have an identifiable cause other than study drug 
administration, 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 

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the first unformed stool with 2 mg po 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 
in 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 regimen (16 mg, or less if there is resolution of the symptoms, of 
loperamide in a 24-hour period).
5.3.3
Neutropenia
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 
administered prophylactically.  If necessary, they may be administered according to 
accepted American Society of Clinical Oncology (ASCO) guidelines to allow re-
treatment.
5.3.4
Anemia
Symptomatic anemia should be treated with red blood cell transfusion and is 
recommended if the hemoglobin falls below 8 g/dL.  Use of erythropoietin will follow 
ASCO guidelines.
5.3.5
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.
5.4
Duration of Therapy
In the absence of treatment delays due to adverse event(s), treatment may continue until one of 
the following criteria applies:

Disease progression

Intercurrent illness that prevents further administration of treatment

Significant toxicity occurs despite 2 dose reductions as described in Section 6 or no 
lower dose level exists

Pregnancy

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.
5.5
Duration of Follow Up

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Patients will be followed for 30 days after the last dose is administered or until one of the 
following occurs: patient enrolls on another protocol, patient receives standard of care, or death, 
whichever comes first.  The follow-up will consist of a phone call between Days 27-30 after the 
last dose to evaluate adverse events that were ongoing and any new events that might be 
deemed related to the therapy.  Toxicities felt to be possibly, probably, or definitely related to 
the study drugs that have not resolved or stabilized by Day 30 post-treatment will be followed 
until stabilization or resolution via phone calls as clinically indicated.  Whenever possible, 
Phase II patients will be followed via phone calls until they progress or start another treatment 
for their cancer. 
5.6
Criteria for Removal from Study
Patients will be removed from study for one of the following reasons: completed 30-day follow-
up period or toxicities are unresolved but stabilized.  The reason for study removal and the date 
the patient was removed must be documented in the medical record and communicated to 
Central Registration per Section 4.
6. DOSING DELAYS/DOSE MODIFICATIONS
Non-hematologic toxicities (at least possibly related to therapy) should have resolved to 
≤ Grade 1 or baseline prior to starting the next cycle or receiving the next dose of TRC102 or 
TMZ within a cycle.  Grade 2 electrolyte abnormalities, grade 2 nausea/vomiting/diarrhea in the 
absence of maximal palliative support, or any grade alopecia, will not result in holding or 
delaying study drugs. 
Hematologic toxicities (at least possibly related to therapy) should have resolved to ≤ Grade 1 
for thrombocytopenia, ≤ Grade 2 for neutropenia and anemia prior to resuming therapy except 
any grade lymphopenia, leucopenia in the absence of grade 3 neutropenia.
Even though study drug administration is allowed at these lower grades, every effort should be 
made to correct the abnormal lab values to normal if possible.  If the potassium level is grade 2 
or greater and/or if the calcium, magnesium, and/or phosphate are grade 3 or higher, an EKG 
must be performed and appropriate action taken based on the results.
Treatment may be delayed for a maximum of 2 weeks for toxicities.  In case toxicities do not 
resolve as stated, the patient will not receive further therapy on this protocol and will be 
followed for resolution of toxicities.  Start of next cycle may be delayed for up to 1 week to 
accommodate scheduling conflicts.  Dose modifications are intended for within-cycle and start-
of-next-cycle changes. 
A maximum of 2 dose reductions will be allowed before patient is taken off treatment.  Patients 
who require a dose reduction will not have the dose re-escalated.  For patients who had one dose 
reduction without significant change in their toxicities (i.e., fatigue), they could be dose reduced 
to 4 days of the same dose level instead of dropping to the next lower dose level.

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Patients will be carefully monitored for evidence of liver toxicity and toxicity will be graded per 
CTCAE criteria (section 7.2).  For patients who develop grade 3 LFT abnormalities, study drugs 
will be held and the benefits and risks of continuing treatment on a reduced dose will be 
discussed with the patient by the study team. 
6.1  Dose Reduction
Grade 2 Drug-related toxicities: No changes will be made to the dose of TRC102/TMZ for 
Grade 2 toxicities.
Grade 3-4 Drug-related non-hematologic toxicities (except alopecia): Doses of 
TRC102/TMZ will be held until toxicities recover to ≤ Grade 1 prior to re-initiating treatment at 
the lower dose level.  Electrolyte abnormalities will not require dose reduction if resolution to 
Grade 2 or less is documented.  Dose modifications for nausea, vomiting, and diarrhea will be 
made only if they are refractory to treatment (See Section 5.2). 
Grade 3-4 Drug-related thrombocytopenia: Dose of TRC102/TMZ will be held until it has 
resolved to ≤ Grade 1 prior to re-initiating treatment at the lower dose level.
Grade 3-4 Drug-related neutropenia: Dose of TRC102/TMZ will be held until it has resolved 
to ≤ Grade 2 prior to re-initiating treatment at the lower dose level.
A drop in Hgb ≥3.0 g/dL over one week that results in grade 3 anemia: Dose of TRC102/TMZ 
will be held until it has resolved to ≤ Grade 2 prior to re-initiating treatment at the lower dose 
level.
Any grade lymphopenia, leucopenia in the absence of at least grade 3 neutropenia: Dose of 
TRC102/TMZ will not be held or modified.
Definition of Complete Course: A cycle is defined as 28 days, with D1 starting the first day of 
TRC102/TMZ administration.  If a patient receives 90% of scheduled treatments of 
TRC102/TMZ and remains in the study until D28, the patient will be considered to have 
completed a cycle of therapy.  Patients who do not complete a cycle of therapy for reasons other 
than toxicity will be replaced.  Patients who do not complete one cycle of therapy due to 
toxicity will not be replaced.  All patients should have completed one cycle of therapy (unless 
removed from the study due to toxicity), at which point they will be evaluable.
7. 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 7.1) and the characteristics of an observed AE (Section 7.2) will 
determine whether the event requires expedited reporting (via CTEP-AERS) in addition to 
routine reporting.  
7.1
Comprehensive Adverse Events and Potential Risks List(s) (CAEPRs)
Comprehensive Adverse Events and Potential Risks list (CAEPR) for Methoxyamine 
hydrochloride (TRC102, NSC 3801)
The Comprehensive Adverse Events and Potential Risks list (CAEPR) provides a single list of reported 

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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 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.
The CAEPR does not provide frequency data; refer to the Investigator's Brochure for this information. 
Below is the CAEPR for Methoxyamine hydrochloride (TRC102).
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 1.1, April 25, 20191
Adverse Events with Possible
Relationship to Methoxyamine hydrochloride (TRC102) 
(CTCAE 5.0 Term)
 Specific Protocol Exceptions to 
Expedited Reporting (SPEER) 
BLOOD AND LYMPHATIC SYSTEM DISORDERS
Anemia
Anemia (Gr 2)
GASTROINTESTINAL DISORDERS
Diarrhea
Diarrhea (Gr 1)
Mucositis oral
Mucositis oral (Gr 1)
Nausea
Nausea (Gr 2)
Vomiting
Vomiting (Gr 2)
GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS
Fatigue
Fatigue (Gr 2)
Fever
Fever (Gr 1)
INVESTIGATIONS
Blood bilirubin increased
Blood bilirubin increased (Gr 1)
Haptoglobin decreased
Haptoglobin decreased (Gr 1)
METABOLISM AND NUTRITION DISORDERS
Anorexia
Anorexia (Gr 1)
SKIN AND SUBCUTANEOUS TISSUE DISORDERS
Pruritus
Pruritus (Gr 1)
Rash maculo-papular
Rash maculo-papular (Gr 1)
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.
Adverse events reported on Methoxyamine hydrochloride (TRC102) trials, but for which there is 
insufficient evidence to suggest that there was a reasonable possibility that Methoxyamine 
hydrochloride (TRC102) caused the adverse event:

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BLOOD AND LYMPHATIC SYSTEM DISORDERS - Hemolysis
GASTROINTESTINAL DISORDERS - Constipation
IMMUNE SYSTEM DISORDERS - Allergic reaction
INVESTIGATIONS - Alanine aminotransferase increased; Aspartate aminotransferase increased; 
Creatinine increased; Lymphocyte count decreased; Neutrophil count decreased; Platelet count 
decreased; White blood cell decreased
METABOLISM AND NUTRITION DISORDERS - Hypocalcemia
NERVOUS SYSTEM DISORDERS - Dysgeusia
PSYCHIATRIC DISORDERS - Psychosis
RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS - Dyspnea
VASCULAR DISORDERS - Thromboembolic event
Animal Data: The following toxicities have been observed in animal studies with Methoxyamine 
hydrochloride (TRC102):
Dogs
BLOOD AND LYMPHATIC SYSTEM DISORDERS - Bone marrow hypercellularity; Epididymal 
cellular debris; Small thymus
GASTROINTESTINAL DISORDERS - Abnormal excreta
INVESTIGATIONS - Increased reticulocytes, Lymphocytes increased
NERVOUS SYSTEM DISORDERS - Convulsions; Tremors
REPRODUCTIVE SYSTEM AND BREAST DISORDERS - Seminiferous tubule degeneration
Rats
BLOOD AND LYMPHATIC SYSTEM DISORDERS - Enlarged or Swollen spleens; Increased 
spleen weight
INVESTIGATIONS - Increased monocytes; Increased reticulocytes; Lymphocytes increased
Note: Methoxyamine hydrochloride (TRC102) 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.
7.1.1
Adverse Event Lists for TMZ
The most common adverse reactions (≥10% incidence) are alopecia, fatigue, nausea, 
vomiting, headache, constipation, anorexia, convulsions, rash, hemiparesis, diarrhea, 
asthenia, fever, dizziness, coordination abnormal, viral infection, amnesia, and insomnia.  
The most common Grade 3 to 4 hematologic laboratory abnormalities (≥10% incidence) 
that have developed during treatment with temozolomide are lymphopenia, 
thrombocytopenia, neutropenia, and leucopenia which requires monitoring absolute 
neutrophil count (ANC) and platelet count prior to dosing and throughout treatment.  
Cases of myelodysplastic syndrome and secondary malignancies, including myeloid 
leukemia, have been observed.  All patients, particularly those receiving steroids, should 
be observed closely for the development of lymphopenia and PCP.  
Cases of hepatic injury, including fatal hepatic failure, have also been observed in 
patients enrolled on temozolomide studies; it was noted that liver toxicity may occur 

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several weeks or more after initiation of treatment or after temozolomide 
discontinuation.
Refer to the package insert for the comprehensive list of adverse events.
7.2
CTEP Reporting Requirements
7.2.1
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 4.0 will be utilized until March 31, 2018 for AE 
reporting.  CTCAE version 5.0 will be utilized for AE reporting beginning April 
1, 2018.  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 web site 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm.
  

‘Expectedness’: AEs can be ‘Unexpected’ or ‘Expected’ (see Section 7.1 above) 
for expedited reporting purposes only.  ‘Expected’ AEs (the ASAEL) are bold 
and italicized in the CAEPR (Section 7.1).

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.
7.2.2
Routine Adverse Event Reporting
All Adverse Events must be reported in routine study data submissions.  AEs 
reported through CTEP-AERS must also be reported in routine study data 
submissions.
7.2.3
Expedited Adverse Event Reporting
7.2.3.1 Expedited AE reporting for this study must use CTEP-AERS (CTEP Adverse 
Event Reporting System), accessed via the CTEP Web site 
(http://ctep.cancer.gov).  The reporting procedures to be followed are 
presented in the “NCI Guidelines for Investigators: Adverse Event Reporting 
Requirements for DCTD (CTEP and CIP) and DCP INDs and IDEs” which 
can be downloaded from the CTEP Web site (http://ctep.cancer.gov).  These 
requirements are briefly outlined in the tables below.
In the rare occurrence when Internet connectivity is lost, a 24-hour 
notification is to be made to CTEP by telephone at 301-897-7497.  Once 

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Internet connectivity is restored, the 24-hour notification phoned in must be 
entered electronically into CTEP-AERS by the original submitter at the site.
7.2.3.2 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 causality, unless as noted below.  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. 
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 CTEP-
AERS 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
Not resulting in Hospitalization 
≥ 24 hrs
Not required
24-Hour 5 
Cale
ndar 
Days

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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 reported via CTEP-AERS 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 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
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
7.2.3.3 Protocol-specific expedited AE reporting exclusions
For this protocol only, certain AEs/grades are exceptions to the Expedited 
Reporting Guidelines and do not require expedited reporting (i.e., CTEP-
AERS).  These are: any grade lymphopenia, any grade alopecia, Grade 2 
electrolyte (sodium, potassium, phosphorous, magnesium) abnormalities, 
Grade 2 anemia, Grade 2 hypoalbuminemia, Grade 2 hyperglycemia, Grade 
2 INR, Grade 2 PTT, and Grade 2 hyperuricemia will NOT be reported 
through CTEP-AERS but will be reported in the routine data submissions.
7.2.3.4 Pregnancy, Fetal Death, and Death Neonatal
NOTE: When submitting CTEP-AERS reports for “Pregnancy”, “Pregnancy 
loss”, or “Neonatal loss”, the Pregnancy Information Form should be 
completed and faxed along with any additional medical information to 301-
230-0159.  The potential risk of exposure of the fetus to the investigational 
agent(s) or chemotherapy agent(s) should be documented in the 
“Description of Event” section of the CTEP-AERS report.
7.2.3.5 Pregnancy
• Because patients who become pregnant on study risk intrauterine exposure 
of the fetus to agents which may be teratogenic, DCTD/DCP is requesting 
that pregnancy should be reported in an expedited manner via CTEP-AERS 
as Grade 3 “Pregnancy, puerperium and perinatal conditions - Other 
(pregnancy)” under the Pregnancy, puerperium and perinatal conditions 
SOC.
• The pregnancy outcome for patients on study should be reported via CTEP-
AERS at the time the outcome becomes known, accompanied by the same 
Pregnancy Report Form used for the initial report.

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7.2.3.6 Pregnancy loss
• Pregnancy loss is define in CTCAE as “Death in utero.”
• Any pregnancy loss should be reported expeditiously, as Grade 4 
“Pregnancy loss” under the Pregnancy, puerperium and perinatal conditions 
SOC.
• A pregnancy loss should NOT be reported as  a Grade 5 event under the 
Pregnancy, puerperium and perinatal conditions SOC, as currently CTEP-
AERS recognizes this event as a patient death.
7.2.3.7 Death Neonatal
• Neonatal death, defined in CTCAE as “A disorder characterized by cessation 
of life occurring during the first 28 days of life” that is felt by the 
investigator to be at least possibly due to the investigational 
agent/intervention, should be reported expeditiously.
• A neonatal death should be reported expeditiously as Grade 4 “Death 
neonatal” under the General disorders and administration SOC.
• Neonatal death should NOT be reported as Grade 5 “Death neonatal” under 
the General disorders and administration SOC.  If reported as such, the 
CTEP-AERS interprets this as a death of the patient being treated.
7.2.4
Information regarding transfusion requirements in the first 3 cycles of treatment 
will be reported to CTEP. It will not require expedited reporting.
7.2.5
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 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. 

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7.2.6
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 reporting via CDUS unless otherwise specified. 
7.3
NIH Reporting Requirements 
7.3.1
Definitions
Please refer to definitions provided in Policy 801: Reporting Research Events 
(https://irbo.nih.gov/confluence/display/IRBO/Policies+and+SOPs).
7.3.2
OHSRP Office of Compliance and Training / IRB Reporting 
Please refer to the reporting requirements in Policy 801: Reporting Research 
Events and Policy 802 Non-Compliance Human Subjects Research found at 
https://irbo.nih.gov/confluence/display/IRBO/Policies+and+SOPs
Note: Only IND Safety Reports that meet the definition of an unanticipated 
problem will need to be reported per these policies.
7.3.3
IRB Requirements for PI Reporting at Continuing Review
Please refer to the reporting requirements in Policy 801: Reporting Research 
Events found at 
https://irbo.nih.gov/confluence/display/IRBO/Policies+and+SOPs 
7.3.4
NCI Clinical Director Reporting 
Problems expeditiously reported to the OHSRP/IRB in iRIS will also be reported 
to the NCI Clinical Director.  A separate submission is not necessary as reports 
in iRIS will be available to the Clinical Director.
In addition to those reports, all deaths that occur within 30 days after receiving a 
research intervention should be reported via email to the Clinical Director unless 
they are due to progressive disease.  
To report these deaths, please send an email describing the circumstances of the 
death to the Clinical Director/designee at NCICCRQA@mail.nih.gov within one 
business day of learning of the death.

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8.
PHARMACEUTICAL INFORMATION
8.1
TRC102 (NSC 3801)
Chemical Name:  Methoxyamine hydrochloride
Other Names:  Methoxylamine HCl
Classification:  Biochemical inhibitor of the BER pathway 
Molecular Formula: CH5NO•HCl
M.W.: 83.52 Daltons
Approximate Solubility:   At ambient temperature, TRC102 is freely soluble in water, 
sparingly in ethanol (70 mg/mL), and slightly in DMSO (140 mg/mL). 
Mode of Action: TRC102 has the ability to interrupt the process of base excision repair 
(BER) by binding to apurinic/apyrimidinic sites produced during the initial step of the 
BER pathway.  These sites are substrates for topoisomerase II (topo II); an enzyme that 
cleaves damaged DNA.  TRC102 has demonstrated the ability to potentiate the activity 
of the alkylating agents temozolomide and carmustine, and antimetabolite agents 
fludarabine and pemetrexed, in murine models of human cancer.  Therefore, TRC102 
may be able to potentiate the activity of alkylating and antimetabolite chemotherapy in 
patients.
Description:  TRC102, a white, crystalline solid, is the hydrochloride salt of 
methoxyamine.
How Supplied: TRC102 is supplied by IriSys, Inc. (white capsules) or the 
Developmental Therapeutics Program, DCTD/NCI (pink capsules) and distributed by 
the Pharmaceutical Management Branch, CTEP/DCTD/NCI.  Each opaque, size 2, hard 
gelatin capsules contain 25 mg of Methoxyamine HCl powder, microcrystalline 
cellulose, crospovidone, sodium starch glycolate, colloidal silicon dioxide, and talc.  
Each HDPE bottle has a child-resistant screw cap and contains 30 capsules.
Storage: Store bottles of TRC102 at refrigerated temperature (2-8°C). 
Stability:  Shelf life surveillance of the intact bottles in ongoing.
Route of Administration:  Oral
Method of Administration:  Capsules are taken in the morning after patients have fasted 
for at least two hours.  Take with 8 ounces of water.  Capsules should be swallowed whole, 
do not chew or crush.  Patients should refrain from eating or drinking for one hour 
following TRC102 dosing.
Elimination: There have been no data published on excretion or metabolism of TRC102.  

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Based on the chemical structure of such a small molecule, no CYP interaction is expected
8.2
Temozolomide 
Temozolomide is commercially available.  Please refer to the FDA-approved package 
insert for more information about this drug.  Full prescribing information is available at: 
http://www.temodar.com/temodar/index.do
Chemical Name: 3,4-dihydro-3-11 methyl-4-oxoimidazo[5,1-d]-as-tetrazine-8-
carboxamide.
Other Names: Temodar®, Temodal, Temcad
Molecular Formula: C6H6N6O2 (M.W: 194.15)
Mode of Action: Temozolomide is not directly active but undergoes rapid nonenzymatic 
conversion at physiologic pH to the reactive compound MTIC.  The cytotoxicity of 
MTIC is thought to be primarily due to alkylation of DNA.  Alkylation (methylation) 
occurs mainly at the O6 and N7 positions of guanine.
How Supplied: Capsules are supplied in amber glass bottles with child resistant 
polypropylene caps containing the following capsule strengths: 5 mg; 20 mg; 100 mg; 
140 mg; 180 mg; 250 mg.  The dose of temozolomide is based on BSA then rounded off 
to the nearest 5 mg.
Storage: Store at 25°C (77°F); excursions permitted to 15°-30°C (59°-86°F).
Stability: The molecule is stable at acidic pH (<5), and labile at pH >7
Route of Administration: Oral: Take each day’s dose of capsules at one time, with a full 
glass of water.  They should be swallowed whole and never chewed.  If capsules are 
vomited do not take a second dose.
Potential Drug Interaction: Administration of valproic acid decreases oral clearance of 
temozolomide by about 5%.
Availability:  Temozolomide is commercially available
8.3
Agent Ordering and Agent Accountability
8.3.1
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 

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(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.
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 an active 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.
8.3.2
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.
8.3.3
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 an active registration status.  Questions about IB access 
may be directed to the PMB IB Coordinator via email.
8.3.4
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 

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. BIOMARKER, CORRELATIVE, AND SPECIAL STUDIES

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9.1
Pharmacokinetics (Phase I only)
Blood samples for PK analyses (optional) will be collected prior to drug administration and 1, 2, 
3, 4, 8, 12, and 24 hours post-dosing on cycle 1 day 1 in both the escalation and expansion 
phases and on cycle 1 day 5, prior to dosing on that day in the expansion phase only.  Based on 
results from initial measurements, sampling times may be adjusted, but neither the total number 
of samples nor the total amount of blood drawn per patient will be increased.  Samples will be 
collected in lavender-top (EDTA) tubes with 2 mL of blood per sample and stored refrigerated 
(2-8°C).  All samples will be centrifuged and plasma will be stored at -70oC for analysis as 
described in Appendix E.  Samples will be analyzed using a validated LC-MS or LC-MS/MS 
method in human plasma.  
Urine (optional) will be collected prior to drug administration on cycle 1 day 1, and kept 
separate and then treat as described in Appendix E.  Urine will then be collected at every void 
from 0 to 24 hours post-treatment on day 1 in cycle 1 for PK analysis.  Start urine collection 
immediately after study drug is administered and store refrigerated (2-8°C) acidified as 
described in Appendix E.  
Send PK samples to the following lab contact:
Laboratory Contact:
Tracy W. Webb
Office of the Associate Director/DTP/NCI
FNLCR
Boyles Street
Building 1047, Room 8
Frederick, MD 21702
301-846-7402; webbtw@mail.nih.gov
9.2
Pharmacodynamics
Tumor tissue: Evaluation of drug effect on DNA damage response will be performed by 
immunofluorescence assay for measurement of H2AX levels and other biomarkers of 
DNA damage response/repair.  Epithelial-mesenchymal transition (EMT), MGMT, and 
levels of cCasp3 (apoptosis) will also be measured.  Any remaining tumor tissue may be 
formalin-fixed and paraffin-embedded and sent to the laboratory of Dr. Stanton Gerson 
(Case Western Reserve University) for IHC assessment of the enzyme N-methylpurine 
DNA glycosylase (MPG).  No patient data will accompany any samples sent to Dr. Gerson’s 
lab.
9.2.1
Laboratory Contact
At least 24 hours prior to tumor biopsy or blood sample collection, the research nurse 
will contact the NCI Phase I/II PK/PD Support Group in NIH Building 10: 
E-mail (preferred): NCIPK-PDsupportgroup@mail.nih.gov 

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Pager (preferred): 102-12798 
Phone: 240-858-3963
Fax: 301-480-5871.  
For biopsies, tubes pre-labeled with the information specified in Section 9.2.4, biopsy 
date, and site of tissue biopsy will be provided.  Initial processing and shipping of the 
samples will be completed as described below.  If a patient had a biopsy within the past 
3 months and had no treatment since that biopsy, an attempt will be made to obtain and 
process that tissue as the baseline biopsy.  
9.2.2
Blood Collection for CTC Studies (Optional)
At each time point, whole blood (at least 7.5 mL) will be collected aseptically by 
venipuncture or from a venous port into one 10 mL Streck tube (catalog number 
218962).  One 10 mL RareCyte tube (catalog number 24-1070-005) is also acceptable. 
Tubes must be inverted 8 times to ensure adequate mixing of the additive.  Blood 
samples for CTCs will be collected from patients at the following time points:
Phase I

Cycle 1 prior to treatment 

Cycle 1, day 1: 8 hrs after drug in both the escalation and expansion phases 

Cycle 1, day 5 (±1 day in case of scheduling difficulties): before drug in the 
expansion phase only

Cycle 2, day 1: before drug in both the escalation and expansion phases

Day 1 of all subsequent cycles: before drug in both the escalation and expansion 
phases

At time of disease progression
Phase II

Cycle 1 prior to treatment 

Cycle 1 day 4 or 5 before drug (on the same day as the post-dose biopsy) 

Day 1 (±1 day) of all subsequent cycles before drug 

At time of disease progression
Blood for CTC analysis will be shipped to the PADIS laboratory on the day it is 
collected.  Arrangement will be made for pickup with the CSP courier service (301-846-
5893) and the receiving laboratory will be notified by email of the shipment 
(NCIPDSupportPADIS@mail.nih.gov).  Because of the 48-hour window of CTC sample 
stability, no CTC specimens should be collected or shipped on Friday afternoon.  
Samples will be sent to:
Attn: PADIS CTC Laboratory
Frederick National Laboratory for Cancer Research
Leidos Biomedical Research, Inc.
1050 Boyles Street
Building 425, Room 102
Frederick, MD 21702

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Phone: 301.846.4711
NCIPDSupportPADIS@mail.nih.gov
9.2.2.1
Measurement of Circulating Tumor Cells (CTC) in Peripheral Blood
PADIS, Frederick National Lab, is developing and validating the process to isolate 
circulating tumor cells using the ApoStreamTM technology platform which uses size 
and cell surface charge to isolate cancer cells; this platform is of particular interest in 
that it can detect CTCs from patients with cancer of mesenchymal origin, such as 
sarcomas.  
9.2.3
Tumor Biopsies
9.2.3.1
Timing of tumor biopsies
Biopsies will be optional during the escalation phase and mandatory in up to 15 
additional patients enrolled at the MTD during the Phase I expansion phase and 
during the Phase II portion of the study (patients with NSCLC will not be biopsied).  
Biopsies will be collected:

before drug administration on study (baseline), and

post-treatment:
o
Phase I: on cycle 1, day 5 (± 1 day in case of scheduling difficulties), 
3-4 hrs after drug
o
Phase II: on cycle 1, day 4 or 5, 3-4 hrs after drug 

OPTIONAL—One optional tumor biopsy may be collected:
o
A restaging follow-up biopsy may be performed on day 1 ( 2 days) 
of the cycle following any restaging at which a 10-19% increase in 
tumor volume is observed (according to RECIST criteria) if the 
patient has been on study for at least 4 cycles
OR
o
A progression tumor biopsy may be performed at the time of disease 
progression
  
9.2.3.2
 Biopsy Procedure
Serial tumor biopsies will be obtained by the Interventional Radiology team by a 
percutaneous approach, a dermatologist for skin lesions, or an ENT for lesions that 
are easily biopsiable through ENT exam.  If the patient has skin lesions or peripheral 
lymph nodes that are easily accessible by the dermatologist or surgeon and could be 
done under local anesthesia with < 5% risk of complications, this will be acceptable 
if agreeable with the patient and investigator.  If considered feasible and safe by the 
biopsy team, up to five core biopsies ≥18 gauge in diameter and ≥1 cm in length, or 
equivalent, will be obtained during each procedure.  It is estimated that there will be 
between 2 million–5 million cells from each biopsy.  If a site is deemed appropriate 
for biopsy with minimal risk to the participant by agreement between the 
investigators and the biopsy team, an attempt for biopsy will be made.  If possible, 
the lesion from which each biopsy is taken will be documented.

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The use of imaging to facilitate biopsies will be decided by members of the biopsy 
team and may include ultrasound, CT scan, or MRI.  Should a CT scan be needed for 
biopsy, the number of scans for each procedure will be limited to the minimum 
number needed to safely obtain a biopsy.  Tumor biopsies and local anesthesia will 
be administered only if they are considered to be of low risk to the participant, as 
determined by the investigators and the biopsy team.  In the expansion cohort, if the 
participant chooses not to undergo tumor biopsy after the initial attempt, he/she will 
still remain in the study and receive study medication, and all the other correlative 
studies will be performed.
Tumor biopsies are optional during the escalation phase but mandatory for the 15 
additional patients enrolled at the MTD during the expansion phase and Phase II portion 
of the study.  Baseline biopsies will be performed following patient enrolling on study or 
if the patient had a biopsy within the past 3 months and had no treatment since that 
biopsy, an attempt will be made to obtain and process that tissue as baseline biopsy.  If 
the tissue could not be used for planned PD studies, a biopsy will be performed.  If an 
initial attempt at biopsy is unsuccessful, the patient will be given an option to proceed 
with a repeated attempt.  A separate consent form must be signed for each biopsy 
procedure, so patients may choose not to undergo subsequent biopsies.  If the baseline 
biopsy is unsuccessful or the patient refuses to undergo subsequent biopsies, no further 
biopsies will be performed but the patient will remain on study, receive study 
medication, and other correlative studies will be performed.  The patient will be replaced 
in the accrual scheme of the expansion phase.  
9.2.3.3
Solid Tumor Biopsy Processing
Up to five tissue cores, or equivalent, will be collected and flash frozen in liquid 
nitrogen and submitted for evaluation of DNA damage markers, and H&E pathology 
evaluation.  The frozen biopsy specimens are transferred to PADIS, where the 
biopsy samples are stored at -80°C until processing.  Refer to SOP340507 
(https://dctd.cancer.gov/ResearchResources/biomarkers/docs/par/SOP340507_Biops
y_Frozen.pdf) for detailed instructions on biopsy collection and handling.  The 
sample shipping manifest and batch record are included in SOP340507.  Biopsy 
samples will be analyzed for PD and genomic analyses based on the assay 
prioritization shown in Figure 4 and the discretion of the PI. 
Biopsies will be shipped on dry ice to:
ATTN: PADIS IQC Lab
Frederick National Laboratory for Cancer Research
Leidos Biomedical Research, Inc.
1050 Boyles Street
Building 425, Room 105
Frederick, MD 21702
Phone: 301.846.1951 or 301.846.6747
NCIPDSupportPADIS@mail.nih.gov

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Shipment should be by CSP Courier and may be arranged by contacting Mike 
Johnston, FNLCR, Tel.: 301-846-5893
Note: Per the discretion of the PI, restaging follow-up or progression biopsies may be 
collected per SOP to be used for analyses other than those listed here, including 
analyses carried out as a part of other DTC clinical trials.
9.2.3.4
Archival Tissue Submission
Archival tumor tissue submitted as a baseline specimen must have been collected 
within 3 months prior to patient registration, and the patient must not have received 
any intervening cancer therapy since collection of the specimen.  Archival tissue 
must be collected and processed according to SOP340507 
(https://dctd.cancer.gov/ResearchResources/biomarkers/docs/par/SOP340507_Biops
y_Frozen.pdf), including flash-freezing in liquid nitrogen, minimal cold ischemia 
time (< 5 minutes), and shipment on dry ice.
Please send an email to FNLCR PD Specimen Central Receiving 
(NCIPDSupportPADIS@mail.nih.gov ) to advise that archival tissue is being 
prepared for shipment.  State “Protocol Name PD Specimens Ready for Shipment” 
in the subject line.  If needed, FNLCR PD Central Receiving can be contacted 
directly at 301-846-1951 or 301-846-6747.
9.3
Exploratory Genomic Analyses
As of Amendment R (5/12/2018), the MoCha laboratory will investigate the occurrence 
of tumor genomic alterations potentially associated with sensitivity and/or the 
development of resistance to TRC102 and temozolomide, such as the status of mismatch 
repair genes or the development of increased mutational load and neoantigen production.  
Whole exome (WES) and whole transcriptome (RNASeq) genomic analysis of tumor 
samples collected before and during treatment and upon progression will be performed 
and compared with data from germline tissue to identify somatic variants, which will be 
used to determine tumor mutational load and identify neoantigens.
9.3.1
Tumor Genomic Analyses
Tumor biopsies will be collected and processed as detailed in Section 9.2.3.3.
9.3.1.1
Baseline and Cycle 1, Day 4/5 Biopsies
Paired tumor biopsy tissues collected at baseline and after treatment on 
cycle 1, day 4 or 5 will be used for exploratory targeted (OCAv3) and 
WES analyses only if there is sufficient tissue remaining after PD 
biomarker analyses have been completed (Figure 4). Tissue 
availability will be tracked in real time using Labmatrix. Although the 
MoCha Laboratory is CLIA-certified, these sequencing studies will 
not be done per CLIA specifications as these data will not be returned 
to the patients or used for clinical decision making.

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9.3.1.2
Restaging Follow-Up and Progression Biopsies 
Tumor tissue collected at the (optional) restaging follow-up or 
progression biopsy will be analyzed by the CLIA-certified OCAv3 assay 
and results will be returned to patients in the form of the Oncomine 
report.  If additional tumor tissue is available after the OCAv3 analysis, 
exploratory, non-CLIA certified genomic analyses such as WES may be 
conducted but the results will not be returned to patients or used for 
clinical decision making (Figure 4).  Note: Per the discretion of the PI, 
restaging follow-up or progression biopsies may be used for other 
analyses, including analyses carried out as a part of other DTC clinical 
trials, providing that the tissue is collected per SOP for those analyses.
9.3.2
Germline Genomic Analysis
One whole blood sample (10-mL Streck tube) will be collected before drug 
administration on cycle 1 day 1 (or another time while the patient is on study) 
and mononuclear cells isolated for nucleic acid extraction and exploratory WES 
and RNAseq to compare somatic variant calls, mutational load, MMR 
inactivation, and other potential markers of drug sensitivity or resistance to the 
tumor biopsy specimen, the results of which will not be returned to patients 
(Figure 4).  
Blood specimens for germline genomic analysis will be shipped at ambient 
temperature to: 
ATTN: Gloryvee Rivera/Lindsay Dutko
MoCha Histology Lab
Frederick National Laboratory for Cancer Research
Leidos Biomedical Research, Inc.
1050 Boyles Street
Building 321 Room 107
Frederick, MD 21702
Shipment Notification:
MoChaSampleReceiving@nih.gov
Samples should be labeled with only the unique patient ID. Do NOT include 
patient identifiers (e.g., medical record number, patient name, or initials) 
with the samples.

CTEP #9483
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Fig. 4. Summary of Planned Tumor and Germline Genomic Analyses:
9.3.3
Privacy Considerations
As patient clinical response data (both for this study and, potentially, after a 
patient is off-study) will be required for comparison to sequencing results, de-
identifying the samples is not feasible.  A Certificate of Confidentiality has been 
obtained to help protect the privacy of all study participants.  The informed 
consent document for this protocol after Amendment R (5/12/2018) contains 
language informing patients about the performance of genetic studies, and 
patients will have the option to choose whether they wish to take part in these 
studies.
9.3.4
Management of Results and Genetic Counseling
Whole-exome sequencing (WES) of tumor and blood performed for research 
purposes can detect non-ambiguous germline variants, which may raise health 
and privacy implications for the patient and his or her family.  WES will not be 
validated for clinical use, and no clinical decisions can be made based on its 
results.  
The results of the CLIA-certified OCAv3 targeted sequencing assay on restaging 
follow-up or progression tumor biopsy tissue at MoCha (CLIA certification 
#21D2097127) will be returned to patients in the Oncomine report.

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9.4
Sample Collection and Processing
Biospecimens will be collected and processed using validated SOPs that will ensure both 
specimen quality and patient confidentiality pursuant to informed consent provisions.  
Information about each specimen (e.g., blood, tumor biopsy, circulating tumor cells, per 
specific protocol) will be recorded on a PK/PD collection worksheet included in 
Appendix D. 
Using a computerized inventory system and a backup hardcopy process, all specimen 
collection and processing steps will be documented and the specific location of each 
specimen will be tracked.  Each new specimen collected will be assigned a unique 
barcode identifier that can be linked to the original specimen collected and other relevant 
information within the inventory system.  To ensure patient confidentiality, only 
containers used for the initial specimen collections will be labeled with patient 
identifiers.  
Only the barcode identifier will be applied to all subsequent specimen containers.  When 
specimens are processed and aliquoted, no patient information will be included on the 
new containers.  Original specimen containers will be discarded.  Only barcode-labeled 
specimens without patient identifiers will be shipped for analysis and/or storage.  
Specimen labels will indicate: CTEP protocol number, unique patient accession number, 
3-digit sample number (see list below), collection time, and total volume collected, as 
appropriate.  Samples from sets of at least three patients will be grouped for scientific 
analysis.
Standardized 3-digit sample collection numbers:
000 series: blood for germline WES
100 series: urine for PK
200 series: blood for PK
300 series: blood for PD
400 series: blood for circulating tumor cells (CTCs)
500 series: tumor biopsies
The inventory process contains other security provisions sufficient to safeguard patient 
privacy and confidentiality.  Access to the inventory system and associated documents 
will be restricted to appropriate individuals.  Requests to use specimens stored in the 
repository must be approved.  The only patient information available in the inventory 
system will be the patient sex, diagnosis, and level of informed consent given.  SOPs 
ensure that any changes in informed consent made by a patient and relayed to the PI will 
be reflected in the inventory system to ensure that specimens are destroyed as 
appropriate.  All laboratory personnel will be trained to adhere to SOPs and will be 
monitored for high-quality performance.
Any new use of these samples will require prospective IRB review and approval.  
Access to these samples will only be granted following IRB approval of an additional 
protocol, granting the rights to use the material.

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If at any time, a patient withdraws from the study and does not wish for their existing 
samples to be utilized, the individual must provide a written request.  Following receipt 
of this request, the samples will be destroyed (or returned to the patient, if so requested), 
and reported as such to the IRB.  Any samples lost (in transit or by a researcher) or 
destroyed due to unknown sample integrity (i.e., broken freezer allows for extensive 
sample thawing, etc.) will be reported as such to the IRB.
9.4.1
Human Data Sharing Plan
What data will be shared? 
We will share human data generated in this research for future research as follows:
X De-identified data in an NIH-funded or approved public repository 
X Identified data in BTRIS (automatic for activities in the Clinical Center) 
X De-identified or identified data with approved outside collaborators under appropriate 
agreements 
How and where will the data be shared? 
Data will be shared through: 
X An NIH-funded or approved public repository: clinicaltrials.gov
X BTRIS (automatic for activities in the Clinical Center) 
X Approved outside collaborators under appropriate individual agreements 
X Publication and/or public presentations 
When will the data be shared? 
X At the time of publication or shortly thereafter
10. STUDY CALENDAR
Baseline evaluations are to be conducted within 8 days prior to start of protocol therapy.  Scans 
and x-rays 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.  Start of next cycle may be delayed for up to 1 week to 
accommodate scheduling conflicts.  History and physical examination and laboratory 
evaluations can be performed up to 1 week before the start of the next cycle.
Pre-
Study
C1
W1
C1
W2k
C1 
W3k
C1 
W4k
C2
 W1k
C2
W3
C3
 W1k
C3   
W3k on
Off 
Treatmentj
TRC102/TMZa
X
X
X
Informed consent
X
Demographics
X
Medical history
X
X
X
Concurrent meds
X
X------------------------------------------------------------------------------------X
Physical examb
X
X
X
X
X

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Vital signs
X
X
X
X
Height
X
Weight
X
X
X
X
Performance 
status
X
X
X
X
CBC w/diff, pltsc
X
X
X
X
X
Serum chemistryc
X
X
X
X
X
Anemial
X
X
X
PT, INR, PTTd
X
β-HCGe
X
AE evaluation
X------------------------------------------------------------------------------------X
X
Tumor 
measurements
X
Tumor measurements are repeated every 2 cycles (3 cycles for patients 
on study more than 1 year).  Documentation (radiologic) must be 
provided for patients removed from study for progressive disease.
Tumor biopsyf
X
X
PK bloodg
X
Circulating tumor 
cellsh
X
X
X
ECGi
X
a: TRC102 and TMZ will be administered once daily, days 1-5, in 28 day cycles.        
b: Physical exam at the Clinical Center should be performed at the start of each cycle (up to 1 week 
before start of a new cycle).  Start of the next cycle may be delayed for up to 1 week to accommodate 
scheduling conflicts.
c: Serum chemistry (albumin, total bilirubin, calcium, creatinine, phosphorus, magnesium, potassium, 
sodium, SGOT [AST], SGPT [ALT], LDH); CBC w/diff, platelets up to week 3 during cycle 1 (Phase I 
only), and during week 1 in cycle 2 and subsequent cycles (up to 1 week before start of a new cycle) 
(Phase I and II).  If clinically indicated, labs may be obtained more frequently. 
d. PT/INR, PTT as clinically indicated.
e: Serum pregnancy test (women of childbearing potential) within 8 days prior to enrollment and as 
clinically indicated.
f: Tumor biopsies (mandatory only in Phase I expansion cohort and Phase II where appropriate) will be 
performed in patients who have disease amenable to biopsy prior to treatment and 3-4 hours after the 
5th dose of TRC102 on C1D5 (±1 day in case of scheduling difficulties; day 4 or 5, 3-4 hrs after drug 
for patients on Phase II reflecting likely start day/weekend timing).  One optional biopsy at progression 
or on day 1 (± 2 days) of the cycle following any restaging at which a 10-19% increase in tumor 
volume is observed (according to RECIST criteria) for patients who have been on study for at least 4 
cycles may also be obtained.
g: Phase I only: PK samples (optional) will be performed at the following time points:  C1D1: Pre-dose, 
at 1, 2, 3, 4, 8, 12, and 24 hours after the first dose in both the escalation and expansion phases; C1D5 
pre-dose in the expansion phase only.  Additional PK sampling will be optional (but encouraged). 
h: Circulating tumor cells (optional) will be collected as described in Section 9.2 
i: ECG will be performed at baseline for evaluation of QTc and as clinically indicated thereafter.
j: Off-study evaluation.
k: ±1 day in case of scheduling difficulties

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l: refer to Appendix F
11. MEASUREMENT OF EFFECT
11.1
Antitumor Effect – Solid Tumors
Although response is not the primary endpoint of this trial, patients with measurable disease 
will be assessed by standard criteria.  For the purposes of this study, patients should be re-
evaluated for response every 8 weeks (every 2 cycles; 3 cycles for patients on study for a year).  
In addition to a baseline scan, confirmatory scans should also be obtained at least 4 weeks 
following initial documentation of objective response.
Response and progression will be evaluated in this study using the new international criteria 
proposed by the revised Response Evaluation Criteria in Solid Tumors (RECIST) guideline 
(version 1.1) [Eur J Ca 45:228-247, 2009].  Changes in the largest diameter (unidimensional 
measurement) of the tumor lesions and the shortest diameter in the case of malignant lymph 
nodes are used in the RECIST criteria.
11.1.1 Definitions
Evaluable for toxicity:  All patients will be evaluable for toxicity from the time of their 
first treatment with study drugs.
Evaluable for objective response:  All patients who received study drug will be 
considered evaluable for assessment of 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.)
Evaluable Non-Target Disease Response:  Patients who have lesions present at baseline 
that are evaluable but do not meet the definitions of measurable disease, have received at 
least one cycle of therapy, and have had their disease re-evaluated will be considered 
evaluable for non-target disease.  The response assessment is based on the presence, 
absence, or unequivocal progression of the lesions. 
11.1.2 Disease Parameters
Measurable disease:  Measurable lesions are defined as those that can be accurately 
measured in at least one dimension (longest diameter to be recorded) as >20 mm by 
chest x-ray or as ≥10 mm with CT scan, MRI, or calipers by clinical exam.  All tumor 
measurements must be recorded in millimeters (or decimal fractions of centimeters).
Malignant lymph nodes:  To be considered pathologically enlarged and measurable, a 
lymph node must be ≥15 mm in short axis when assessed by CT scan (CT scan slice 

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thickness recommended to be no greater than 5 mm).  At baseline and in follow-up, only 
the short axis will be measured and followed.
Non-measurable disease:  All other lesions (or sites of disease), including small lesions 
(longest diameter <10 mm or pathological lymph nodes with ≥10 to <15 mm short axis), 
are considered non-measurable disease.  Bone lesions, leptomeningeal disease, ascites, 
pleural/pericardial effusions, lymphangitis cutis/pulmonitis, inflammatory breast 
disease, and abdominal masses (not followed by CT or MRI), are considered as non-
measurable.
Note:  Cystic lesions that meet the criteria for radiographically defined simple cysts 
should not be considered as malignant lesions (neither measurable nor non-measurable) 
since they are, by definition, simple cysts.
‘Cystic lesions’ thought to represent cystic metastases can be considered as measurable 
lesions, if they meet the definition of measurability described above.  However, if non-
cystic lesions are present in the same patient, these are preferred for selection as target 
lesions.
Target lesions:  All measurable lesions up to a maximum of 2 lesions per organ and 5 
lesions in total, representative of all involved organs, should be identified as target 
lesions and recorded and measured at baseline.  Target lesions should be selected on the 
basis of their size (lesions with the longest diameter), be representative of all involved 
organs, but in addition should be those that lend themselves to reproducible repeated 
measurements.  It may be the case that, on occasion, the largest lesion does not lend 
itself to reproducible measurement in which circumstance the next largest lesion which 
can be measured reproducibly should be selected.  A sum of the diameters (longest for 
non-nodal lesions, short axis for nodal lesions) for all target lesions will be calculated 
and reported as the baseline sum diameters.  If lymph nodes are to be included in the 
sum, then only the short axis is added into the sum.  The baseline sum diameters will be 
used as reference to further characterize any objective tumor regression in the 
measurable dimension of the disease.
Non-target lesions:  All other lesions (or sites of disease) including any measurable 
lesions over and above the 5 target lesions should be identified as non-target lesions 
and should also be recorded at baseline.  Measurements of these lesions are not required, 
but the presence, absence, or in rare cases unequivocal progression of each should be 
noted throughout follow-up. 
11.1.3 Guidelines for Evaluation of Measurable Disease
All measurements should be taken and recorded in metric notation using a ruler or 
calipers.  All baseline evaluations should be performed as closely as possible to the 
beginning of treatment and never more than 4 weeks before the beginning of the 
treatment.

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The same method of assessment and the same technique should be used to characterize 
each identified and reported lesion at baseline and during follow-up.  Imaging-based 
evaluation is preferred to evaluation by clinical examination unless the lesion(s) being 
followed cannot be imaged but are assessable by clinical exam.
Clinical lesions:  Clinical lesions will only be considered measurable when they are 
superficial (e.g., skin nodules and palpable lymph nodes) and 10 mm diameter as 
assessed using calipers (e.g., skin nodules).  In the case of skin lesions, documentation 
by color photography, including a ruler to estimate the size of the lesion, is 
recommended. 
Chest x-ray:  Lesions on chest x-ray are acceptable as measurable lesions when they are 
clearly defined and surrounded by aerated lung.  However, CT is preferable. 
Conventional CT and MRI:  This guideline has defined measurability of lesions on CT 
scan based on the assumption that CT slice thickness is 5 mm or less.  If CT scans have 
slice thickness greater than 5 mm, the minimum size for a measurable lesion should be 
twice the slice thickness.  MRI is also acceptable in certain situations (e.g., for body 
scans).  
As with CT, if an MRI is performed, the technical specifications of the scanning 
sequences used should be optimized for the evaluation of the type and site of disease.  
Furthermore, as with CT, the modality used at follow-up should be the same as was used 
at baseline and the lesions should be measured/assessed on the same pulse sequence.  It 
is beyond the scope of the RECIST guidelines to prescribe specific MRI pulse sequence 
parameters for all scanners, body parts, and diseases.  Ideally, the same type of scanner 
should be used and the image acquisition protocol should be followed as closely as 
possible to prior scans.  Body scans should be performed with breath-hold scanning 
techniques, if possible.
PET-CT:  At present, the low dose or attenuation correction CT portion of a combined 
PET-CT is not always of optimal diagnostic CT quality for use with RECIST 
measurements.  However, if the site can document that the CT performed as part of a 
PET-CT is of identical diagnostic quality to a diagnostic CT (with IV and oral contrast), 
then the CT portion of the PET-CT can be used for RECIST measurements and can be 
used interchangeably with conventional CT in accurately measuring cancer lesions over 
time.  Note, however, that the PET portion of the CT introduces additional data which 
may bias an investigator if it is not routinely or serially performed.  
Ultrasound:  Ultrasound is not useful in assessment of lesion size and should not be used 
as a method of measurement.  Ultrasound examinations cannot be reproduced in their 
entirety for independent review at a later date and, because they are operator dependent, 
it cannot be guaranteed that the same technique and measurements will be taken from 
one assessment to the next.  If new lesions are identified by ultrasound in the course of 
the study, confirmation by CT or MRI is advised.  If there is concern about radiation 
exposure at CT, MRI may be used instead of CT in selected instances.

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Endoscopy, Laparoscopy:  The utilization of these techniques for objective tumor 
evaluation is not advised.  However, such techniques may be useful to confirm complete 
pathological response when biopsies are obtained or to determine relapse in trials where 
recurrence following complete response (CR) or surgical resection is an endpoint.
Tumor markers:  Tumor markers alone cannot be used to assess response.  If markers are 
initially above the upper normal limit, they must normalize for a patient to be considered 
in complete clinical response.  Specific guidelines for both CA-125 response (in 
recurrent ovarian cancer) and PSA response (in recurrent prostate cancer) have been 
published [JNCI 96:487-488, 2004; J Clin Oncol 17, 3461-3467, 1999; J Clin Oncol 
26:1148-1159, 2008].  In addition, the Gynecologic Cancer Intergroup has developed 
CA-125 progression criteria which are to be integrated with objective tumor assessment 
for use in first-line trials in ovarian cancer [JNCI 92:1534-1535, 2000].
Cytology, Histology:  These techniques can be used to differentiate between partial 
responses (PR) and complete responses (CR) in rare cases (e.g., residual lesions in tumor 
types, such as germ cell tumors, where known residual benign tumors can remain).
The cytological confirmation of the neoplastic origin of any effusion that appears or 
worsens during treatment when the measurable tumor has met criteria for response or 
stable disease is mandatory to differentiate between response or stable disease (an 
effusion may be a side effect of the treatment) and progressive disease.
FDG-PET:  While FDG-PET response assessments need additional study, it is 
sometimes reasonable to incorporate the use of FDG-PET scanning to complement CT 
scanning in assessment of progression (particularly possible 'new' disease).  New lesions 
on the basis of FDG-PET imaging can be identified according to the following 
algorithm: 
a. Negative FDG-PET at baseline, with a positive FDG-PET at follow-up is a sign 
of PD based on a new lesion.
b. No FDG-PET at baseline and a positive FDG-PET at follow-up:  If the positive 
FDG-PET at follow-up corresponds to a new site of disease confirmed by CT, 
this is PD.  If the positive FDG-PET at follow-up is not confirmed as a new site 
of disease on CT, additional follow-up CT  scans are needed to determine if there 
is truly progression occurring at that site (if so, the date of PD will be the date of 
the initial abnormal FDG-PET scan).  If the positive FDG-PET at follow-up 
corresponds to a pre-existing site of disease on CT that is not progressing on the 
basis of the anatomic images, this is not PD.
c. FDG-PET may be used to upgrade a response to a CR in a manner similar to a 
biopsy in cases where a residual radiographic abnormality is thought to represent 
fibrosis or scarring.  The use of FDG-PET in this circumstance should be 
prospectively described in the protocol and supported by disease-specific 
medical literature for the indication.  However, it must be acknowledged that 
both approaches may lead to false positive CR due to limitations of FDG-PET 
and biopsy resolution/sensitivity.

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Note:  A ‘positive’ FDG-PET scan lesion means one which is FDG avid with an uptake 
greater than twice that of the surrounding tissue on the attenuation corrected image.
11.1.4 Response Criteria
11.1.4.1
Evaluation of Target Lesions
Complete Response (CR): Disappearance of all target lesions.  Any pathological 
lymph nodes (whether target or non-target) must have 
reduction in short axis to <10 mm.
Partial Response (PR):
At least a 30% decrease in the sum of the diameters of 
target lesions, taking as reference the baseline sum 
diameters.
Progressive Disease (PD): At least a 20% increase in the sum of the diameters of 
target lesions, taking as reference the smallest sum on 
study (this includes the baseline sum if that is the smallest 
on study).  In addition to the relative increase of 20%, the 
sum must also demonstrate an absolute increase of at 
least 5 mm.  (Note:  the appearance of one or more new 
lesions is also considered progressions).
Stable Disease (SD):
Neither sufficient shrinkage to qualify for PR nor 
sufficient increase to qualify for PD, taking as reference 
the smallest sum diameters while on study.
11.1.4.2
Evaluation of Non-Target Lesions
Complete Response (CR): Disappearance of all non-target lesions and normalization 
of tumor marker level.  All lymph nodes must be non-
pathological in size (<10 mm short axis).
Note:  If tumor markers are initially above the upper normal limit, they must 
normalize for a patient to be considered in complete 
clinical response.
Non-CR/Non-PD:
Persistence of one or more non-target lesion(s) and/or 
maintenance of tumor marker level above the normal 
limits.
Progressive Disease (PD): Appearance of one or more new lesions and/or 
unequivocal progression of existing non-target lesions.  
Unequivocal progression should not normally trump 
target lesion status.  It must be representative of overall 
disease status change, not a single lesion increase.    

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Although a clear progression of “non-target” lesions only is exceptional, the opinion 
of the treating physician should prevail in such circumstances, and the progression 
status should be confirmed at a later time by the review panel (or Principal 
Investigator).
11.1.4.3
Evaluation of Best Overall Response
The best overall response is the best response recorded from the start of the 
treatment until disease progression/recurrence (taking as reference for progressive 
disease the smallest measurements recorded since the treatment started).  The 
patient's best response assignment will depend on the achievement of both 
measurement and confirmation criteria.
For Patients with Measurable Disease (i.e., Target Disease)
Target 
Lesions
Non-Target 
Lesions
New 
Lesions
Overall 
Response
Best Overall Response when 
Confirmation is Required*
CR
CR
No
CR
≥4 wks. Confirmation**
CR
Non-CR/Non-PD
No
PR
CR
Not evaluated
No
PR
PR
Non-CR/Non-
PD/not evaluated
No
PR
≥4 wks. Confirmation**
SD
Non-CR/Non-
PD/not evaluated
No
SD
Documented at least once ≥4 
wks. from baseline**
PD
Any
Yes or No
PD
Any
PD***
Yes or No
PD
Any
Any
Yes
PD
no prior SD, PR or CR
*
See RECIST 1.1 manuscript for further details on what is evidence of a new lesion.
**
Only for non-randomized trials with response as primary endpoint.
***
In exceptional circumstances, unequivocal progression in non-target lesions may be accepted as 
disease progression.
Note:
Patients with a global deterioration of health status requiring discontinuation of treatment 
without objective evidence of disease progression at that time should be reported as 
“symptomatic deterioration.”  Every effort should be made to document the objective 
progression even after discontinuation of treatment.
For Patients with Non-Measurable Disease (i.e., Non-Target Disease)
Non-Target Lesions
New Lesions
Overall Response
CR
No
CR
Non-CR/non-PD
No
Non-CR/non-PD*
Not all evaluated
No
not evaluated
Unequivocal PD
Yes or No
PD
Any
Yes
PD

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*  ‘Non-CR/non-PD’ is preferred over ‘stable disease’ for non-target disease since SD is increasingly 
used as an endpoint for assessment of efficacy in some trials so to assign this category when no lesions 
can be measured is not advised
11.1.5 Duration of Response
Duration of overall response:  The duration of overall response is measured from the 
time measurement criteria are met for CR or PR (whichever is first recorded) until the 
first date that recurrent or progressive disease is objectively documented (taking as 
reference for progressive disease the smallest measurements recorded since the 
treatment started).
The duration of overall CR is measured from the time measurement criteria are first met 
for CR until the first date that progressive disease is objectively documented.
Duration of stable disease:  Stable disease is measured from the start of the treatment 
until the criteria for progression are met, taking as reference the smallest measurements 
recorded since the treatment started, including the baseline measurements. 
12. DATA REPORTING / REGULATORY REQUIREMENTS
Adverse event lists, guidelines, and instructions for AE reporting can be found in Section 
7.0 (Adverse Events: List and Reporting Requirements).
12.1
Data Reporting
12.1.1 Method
This study will be monitored by the Clinical Trials Monitoring Service (CTMS).  Data 
will be collected in the Center for Cancer Research C3D database and will be 
transmitted to CTMS electronically at least once every 2 weeks.
Note: All adverse events that have occurred on the study, including those reported 
through CTEP-AERS, must be reported via the monitoring method identified above.
12.1.2 Responsibility for Data Submission
N/A
12.2
CTEP Multicenter Guideline
N/A
12.3
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 

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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 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. 

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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 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:  E-mail: 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.
12.4
Genomic Data Sharing Plan
The NIH Genomic Data Sharing (GDS) Policy does not apply to this protocol as we will 
be performing exploratory genetic analysis of no more than 80 patient samples; 
therefore, this study does not meet GDS criteria, and a GDS plan is not warranted.
13. STATISTICAL CONSIDERATIONS
13.1
Study Design
The Phase I portion of the study will use a standard 3+ 3 design without intra-patient dose 
escalation.  The MTD or recommended Phase II dose is the dose level at which no more than 1 
of 6 patients experience DLT during the first cycle of treatment, and the dose below that at 
which at least 2 (of ≤ 6) patients have DLT as a result of the drug.
The Phase II portion will follow the Simon two-stage minimax design and will consist of three 
separately evaluated cohorts: patients with NSCLC, colon, and granulosa cell ovarian cancer.  
For NSCLC and colon, the maximum cohort size would be set at 25 evaluable patients.  If at 
least 5 responses (at least 20%) are observed among the 25 evaluable patients, this regimen 
would be considered worthy of further testing in this disease.  If no more than 1 response (no 
more than 6%) is observed among the initial 16 patients, the cohort would be terminated early 
and declared negative.  This design yields at least 90% power to detect a true response rate of at 

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least 30%.  It yields at least .90 probability of a negative result if the true response rate is no 
more than 10%, with at least .52 probability of early negative stopping.  For granulosa cell 
ovarian cancer, maximum cohort size would be set at 15 evaluable patients.  If at least 4 
responses (at least 27%) are observed among the 15 evaluable patients, this regimen would be 
considered worthy of further testing in this disease.  If no responses are observed among the 
initial 8 patients, the cohort would be terminated early and declared negative.  This design 
yields at least 90% power to detect a true response rate of at least 40%.  It yields at least .945 
probability of a negative result if the true response rate is no more than 10%, with at least .43 
probability of early negative stopping.
13.2
Sample Size/Accrual Rate
The Phase I portion of the study is designed to have 8 dose escalation cohorts, with a standard 
design using 3 patients per cohort, unless DLT is noted, in which point up to 6 patients may be 
enrolled in a cohort.  In addition, up to an additional 15 patients will be enrolled at the MTD to 
further define the dose and evaluate PD studies at this dose level; with that number, and a tumor 
biopsy QA criteria failure rate of 50% with respect to paired (pre- and post-dose) biopsies, we 
have an 85% likelihood of having at least 6 usable PD samples, and 95% likelihood of having at 
least 5 usable samples.  Biopsy tissue quality will be monitored and accrual will stop once we 
have obtained 6 usable paired samples.  To allow for a small number of patients who may not 
be evaluable, the accrual ceiling for this trial is set at 65 (55 patients plus 10 healthy 
volunteers).  
The Phase II portion of the study is designed to have up to 65 evaluable patients, and to allow 
for a small number of patients who may not be evaluable, the accrual ceiling for the Phase II 
portion is set at 75 patients.  It is anticipated that 2-3 patients per month may be enrolled onto 
this study.  Depending on whether the Phase I expansion and Phase II cohorts accrue to 
completion, it is expected that approximately 4 years will be required to accrue the number of 
patients necessary to complete the entire trial. 
14. HUMAN SUBJECTS PROTECTIONS
14.1
Justification for Subject Selection
This study will be open to all individuals regardless of gender, ethnicity, or race, provided that 
the aforementioned inclusion and exclusion criteria are met.  Patients for this study will be 
recruited through internal referral, our physician referral base, and through various cancer 
information hotlines (i.e., Clinical Studies Support Center, 1-800-4Cancer).  To date, there is no 
information that suggests that differences in drug metabolism or effect on tumor would be 
expected in one ethnic group compared to another.  Efforts will be made to extend accrual to 
each representative population, but a balance must be struck between participant safety 
considerations and limitations on the number of individuals exposed to potentially ineffective 
treatments on the one hand and the need to explore racial/ethnic aspects of clinical research on 
the other hand.  If differences in outcome that correlate to ethnic identity are noted, a follow-up 
study may be written to investigate those differences more fully.

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Due to lack of knowledge of the effects of TRC102 on the fetus or infants, as well as the 
possibility of teratogenic effects, pregnant and nursing women will be excluded from this trial.  
Patients with unstable or serious medical conditions are excluded due to the possibility that 
TRC102 may worsen their condition and the likelihood that the underlying condition may 
obscure the attribution of adverse events to TRC102.  HIV-positive patients on combination 
antiretroviral therapy are excluded from the study because of possible PK interactions with 
TRC102.
14.1.1 Participation of Children
This study includes patients 18 years of age and older.  Because insufficient dosing or 
adverse event data are currently available on the use of TRC102 in patients <18 years of 
age, children are excluded from this study, but may be eligible for future pediatric trials.  
Studies will be performed in patients <18 years of age when it is appropriate to do so.
14.2
Evaluation of Benefits and Risks/Discomforts
There may or may not be any clinical benefit to a patient from participation in this trial.  Their 
participation will benefit future cancer patients.  Potential risks include the possible occurrence 
of any of a range of side effects that are listed in the consent document.  The procedure for 
protecting against or minimizing risks will be to medically evaluate patients as described in 
Sections 5 and 6.  Although no compensation is available, any injury will be fully evaluated and 
treated in keeping with the benefits or care to which participants are entitled under applicable 
regulations.
14.3
Consent and Assent Process and Documentation
An associate or principal investigator on the trial will inform patients of the purpose, 
alternatives, drug administration plan, research objectives, and follow-up of this trial.  The 
patient will be provided an IRB-approved consent for review and signature and his/her 
questions will be answered.  After a decision is made to enroll into the study, a signature will be 
obtained from the patient.  The original signed consent goes to Medical Records; a copy will be 
placed in the research record.
All patients must have a signed informed consent form and an on-study (confirmation of 
eligibility) form filled out and signed by a participating investigator before entering on study.
Please note: As of Amendment R (5/12/18) healthy adult volunteers will no longer be recruited 
to provide blood for this study. Healthy volunteers 18 years and older will give informed 
consent prior to undergoing blood collection.  The study investigator will emphasize that there 
will be no benefit to them for entering into this study.  Volunteers will be apprised of the risks 
of the study and be informed that the study is for research purposes only.  Volunteers will be 
encouraged to ask questions and will sign the consent only if they choose to enter the study.  
The volunteer will sign the consent with a nurse or non-study participant as a witness while at 
NIH, prior to undergoing any study procedure.

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14.3.1 Participation of subjects unable to give consent
Adults unable to give consent are excluded from enrolling in the protocol.  However, re-
consent may be necessary and there is a possibility, though unlikely, that subjects could 
become decisionally impaired.  For this reason and because there is a prospect of direct 
benefit from research participation (i.e., long-term stabilization and/or improvement in 
the pain and physical impairment cause by cancer), all subjects ≥ age 18 at the NCI only 
will be offered the opportunity to fill in their wishes for research and care, and assign a 
substitute decision maker on the “NIH Advance Directive for Health Care and Medical 
Research Participation” form so that another person can make decisions about their 
medical care in the event that they become incapacitated or cognitively impaired during 
the course of the study. Note: The PI or AI will contact the NIH Ability to Consent 
Assessment Team for evaluation.  For those subjects that become incapacitated and do 
not have pre-determined substitute decision maker, the procedures described in OHSRP 
Policy 403 for appointing a surrogate decision maker for adult subjects who are (a) 
decisionally impaired, and (b) who do not have a legal guardian or durable power of 
attorney, will be followed.
14.3.2 Informed Consent of non-English speaking subjects
If there is an unexpected enrollment of a research participant for whom there is no 
translated extant IRB approved consent document, the principal investigator and/or 
those authorized to obtain informed consent will use the Short Form Oral Consent 
Process as described in MAS Policy M77-2, OHSRP SOP 12, 45 CFR 46.117 (b) (2).  
The summary that will be used is the English version of the extant IRB approved 
consent document.  Signed copies of both the English version of the consent and the 
translated short form will be given to the subject or their legally authorized 
representative, and the signed original will be filed in the medical record.
Unless the PI is fluent in the prospective subject’s language, an interpreter will be 
present to facilitate the conversation (using the Short Form process).  Preferably, 
someone who is independent of the subject (i.e., not a family member) will assist in 
presenting information and obtaining consent.  Whenever possible, interpreters will be 
provided copies of the relevant consent documents well before the consent conversation 
with the subject (24 to 48 hours if possible).
14.3.3 As of Amendment R (5/12/2018), patients who have come off study but are still able to 
consent may be re-consented to the most recent protocol consent document if they wish 
to allow tumor tissue collected while on the trial to be used for the additional genomic 
analyses added in Amendment R (Section 9.3).
14.4
Procedure for Protecting Against or Minimizing Any Potential Risks

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All care will be taken to minimize side effects, but they can be unpredictable in nature and 
severity.  This study may involve risks to patients, which are currently unforeseeable.  All 
patients will be monitored for side effects from taking study medication.  
This study requires up to 3 CT-guided research tumor biopsies and up to 7 CT scans per year.  
These procedures confer radiation exposure at an effective dose of up to 10.1 rem per year.  
This dose is above NIH RSC guidelines of 5.0 rem per year in adults.
14.5
Patient Advocate
The patients’ rights representative is available to patients receiving treatment on this protocol at 
the NIH Clinical Center at (301) 496-2626 in Building 10 of the Clinical Research Center, 
Room 1-3521, on the Bethesda NIH campus.  Patients will be informed that they can contact the 
study PI or RN at any time with questions about their medical care, and that the patients’ rights 
representative is also available to answer non-medical questions about the study.

CTEP #9483
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REFERENCES
1.
TRC102 Investigator Brochure v2.0, Sept 26, 2012.
2.
Yan, L., et al., Combined Treatment with Temozolomide and Methoxyamine: Blocking 
Apurininc/Pyrimidinic Site Repair Coupled with Targeting Topoisomerase IIÎ. Clinical 
Cancer Research, 2007. 13(5): p. 1532-1539.
3.
Liu, L., Y. Nakatsuru, and S.L. Gerson, Base Excision Repair as a Therapeutic Target in 
Colon Cancer. Clinical Cancer Research, 2002. 8(9): p. 2985-2991.
4.
Liu, L., et al., Blockage of abasic site repair enhances antitumor efficacy of 1,3-bis-(2-
chloroethyl)-1-nitrosourea in colon tumor xenografts. Molecular Cancer Therapeutics, 
2003. 2(10): p. 1061-1066.
5.
Weiss, G., et al. Final results from a phase I study of oral TRC102 (methoxyamine HCl), 
an inhibitor of base-excision repair, to potentiate the activity of pemetrexed in patients 
with refractory cancer
in ASCO. 2010.
6.
Anthony, S., et al. A phase I study of daily oral TRC102 (methoxyamine) to enhance the 
therapeutic effects of pemetrexed in patients with advanced refractory cancer. in ASCO. 
2009.
7.
Schumer, S.T. and S.A. Cannistra, Granulosa cell tumor of the ovary. J Clin Oncol, 
2003. 21(6): p. 1180-9.
8.
Kaye, S.B. and E. Davies, Cyclophosphamide, adriamycin, and cis-platinum for the 
treatment of advanced granulosa cell tumor, using serum estradiol as a tumor marker. 
Gynecol Oncol, 1986. 24(2): p. 261-4.
9.
Rey, R.A., et al., Antimullerian hormone as a serum marker of granulosa cell tumorsof 
the ovary: comparative study with serum alpha-inhibin and estradiol. Am J Obstet 
Gynecol, 1996. 174(3): p. 958-65.
10.
Rodgers, K.E., et al., Follicle regulatory protein: a novel marker for granulosa cell 
cancer patients. Gynecol Oncol, 1990. 37(3): p. 381-7.
11.
Silverman, L.A. and S.E. Gitelman, Immunoreactive inhibin, mullerian inhibitory 
substance, and activin as biochemical markers for juvenile granulosa cell tumors. J 
Pediatr, 1996. 129(6): p. 918-21.
12.
Koukourakis, G.V., et al., Granulosa cell tumor of the ovary: tumor review. Integr 
Cancer Ther, 2008. 7(3): p. 204-15.
13.
Fujita, F., et al., A recurrent granulosa cell tumor of the ovary 25 years after the initial 
diagnosis: A case report. Int J Surg Case Rep, 2015. 12: p. 7-10.
14.
Ayhan, A., et al., Prognostic factors in adult granulosa cell tumors of the ovary: a 
retrospective analysis of 80 cases. J Gynecol Oncol, 2009. 20(3): p. 158-63.
15.
Bjorkholm, E. and C. Silfversward, Prognostic factors in granulosa-cell tumors. 
Gynecol Oncol, 1981. 11(3): p. 261-74.
16.
Wolf, J.K., et al., Radiation treatment of advanced or recurrent granulosa cell tumor of 
the ovary. Gynecol Oncol, 1999. 73(1): p. 35-41.
17.
Zambetti, M., et al., cis-platinum/vinblastine/bleomycin combination chemotherapy in 
advanced or recurrent granulosa cell tumors of the ovary. Gynecol Oncol, 1990. 36(3): 
p. 317-20.
18.
Gershenson, D.M., et al., Treatment of poor-prognosis sex cord-stromal tumors of the 
ovary with the combination of bleomycin, etoposide, and cisplatin. Obstet Gynecol, 

CTEP #9483
Clinical Center #: 13-C-0118
64
1996. 87(4): p. 527-31.
19.
Tresukosol, D., et al., Recurrent ovarian granulosa cell tumor: a case report of a 
dramatic response to Taxol. Int J Gynecol Cancer, 1995. 5(2): p. 156-159.
20.
Schwartz, P.E., et al., Tamoxifen-induced increase in cytosol progestin receptor levels in 
a case of metastatic endometrial cancer. Gynecol Oncol, 1983. 16(1): p. 41-8.
21.
DeSantis, C.E., et al., Cancer treatment and survivorship statistics, 2014. CA Cancer J 
Clin, 2014. 64(4): p. 252-71.
22.
Voena, C. and R. Chiarle, Advances in cancer immunology and cancer immunotherapy. 
Discov Med, 2016. 21(114): p. 125-33.
23.
Stratton, M.R., P.J. Campbell, and P.A. Futreal, The cancer genome. Nature, 2009. 
458(7239): p. 719-24.
24.
Fearon, E.R., Molecular genetics of colorectal cancer. Annu Rev Pathol, 2011. 6: p. 
479-507.
25.
Martini, M., et al., Targeted therapies: how personal should we go? Nature Reviews 
Clinical Oncology, 2012. 9(2): p. 87-97.
26.
Torkamani, A., G. Verkhivker, and N.J. Schork, Cancer driver mutations in protein 
kinase genes. Cancer Lett, 2009. 281(2): p. 117-27.
27.
Llosa, N.J., et al., The Vigorous Immune Microenvironment of Microsatellite Instable 
Colon Cancer Is Balanced by Multiple Counter-Inhibitory Checkpoints. Cancer 
Discovery, 2015. 5(1): p. 43-51.
28.
Fernandez-Capetillo, O., et al., H2AX: the histone guardian of the genome. DNA Repair 
(Amst), 2004. 3(8-9): p. 959-67.
29.
Rogakou, E.P., et al., DNA double-stranded breaks induce histone H2AX 
phosphorylation on serine 139. J Biol Chem, 1998. 273(10): p. 5858-68.
30.
Paull, T.T., et al., A critical role for histone H2AX in recruitment of repair factors to 
nuclear foci after DNA damage. Curr Biol, 2000. 10(15): p. 886-95.
31.
Bassing, C.H. and F.W. Alt, H2AX may function as an anchor to hold broken 
chromosomal DNA ends in close proximity. Cell Cycle, 2004. 3(2): p. 149-53.
32.
Redon, C., et al., Histone H2A variants H2AX and H2AZ. Current Opinion in Genetics 
& Development, 2002. 12(2): p. 162-169.
33.
Celeste, A., et al., Genomic instability in mice lacking histone H2AX. Science, 2002. 
296(5569): p. 922-7.
34.
Kinders, R.J., et al., Development of a Validated Immunofluorescence Assay for Î³H2AX 
as a Pharmacodynamic Marker of Topoisomerase I Inhibitor Activity. Clinical Cancer 
Research, 2010. 16(22): p. 5447-5457.
35.
Engelward, B.P., et al., Repair-deficient 3-methyladenine DNA glycosylase homozygous 
mutant mouse cells have increased sensitivity to alkylation-induced chromosome 
damage and cell killing. EMBO J, 1996. 15(4): p. 945-52.
36.
Agnihotri, S., et al., Alkylpurine-DNA-N-glycosylase confers resistance to temozolomide 
in xenograft models of glioblastoma multiforme and is associated with poor survival in 
patients. J Clin Invest, 2012. 122(1): p. 253-66.
37.
Germano, G., et al., Inactivation of DNA repair triggers neoantigen generation and 
impairs tumour growth. Nature, 2017. 552(7683): p. 116-120.
38.
Gordon, M., et al., A phase 1 study of TRC102, an inhibitor of base excision repair, and 
pemetrexed in patients with advanced solid tumors. Investigational New Drugs, 2013. 
31(3): p. 714-723.

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39.
Evelo, C.T.A., et al., Two Mechanisms for Toxic Effects of Hydroxylamines in Human 
Erythrocytes: Involvement of Free Radicals and Risk of Potentiation. Blood Cells, 
Molecules, and Diseases, 1998. 24(3): p. 280-295.
40.
Dressler, L.G. and S.F. Terry, How will GINA influence participation in 
pharmacogenomics research and clinical testing? Clin Pharmacol Ther, 2009. 86(5): p. 
472-5.
41.
Kass, N.E., et al., Medical privacy and the disclosure of personal medical information: 
the beliefs and experiences of those with genetic and other clinical conditions. Am J 
Med Genet A, 2004. 128A(3): p. 261-70.
42.
Tabor, H.K., et al., Genomics really gets personal: how exome and whole genome 
sequencing challenge the ethical framework of human genetics research. Am J Med 
Genet A, 2011. 155A(12): p. 2916-24.
43.
Berg, J.S., M.J. Khoury, and J.P. Evans, Deploying whole genome sequencing in clinical 
practice and public health: meeting the challenge one bin at a time. Genet Med, 2011. 
13(6): p. 499-504.
44.
Caulfield, T., et al., Research ethics recommendations for whole-genome research: 
consensus statement. PLoS Biol, 2008. 6(3): p. e73.
45.
Kalia, S.S., et al., Recommendations for reporting of secondary findings in clinical 
exome and genome sequencing, 2016 update (ACMG SF v2.0): a policy statement of the 
American College of Medical Genetics and Genomics. Genet Med, 2017. 19(2): p. 249-
255.

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

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APPENDIX B: INFORMATION ON POSSIBLE DRUG INTERACTIONS
Information on Possible Interactions with Other Agents for Patients and Their Caregivers 
and Non-Study Healthcare Team
The patient ____________________________ is enrolled on a clinical trial using the 
experimental agent TRC102 and temozolomide.  This clinical trial is sponsored by the National 
Cancer Institute.  This form is addressed to the patient, but includes important information for 
others who care for this patient. 
TRC102 and temozolomide may interact with other drugs that are processed by your body.  
Because of this, it is very important to tell your study doctors about all of your medicine before 
you start this study.  It is also very important to tell them if you stop taking any regular 
medicine, or if you start taking a new medicine while you take part in this study.  When you talk 
about your medicine with your study doctor, include medicine you buy without a prescription at 
the drug store (over-the-counter remedy), or herbal supplements. 
Many health care prescribers can write prescriptions.  You must also tell your other prescribers 
(doctors, physicians’ assistants, or nurse practitioners) that you are taking part in a clinical trial. 

CTEP #9483
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APPENDIX C: PATIENT’S MEDICATION DIARY
INSTRUCTIONS
1. Complete one form for each cycle of treatment.
2. Swallow study drugs whole with a full glass of water either 1 hour before or 2 hours after a 
meal.  Do not chew or open the capsules.  If capsule is broken and the powder of the 
capsules gets on skin, wash the exposed area with as much water as necessary.  Inform 
investigator or nurse if that occurs.
3. Record the date and time you took the drugs. 
4. If you have any comments or notice any side effects, please record them in the Comments 
column.
5. Please bring this form and your bottle of drugs when you return for your appointment.  
6. In case of errors, please place a single slash mark through the error and initial it.  Please do 
not white out any error or scribble it out with ink.  Please do not write the correct 
information directly over the error, but on a separate line next to the error.

CTEP #9483
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APPENDIX C: PATIENT’S MEDICATION DIARY
Today’s Date _____________________Cycle #______  TRC102 Dose_______mg TMZ Dose_______mg
Patient Name_____________________(initials acceptable)
 Patient Study ID __________________
Day
Date
Time of dose
Number of Capsules Taken
Comments
AM
PM
TRC102
Temozolomide
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
Patient’s signature: ___________________________________________________________________

APPENDIX D: PK/PD COLLECTION WORKSHEETS
PHASE I RESEARCH URINE SAMPLE COLLECTION SHEET: CYCLE 1 DAY 1
CTEP Protocol 
P9483
Site:
Research Nurse: 
Dose level:
TRC102 Dose: 
Phone:                  Pager: 
Patient ID:
TMZ Dose:
Page ----------------------------for 
sample pick-up 
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
Phone: 301-768-2749
PLEASE ENTER DATE, TIME, and VOLUME OF EACH VOID FOR 24 HOURS POST DRUG ADMINISTRATION
KEEP SAMPLES REFRIGERATED UNTIL PICKED UP
Date
Time
Volume of 
Void
Record comments (i.e., if collection missed), and sign each time 
you collect a sample
Pre-Drug Administration (SAMPLE #100)
Administer TRC102 and Temozolomide, Time:
DAY 1 Start 24 hour urine collection immediately after study drug administration, keep on ice.  Record volume of each void on this sheet 
and record total 24 hour volume at the end of the collection.  At the end of 24 hours, retain 10 mL aliquot only, place on ice and discard 
remainder of urine.  Charge urine collection container with acid as specified in Appendix E.
Date
Time 
(start of 24 
hour 
collection 
time)
Volume of 
Void
24-hour urine collection
Record comments (i.e., if collection missed), and sign each time 
you collect a sample
Volume 1
Volume 2
Volume 3
Volume 4
Volume 5
Volume 6
Volume 7
Volume 8
Volume 9
Total (24-hour 
volume)
10 mL aliquot obtained
(please check box)
Sample #101 (10 mL retained from total volume)
End 24-hour Urine Collection Time:
Please Send a Copy of this sheet with the last specimen

CTEP #9483
Clinical Center #: 13-C-0118
71
Date: 
PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 1
CTEP Protocol P9483
Ht: 
Page -------------------------for 
Sample Pick-up
Lab phone: 301-451-1169
Research Nurse:  
Dose level:
TRC102 Dose:
Wt:  
Phone:                  Pager:  
Patient ID:
TMZ Dose: 
BSA:  
PI: A. P. Chen, MD      
*****ONLY PK SAMPLES GO ON ICE******
Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal 
Time
Actual 
Time
Record comments (i.e., if collection missed), and 
sign each time you collect a sample
Day 1
Prior to drug 
administration
PK 200 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
-----
 
Day 1
Prior to drug 
administration
PD 400 4mL NaHep
Label tube: drug, draw date and time 
Administer TMZ and TRC102, Time:
Day 1
1 hour post 
dose
PK 201 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
Day 1
2 hours post 
dose
PK 202 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
 
 
Day 1
3 hours post 
dose
PK 203 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
 
Day 1
4 hours post 
dose
PK 204 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
 
 
 
Continued over

CTEP #9483
Clinical Center #: 13-C-0118
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Day
Time
Instructions
Ideal 
Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 1
8 hours post 
dose
PK 205 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
 
 
Day 1
8 hours post 
dose
PD 401  4 mL NaHep
Label tube: drug, draw date and time 
Day 1
12 hours post 
dose
PK 206 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
Day 2
24 hours post 
dose
PK 207 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE

CTEP #9483
Clinical Center #: 13-C-0118
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Date: 
PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 5
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level:
TRC102 Dose:
Wt:  
Phone:                 Pager: 
Patient ID:
TMZ Dose: 
BSA:  
Page -------------------------
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
*****ONLY PK SAMPLES GO ON ICE******
Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal 
Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 5
Prior to dosing
PK 208 EDTA (lavender top) 2 mL
Label tube: drug, draw date and time
PLACE ON ICE
Day 5
Prior to dosing
PD 402 4 mL NaHep
Label tube: drug, draw date and time 

CTEP #9483
Clinical Center #: 13-C-0118
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Date: 
PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 2 Day 1 
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page ______________ 
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
*****ONLY PK SAMPLES GO ON ICE******
 Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 1
Prior to 
dosing
PD 403 4 mL NaHep
Label tube: drug, draw date and time 
Date: 
PHASE I PK/PD SAMPLE COLLECTION SHEET: Day 1 each cycle/as 
determined by PI
Cycle number: ______
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                 Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page                                     
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
*****ONLY PK SAMPLES GO ON ICE******
      Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 1
Prior to dosing
PD 40X 4 mL NaHep
Label tube: drug, draw date and time 

CTEP #9483
Clinical Center #: 13-C-0118
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Date: 
PHASE II PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 4 or 5 (day 
of biopsy) 
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page ______________ 
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
 Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 4/5
Prior to 
dosing
401 10 mL Streck
Label tube: drug, draw date and time 
Date: 
PHASE II PK/PD SAMPLE COLLECTION SHEET: Day 1 (±1 day) each 
Date: 
PHASE II PD SAMPLE COLLECTION SHEET: Cycle 1 Day 1
CTEP Protocol P9483
Ht: 
Research Nurse:  
Dose level:
TRC102 Dose:
Wt:  
Phone:                  Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page -------------------------
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal 
Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 1
Prior to drug 
administration
400 10 mL Streck
Label tube: drug, draw date and time 

CTEP #9483
Clinical Center #: 13-C-0118
76
cycle/as determined by PI
Cycle number: ______
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                 Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page                                     
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
      Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day 1
Prior to dosing
40__ 10 mL Streck
Label tube: drug, draw date and time 
Date: 
PHASE II PK/PD SAMPLE COLLECTION SHEET: Day of restaging follow-
up or progression biopsy 
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page ______________ 
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      
 Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Ideal Time
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Day X
Prior to 
dosing
40__ 10 mL Streck
Label tube: drug, draw date and time 
Date: 
PHASE II PK/PD SAMPLE COLLECTION SHEET: Optional blood for 
germline WES, collected any time while on trial
CTEP Protocol P9483
Ht: 
Research Nurse: 
Dose level: 
TRC102 Dose:
Wt:  
Phone:                Pager:  
Patient ID:
TMZ Dose: 
BSA:  
Page ______________ 
for Sample Pick-up
Lab phone: 301-451-1169
PI: A. P. Chen, MD      

CTEP #9483
Clinical Center #: 13-C-0118
77
 Phone: 301-768-2749
 PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION
Day
Time
Instructions
Actual 
Day
Actual 
Time
Record comments (i.e., if collection missed), 
and sign each time you collect a sample
Any day
Any time
001 10 mL Streck
Label tube: drug, draw date and time 

APPENDIX E: PK SAMPLE HANDLING
PLASMA
The 2-mL blood sample is immediately placed on ice and centrifuged within 30 minutes of 
collection to separate the plasma.  The plasma is divided between two tubes, one of which is 
labeled "A" (which is frozen and stored at -70°C until analyzed) and the other is labeled "B," 
which contains 25 µL of commercially available 1N hydrochloric acid (HCl): sample "B" is then 
mixed by vortex action, frozen, and stored at -70°C until analysis.
URINE
A pre-dose void is collected, its volume measured and recorded, and then a 10-mL aliquot of 
urine is removed to which 0.4 mL of 1N hydrochloric acid is added, mixed, frozen and stored at
 -70°C until analysis.
The plastic container used for the collection/storage of all voids over the 24-hr collection period 
is first charged with 25 mL of commercially available 1N HCl prior to the introduction of any 
patient urine.  An additional 25 mL of 1N HCl is added for each 500 mL of urine collected.  The 
contents of the container are mixed by swirling.  
Each void volume is first measured before adding to the container.  
At the conclusion of collecting all voids, the contents of the container are mixed by swirling, and 
then a 10-mL aliquot is removed, frozen, and stored at -70°C until analysis. 
Send plasma and urine samples to:
Tracy W. Webb
Office of the Associate Director/DTP/NCI
FNLCR
Boyles Street
Building 1047, Room 8
Frederick, MD 21702
301-846-7402
webbtw@mail.nih.gov

APPENDIX F: ERYTHROCYTE ANALYSIS 
Please note: As of Amendment R (5/12/18), we will no longer perform this analysis of the 
mechanisms of extravascular hemolysis.
Blood will be collected for the analysis below in collaboration with the hematology attending 
consult team.  These studies are optional.  Blood volumes may be changed at the PI/consult 
team’s discretion.  Additional studies (Folate, Vitamin B12, Erythropoietin, TSH) may be done 
as clinically indicated based on baseline Hgb values.  
Blood collection tubes, volumes, and service for each test can be found at the Dept. of Transfusion 
Medicine’s Test Guide:  http://cclnprod.cc.nih.gov/dlm/testguide.nsf/
When patients develop Hgb level decrement of more than 2 g/dL of baseline during cycle 
follow-up, contact the hematology consult team for official consultation (Dr. Kazusa Ishii, 
kazusa.ishii@nih.gov; 301-496-5093); the hematology attending consult team will develop a 
reference guide/SOP.  The hematology consult team will order necessary labs, review the 
peripheral blood smear, and obtain systematic relevant hematological history at the time of 
consultation.
Laboratory testing/ timing
1) Baseline (within 72 hours prior to enrollment)

CBC with WBC differential

Reticulocyte count

Peripheral blood smear evaluation

LDH

LFTs

Haptoglobin

Ferritin, iron study

DAT (Direct Coombs test)

Renal function

Urinalysis with microscopy, urine urobilinogen

G6PD

Spleen size evaluation (if CT is done recently per protocol, there is no need to repeat.  If 
not, then US spleen)

Osmotic fragility test

RBC enzyme evaluation

Methemoglobin, venous
2) C1D7 +/- 2 days

CBC with diff

Reticulocyte count

Peripheral blood smear

LDH

CTEP #9483
Clinical Center #: 13-C-0118
80

LFTs

Haptoglobin

DAT

Ferritin, iron study

Urinalysis with microscopy, urine urobilinogen

RBC enzyme evaluation 

Osmotic fragility test 

Methemoglobin, venous 
3) C1D14 +/- 2 days, C1D21 +/- 2 days

CBC with diff

Reticulocyte count

Peripheral blood smear

LDH

LFTs

Haptoglobin

DAT

Urinalysis with microscopy, urine urobilinogen