Clinical trial NCT01441115
Conditions: Cancer Neoplasm Metastasis Radiation Oncology Neoplasm Metastasis
Keywords: Metastasis Cancer Radiation Therapy Locally Advanced Palliation Metastatic Cancer
Sponsor: National Cancer Institute (NCI)
Start date: 2011-09-06; Completion date: 2013-04-24
Enrollment: 2 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
Background:

\- ECI301 is a drug that may help make cancer cells more visible to the immune system after radiation. The drug may also help the immune system destroy the cancer at sites that have not received radiation therapy. Researchers want to study ECI301 in people with advanced cancer or cancer that has spread in the body (metastatic).

Objectives:

\- To test ECI301 with radiation therapy for advanced or metastatic cancer.

Eligibility:

\- People at least 18 years of age with either metastatic or advanced cancer that may benefit from radiation therapy.

Design:

* Participants will be screened with a medical history and physical exam. They will also have blood and urine tests, and imaging studies.
* All participants will have radiation therapy 5 days a week for 2 weeks.
* They will have different doses of ECI301 to test its safety and effectiveness. ECI301 will be given in a vein during the second week of radiation therapy. Frequent blood tests and imaging studies will monitor the treatment.
* After participants have ECI301, tumor samples may be taken from the site that had radiation and another site that did not have radiation.
* Follow-up visits will include blood tests and imaging studies.

Detailed description:
Background:

* Patients with metastatic or locally advanced cancer frequently require palliative radiotherapy to relieve symptoms; however, progression of disease is frequent in patients with extended survival
* Radiation results in tumor cell death which can result in increased dendritic cell activation and trafficking
* ECI301 is a derivative of Macrophage Inflammatory Protein-1 alpha, a 70 amino acid chemokine that is a ligand for C-C chemokine receptor type 1 (CCR1) and C-C chemokine receptor type 5 (CCR5), the chemokine receptors of immature dendritic cells.
* ECI301 has been shown to enhance the effect of radiotherapy in animal models.

Objectives:

* The primary objective is to determine the maximum tolerated dose (MTD) of ECI301 delivered in combination with 30 Gray (Gy) of external beam radiation to patients with metastatic or locally advanced cancer.
* The secondary objectives are:

  * To describe the safety and tolerability of ECI301 delivered in combination with 30 Gy of external beam radiation to patients with metastatic or locally advanced cancer
  * To evaluate the humoral and cellular immune responses by:

    * Measurement of circulating precursor dendritic cells before and after the completion of ECI301
    * Measurement of circulating MIP-alpha before and after the completion of ECI301
    * Assessment of T-lymphocyte quantitative and qualitative changes by flow cytometry and assays for interferon (IFN-gamma) production
  * To define pharmacologic parameters following the intravenous dose of ECI301
  * To determine if neutralizing anti-EC301 antibodies occur after treatment
  * To describe the response at the radiated site and distant sites after radiation in combination with ECI301

Eligibility:

* Age \>18 years.
* Eastern Cooperative Oncology Group (ECOG) performance status \<2.
* Life expectancy of greater than 3 months
* Histologically confirmed metastatic or locally advanced cancer for which radiotherapeutic management would be appropriate
* No recent history of myocardial infarction or unstable angina

Design:

* This is a Phase I trial to determine the maximum tolerated dose of ECI301 in combination with external beam radiation therapy in patients with locally advanced or metastatic solid tumors.
* Patients will be treated with radiation therapy in a standard manner with ECI301 given daily during radiation. The dose of ECI301 will be escalated over the course of the trial to determine the maximum tolerated dose (MTD of daily ECI301 in combination with radiotherapy.
* We anticipate that accrual to this trial of 30 patients will take approximately 2 years.

Arms: arm 1: ECI301 Dose level 1 - 25 ug/kg and Radiation for Advanced or Metastatic Cancer
Interventions: intervention 1: Radiation Therapy intervention 2: ECI301
Intervention descriptions: intervention 1: None intervention 2: None

Protocol documents:
Prot_SAP_000.pdf: 
Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Protocol Number: 11-C-0248B 
Phase I trial of ECI301 in Combination with Radiation in Patients with Advanced or 
Metastatic Cancer  
 
Principal Investigator: 
Deborah Citrin, M.D. 1 
 
Lead Associate Investigator:  Karl Haglund, M.D., Ph.D.1 
 
Associate Investigators: 
Kevin Camphausen, M.D. 1 
Aradhana Kaushal, M.D. 1 
DeeDee Smart, M.D., Ph.D 1 
Giuseppe Giaccone, M.D., Ph.D.2 
Arun Rajan, M.D. 2 
Bradford Wood, M.D.3 
Aradhana Venkatesan, M.D.3 
Sharon Smith, RN1 
Debbie McNally, RN1 
Jane Trepel, Ph.D.2 
*Joanna Shih, Ph.D.4  
*William Ershler, MD5 
 
Judith Starling, R. Ph.6 
Chyi-Chia Richard Lee, M.D., Ph.D.7 
 
1Radiation Oncology Branch, NCI 
Bldg. 10CRC/ Rm. B23500 
Bethesda, MD 20892 
Phone (301) 496-5457 
Fax: (301) 480-5439 
 
2 Medical Oncology Branch, NCI 
3 Department of Diagnostic Radiology, CC, NIH  
4 Biometric Research Branch, DCTD, NCI 
5 Scientific Director, Institute for Advanced Studies in Aging, Gaithersburg, MD 
6 Clinical Center Pharmacy 
7 Dermatopathology and Surgical Pathology, NCI 
*Associate investigators without patient contact 
 
IND Sponsor:  
 
National Cancer Institute, Bethesda, MD 
Drug Name:  
 
ECI301 
IND Number: 
 
112789 
Manufacturer: 
 
ECI, Inc., Tokyo, Japan 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 2 of 60 
PRECIS 
BACKGROUND: 
• Patients with metastatic or locally advanced cancer frequently require palliative 
radiotherapy to relieve symptoms; however, progression of disease is frequent in patients 
with extended survival 
• Radiation results in tumor cell death which can result in increased dendritic cell 
activation and trafficking 
• ECI301 is a derivative of Macrophage Inflammatory Protein-1α, a 70 amino acid 
chemokine that is a ligand for CCR1 and CCR5, the chemokine receptors of immature 
dendritic cells. 
• ECI301 has been shown to enhance the effect of radiotherapy in animal models. 
Objectives: 
• The primary objective is to determine the maximum tolerated dose (MTD) of ECI301 
delivered in combination with 30 Gy of external beam radiation to patients with 
metastatic or locally advanced cancer. 
• The secondary objectives are: 
o To describe the safety and tolerability of ECI301 delivered in combination with 30 
Gy of external beam radiation to patients with metastatic or locally advanced cancer 
o To evaluate the humoral and cellular immune responses by: 
 Measurement of circulating precursor dendritic cells before and after the 
completion of ECI301 
 Measurement of circulating MIP-α before and after the completion of ECI301 
 Assessment of T-lymphocyte quantitative and qualitative changes by flow 
cytometry and assays for IFNγ production 
o To define pharmacologic parameters following the intravenous dose of ECI301 
o To determine if neutralizing anti-EC301 antibodies occur after treatment 
o To describe the response at the radiated site and distant sites after radiation in 
combination with ECI301 
Eligibility: 
• Age >18 years. 
• ECOG performance status <2.  
• Life expectancy of greater than 3 months 
• Histologically confirmed metastatic or locally advanced cancer for which 
radiotherapeutic management would be appropriate  
• No recent history of myocardial infarction or unstable angina 
Design: 
• This is a Phase I trial to determine the maximum tolerated dose of ECI301 in 
combination with external beam radiation therapy in patients with locally advanced or 
metastatic solid tumors.  
• Patients will be treated with radiation therapy in a standard manner for two weeks with 
ECI301 given daily following radiation during the second week.  The dose of ECI301 
will be escalated over the course of the trial to determine the MTD of daily ECI301 in 
combination with radiotherapy.   
• We anticipate that accrual to this trial of 30 patients will take approximately 2 years. 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 3 of 60 
Table of Contents 
PRECIS ........................................................................................................................................... 2 
1  INTRODUCTION ...................................................................................................................... 5 
1.1 Study Objectives: ................................................................................................................ 5 
1.2 Background and Rationale: ................................................................................................. 6 
1.2.1 Radiotherapy for palliation of metastases ................................................................ 6 
1.2.2 Modulating the effects of radiation .......................................................................... 6 
1.2.3 ECI301 ...................................................................................................................... 7 
1.2.4 Summary ................................................................................................................ 17 
2 ELIGIBILITY ASSESSMENT AND ENROLLMENT .......................................................... 17 
2.1 Eligibility Criteria ............................................................................................................. 17 
2.1.1 Inclusion Criteria .................................................................................................... 17 
2.1.2 Exclusion Criteria ................................................................................................... 18 
2.2 Screening Evaluation ........................................................................................................ 18 
2.2.1 Clinical Evaluation ................................................................................................. 18 
2.2.2 Laboratory Evaluation ............................................................................................ 19 
2.2.3 Radiographic Evaluation ........................................................................................ 19 
2.2.4 Pathologic Review: ................................................................................................ 19 
2.3 Registration Procedures .................................................................................................... 19 
3 STUDY IMPLEMENTATION ................................................................................................ 20 
3.1 Study Design: .................................................................................................................... 20 
3.1.1 Dose Limiting Toxicity .......................................................................................... 20 
3.1.2 Dose Escalation ...................................................................................................... 20 
3.2 Study Drug Administration: .............................................................................................. 22 
3.3 Dose Modification for Toxicities: ..................................................................................... 22 
3.4 Protocol therapy and evaluation........................................................................................ 22 
3.4.1 Pretreatment studies ............................................................................................... 22 
3.4.2 On treatment studies and therapy ........................................................................... 22 
3.4.3 Follow-up studies ................................................................................................... 23 
3.5 Study Calendar .................................................................................................................. 24 
3.6 Radiation Therapy Guidelines: ......................................................................................... 25 
3.6.1 Simulation. ............................................................................................................. 25 
3.6.2 Treatment Planning. ............................................................................................... 25 
3.6.3 Treatment delivery. ................................................................................................ 25 
3.6.4 Criteria for Removal from Protocol Therapy ......................................................... 25 
3.6.5 Off Study criteria .................................................................................................... 25 
4 SUPPORTIVE CARE .............................................................................................................. 26 
5 BIOSPECIMEN COLLECTION ............................................................................................. 26 
5.1 Correlative Studies for Research/Pharmacokinetic Studies:............................................. 26 
5.2 Sample Storage, Tracking, and Disposition ...................................................................... 27 
6 DATA COLLECTION AND EVALUATION ........................................................................ 28 
6.1 Data Collection ................................................................................................................. 28 
6.2 Measurement of Effect and Response Criteria ................................................................. 28 
6.2.1 Antitumor Effect – Solid Tumors ........................................................................... 28 
6.3 Toxicity Criteria ................................................................................................................ 33 
7 SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING PLAN 33 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 4 of 60 
7.1 Definitions......................................................................................................................... 33 
7.1.1 Adverse Event ........................................................................................................ 33 
7.1.2 Attribution: ............................................................................................................. 34 
7.1.3 Suspected adverse reaction ..................................................................................... 34 
7.1.4 Unexpected adverse reaction .................................................................................. 34 
7.1.5 Serious .................................................................................................................... 35 
7.1.6 Disability ................................................................................................................ 35 
7.1.7 Life-threatening adverse drug experience .............................................................. 35 
7.1.8 Protocol Deviation (NIH Definition) ..................................................................... 35 
7.1.9 Protocol Violation (NIH Definition) ...................................................................... 35 
7.1.10 Unanticipated Problem ........................................................................................... 35 
7.2 NCI-IRB Reporting ........................................................................................................... 36 
7.2.1 
NCI-IRB Expedited Reporting of Adverse Events, Unanticipated Problems, and Deaths 36 
7.2.2 
NCI-IRB Requirements for PI Reporting of Adverse Events at Continuing Review ....... 36 
7.2.3 NCI-IRB Reporting of IND Safety Reports ........................................................... 36 
7.3 FDA Reporting Criteria .................................................................................................... 36 
7.3.1 IND Safety Reports to the FDA (Refer to 21 CFR 312.32) ................................... 36 
7.3.2 FDA Annual Reports (Refer to 21 CFR 312.33).................................................... 37 
7.3.3 Expedited Adverse Event Reporting Criteria to the IND Manufacturer (ECI, Inc) 37 
7.4 Data and Safety Monitoring Plan ...................................................................................... 37 
7.4.1 Principal Investigator/Research Team .................................................................... 37 
7.4.2 Sponsor Monitoring Plan ........................................................................................ 38 
8 STATISTICAL SECTION ....................................................................................................... 38 
8.1 Study Design/Endpoints.................................................................................................... 38 
8.2 Sample size/Accrual Rate ................................................................................................. 38 
8.3 Analysis of secondary endpoints ...................................................................................... 38 
9 HUMAN SUBJECTS PROTECTIONS ................................................................................... 39 
9.1 Rationale for Subject Selection:........................................................................................ 39 
9.2 Participation of Children: .................................................................................................. 40 
9.3 Evaluation of Benefits and Risks/Discomforts: ................................................................ 40 
9.4 Risks/Benefits Analysis: ................................................................................................... 40 
9.5 Consent and Assent Process and Documentation: ............................................................ 41 
10 PHARMACEUTICAL INFORMATION: ............................................................................... 41 
10.1 ECI301 .............................................................................................................................. 41 
10.1.1 Source: .................................................................................................................... 41 
10.1.2 Toxicity .................................................................................................................. 41 
10.1.3 Formulation and preparation .................................................................................. 46 
10.1.4 Stability and storage ............................................................................................... 46 
10.1.5 Administration procedures ..................................................................................... 46 
10.1.6 Incompatibilities ..................................................................................................... 47 
11 REFERENCES ......................................................................................................................... 48 
12 APPENDICES .......................................................................................................................... 50 
12.1 Appendix I :  Acute RTOG Toxicity Scoring ................................................................... 50 
12.2 Appendix II:  Late RTOG Toxicity Scoring ..................................................................... 56 
12.3 Appendix III.  Excluded Immunosupressive Medications ................................................ 60 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 5 of 60 
1  INTRODUCTION 
Metastases are a common complication of the majority of solid tumors including malignancies of 
the breast, prostate, lung, head and neck, and gastrointestinal tract.  Metastatic disease can cause 
local symptoms such as pain, mass effect (airway obstruction, spinal cord compression), and 
pathologic fracture.  Therapy for metastatic disease is usually palliative with the goal of 
ameliorating symptoms.  Often, a short course of radiation such as 250 cGy fractions to a total 
dose of 3750 cGy (three weeks) is delivered with the intention of providing palliation for these 
symptoms.  Although these doses provide palliation in the majority of patients these are not 
considered curative or sterilizing doses.  Patients diagnosed with locally advanced or metastatic 
cancer, with a few exceptions, have a poor prognosis regardless of the therapy they receive, with 
the vast majority of patients succumbing to their disease.  This setting offers the unique 
opportunity to test an agent that may enhance local therapy or result in an abscopal effect due to 
immune activation.   
Radiation is known to result in a number of changes in the tumor bed which may enhance the 
efficacy of immunotherapy, including tumor cell necrosis, upregulation of MHC class I 
molecules, expression of adhesion molecules on tumor vasculature, and expression of a variety 
of proinflammatory cytokines and chemokines.[1, 2]  Chemokines play a major role in the 
recruitment of leukocytes to sites of infection and activation of these cells to enhance local 
inflammation [3-5].  One such chemokine MIP-1α, is produced by macrophages and binds to 
specific dendritic cell (DC) chemokine receptors (CCR1 and CCR5).  Dendritic cells are potent 
antigen presenting cells that perform a vital role in the initiation and regulation of both innate 
and adaptive immunity including anti-tumor responses.  Tissue dendritic cells internalize and 
process locally released antigens, migrate to draining lymph nodes, and present antigenic 
peptides in association with co-stimulatory molecules leading to the induction of helper and 
cytotoxic T cell responses.  ECI301 or eMIP is a derivative of MIP-1α with favorable 
pharmacologic properties and an excellent safety profile in lower species.   
We propose a phase I study of ECI301 in combination with concurrent RT directed at sites of 
locally advanced disease or metastases to define the maximum tolerated dose of this combination 
and to describe any toxicities observed.  Correlative assays and optional biopsies will allow 
investigation of any biologic effect of ECI301 delivered in this setting. We hypothesize that the 
addition of ECI301 will enhance radiotherapeutic effect locally in a variety of histologies while 
increasing immune recognition of tumor at distant sites with acceptable toxicity.    
1.1 
STUDY OBJECTIVES: 
• The primary objective is to determine the maximum tolerated dose (MTD) of ECI301 
delivered in combination with 30 Gy of external beam radiation to patients with 
metastatic or locally advanced solid tumors. 
• The secondary objectives are 
o To describe the safety and tolerability of ECI301 delivered in combination with 30 
Gy of external beam radiation to patients with metastatic or locally advanced 
cancer. 
o To evaluate the humoral and cellular immune responses by 
 Measurement of circulating precursor dendritic cells before and after the 
completion of ECI301 
 Measurement of circulating MIP-α before and after the completion of ECI301 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 6 of 60 
 Assessment of T-lymphocyte quantitative and qualitative changes by flow 
cytometry and assays for IFNγ production 
o To define pharmacologic parameters following the intravenous dose of ECI301 
o To determine if neutralizing anti-EC301 antibodies occur after treatment 
o To describe the response at the radiated site and distant sites after radiation in 
combination with ECI301 
1.2 
BACKGROUND AND RATIONALE: 
1.2.1 Radiotherapy for palliation of metastases 
Metastatic disease is a common occurrence in patients with cancer. Common sites of metastasis 
include the lung, bone, brain, and liver.  Symptoms of metastatic disease are dependent on the 
location of the metastasis, the rate of growth, and the proximity to critical structures.  For 
example, bone metastases may lead to pain or an increased risk of fracture.  Visceral metastases 
may lead to end organ dysfunction due to obstruction, mass effect, or replacement of 
parenchyma.  The therapy chosen for metastatic disease often is dependent on the site of 
metastasis, the overall disease status, and the histology. 
Radiation has been used extensively for the management of metastases at a variety of sites.  One 
common indication for palliative radiotherapy includes treatment of bone metastases causing 
pain or bone destruction.  Visceral and soft tissue metastases are also frequently treated with 
radiation to provide local symptom relief or prevent impending oncologic emergencies or 
urgencies such as cord compression from spinal metastases, superior vena cava syndrome from 
lung or mediastinal metastases, airway compromise from pulmonary or mediastinal metastases, 
obstructive jaundice or uncontrolled capsular pain from hepatic metastases, renal failure due to 
an obstructing lesion, deep venous thrombosis as a result of bulky malignant adenopathy, and a 
variety of other end-organ dysfunctions related to soft tissue or visceral metastases.   
The goal of therapy usually depends on the presenting signs and symptoms, the extent of 
systemic disease, and the systemic therapeutic options.  Frequently, a short course of 
hypofractionated radiation is delivered to maximize pain or symptom relief while minimizing the 
amount of time required for therapy.  Unfortunately, re-treatment due to progression or increase 
in metastatic burden distant to the irradiation site occurs often in patients with relatively longer 
survivals.  In one series of patients treated with radiation for prostate cancer bone metastases, as 
many as 58.6% required re-treatment at some point in their course with 22% of these patients 
requiring re-treatment of the spine [6].  In the majority of randomized trials evaluating various 
radiation fractionation schemes, re-treatment rates for bone metastases for pain or other 
indications approaches a disappointing 20-42% with most conventional doses [7].  Radiation has 
been studied extensively in this setting with local control frequently used as the primary endpoint 
in most series, even in the face of varying systemic therapies. 
1.2.2 Modulating the effects of radiation 
Radiation modifiers are agents that alter the effect of radiation when delivered in proximity to 
the radiation dose. A classic radiation sensitizer is a radiation modifier that has minimal toxicity 
and enhances radiation response selectively in tumor cells compared to normal tissue.  A number 
of agents have been evaluated as radiation modifiers, but currently only cetuximab, a monoclonal 
antibody to the endothelial growth factor receptor (EGFR), is approved for clinical use as a 
radiation modifier.  A number of chemotherapies are used to enhance the effects of radiation in 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 7 of 60 
the clinic, including cisplatin and 5-fluorouracil.  Although these agents are used to enhance 
radiation effects, they are also known to both potentiate radiation toxicity and to result in their 
own significant toxicities which preclude their use as radiation modifiers in the palliative setting.   
Recently, interest has increased in identifying agents or strategies that can exploit the effects of 
radiation on the tumor and tumor microenvironment to increase the efficacy of immunotherapy 
for cancer.  An understanding of the mechanisms by which radiation can alter the immune 
response is critical to the development of strategies combining radiation and immunomodulatory 
drugs.  Importantly, the immune effects of irradiation are quite different depending on the 
volume irradiated (total body versus localized). 
The possible mechanisms by which radiation can enhance immunotherapy are multiple.  Total 
body irradiation has been used as a method of immune suppression for adoptive cell therapies.[8]  
This approach requires delivery of lethal doses of total body irradiation and intensive preparative 
regimens that prepare the host for reconstitution with infused cells, such as tumor infiltrating 
lymphocytes.  A number of other immune modulating effects of total body irradiation have been 
elucidated that may contribute to the superior efficacy of total body irradiation compared to 
alternative preparative regimens.[9, 10]  This therapy has been pioneered and developed by 
investigators at NCI. 
Conversely, localized irradiation has been proposed as a mechanism to minimize the 
immunosuppressive effects of radiation by leaving the majority of host cells unirradiated while 
generating a more immunogenic tumor bed.[1, 2, 11]  Radiation results in tumor cells necrosis 
and increased expression of MHC Class I receptors in the irradiated field.[12, 13]  Additionally, 
the radiation induced expression of pro-inflammatory cytokines and adhesion molecules can 
result in the influx of antigen presenting cells, such as dendritic cells and macrophages.[2, 14]  
Recruited antigen presenting cells are then able to process antigens from necrotic tumor and 
mature into effector T cells, which may be more efficient in the irradiated field.  Mature T-cells 
are able to travel distantly and affect immune responses at distant disease sites.  Importantly, 
localized irradiation avoids delivery of sterilizing doses to the majority of the bone marrow, 
hence minimizing the immunosuppressive effects of therapy.  The immune activating effects of 
localized irradiation have recently been exploited in studies of immune modulating drugs and in 
vaccine based therapies.[15-18]   
1.2.3 ECI301 
Chemokines play a major role in the recruitment of leukocytes to sites of infection and activation 
of these cells to enhance local inflammation [3-5]. One such chemokine MIP-1α, is produced by 
macrophages and binds to specific dendritic cell (DC) chemokine receptors (CCR1 and CCR5). 
Dendritic cells are potent antigen presenting cells that perform a vital role in the initiation and 
regulation of both innate and adaptive immunity including anti-tumor responses. Tissue dendritic 
cells internalize and process locally released antigens, migrate to draining lymph nodes, and 
present antigenic peptides in association with co-stimulatory molecules leading to the induction 
of helper and cytotoxic T cell responses. ECI301 or eMIP is a derivative of MIP-1α with 
favorable pharmacologic properties and an excellent safety profile in lower species (see below). 
1.2.3.1 
Pre-Clinical experience with ECI301 
When administered to laboratory animals, it was found that MIP-1α and its functional derivatives 
have an ability to attract DCs to local sites with inflammation or local cancer and can recruit 
dendritic progenitor cells into the blood at higher magnitude.[19]  And when MIP-1α was 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 8 of 60 
induced at the local site of inflammation in mouse model, dendritic cells accumulated to induce 
antigen specific immune response.[20]  As described above, radiation treatment of tumors is 
known to induce inflammation in the irradiated field and to recruit tumor specific T cells and 
dendritic cells, which seem to play an important role in remission of tumors.[1, 21] 
It has been hypothesized that the proinflammatory properties of MIP-1α could be exploited to 
enhance systemic immunity in a number of clinical conditions. For the purpose of pre-clinical 
evaluations, Effector Cell Institute (Tokyo) re-engineered MIP-1α to generate a biologically 
active derivative with fewer tendencies to form aggregates at physiologic pH and ion 
concentrations. The derivative, ECI301 is a 69 amino acid variant of MIP1 α which has an Asp 
residue substituted for Ala at position 26 of MIP1 α. Recombinant ECI301 was produced in S. 
pombe and found to incorporate the properties of increased solubility and stability while 
retaining full biologic activity and was used in preclinical studies.[22] 
No noticeable toxicity was found in mice as a result of the I.V. administration of ECI301.[22] 
There have been studies reported using the British Biotech product (BB-10010), the same active 
variant of human MIP-1α as ECI301. Recombinant BB-10010 was produced using a budding 
yeast expression system. The biological activity of this recombinant protein was confirmed in 
receptor binding, calcium mobilization and biological assays in vitro and in vivo. The concept in 
the development of BB-10010 was to use the agent as an inhibitor of hematopoetic stem cell 
proliferation at the time of delivery of systemic chemotherapy to minimize hematologic toxicity 
and to enhance the number of peripheral blood stem cells for transplantation.  Its 
myelosuppressive effect was investigated in several clinical trials of patients receiving 
chemotherapy but the myeloprotective effect was therapeutically insufficient.[23-25]   
In a phase I clinical study by E. Marshall et al, 9 patients with advanced breast cancer and 40 
normal healthy volunteers received escalating doses of BB-10010 from 0.1- 300 μg/kg i.v or 
subcutaneous routes, and there was no significant toxicity observed at any dose level. Treatment 
was associated with a dose-related increase in monocyte count and no measurable effects were 
noted on other leukocyte subsets or on circulating progenitor cell numbers. Pharmacokinetic 
analysis revealed that serum concentrations of BB-10010 were detectable using doses of > or = 
10 µg/kg i.v. or > or = 30 µg /kg S.C., and that a single S.C. injection resulted in sustained 
plasma levels over a 24 h period. In all cases, BB-10010 was extremely well tolerated with no 
significant toxicity observed at any dose level and a maximum tolerated dose was not 
defined.[26] 
In another phase II study of BB-10010 conducted by Dr Bernstein et al., use of human MIP-α in 
patients with malignant lymphoma and breast cancer receiving high dose etoposide and 
cyclophosphamide, the compound was well tolerated by the patients. BB-10010 was extremely 
well tolerated without any notable toxicity related to the compound.[27] 
1.2.3.2 
Toxicology studies of ECI301 
Single dosing studies were conducted to evaluate the acute toxicity of IV (bolus) administration 
of ECI301 in mice and to support further toxicity studies. A single dose of 0 (PBS; non-treated 
[NT], n=2), 1, 10, or 300 mg/kg ECI301 (n=3/dose) was administered to 5-week-old male ICR 
mice. All mice were fasted prior to dosing and were observed for 4 hours post dose for signs of 
general condition and mortality. Body weight measurements were taken on Days -1, 0, 1, and 2. 
Animals were sacrificed 2 days after dosing. Parameters evaluated included: hematology for 
albumin/globulin ratios, blood urea nitrogen, creatinine, lactic dehydrogenase, aspartate 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 9 of 60 
aminotransaminase, alanine aminotransaminase, and inorganic phosphorus of the isolated 
plasma. Urine was analyzed for glucose, urobilinogen, and pH. Histological assessment of brain, 
lung, liver, kidney, thymus, intestine, and colon was conducted. No animals died following 
administration of single doses of ECI301 up to 300 mg/kg. There were no significant treatment-
related differences in clinical signs, body weights, plasma examination, histology, or organ 
weights in any dose group. Protein levels in urine showed minor elevations in the 10-mg/kg and 
300-mg/kg groups. There were no other observed changes. These results indicate that a single 
dose of ECI301 of up to 300 mg/kg did not cause any overt toxicity during observations for 2 
days post-treatment. It was concluded that a single dose of ECI301, up to 300 mg/kg, was well 
tolerated, and therefore the NOAEL was 300 mg/kg for this study. [28] 
A single dose study was conducted in accordance with GLP to evaluate the acute toxicity of IV 
administration of ECI301 in rats. A single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.5, or 
5 mg/kg ECI301 was administered IV (bolus) to Crl:CD® (SD) IGS BR rats (5/sex/dose). All 
rats were observed daily for 14 days after dosing for general clinical condition, signs of toxicity, 
and mortality. All animals were subjected to necropsy and macroscopic investigation. An 
additional group of animals was administered a single dose of 0.05, 0.5, or 5 mg/kg ECI301 IV 
for toxicokinetic analysis (6/sex/dose). Animals were bled at 5, 15, and 30 minutes, and 1, 2, and 
4 hours postdose for toxicokinetic analysis. There were no treatment-related deaths for this 
study. There were no significant treatment-related clinical signs or effects on body weight. Organ 
weights were not affected, and there were no treatment-related macroscopic findings. The 
NOAEL was determined to be greater than the 5-mg/kg ECI301 used in this study. The 
toxicokinetic results indicated that the rate and extent of systemic exposure of rats to ECI301 
appeared to be characterized by nonlinear (dose-dependent) kinetics over the dose range. 
Increasing the dose of ECI301 above 0.05 mg/kg was considered likely to result in a higher 
systemic exposure than would be predicted from a linear relationship, which is consistent with 
the possibility of a capacity-limited process for ECI301 elimination. There were no differences 
between the sexes for systemic exposure. It was concluded that a single dose of ECI301 at 5 
mg/kg did not cause any toxicological effects. [28] 
Repeated dosing studies of ECI301 in 5- to 7-week-old ICR mice were conducted as a non-GLP 
investigation to evaluate the potential toxicity of repeat IV administration. ECI301 was 
administered IV at 0 (PBS; 2/dose), 1, 10, or 100 mg/kg/day (3/dose) for two 5-day dosing 
cycles, split by a 3-day off-dose period. An additional group receiving bovine serum albumin 
(BSA; 100 mg/kg) was added to serve as an additional control group for using a different 
protein. Animals were observed for 4 hours post-dose on Day 3 and Day 13 for signs of general 
condition and mortality. Animals were autopsied 24 hours following the last dose. Parameters 
evaluated included daily body weights, urinalysis, and hematology. Organ weights and histology 
of brain, lung, liver, kidney, thymus gland, and colon tissue were conducted. There was no 
treatment-related mortality. There were no treatment-related effects on body weights or any of 
the hematological parameters measured. The protein in urine increased slightly in 1 of 3 animals 
in the 10-mg/kg group and 1 of 3 animals in the 100-mg/kg group. At 100 mg/kg ECI301, there 
was an increase in lung and spleen weights. Histology of the kidney showed an increase in the 
adhesion of glomerular basement membrane and renal glomeruli, and disruption of the 
glomerulus structure in all animals in the group receiving ECI301 at 100 mg/kg, and 1 of 3 
animals in the 10-mg/kg group. It is unclear if these results are due to administration of ECI301 
or to immune complex deposition. In spleen samples from both BSA and ECI301 (100 mg/kg), 
the boundary between the red and white pulp was unclear. There was structural damage to the 


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Page 10 of 60 
white pulp, and the central artery was unclear. There were no treatment-related changes in any of 
the other organs or tissues. This experiment demonstrated that repeated doses of ECI301, to a 
maximum total dose of 1000 mg/kg (1000 times the effective dosage), had no critical effect on 
the kidney, but abnormal tissue images and urine protein elevations were observed that indicated 
some renal damage. Further study is required to determine if this damage is attributable to 
ECI301 or another protein. Data from this non-GLP study supported further study of repeat 
doses over 2 to 4 weeks.[28] 
A separate study was conducted in accordance with GLP to evaluate the systemic toxicity and 
toxicokinetics of IV administration of ECI301 alone or in combination with docetaxel in rats for 
three 5-day dosing cycles. A single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.15, or 1.5 
mg/kg/day ECI301, 12.5 mg/kg docetaxel, or a 0.05-/12.5- mg/kg/day combination of ECI301 
and docetaxel, was administered IV (bolus) to Crl: CD® (SD) IGS BR rats daily for 5 days. 
After 51 days, 10 animals/sex/dose were sacrificed and an additional 5 animals/sex/dose were 
sacrificed following a 10-day recovery period. All rats were observed daily after dosing for 
general clinical condition, signs of toxicity, and mortality. Additionally, food consumption, 
ophthalmology, hematology, blood chemistry, urinalysis, anti-ECI301 antibody assessments, 
peripheral blood immunophenotyping, and organ weights were analyzed. All animals were 
subject to necropsy with macroscopic and microscopic pathology. An additional group of 
animals for each group (6/sex/dose) except control (3/sex/dose) was bled for 
toxicokineticanalysis of ECI301 and docetaxel plasma levels. Animals were bled at 5, 15, and 30 
minutes, and 1, 2, and 4 hours post dose for toxicokinetic analysis. There were no deaths in this 
study attributable to ECI301 treatment alone. Six animals died following docetaxel treatment. 
There were no treatment-related clinical signs or any effect upon bodyweight and food 
consumption. Similarly, there were no treatment-related ophthalmoscopic findings or any effect 
upon the composition of the urine. The hematological examination indicated a slight increase of 
lymphocyte count after the first treatment cycle, resulting in an increased total leukocyte count, 
in males given 0.15 or 1.5 mg/kg, with similar trends also occurring at all doses in females after 
Cycle 3. This was not evident in the males after Cycle 3 or in the females at the end of the 
recovery period. Biochemical changes in the blood plasma were confined to a reduction of 
glucose concentrations after Cycles 1 and 3 in males given 1.5 mg/kg and after Cycle 3 in males 
given 0.15 mg/kg and a small increase of plasma urea after Cycle 3 in males given 1.5 mg/kg. 
An analysis of organ weights after the completion of the third dosing cycle indicated a small 
reduction of spleen and thymus weight in males given 0.05 mg/kg, which was attributed to 
treatment since this dose was at the maximum biological effect level for ECI301. There were no 
treatment-related macroscopic or histopathological findings. The plasma samples taken from the 
control animals were below the limit of quantification in all cases, demonstrating that there was 
no quantifiable contamination with ECI301 in these animals.[28] 
The toxicokinetic investigations indicated that the extent of systemic exposure of rats to ECI301 
appeared to be characterized by non-linear (dose-dependent) kinetics over the dose range 0.05 to 
1.5 mg/kg/day on Days 1 and 47. Increasing the dose of ECI301 above 0.05 mg/kg/day resulted 
in a higher systemic exposure than would be predicted from a linear relationship, indicative of 
limited elimination capacity. In general, there were no differences in the systemic exposure of 
males and females to ECI301 and there was no accumulation after repeated doses. One female 
given 0.05 mg/kg ECI301 showed a positive anti-ECI301 antibody response. All other animals 
showed negative responses. Immunophenotyping after completion of Cycle 3 indicated, in 
animals given 1.5 mg/kg, an increased percentage of CD4+ T cells and the males showed a 


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corresponding decrease in the percentage of CD8+ T cells, although the total numbers for all 
lymphocyte subsets studied were not affected. The numbers of NK lymphocytes were reduced in 
females given 1.5 mg/kg. Following recovery, males given 1.5 mg/kg still showed a reduced 
percentage of CD4+ lymphocytes and an increased percentage of CD8+ lymphocytes. It was 
concluded that IV (bolus) administration of ECI301 to CD rats at doses up to 1.5 mg/kg during 
three 5-day cycles of treatment, each separated by 16 days, was well tolerated, was not 
immunogenic, and caused no toxicologically significant findings. Docetaxel treatment 2 days 
after each cycle in which animals were given ECI301 at 0.05 mg/kg (a pharmacologically active 
dose) at 12.5 mg/kg in the first cycle in both sexes and the second cycle in males, subsequently 
reduced to 6.25 mg/kg for the remaining cycles, caused a range of findings that were attributable 
to its anti-mitotic activity. Compared with animals given docetaxel on the same study days, but 
with no ECI301 treatment, all differences in the response to docetaxel were attributed to normal 
biological variation in response to treatment and, consequently, there was no evidence that 
ECI301 modulated the effects of docetaxel treatment.[28] 
A separate study was conducted to evaluate the potential for systemic toxicity of 3 different 
doses of ECI301, when administered IV to common marmosets for a 5-day dosing period with an 
18-day observation period, to confirm doses to be used in the main toxicity study. Common 
marmosets (Callithrix jacchus) received 0.05, 0.15, or 1.5 mg/kg ECI301 (1/sex/group) IV bolus, 
once a day, for 5 days. Animals were observed for 18 days for clinical condition, injection-site 
reactions, and body weight. Hematology, blood chemistries, organ weight, and macroscopic 
pathology investigations were undertaken. There were no treatment-related deaths during this 
study. There were no treatment-related clinical signs of toxicity, and body weights were 
unaffected. There was no effect of treatment on organ weights or macropathology. All animals 
had reduced hematocrit, hemoglobin concentration, and/or erythrocyte count on Day 9 and/or 
Day 22, often accompanied by increased reticulocyte count. Mean cell volume was increased in 
the male receiving 0.05 mg/kg, the female receiving 0.5 mg/kg, and both animals receiving 1.5 
mg/kg. Mean cell hemoglobin concentration was decreased in the male receiving 0.05 mg/kg and 
the female receiving 1.5 mg/kg. Anisocytosis was reported on the blood film for all females. 
There was no strong trend with dosage, and it is possible that these changes may have been 
caused by repeated blood sampling. However, with the absence of control animals on this study, 
the possibility that these changes were due to treatment with ECI301 cannot be excluded. An 
increase in monocyte count occurred on Day 9 in the female marmosets receiving 0.05 mg/kg or 
0.15 mg/kg. These values normalized by Day 22. Minor blood chemistry changes occurred on 
Day 9 and Day 23, but these changes were small and were not dose related in degree. All animals 
showed reduced alkaline phosphatase activity on Day 23. Creatinine concentrations were 
reduced on Day 23 in the males given 0.05 mg/kg and in animals given 0.15 mg/kg or 1.5 mg/kg. 
Total triglyceride concentrations were high on Day 9 and Day 23 for all animals except males 
given 1.5 mg/kg. In the animals given 0.05 mg/kg and the females given 0.15 mg/kg, there was a 
reduction of total cholesterol level. All animals showed an increase of sodium concentration on 
Day 9 and, at 0.15 and 15 mg/kg, this was associated with increased chloride concentration. 
Potassium concentrations were low on Day 9 in all animals, and in the males, this persisted to 
Day 23. Total protein concentrations were low on Day 23 in animals given 0.05 mg/kg or 0.15 
mg/kg, but not in those given 1.5 mg/kg, and most showed an increase in the albumin to globulin 
ratio. It was concluded that the IV administration of ECI301 to common marmosets for a single 
5-day dosing period was well tolerated up to doses of 1.5 mg/kg. There were no toxicologically 


Abbreviated Title: Phase 1 ECI301 with radiation 
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Page 12 of 60 
significant findings, and therefore it was concluded that 1.5 mg/kg was a suitable dose for future 
studies in marmosets.[28] 
Study EFC0010 was conducted to evaluate the systemic toxicity and toxicokinetics of IV 
administration of ECI301 to Callithrix jacchus, the common marmoset for three 5-day cycles. A 
single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.15, or 1.5 mg/kg ECI301 was 
administered IV (bolus) to marmosets (3/sex/dose). Radiation with chemotherapy is standard of 
care regimen for treatment of cancer. Docetaxel is one of the common chemotherapy drugs; 
however, significant AEs such as suppression of bone marrow function are associated with the 
administration of docetaxel. In phase 1 clinical study, ECI301 is expected to be administered to 
cancer patients in combination with radiation treatment. However, it may be necessary for 
docetaxel to be administered to patients in addition to ECI301. Therefore, toxicity data of 
ECI301 in combination with docetaxel were found to be necessary. The design of the study was 
as indicated below and also included 2 groups that were dosed with docetaxel, one in 
combination with ECI301. All animals were observed daily throughout the study period for 
general clinical condition, injection-site observations, body weight, ophthalmology, 
electrocardiography, blood pressure, hematology, blood chemistry, urinalysis, peripheral blood 
immunophenotyping, organ weights, macroscopic and microscopic pathology, and mortality. An 
additional group of animals for each group except control (2/sex/dose) were bled for 
toxicokinetic analysis. Animals were bled at 5, 15, and 30 minutes, and 1, 2, and 4 hours 
postdose for toxicokinetic analysis of plasma concentrations of ECI301 or docetaxel, and anti-
ECI301antibody assessment. Two animals were sacrificed prematurely during this study. Neither 
death was attributed to ECI301 treatment, but 1 of them was determined to be possibly due to 
docetaxel treatment. There were no clinical signs that were attributable to treatment, and there 
was no effect upon bodyweight. One male receiving 0.15 mg/kg was killed on Day 59 due to 
gradual decline in its general clinical condition. The macroscopic and histopathological 
examinations did not identify any finding that would account for the ill health of this animal and, 
consequently, this death was not attributed to treatment. There were no treatment-related 
ophthalmoscopic findings and no effect upon the electrophysiology of the heart or blood 
pressure. There were no hematological findings or changes in blood plasma or urinary 
composition that were attributable to treatment. There were no toxicologically significant organ 
weight differences after 3 cycles of dosing with ECI301 and no macroscopic or histopathological 
findings that were due to treatment.[28] 
The plasma samples taken from the control animals were below the limit of quantification in all 
cases, demonstrating that there was no quantifiable contamination with ECI301 in these animals. 
Mean plasma concentrations (Cmax) and ECI301 concentration-time curves estimated up to 4 
hours post dose (AUC4) on Day 1 and 47 are also shown. The toxicokinetic investigations 
indicated that the extent of systemic exposure of marmosets to ECI301 appeared to be 
characterized by non-linear (dose-dependent) kinetics over the dose range 0.05 to 1.5 mg/kg on 
Days 1 and 47. Increasing the dose of ECI301 above 0.15 mg/kg was considered likely to result 
in a higher systemic exposure than would be predicted from a linear relationship, which is 
consistent with the possibility of a capacity limited process for the elimination of ECI301. In 
general, there were no differences in the systemic exposure of male and female marmosets to 
ECI301, and there was no accumulation after repeated doses. All animals showed negative anti-
ECI301 antibody responses. Immunophenotyping after completion of Cycle 3 indicated no 
significant change at any dose.[28] 


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The toxicokinetic investigations indicated that the co-administration of ECI301 with docetaxel 
generally appeared to have little effect on the systemic exposure of male and female marmosets 
to docetaxel. The co-administration of ECI301 with docetaxel did not elicit an anti-ECI301 
antibody response in any animal. Peripheral blood immunophenotyping indicated a reduction in 
the number of CD3+ T lymphocytes in males and females given docetaxel alone and a reduction 
in CD20+ lymphocytes in the males. These males also showed an increase in the percentage of 
CD3+ lymphocytes and a corresponding decrease in the percentage of CD20+ B lymphocytes. 
This T/B cell ratio change was not seen in the females. A reduction in the number and percentage 
of CD14+ monocytes was also seen in the males, but not in the females. It was concluded that IV 
(bolus) administration of ECI301 to common marmosets at doses up to 1.5 mg/kg during three 5-
day cycles of treatment, each separated by 16 days, was well tolerated, was not immunogenic, 
and caused no toxicologically significant findings. Docetaxel treatment at 3 mg/kg, 2 days after 
each cycle in which animals were given ECI301 at 0.05 mg/kg (a pharmacologically active 
dose), caused several findings that were attributable to its anti-mitotic activity. There was no 
evidence in this study that ECI301 modulated adversely the effect of docetaxel treatment, but 
ECI301 coadministration did appear protective with regard to the effect upon lymphocyte 
populations in the peripheral blood.[28] 
1.2.3.3 
Preclinical experience with ECI301 as a radiation modifier 
K. Shiraishi et al used murine tumor models to demonstrate that administration of the MIP-1α 
derivative ECI301 after radiation of a primary subcutaneous tumor nodule enhanced not only the 
anti-tumor efficacy of radiation at this site, but also promoted anti-tumor immunity at distant 
sites, referred to as an abscopal effect (figure 1, figure2).[22] To determine this, mice were 
implanted with colon 26 adenocarcinoma cells in the right flank or both flanks and when the 
tumor size reached 10 mm, local irradiation with 6 Gy was delivered to the right flank tumor. 
After 20 hrs, 2 μg per mouse of ECI301 was administered for 3- 5 consecutive days. This study 
showed that a combination of radiotherapy and ECI301 I.V. administration effectively prevented 
tumor growth at the irradiated site and also promoted the abscopal effect (prevention of tumor 
growth at the non-irradiated site).  Prevention of tumor growth was observed with various mouse 
tumors including Lewis lung cancer, Meth A fibrosarcoma and Colon 26 adenocarcinoma 
suggesting that the effect was not restricted to a specific tumor type.   Perhaps most importantly, 
rechallenge with the same tumor type in mice exhibiting a complete response to ECI301 and 
irradiation resulted in no tumor growth. 


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Figure 1. Effect of ECI301administration on Colon26 or LLC tumor growth at the irradiated (B and D) and nonirradiated 
sites (A and C). A and B, time course of tumor growth. Colon26 cells were inoculated in the left (1 x105 cells; A) or the right 
flanks (2 x 105 cells; B) of BALB/c mice and 18 d after inoculation; only the right side was exposed to radiation. ECI301was 
given at 1, 8, and 15 d starting 20 h after irradiation (⁪). Mice with radiation treatment only (О) or without treatment (●) 
served controls. Significant differences from the control group: *, P < 0.05; **, P < 0.01 (ANOVA). C and D, time course of 
tumor growth. LLC cells were inoculated in the right (4 _105 cells; D) and left flanks (2 x 105 cells; C) of C57BL/6 mice and 
18 d after inoculation, only the right side was exposed to radiation. ECI301 (E, 0.08; 4, 0.4; 5 2 μg per mouse) was given at1, 
8, and 15 d starting 20 h after irradiation. Results of mice with radiation treatment only (О), without treatment (●), or 
ECI301administration (2 μg per mouse) without irradiation () were used as controls. Points, mean tumor volume; bars, SE 
(n = 8). Inset, columns, mean tumor volumes of eight mice 12 d after irradiation; bars, SE. Significant differences from 
control group: *, P < 0.05; **, P < 0.01 (ANOVA). Reproduced from Shiraishi et al.  [22]. 
 


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Figure 2.  Effect of daily ECI301 administration on tumor growth.   Effect of ECI301administration on different tumors 
grown at the irradiated (B and D) and non-irradiated sites (A and C). A and B, time course of tumor growth in the left flank 
(MethA, non-irradiated site; A) and the right flank (Colon26, irradiated site; B). Inoculum sizes of MethA and Colon26 cells 
were 1x105 and 2 x 105, respectively. ECI301 (2 μg per mouse) was given weekly starting from day 1after irradiation 
(⁪).Mice with radiation treatment only (О) and without treatment (●). Mean tumor volume in eight mice are presented. 
Significant differences from control group: **, P < 0.01 (ANOVA). A similar time course of tumor growth of MethA cells 
without treatment (●) was observed after ECI301administration without irradiation. C and D, time course of tumor growth in 
the left flank (Colon38, non-irradiated site; C) and the right flank (LLC, irradiated site; D). Inoculum size of LLC cells was 4 
x105, and Colon38 was 2 mm3. ECI301(2 μg per mouse) was given weekly starting from day 1after irradiation (⁪). Mice 
with radiation treatment only (О) and without treatment (●). Significant differences from control groups: **, P < 0.01 
(ANOVA). Reproduced from Shiraishi et al.  [22]. 
 
The enhancement in tumor response to irradiation was accompanied by an increase in infiltration 
of CD4+ and CD8+ lymphocytes into tumor sites at assessed by immunohistochemistry.  
Additional mechanistic studies completed in the radiation in combination with ECI301 studies 
included specific immune cell depletion via systemic delivery of CD4, CD8, and NK1.1 
antibodies.  Depletion of CD8+ cells reduced the benefit of ECI301 with radiation at both the 
irradiated site and at the unirradiated site (non-significant at the unirradiated site).  Depletion of 
CD4+ or NK1.1 cells decreased the effect of combination therapy at the unirradiated site.  
Suppression of Natural Killer Cells did not alter the efficacy of the combined treatment. 


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Additional unpublished studies of ECI301 in combination with radiation have been completed 
with fractionated radiation and repetitive dosing to determine if delivering ECI301 concurrently 
with irradiation results in similar responses locally.  The theoretical concern is that radiating the 
tumor bed repeatedly during dosing of ECI301 could impair the immune activation.  Studies in 
BALB/c mice found that delivery of ECI301 during a portion of a fractionated treatment 
schedule in fact resulting in similar efficacy as that seen in the unfractionated model (Figure 3).   
 
 
Figure 3.  Effects of ECI301 administered concurrent to fractionated irradiation.  BALB/c mice were 
implanted with colon26 cells in the right flank.  A)  On the day of treatment animals were randomized to no 
treatment, radiation alone, or fractionated radiation with the combination of ECI301 in one of two 
schedules.  B)  Tumor growth was inhibited in the groups receiving twice weekly and five times weekly 
dosing of ECI301.  Improved efficacy was observed in the five times weekly dosing compared to the twice 
weekly dosing regimen.  Confidential data provided by ECI. Inc. 
1.2.3.4 
Dose and Schedule Selection 
The starting dose in this protocol is based on the results of toxicology studies conducted in rats 
and marmosets which established the no observable adverse effect level (NOAEL) for ECI301 at 
0.15 mg/kg. The conversion factor from these species to humans is 0.16 which makes the human 
equivalent dose (HED) 0.024 mg/kg. The standard “safety factor” of 10 establishes the MRSD at 
0.0024 mg/kg.  
Preclinical studies demonstrated that a dose between 2μg and 3μg/mouse in combination with 
radiation therapy inhibited tumor growth. Given that the body weight of mice is 30g, the dosage 
between 2μg and 3μg is approximately 70μg/kg to 100μg/kg. Therefore it is predicted that the 
optimal dosage for obtaining the anti-cancer effect would be lower than 100μg/kg in mice. Thus 
it is proposed that 25 μg /kg, 50 μg /kg, 82.5 μg /kg, 125 μg /kg or 175 μg/kg will be 
intravenously administered for 5 consecutive days for one week to the patients in this phase I 


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study to assess the effect and safety of the combination treatment with radiation using dose-
escalating method. 
Daily dosing of ECI301 has been selected based on the preclinical data above that suggests 
optimal efficacy with daily dosing compared to less frequent dosing regimens.  Repeated dosing 
experiments with irradiation lasted for one week.  A theoretical concern of combining radiation 
with an immune adjuvant is that effector T cells could migrate into the radiation field and be 
lethally irradiated.  Based on the available literature relating to vaccines and antigen dependent 
T-cell proliferation, a minimum of a 5-8 day window after is likely to be required for the 
development and expansion of effector T-cells.[29, 30]  Therefore, we have designed the study 
to include one week of concurrent therapy with ECI301 during the final week of radiation 
delivery.  Finally, animal models have confirmed that up to five daily doses of ECI301 following 
irradiation can result in measurable immune responses and abscopal effect. 
1.2.4 Summary 
In this planned clinical trial, we will attempt to establish the safety and tolerability of intravenous 
ECI301 (which has the same amino acid sequence as BB-10010) in combination with radiation 
treatment in patients with solid malignancies for whom radiation therapy alone would constitute 
standard of care. In correlative translational studies we will perform pharmacokinetic (PK) 
analysis of MIP-1α serum levels, determine precursor dendritic cell (DC) frequencies before and 
after ECI301 treatment, and measure levels of proinflammatory chemokines, in vitro studies of T 
cell functions and its consequences after completion of ECI301. 
2 
ELIGIBILITY ASSESSMENT AND ENROLLMENT 
2.1 
ELIGIBILITY CRITERIA 
2.1.1 Inclusion Criteria 
2.1.1.1 
Age >18 years. 
2.1.1.2 
ECOG performance status <2.  
2.1.1.3 
Life expectancy of greater than 3 months 
2.1.1.4 
Histologically confirmed cancer 
2.1.1.5 
Extracranial metastatic cancer or locally advanced cancer for which palliative 
radiotherapeutic management would be appropriate (no more than two sites will be 
treated on this trial) 
2.1.1.6 
Patients must have measurable or evaluable disease at the site(s) requiring radiation 
2.1.1.7 
Adequate marrow and organ function defined as  
o absolute neutrophil count (ANC) > 1.5 x 109/L,  
o platelet count > 100 x 109,  
o hemoglobin >9 g/L. 
o creatinine clearance > 60 mL/min/1.73 m2 for patients with creatinine levels 
above institutional normal 
o serum bilirubin <1.5 x upper limit reference range (ULRR), 
o alanine aminotransferase (ALT), aspartate aminotransferase (AST), or  
alkaline phosphatase (ALP) <2.5 x the ULRR (<5 x the ULRR in the presence 
of liver metastases) 


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2.1.1.8 
Female patients of child bearing potential must either be surgically sterile to prevent 
pregnancy, be at least 1-year post-menopausal, or have had no menses for 12 months, 
or agree to use reliable methods of contraception (oral contraceptives, barrier 
methods, approved contraceptive implant, long-term injectable contraception, copper 
banded intrauterine device, tubal ligation or abstinence) from time of screening until 
4 weeks after discontinuing study treatment.  It is not known whether ECI301 has the 
capacity to induce hepatic enzymes so hormonal contraceptives should be combined 
with a barrier method of contraception. 
2.1.1.9 
Male patients must agree to use barrier contraception (i.e. condoms) and refrain from 
donating sperm from the start of dosing until 16 weeks after discontinuing study 
treatment. If male patients wish to father children they should be advised to arrange 
for freezing of sperm prior to the start of study treatment. 
2.1.2 Exclusion Criteria 
2.1.2.1 
Pregnant or lactating females 
2.1.2.2 
Contraindications to radiotherapy (i.e. prior radiotherapy to the intended treatment 
site) 
2.1.2.3 
Untreated or previously treated but progressive intracranial metastases (Patients with 
previously treated intracranial metastases should have no clinical evidence of 
progression and be at least 4 weeks from therapy for intracranial metastases) 
2.1.2.4 
Need for emergent radiotherapy (defined as need for radiotherapy within 24 hours of 
consultation at the judgment of the treating radiation oncologist) 
2.1.2.5 
Active treatment with immunosuppressive therapy and subjects taking systemic 
corticosteroid therapy for any reason including replacement therapy for 
hypoadrenalism 
2.1.2.6 
Chemotherapy, radiation therapy, Tamoxifen or investigational therapy during the 4 
weeks prior to initiation of protocol therapy 
2.1.2.7 
History of rheumatoid arthritis, systemic lupus erythematosus, Sjögrens disease, 
sarcoidosis, vasculitis, polymyositis, temporal arteritis or any other autoimmune 
disease 
2.1.2.8 
History of organ transplant 
2.1.2.9 
HIV, Hepatitis B, or Hepatitis C positivity 
2.1.2.10 
Uncontrolled intercurrent illness including, but not limited to, ongoing or active 
infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac 
arrhythmia, or psychiatric illness/social situations that would limit compliance with 
study requirements 
2.1.2.11 
Use of excluded immune modulating medications within 4 weeks prior to protocol 
therapy, or requirement for concurrent use (Appendix III). 
2.2 
SCREENING EVALUATION 
2.2.1 Clinical Evaluation 
• A complete history and physical examination will be completed by the PI or an 
associate investigator.  This will include a detailed history including comorbidities 
and prior therapy received, measurements of measurable disease to be targeted with 


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radiation, signs, and symptoms caused by the cancer, determination of ECOG 
performance status. 
• Vital signs: Including height and weight 
• ECG within one week of study entry 
2.2.2 Laboratory Evaluation 
Pre- treatment blood tests should be performed within one week of study entry.   
• Hematology: complete blood count, differential, platelet count, PT, PTT 
• Chemistries: LDH, SGOT, SGPT, alkaline phosphatase, bilirubin (total and direct), 
BUN, serum creatinine, serum electrolytes, calcium, magnesium, phosphorus, uric 
acid, albumin. 
• 24 hour urine for measurement of creatinine clearance and urine protein 
• Urinalysis 
• Urine Pregnancy Test: required for females of childbearing potential 
• Viral serologies: HIV screening; Hepatitis B and C screening within three weeks of 
study entry 
2.2.3 Radiographic Evaluation 
• CT will be the preferred imaging modality for metastatic sites of non-bone location.  
CT of the metastatic site to be treated within 3 weeks of entry into the protocol can be 
used as the baseline evaluation.  If scans were obtained from another institution, 
copies should be produced and maintained on file with the PI. 
• For bone lesions with or without soft tissue extension, MRI and plain films of the site 
to be treated will be obtained as the baseline study. 
• Additional complementary imaging studies may be obtained for baseline 
measurements if CT or MRI is felt to be insufficient (i.e. Ultrasound). 
2.2.4 Pathologic Review:   
Outside report confirming metastatic malignancy will be acceptable for study entry.  In certain 
cases, for example a patient with pathologically documented prostate cancer and elevated PSA 
after definitive local therapy with radiographic evidence of metastases, pathologic confirmation 
of metastasis disease is not required.  NIH review of pathologic material will be completed in all 
patients enrolled. 
2.3 
REGISTRATION PROCEDURES 
Authorized staff must register an eligible candidate with NCI Central Registration Office (CRO) 
within 24 hours of signing consent. A registration Eligibility Checklist from the web site 
(http://camp.nci.nih.gov/ccr/welcome.htm) must be completed and faxed to 301-480-0757. After 
confirmation of eligibility at Central Registration Office, CRO staff will call pharmacy to advise 
them of the acceptance of the patient on the protocol prior to the release of any investigational 
agents. Verification of Registration will be forwarded electronically via e-mail to the research 
team. A recorder is available during non-working hours.  


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3 
STUDY IMPLEMENTATION 
3.1 
STUDY DESIGN:   
This is a Phase I trial of ECI301 delivered concurrently with palliative radiotherapy.  Radiation 
will be delivered at a dose of 3 Gy per daily fraction, Monday through Friday, to a total dose of 
30 Gy delivered in two weeks.  As many as two extracranial sites may be treated with 
radiotherapy simultaneously on this protocol.  ECI301 will be delivered as an IV infusion during 
week 2 on the day of irradiation approximately one to two hours after the radiation treatment.  
Escalation will begin at the 25 μg/kg dose level and will proceed in cohorts of 3-6 patients.  The 
MTD cohort will be expanded to six patients.  Optional tumor biopsies will be obtained prior to 
the start of therapy and at one week after completion of radiation.  Patients will be followed until 
6 months after completion of treatment to evaluate response. 
3.1.1 Dose Limiting Toxicity 
• Toxicity for the purposes of dose escalation will be assessed during the active 
treatment phase and for two weeks after the completion of therapy.  Toxicity will be 
graded according to the National Cancer Institute CTCAE (version 4.0) and the 
RTOG acute toxicity scoring scale.  
• Dose-limiting toxicity (DLT) will be defined as follows:  
o Any grade 3 or greater non-hematologic toxicity is a DLT 
o Any grade 3 neutropenia or thrombocytopenia is a DLT 
o Any grade 4 anemia is a DLT 
o Nausea, vomiting, diarrhea, tumor pain, or pre-existing hyponatremia, 
dyselectrolytemia, or orthostatic hypotension has been optimally treated with 
anti-emetics, anti-diarrheal, analgesics, or hydration and which persists for 
over 48 hours despite maximal medical therapy is considered a DLT 
o Toxicity requiring a cumulative radiation treatment delay of 4 or more days 
will be considered a DLT.   
o In addition, grade 3 or higher acute RTOG toxicity will be considered a dose-
limiting toxicity (Appendix I).   
• Grade 3 or higher RTOG toxicities attributable to disease progression or pre-existing 
medical condition will not be considered a DLT.   
• No additional study drug will be delivered after a DLT.   
• Toxicity will be recorded with the RTOG Scoring Scale in patients undergoing follow 
up at time points of weeks 3, 4, 6 and 12 weeks (acute scoring scale – Appendix I) 
and 6 months (late scoring scale - Appendix II). 
3.1.2 Dose Escalation   
Radiation will begin on day 1 for all dose regimens.  ECI301 will be delivered daily 
approximately one to two hours after irradiation (week 2).  Every effort will be made to have the 
time of day of these treatments be consistent.  
Dose escalation will begin at 25 µg/kg daily. 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 21 of 60 
 
Dose Escalation Schedule  
Dose Level 
Daily Dose of ECI301 
Level 1 
25 μg/kg 
Level 2 
50 μg/kg 
Level 3 
82.5 μg/kg 
Level 4 
125 μg/kg 
Level 5 
175 μg/kg 
Dose escalation will proceed in cohorts of 3–6 patients based on the rules outlined in the table 
below.  DLT will be evaluated during the active treatment phase and in the first three weeks after 
treatment.  Dose escalation will not proceed until all patients have concluded 2 weeks of follow 
up in the prior dose level to allow complete documentation of DLTs.  The MTD or 
recommended Phase II dose is the highest dose level at which no more than 1 of 6 patients 
experience DLT during treatment or the first three weeks after treatment, and the dose below that 
at which at least 2 (of ≤6) patients have DLT as a result of the drug.  If a patient did not 
experience DLT and did not finish treatment, he or she will not be evaluable for toxicity and will 
be replaced in the study. 
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. 


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Version Date: November 1, 2011 
Page 22 of 60 
3.2 
STUDY DRUG ADMINISTRATION: 
ECI301 will be prepared by the Clinical Center Pharmacy and provided to the Day Hospital or 
inpatient unit.  The pharmacy will provide a single syringe of ECI301 appropriate for 
intravenous bolus infusion.  ECI301 will be delivered as a daily infusion approximately one to 
two hours after irradiation during week 2, Monday through Friday. The infusion will be 
delivered in the Clinical Center Day Hospital or inpatient unit.  The infusion will be delivered 
after completion of radiation.  Every effort will be made to have the time of day of these 
treatments and sample collections be consistent. The bolus infusion will be administered at a rate 
of approximately 1 mL/second. Vital signs (temperature, heart rate, blood pressure, respirations 
and pulse oximetry) will be assessed on Week 2 Day 1 prior to infusion and at 15 minutes, 30 
minutes, 1, 2, 4, 6, and 8 hours following the dose. On subsequent days, Week 2, Days 2-5, vital 
signs will be assessed prior to ECI301 infusion and an hour following.  
There is no required or recommended premedication prior to infusion.  
3.3 
DOSE MODIFICATION FOR TOXICITIES: 
There will be no dose modifications for toxicity.  If a patient experiences a DLT they will be 
removed from the study drug but will continue to receive radiotherapy as deemed appropriate by 
the PI. 
3.4 
PROTOCOL THERAPY AND EVALUATION 
3.4.1 Pretreatment studies (Baseline) 
(to be obtained within one week of initiation of protocol therapy unless otherwise noted) 
• Clinical evaluation, vital signs 
• Laboratory:  if done at screening, repeat only if greater than a week has elapsed 
prior to start of treatment. 
• CBC with differential, lymphocyte phenotype TBNK, serum chemistry with 
electrolytes, BUN, Creatinine, liver function tests, PT, PTT, Urinalysis 
• Adverse Event/Toxicity assessment 
• Optional biopsy 
• Research laboratory studies. Every effort will be made to have the time of day of 
these collections be consistent. 
o 6 mL plasma EDTA tube 
o three 8 mL CPT tubes  
3.4.2 On treatment studies and therapy 
• Clinical evaluation, vital signs, assessment of adverse events weekly (CTC and 
acute RTOG) 
• Laboratory:  CBC with differential, lymphocyte phenotype TBNK, serum 
chemistry with electrolytes, BUN, Creatinine, liver function tests, PT, PTT, 
(Week 1Day 4 or 5; Week 2: Day 1 and 5) 
•  Urinalysis Week 2 Day 1 and 5 
• Electrocardiogram Week 2 Day1 and Day 5 approximately 45 minutes after 
ECI301 
• Radiotherapy:  Radiotherapy will be delivered M-F week 1 and week 2, as 
described in section 3.6.  


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 23 of 60 
• ECI301 will be delivered as a bolus infusion daily, approximately one to two 
hours after irradiation during week 2, as described in 3.2.   
• 
Vital signs (temperature, heart rate, blood pressure, respirations, pulse oximetry) 
will be assessed on Week 2 Day 1 prior to ECI301 infusion and at 15 minutes, 30 
minutes, 1, 2, 4, 6, and 8 hours following the dose. On subsequent days, Week 2, 
Days 2-5, vital signs will be assessed prior to ECI301 infusion and an hour 
following.  
• Research laboratory studies (Week 1: Day 4 or 5; Week 2 Day 1 and Day 5). 
Every effort will be made to have the time of day of these collections be 
consistent.   
o 6 mL plasma EDTA tube  
o three 8 mL CPT tubes (NOT needed on Week 2 Day 1) 
• Pharmacokinetics:  Week 2, Day 1and Day 4 at time 0, 15 minutes, 30 minutes, 1, 
2, 4, 6, 8, and 24 hours after the last dose of ECI301.   Plasma will be drawn in 3 
mL EDTA tubes Follow-up studies 
3.4.3 Follow-up studies 
• Week 3 and 4 (week 1 and 2 after completion of therapy) 
o Clinical evaluation and vital signs, assessment of adverse events(CTC and 
acute RTOG) 
o Laboratory:  CBC with differential, lymphocyte phenotype TBNK, serum 
chemistry with electrolytes, BUN, Creatinine, liver function tests , PT, PTT, 
urinalysis 
o Optional biopsy (week 3 only) 
o Research laboratory. Every effort will be made to have the time of day of 
these collections be consistent:   
o 6 mL plasma EDTA tube  
o three 8 mL CPT tubes (Not needed on Week 4). 
• Week 6, week 12, and 6 months 
o Clinical evaluation and vital signs,  RTOG toxicity scoring (acute scoring at 6 
weeks and 12 weeks, late scoring at 6 months 
o Laboratory:  CBC with differential, serum chemistry with electrolytes, BUN, 
Creatinine, liver function tests 
o Research labs (at week 6 only). Every effort will be made to have the time of 
day of these collections be consistent. 
o 6 mL plasma EDTA tube  
o 24 hour urine collection for creatinine clearance and protein (week 6 only). 
o Radiographic evaluation of the treated site as appropriate for response 
assessment, (CT or MR the same modality as used for pretreatment 
measurements) 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 24 of 60 
3.5 
STUDY CALENDAR 
Procedure 
Screening/ 
Baseline 
Active Treatment 
Post Therapy Follow-up 
Week 1 
Week 2 
Week 3 
Week 4 
Week 6 
Week 12 
Month 6 
History and PE 
X1 
 
 
 
 
 
 
 
Clinical evaluation and Vital signs 
X 
X 
X 
X 
X 
X 
X 
X 
Performance Score 
X 
 
 
 
 
 
 
 
Pathology Review 
X1 
 
 
 
 
 
 
 
Radiation 
 
X 
X 
 
 
 
 
 
ECI301 
 
 
X 
 
 
 
 
 
Radiological/ Response Assessments 
X1 
 
 
 
 
X 
X 
X 
Adverse Events/Toxicity 
Assessment5 
X 
X 
X 
X 
X 
X 
X 
X 
Concomitant Medications 
X 
X 
X 
X 
X 
X 
X 
X 
ECG 
X 
 
X3 X3 
 
 
 
 
 
Clinical Labs: 
CBC with diff, Acute Care, Hepatic 
& Mineral Panels, LDH, uric acid, 
PT/PTT 
 
X 
 
X2 
 
X2 X2 
 
X 
 
X 
 
X 
 
X 
 
X 
lymphocyte phenotype TBNK 
X 
X2 
X2X2 
X 
X 
 
 
 
urine HcG 
X1 
 
 
 
 
 
 
 
24 hour urine creatinine/protein 
X1 
 
 
 
 
X  
 
 
Urinalysis 
X 
 
X 
X 
X 
 
 
 
HIV, Hep B, Hep C screening 
X1 
 
 
 
 
 
 
 
Research Labs: 
three 8mL CPT tubes   
6 mL EDTA tube 
 
X 
X 
 
X 
X 
 
X4 
XX 
 
X 
X 
 
 
X4 
 
 
X4 
 
 
Pharmakokinetics6 
 
 
X6 X6 
 
 
 
 
 
Optional Biopsies 
X 
 
 
X 
 
 
 
 
1. 
Studies that are required at screening only (i.e., If done at screening, do not need to be repeated at baseline) 
2. 
Clinical laboratory tests: Week 1: Day 4 or 5 and Week 2: Day 1 and 5.  
3. 
ECG within 1 week baseline, Week 2 Day 1  and Day 5 approximately 45 minutes after ECI301 
4. 
Research labs Week 2 Day 1 EDTA tube only; Week 2 Day 5 EDTA and three CPT tubes; Weeks 4and 6 EDTA only 
5. 
CTC and RTOG until week 4, then RTOG only. Use RTOG acute through 12 weeks, then RTOG late. 
6. 
Pharmacokinetic analysis will be performed on Week 2, Day 1 and Day 4 at time 0, 15 minutes, 30 minutes, 1, 2, 4, 6, 8, and 24 hours after the last dose of ECI301.   


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 25 of 60 
3.6 
RADIATION THERAPY GUIDELINES: 
3.6.1 Simulation. 
All patients will undergo CT simulation in the Radiation Oncology Branch.  Positioning will be 
optimized to allow maximal sparing of normal tissues from the treatment field.    
3.6.2 Treatment Planning. 
The radiated target volume will be defined using the simulation CT with fusion of additional 
imaging (i.e. diagnostic CT, PET, MRI) as needed for target delineation.  The radiated target will 
be chosen based on a clinical need for treatment.  The gross tumor volume (GTV) will be defined 
as all evident tumor in the radiation target site on clinical examination and radiographic imaging.  
The clinical target volume (CTV) will be defined as the GTV with a 1 cm margin with 
corrections in margin allowed for normal barriers to tumor spread (i.e. uninvolved bone or skin).  
Maximum planning target volume (PTV) margin on CTV will be limited to 1.5 cm with a 
minimum of 0.5 cm. Three dimensional planning will be used to minimize exposure of 
surrounding normal tissues.  Planning will be optimized such that the planning target volume 
will be encompassed within the 95% isodose line.  Every attempt will be made to ensure that the 
global maximum dose should not exceed 115% of the prescription dose.  Treatment will be 
delivered with 6MV or greater energy photons or 6MeV or higher electrons.  Modifications to 
the treatment volumes may be made with the approval of the Principal Investigator or Lead 
Associate Investigator if there is concern that the tumor is inadequately treated or if a normal 
tissue receives what is considered to be an unsafe dose. 
3.6.3 Treatment delivery. 
External beam radiation therapy will be administered as 30 Gy delivered over two weeks in 3 Gy 
daily fractions (Monday through Friday with the exception of federal holidays) in the Radiation 
Oncology Branch, NCI. Every effort will be made to have the time of day of these treatments be 
consistent.  
3.6.4 Criteria for Removal from Protocol Therapy 
• Dose limiting toxicity 
• Concurrent illness that in the judgment of the PI would make further protocol therapy 
unsafe or inappropriate. 
• Progression of disease to the extent that in the judgment of the PI continuation of 
radiotherapy is inappropriate. 
• Patient refusal of further protocol therapy. 
• Death 
3.6.5 Off Study criteria 
• Completion of protocol therapy and 6 months of follow up. 
• Patient withdrawal of consent for further follow up. 
• Death 
Authorized staff must notify Central Registration Office (CRO) when a subject is taken off-
study. An off-study form from the web site (http://camp.nci.nih.gov/ccr/welcome.htm) main 
page must be completed and faxed to 301-480-0757. 


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Version Date: November 1, 2011 
Page 26 of 60 
4 
SUPPORTIVE CARE 
Any medication that is considered necessary for the patient's safety and well-being, eg, anti-
emetics, opioids, may be given at the discretion of the investigator(s) with the exceptions noted 
below.  Concurrent chemotherapy and concurrent targeted agents may not be delivered to 
patients receiving therapy on this protocol.  Patients may continue to use endocrine therapy 
during protocol therapy (for example androgen suppression or estrogen modulators with the 
exception of tamoxifen).  No other investigational agents, radiotherapy, or chemotherapy may be 
prescribed during protocol therapy or during the three weeks of follow up after therapy.  Growth 
factors should be avoided during active therapy.  The patient must be instructed that no 
additional medication will be allowed without the prior consent of the investigator.  The 
administration of all medication (including investigational products, prescription, non-
prescription or over-the-counter medication) must be recorded in the appropriate sections of the 
patient’s medical record.  Concomitant use of anti-inflammatory drugs including NSAIDs and 
steroids should also be avoided, unless clinically essential.  Concurrent use of 
immunosuppressive medications should be avoided (Appendix III).  Concomitant medications 
should be re-assessed on each weekly on treatment visit and during each post-treatment visit.  
Vaccinations should be avoided during active therapy and during the first four weeks of follow 
up. 
5 
BIOSPECIMEN COLLECTION 
5.1 
CORRELATIVE STUDIES FOR RESEARCH/PHARMACOKINETIC STUDIES: 
Plasma (6 cc EDTAtube) will be collected for cytokine analysis pre-treatment, and on Week 1: 
Day 4 or 5, Week 2: Day 1 and Day 5 and Week 3, week 4 and week6..  Every effort will be 
made to have the time of day of these collections be consistent. Plasma will be transported 
immediately on ice to the laboratory of Dr. Deborah Citrin for processing.  Aliquots of plasma 
will be analyzed for cytokines thought to be involved in the dendritic cell natural killer pathway. 
1 mL aliquots of plasma will be sent to Huntingdon Life Sciences for evaluation of anti-MIP1α 
antibodies.   Samples will be stored at -80°C in Dr. Citrin’s laboratory until complete validation 
of the assay is available 
Pharmacokinetic analysis will be performed on Week 2, Day 1 and Day 4 at time 0, 15 minutes, 
30 minutes, 1, 2, 4, 6, 8, and 24 hours after the last  dose of ECI301. PK samples will be 
collected in 3ml EDTA tubes and transported immediately on ice to the laboratory of Dr. 
Deborah Citrin for processing. 1 mL aliquots of plasma will be sent to Huntingdon Life Sciences 
for PK analysis. Samples will be stored at -80°C in Dr. Citrin’s laboratory until complete 
validation of the assay is available. 
Circulating peripheral mononuclear cells will be collected in 3 CPT tubes (8 ml per tube) pre-
treatment, and on Week 1: Day 4 or 5, Week 2: Day 5 and Week 3. .  CPT tubes will be 
transported immediately on ice to the laboratory of Dr. Deborah Citrin for processing..  
Lymphocytes will be divided into aliquots for cryopreservation and fixation for additional 
studies that will include assessment of dendritic cell precursors and assessment of IFNγ 
secretion.  Aliquots of lymphocytes will be provided to Dr. Jane Trepel for flow cytometry based 
studies. 
Optional core tumor biopsies will be obtained prior to treatment and one week after completion 
of radiation.  These biopsies will be obtained from the target lesion (the radiated metastatic site) 
pretreatment, which will serve as a baseline measurement for all sites.  A core biopsy will be 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 27 of 60 
obtained from both the radiated target lesion and a separate non-irradiated lesion at the one week 
follow up time point.  Biopsies will be performed in collaboration with interventional radiology.  
Standard techniques will be used for percutaneous biopsies which may include CT and / or 
ultrasound guided biopsy. In some cases, such biopsies may be expedited and facilitated with the 
use of navigation tools, such as a laser guide or GPS-electromagnetic tracking of the needle or 
ultrasound, to determine which exact angle the biopsy needle will be inserted. 
Following collection, specimens will be snap frozen, transported to the laboratory of Dr. 
Deborah Citrin, and stored at -80º C until use. 
Deborah Citrin, MD 
Bldg 10, Rm 3B42 
Bethesda, MD 20892 
301-496-5457 
Core biopsies will be evaluated for inflammatory cell infiltration via immunohistochemistry.  
Core biopsies will be processed by the laboratory of pathology and paraffin embedded.  The 
panel of markers tested may include CD3, CD4, CD8, and CD20.  Depending on the number of 
lymphocytes observed, additional studies may include markers such as CD56, TIA, and 
granzyme.  Biopsy studies will be coordinated with Dr. Richard Lee in the department of 
Dermatopathology and Surgical Pathology. 
5.2 
SAMPLE STORAGE, TRACKING, AND DISPOSITION 
• 
Samples will be ordered and tracked through the CRIS Screens.  Should a CRIS screen 
not be available, the NIH form 2803-1 will be completed and will accompany the 
specimen and be filed in the medical record.  Samples will not be sent outside NIH 
without IRB notification and an executed MTA. 
• 
In the lab, all samples will be barcoded, with data entered and stored in the Labmatrix 
data base. This is a secure program, with access to the Labmatrix data base limited to 
defined personnel, who are issued individual user accounts. The program creates a 
unique barcode ID for every sample and sample box, which cannot be traced back to 
patients without Labmatrix data base access.  
• 
The data recorded for each sample includes the patient ID, name, trial name/protocol 
number, time drawn, cycle time point, dose, material type, as well as box and freezer 
location. Patient demographics associated with the clinical center patient number are 
provided in the system. For each sample, there are notes associated with the processing 
method (delay in sample processing, storage conditions on the ward, etc.). 
• 
Barcoded samples are stored in barcoded boxes in a locked freezer at either -20 or -
80°C according to stability requirements.  These freezers are located onsite in the ROB 
lab area of NIH Building 10, B2.  
• 
Samples will be stored until requested by a researcher named on the protocol. All 
requests are monitored and tracked in the Labmatrix data base and are only to be used 
for research purposes associated with this trial (as per the IRB approved protocol) and 
any unused samples must be returned to the ROB. Plasma, serum, urine and blister 
fluid samples collected in the course of this research project may be banked and used in 
the future to investigate new scientific questions related to this study.  However, this 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 28 of 60 
research may only be done if the risks of the new questions were covered in the consent 
document.  
• No germline mutation testing will be performed on any of the samples collected.   
• At the completion of the protocol, the investigator will dispose of all specimens in 
accordance with the environmental protection laws, regulations and guidelines of the 
Federal Government and the State of Maryland. 
• Any loss or unintentional destruction of the samples will be reported to the IRB. 
• Any new use of the samples will require prospective IRB review and approval. 
6 
DATA COLLECTION AND EVALUATION 
6.1 
DATA COLLECTION 
• Clinical data and acquired images will be recorded in a NCI CCR database (C3D). 
• Clinical data collection for participants receiving radiation therapy will include: 
demographic information, pathologic diagnosis, clinical stage, history including prior and 
concurrent therapies, CT reports, lab reports, vital signs, dose of radiation delivered, and 
toxicity assessment. 
• The results of any procedures and/or tests will be included in the patients’ hospital chart 
and/or research records as appropriate.  Data from these records may be used in research 
and scientific publications. 
• Data collected may be used anonymously, for publications not originally specified, 
concerning the natural history of disease processes and long term effects of treatment.  
The data may also be used as the basis for entirely new protocols. 
6.2 
MEASUREMENT OF EFFECT AND RESPONSE CRITERIA 
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 
at 6 and 12 weeks and 6 months.  Lesions treated with radiotherapy will be considered radiated 
target lesions.  As many as two sites of metastatic disease may receive radiotherapy 
simultaneously.  Additional metastatic sites (non-irradiated target lesions), up to 3 additional 
lesions, will be followed for evidence of response. 
Patients who have received complete therapy with ECI301 and radiation will be considered 
evaluable for response.  Patients with objective disease progression prior to completion of 
therapy will also be considered evaluable for response rate.  Target (irradiated and non-
irradiated) lesion measurements will be recorded at each assessment time point for each patient 
and response will be determined.  An analysis of response rate based on a subset of patients must 
explain which patients were excluded and for which reasons.   
6.2.1 Antitumor Effect – Solid Tumors 
Response and progression will be evaluated in this study using the Revised Response Evaluation 
Criteria in Solid Tumors (RECIST) Committee [European Journal Cancer.  45(2):228-247, 
2009].  Changes in only the largest diameter (unidimensional measurement) of the tumor lesions 
are used in the RECIST criteria. 
6.2.1.1 
Definitions 


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Version Date: November 1, 2011 
Page 29 of 60 
Evaluable for
Evaluable 
 toxicity.  All patients will be evaluable for toxicity from the time of their first 
treatment with ECI301. 
for
6.2.1.2 
Disease Parameters 
 objective response.  Only those patients who have measurable disease present at 
baseline, have received a complete treatment, and have had their disease re-evaluated will be 
considered evaluable for response.  These patients will have their response classified according 
to the definitions stated below.  (Note:  Patients who exhibit objective disease progression prior 
to the end of treatment will also be considered evaluable.) 
Measurable disease.   
Tumor lesions: Must be accurately measured in at least one dimension (longest diameter in the 
plane of measurement is to be recorded) with a minimum size of: 
• 10mm by CT scan (CT scan slice thickness no greater than 5 mm. 
• 10mm caliper measurement by clinical exam (lesions which cannot be accurately measured 
with calipers should be recorded as non-measurable). 
• 20mm by chest X-ray. 
Malignant lymph nodes: To be considered pathologically enlarged and measurable, a lymph 
node must be >15mm in short axis when assessed by CT scan (CT scan slice 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, including small lesions (longest diameter <10mm or 
pathological lymph nodes with P10 to <15mm short axis) as well as truly non-measurable 
lesions. Lesions considered truly non-measurable include: leptomeningeal disease, ascites, 
pleural or pericardial effusion, inflammatory breast disease, lymphangitic involvement of skin or 
lung, abdominal masses/abdominal organomegaly identified by physical exam that is not 
measurable by reproducible imaging techniques. 
Special considerations regarding lesion measurability Bone lesions, cystic lesions, and lesions 
previously treated with local therapy: 
Bone lesions: 
• 
Bone scan, PET scan or plain films are not considered adequate imaging techniques to 
measure bone lesions. However, these techniques can be used to confirm the presence or 
disappearance of bone lesions. 
• 
Lytic bone lesions or mixed lytic-blastic lesions, with identifiable soft tissue components, 
that can be evaluated by cross sectional imaging techniques such as CTor MRI can be 
considered as measurable lesions if the soft tissue component meets the definition of 
measurability described above. 
• 
Blastic bone lesions are non-measurable. 
Cystic lesions: 
• 
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 noncystic 
lesions are present in the same patient, these are preferred for selection as target lesions. 
Lesions with prior local treatment: 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 30 of 60 
• 
Tumor lesions situated in a previously irradiated area, or in an area subjected to other loco-
regional therapy, are not considered measurable unless there has been demonstrated 
progression in the lesion.  
Target lesions.   
For the purposes of this protocol, lesions will be considered a radiated target lesion if they are 
encompassed in the radiation treatment field.  Up to two sites will be radiated and considered 
evaluable.  When more than one measurable lesion is present at baseline all non-irradiated 
lesions up to a maximum of three lesions total (and a maximum of two lesions per organ/site) 
should be identified as unirradiated target lesions and will be recorded and measured at baseline. 
Radiated target lesions are chosen based on clinical need for treatment.  Non-irradiated target 
lesions for measurement 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.  
Lymph nodes merit special mention since they are normal anatomical structures which may be 
visible by imaging even if not involved by tumor. Pathological nodes which are defined as 
measurable and may be identified as target lesions must meet the criterion of a short axis of 
>15mm by CT scan. Only the short axis of these nodes will contribute to the baseline sum. The 
short axis of the node is the diameter normally used by radiologists to judge if a node is involved 
by solid tumor. Nodal size is normally reported as two dimensions in the plane in which the 
image is obtained (for CT scan this is almost always the axial plane; for MRI the plane of 
acquisition may be axial, saggital or coronal). The smaller of these measures is the short axis. All 
other pathological nodes (those with short axis >10mm but <15 mm) should be considered non-
target lesions. Nodes that have a short axis <10mm are considered non-pathological and should 
not be recorded or followed. 
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 as noted above, 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. 
All other lesions (or sites of disease) including pathological lymph nodes should be identified as 
non-target lesions and should also be recorded at baseline. Measurements are not required and 
these lesions should be followed as ‘present’, ‘absent’, or in rare cases ‘unequivocal progression’ 
(more details to follow). In addition, it is possible to record multiple nontarget lesions involving 
the same organ as a single item on the case record form (e.g. ‘multiple enlarged pelvic lymph 
nodes’ or ‘multiple liver metastases’). 
Non-target lesions.  All other lesions (or sites of disease) including pathological lymph nodes 
should be identified as non-target lesions and should also be recorded at baseline.  Non-target 
lesions should be outside the radiated field. Measurements are not required and these lesions 
should be followed as ‘present’, ‘absent’, or in rare cases ‘unequivocal progression’. In addition, 
it is possible to record multiple non-target lesions involving the same organ as a single item on 
the case record form (e.g. ‘multiple enlarged pelvic lymph nodes’ or ‘multiple liver metastases’). 


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Version Date: November 1, 2011 
Page 31 of 60 
6.2.1.3 
Methods for Evaluation of Measurable Disease 
All measurements should be taken and recorded in metric notation using a caliper if clinically 
assessed.  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. 
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 should 
always be done rather than 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 
and >10mm diameter as assessed using calipers (e.g. skin nodules). For the case of skin lesions, 
documentation by color photography including a ruler to estimate the size of the lesion is 
suggested. As noted above, when lesions can be evaluated by both clinical exam and imaging, 
imaging evaluation should be undertaken since it is more objective and may also be reviewed at 
the end of the study. 
Chest x-ray  Chest CT is preferred over chest X-ray, particularly when progression is an 
important endpoint, since CT is more sensitive than X-ray, particularly in identifying new 
lesions. However, lesions on chest X-ray may be considered measurable if they are clearly 
defined and surrounded by aerated lung.  
Conventional CT and MRI  CT is the best currently available and reproducible method to 
measure lesions selected for response assessment. This guideline has defined measurability of 
lesions on CT scan based on the assumption that CT slice thickness is 5mm or less. When 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).  
Ultrasound (US)  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. 
Endoscopy, Laparoscopy  The utilization of these techniques for objective tumor evaluation is 
not advised. However, they can be useful to confirm complete pathological response when 
biopsies are obtained or to determine relapse in trials where recurrence following complete 
response or surgical resection is an endpoint. 
Tumor markers  Tumor markers alone cannot be used to assess objective tumor response. If 
markers are initially above the upper normal limit, however, they must normalize for a patient to 
be considered in complete response. Because tumor markers are disease specific, instructions for 
their measurement should be incorporated into protocols on a disease specific basis.  Tumor 
markers will not be used to assess response on this protocol. 
Cytology, Histology  These techniques can be used to differentiate between PR and CR in rare 
cases if required by protocol (for example, residual lesions in tumor types such as germ cell 
tumors, where known residual benign tumors can remain). When effusions are known to be a 
potential adverse effect of treatment, the cytological confirmation of the neoplastic origin of any 


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Page 32 of 60 
effusion that appears or worsens during treatment can be considered if the measurable tumor has 
met criteria for response or stable disease in order to differentiate between response (or stable 
disease) and progressive disease. 
6.2.1.4 
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 diameters of target lesions, taking 
as reference the baseline sum diameters. 
Progressive Disease (PD): At least a 20% increase in the sum of 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 progression). 
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. 
6.2.1.5 
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 (<10mm short axis). 
Incomplete Response/ 
Stable Disease (SD): Persistence of one or more non-target lesion(s) and/or maintenance of 
tumor marker level above the normal limits 
Progressive Disease (PD): 
Unequivocal progression of existing non-target lesions. (Note: the 
appearance of one or more new lesions is also considered progression). 
6.2.1.6 
Evaluation of Best Overall Response 
The best overall response is the best response recorded from the start of the study treatment until 
the end of measurements (up to 6 months after therapy) taking into account any requirement for 
confirmation.  The patient's best response assignment will depend on the achievement of both 
measurement and confirmation criteria. 
Target Lesions 
Non-Target 
Lesions 
New Lesions 
Overall 
Response 
CR 
CR 
No 
CR 
CR 
Non-CR/Non-PD 
No 
PR 
CR 
Not-evaluated 
No 
PR 
PR 
Non-PD or not all 
evaluated 
No 
PR 
SD 
Non-PD or not all 
evaluated 
No 
SD 
Not all evaluated 
Non-PD 
No 
NE 


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Target Lesions 
Non-Target 
Lesions 
New Lesions 
Overall 
Response 
PD 
Any 
Yes or No 
PD 
Any 
PD 
Yes or No 
PD 
Any 
Any 
Yes 
PD 
CR=complete response, PR= partial response, SD=stable disease, 
PD=progressive disease, and NE=not evaluable 
6.2.1.7 
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 recurrent 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.  
6.2.1.8 
Progression-Free Survival 
PFS is defined as the duration of time from start of treatment to time of progression. 
6.3 
TOXICITY CRITERIA 
The following adverse event management guidelines are intended to ensure the safety of each 
patient while on the study. Adverse events occurring during the study will be graded according to 
the NCI Common Terminology Criteria for Adverse Events version 4.0 (CTCAE v4.0) found at: 
http://ctep.cancer.gov/reporting/ctc_v30.html.  In addition, the study will use the RTOG/EORTC 
Morbidity Scoring Scheme to score acute and late effects of radiation treatment.  These criteria 
are available from the RTOG website http://rtog.org/members/toxicity/main.html 
7 
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN 
7.1 
DEFINITIONS 
7.1.1 Adverse Event 
An adverse event is defined as any reaction, side effect, or untoward event that occurs during the 
course of the clinical trial associated with the use of a drug in humans, whether or not the event 
is considered related to the treatment or clinically significant. For this study, AEs will include 
events reported by the patient, as well as clinically significant abnormal findings on physical 
examination or laboratory evaluation. A new illness, symptom, sign or clinically significant 
laboratory abnormality or worsening of a pre-existing condition or abnormality is considered an 
AE.  All AEs must be recorded on the AE case report form. 
 
All AEs, including clinically significant abnormal findings on laboratory evaluations, regardless 
of severity, will be followed until satisfactory resolution. AEs should be reported up to 30 days 
following the last dose of study drug.  AEs that are considered treatment related, expected, 


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Version Date: November 1, 2011 
Page 34 of 60 
continuing, but not resolvable by 30 days after treatment completion (e.g., alopecia) will not be 
followed after the 30-day period.  
 
An abnormal laboratory value will be considered an AE if the laboratory abnormality is 
characterized by any of the following: 
• 
Results in discontinuation from the study 
• 
Is associated with clinical signs or symptoms  
• 
Requires treatment or any other therapeutic intervention 
• 
Is associated with death or another serious adverse event, including hospitalization.  
• 
Is judged by the Investigator to be of significant clinical impact 
• 
If any abnormal laboratory result is considered clinically significant, the investigator will 
provide details about the action taken with respect to the test drug and about the patient’s 
outcome. 
7.1.2 Attribution:  
An assessment of the relationship between the AE and the medical intervention.  CTCAE does 
not define an AE as necessarily “caused by a therapeutic intervention”.  After naming and 
grading the event, the clinical investigator must assign an attribution to the AE using the 
following attribution categories:  
RELATIONSHIP  
ATTRIBUTION  
DESCRIPTION  
Unrelated to investigational 
agent/intervention1 
Unrelated 
The AE is clearly NOT 
related to the intervention 
Unlikely 
The AE is doubtfully related 
to the intervention 
Related to investigational 
agent/intervention1 
Possible 
The AE may be related to the 
intervention  
Probable 
The AE is likely related to the 
intervention 
Definite 
The AE is clearly related to 
the intervention 
1NOTE: AEs listed as “possibly, probably, or definitely” related to the investigational 
agent/intervention are considered to have a suspected “reasonable causal relationship” to 
the investigational agent/intervention (ICH E2A). 
7.1.3 Suspected adverse reaction 
Suspected adverse reaction means any adverse event for which there is a reasonable possibility 
that the drug caused the adverse event. For the purposes of IND safety reporting, ‘reasonable 
possibility’ means there is evidence to suggest a causal relationship between the drug and the 
adverse event. A suspected adverse reaction implies a lesser degree of certainty about causality 
than adverse reaction, which means any adverse event caused by a drug. 
7.1.4 Unexpected adverse reaction 
An adverse event or suspected adverse reaction is considered “unexpected” if it is not listed in 
the investigator brochure or is not listed at the specificity or severity that has been observed; or, 
if an investigator brochure is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current application. 
"Unexpected”, also refers to adverse events or suspected adverse reactions that are mentioned in 


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Version Date: November 1, 2011 
Page 35 of 60 
the investigator brochure as occurring with a class of drugs or as anticipated from the 
pharmacological properties of the drug, but are not specifically mentioned as occurring with the 
particular drug under investigation. 
7.1.5 Serious  
An adverse event or suspected adverse reaction is considered serious if in the view of the 
investigator or the sponsor, it results in any of the following: 
• 
Death 
• 
A life-threatening adverse drug experience 
• 
Inpatient hospitalization or prolongation of existing hospitalization 
• 
Persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions 
• 
A congenital anomaly/birth defect. 
• 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse drug experience when, based upon 
appropriate 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. 
7.1.6 Disability 
A substantial disruption of a person’s ability to conduct normal life functions. 
7.1.7 Life-threatening adverse drug experience 
Any adverse event or suspected adverse reaction that places the patient or subject, in the view of 
the investigator or sponsor, at immediate risk of death from the reaction as it occurred, i.e., it 
does not include a reaction that had it occurred in a more severe form, might have caused death. 
7.1.8 Protocol Deviation (NIH Definition) 
A protocol deviation is any change, divergence, or departure from the study design or procedures 
of a research protocol that is under the investigator’s control and that has not been approved by 
the IRB.  
7.1.9 Protocol Violation (NIH Definition) 
Any change, divergence, or departure from the study procedures in an IRB-approved research 
protocol that has a major impact on the subject’s rights, safety, or well-being and/or the 
completeness, accuracy or reliability of the study data. 
7.1.10 Unanticipated Problem 
Any incident, experience, or outcome that: 
•  Is unexpected in terms of nature, severity, or frequency in relation to  
(a) the research risks that are described in the IRB-approved research protocol and 
informed consent document; Investigator’s Brochure or other study documents, and 
(b) the characteristics of the subject population being studied; AND 
• Is related or possibly related to participation in the research; AND 


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• Places subjects or others at a greater risk of harm (including physical, psychological, 
economic, or social harm) than was previously known or recognized. 
7.2 
NCI-IRB REPORTING 
7.2.1 NCI-IRB Expedited Reporting of Adverse Events, Unanticipated Problems, and Deaths 
The Protocol PI will report to the NCI-IRB: 
• All unexpected serious adverse events that are possibly, probably, or definitely related to 
the research  
• All deaths, except deaths due to progressive disease  
• All Protocol Violations or Deviations 
• All Unanticipated Problems 
Reports must be received by the NCI-IRB within 7 working days of PI awareness via iRIS. 
7.2.2 NCI-IRB Requirements for PI Reporting of Adverse Events at Continuing Review 
The protocol PI will report to the NCI-IRB:  
• 
All Grade 2 unexpected events that are possibly, probably or definitely related to 
the research;  
• 
All Grade 3 and 4 events that are possibly, probably or definitely related to the 
research;  
• 
All Grade 5 events regardless of attribution;  
• 
All Serious Events regardless of attribution.  
NOTE: Grade 1 events are not required to be reported.  
7.2.3 NCI-IRB Reporting of IND Safety Reports 
Only IND Safety Reports that require a sponsor recommended change to the protocol or the 
consent form or in the opinion of the PI increases risks to study participants will need to be 
reported to the NCI IRB. 
7.3 
FDA REPORTING CRITERIA 
7.3.1 IND Safety Reports to the FDA (Refer to 21 CFR 312.32) 
7.3.1.1 
Expedited reporting to the FDA 
The Sponsor will notify FDA  via phone, fax, or email of any unexpected fatal or life-threatening 
suspected adverse reactions as soon as possible but no later than 7 calendar days of initial receipt 
of the information. This will be followed with a written report within 15 days using the 
MedWatch Form 3500a.  
The study Sponsor will notify FDA in writing of any suspected adverse reaction that is both 
serious and unexpected as soon as possible but no later than 15 calendar days after initial receipt 
of the information using the MedWatch Form 3500a. If FDA requests any additional data or 
information, the sponsor must submit it to the FDA as soon as possible, but no later than 15 
calendars days after receiving the request.  
The study Sponsor will also report expeditiously as above: 


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• any findings from clinical, epidemiological, or pooled analysis of multiple studies or any 
findings from animal or in vitro testing that suggest a significant risk in humans exposed 
to the drug 
• clinically important increase in the rate of a serious suspected adverse reaction over that 
listed in the protocol or investigator brochure.  
7.3.1.2 
Exclusions to expedited reporting to the FDA 
The following events will not be reported in an expedited manner but will be included in the 
annual report:  
• Pain due to disease 
• Events associated with vascular access devices not implanted or used on this protocol 
7.3.2 FDA Annual Reports (Refer to 21 CFR 312.33) 
The study Sponsor will submit a brief report annually of the progress of the trial within 60 days 
of the anniversary date that the IND went into effect as indicated in 21CFR 312.33, and any 
associated FDA correspondences regarding the IND annual report.  
7.3.3 Expedited Adverse Event Reporting Criteria to the IND Manufacturer (ECI, Inc) 
The Sponsor will notify ECI, Inc via phone, fax, or email of any unexpected fatal or life-
threatening suspected adverse reactions as soon as possible but no later than 7 calendar days of 
initial receipt of the information. This will be followed with a written report within 15 days.  
The study Sponsor will notify ECI, Inc in writing of any suspected adverse reaction that is both 
serious and unexpected as soon as possible but no later than 15 calendar days after initial receipt 
of the information. 
7.3.3.1 
Exclusions to expedited reporting to the Manufacturer 
The following events will not be reported in an expedited manner but will be included in the 
annual report:  
• Pain due to disease 
• Events associated with vascular access devices not implanted or used on this protocol 
7.4 
DATA AND SAFETY MONITORING PLAN 
7.4.1 Principal Investigator/Research Team  
The clinical research team will meet on a weekly basis when patients are being actively treated 
on the trial to discuss each patient. Decisions about dose level enrollment and dose escalation if 
applicable will be made based on the toxicity data from prior patients.  
All data will be collected in a timely manner and reviewed by the principal investigator or a lead 
associate investigator. Adverse events will be reported as required above. Any safety concerns, 
new information that might affect either the ethical and or scientific conduct of the trial, or 
protocol deviations and violations will be immediately reported to the IRB using iRIS and if 
applicable to the Sponsor.  
The principal investigator will review adverse event and response data on each patient to ensure 
safety and data accuracy. The principal investigator will personally conduct or supervise the 
investigation and provide appropriate delegation of responsibilities to other members of the 
research staff.  


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7.4.2 Sponsor Monitoring Plan 
This trial will be monitored by personnel employed by Harris Technical Services on contract to 
the NCI, NIH. Monitors are qualified by training and experience to monitor the progress of 
clinical trials. Personnel monitoring this study will not be affiliated in any way with the trial 
conduct. 
At least 25% of enrolled patients’ will be randomly selected and monitored at least quarterly, 
base on accrual rate. The patients selected will have 100% source document verification done. 
Additional monitoring activities will include: adherence to protocol specified study eligibility, 
treatment plans, data collection for safety and efficacy, reporting and time frames of adverse 
events to the NCI IRB and FDA, and informed consent requirements. Written reports will be 
generated in response to the monitoring activities and submitted to the Principal investigator and 
Clinical Director or Deputy Clinical Director, CCR, NCI. 
8 
STATISTICAL SECTION 
8.1 
STUDY DESIGN/ENDPOINTS 
Dose escalation of ECI301 will proceed in cohorts of 3-6 patients are started at each dose level.  
DLT will be defined as CTCAE v4.0 any grade 3 or greater non-hematologic toxicity, any grade 
3 neutropenia or thrombocytopenia, any grade 4 anemia and nausea, vomiting, diarrhea, tumor 
pain, or pre-existing hyponatremia, dyselectrolytemia, or orthostatic hypotension has been 
optimally treated with anti-emetics, anti-diarrheal, analgesics, or hydration and which persists for 
over 48 hours despite maximal medical therapy. Toxicity resulting in cumulative radiation 
treatment interruptions of 4 or more days will be considered a DLT.  In addition, grade 3 or 
higher RTOG acute toxicity attributed as being possibly, probably, or definitely related to 
ECI301 is observed, this will be defined as a DLT.  If 0/3 experience first course DLT, the next 
cohort starts one dose level higher.  If 1/3 experience first course DLT, up to 3 more patients are 
started at that same dose level.  If 2 or more experience first course DLT, no further patients are 
started at that dose and the next lower dose level is expanded to a total of 6 patients if only three 
were treated at that dose.  The MTD or recommended Phase II dose is the dose level at which no 
more than 1 of 6 patients experience DLT during treatment and the three weeks after completion. 
8.2 
SAMPLE SIZE/ACCRUAL RATE 
The number of patients required to complete this trial involves 5 cohorts of 3 to 6 patients.  The 
total number of patients that will accrue to this study is therefore between 3 to 30, at a rate of 2 
patients per month. 
No group (except patients with cognitive impairment unable to give informed consent and those 
less than 18 years of age) is being excluded from participation.  Subjects of both genders, from 
all racial and ethnic groups are eligible for this trial.  Efforts will be made to extend accrual to a 
representative population. 
8.3 
ANALYSIS OF SECONDARY ENDPOINTS 
Analysis of inflammatory cell infiltration in tumors, immune cell profiles, and immune cell IFN 
production will be performed.  The statistical analysis will involve only descriptive statistics and 
exploratory graphical representations of the data.  That is, no inferential analysis will be 
conducted.  Exploratory modeling of dose-response relationship in terms of anti-tumor activity 


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Page 39 of 60 
and immunologic changes will be performed. These results of these studies will be for 
descriptive purposes.   
The final PK analyses and evaluation for anti-MIPα antibodies will be the responsibility of ECI, 
Inc. The ECI301 concentration-time data, along with the derived PK parameters, will be listed 
and summarized appropriately.  Additional parameters may be determined if deemed 
appropriate: 
ECI301: Cmax, tmax, AUC(0-12), AUC(0-24), AUC(0-t), Cmin, CL/F 
The PK parameters will be derived using non-compartmental analysis. The maximum plasma 
concentrations (Cmax) and the time to reach the maximum plasma concentrations (tmax) will be 
determined by visual inspection of the plasma concentration-time profiles. The area under the 
plasma concentration-time curve from zero to 8 hours post dose AUC(0-12) and zero to 24 hours 
post-dose AUC(0-24) will be calculated by the linear trapezoidal rule. The total oral clearance 
from plasma (CL/F) after an oral dose will be calculated from the dose divided by the AUC 
during the appropriate dosing interval.   
9 
HUMAN SUBJECTS PROTECTIONS 
9.1 
 RATIONALE 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. For safety reasons, pregnant women and 
children are excluded from this study. 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.) Participants should realize that there is no guarantee of benefit 
to them from participation in this trial. The results of this trial may benefit future cancer patients. 
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. Inclusion of 
Women and Minorities:  This study will be open to all individuals regardless of gender, 
ethnicity, or race provided that the aforementioned inclusion and exclusion criteria are met. 
Pregnant women are excluded from this study because the effects of ECI301 on the developing 
fetus is unknown and because radiation is a known teratogen.  Because there is an unknown but 
potential risk for adverse events in nursing infants secondary to treatment of the mother with 
ECI301, breastfeeding should be discontinued if the mother is treated with ECI301. Participants 
with unstable or serious medical conditions such as uncontrolled diabetes, uncontrolled 
hypertension, symptomatic congestive heart failure, unstable angina pectoris, myocardial 
infarction within the past 6 months, uncontrolled cardiac arrhythmia; or psychiatric illness/social 
situations that would limit compliance with study requirements are excluded due to the 
possibility that the underlying condition may obscure the attribution of effect and adverse events 
and may limit study compliance. HIV-positive patients are ineligible because they harbor an 
incompletely effective immune system which could impair response to ECI301.  Appropriate 
studies will be undertaken in patients receiving combination antiretroviral therapy when 


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Page 40 of 60 
indicated.  Patients with autoimmune diseases or hepatitis are excluded due to the possibility that 
ECI301 could worsen inflammatory processes.   
9.2 
 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 ECI301 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. 
9.3 
EVALUATION OF BENEFITS AND RISKS/DISCOMFORTS: 
Participants will receive treatment with radiotherapy to sites of metastatic disease and the risks of 
this therapy may be increased with the use of ECI301 delivered concurrently as a radiation 
sensitizer.  The risks of radiotherapy to a metastatic site are highly dependent on the area 
receiving treatment with the exception of fatigue and skin reaction.  Patients will be provided 
with a separate consent specific to the area receiving radiotherapy before treatment that will 
detail the specific risks of radiotherapy to that site as is standard practice.  Participants may also 
experience risks from delivery of the therapeutic agent ECI301 relating to the infusion or drug 
reactions.  Infusional risks include line infection and bleeding.  The protocol therapy could 
enhance the effects of radiotherapy and result in more durable palliation.   
In addition to the risks of protocol therapy, participants may undergo optional biopsies.  The 
risks of biopsy are also dependent on the region biopsied and may include bleeding, pain, and 
infection.   
9.4 
RISKS/BENEFITS ANALYSIS: 
This is a first in man study of ECI301.  The side effects of ECI301 infusions in man are 
unknown.  Animal toxicology studies have suggested that ECI301 is considered to have an 
acceptable safety margin at the likely doses to be studied in this Phase I protocol with evidence 
of reduction in circulating monocytes transiently.  Human studies of a similar agent, BB-10010 
showed acute changes in monocyte counts which rapidly resolved and no other observable 
toxicity.  The expected discomforts of infusions of ECI301 are those to be expected from 
everyday infusions:  pain and discomfort from venipuncture.  In this study using ECI301, a 
conservative approach to starting dose has been chosen to minimize patient risk. 
It is possible that ECI301 will enhance tumor sensitivity to radiation or could enhance the 
immune system’s recognition of tumor at unirradiated sites.  This could result in more durable 
palliation and reduction in metastatic burden.  It is also possible that ECI301 could increase the 
effects of radiation on normal tissues that are irradiated by resulting in inflammation.  For this 
reason, we have chosen small margins of additional tissue to include in our treatment volume.  It 
remains a possibility that patients may have a greater degree of acute toxicity from radiotherapy 
than would be expected which could include dermatitis, fatigue, or pain. 
In summary the potential risk: benefit ratio for Phase I studies is considered to be acceptable in 
light of potential benefits, the initial patient eligibility criteria, and proposed monitoring of 
patients. 
Optional biopsies may also result in a risk to the patient.  The risks of biopsy include bleeding, 
infection, pain, and site specific side effects.  Biopsies will only be performed if they are felt to 
be of minimal risk and if the patient consents. 


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9.5 
CONSENT AND ASSENT PROCESS AND DOCUMENTATION: 
• 
The procedures and treatments involved in this protocol, with their attendant risks and 
discomforts, potential benefits, and potential alternative therapies will be carefully 
explained to the treatment subject. A signed, informed consent document will be obtained 
from the subjects by one of the physician investigators.  
• 
Consent forms: The original signed informed consent documents will be kept with patient 
medical records. Central Registration will also retain a copy of the informed consent 
document. A copy of their own signed informed consent documents will also be given to 
study participants.  
• 
Central Registration will ascertain the date of IRB approval before registering the first 
subject.  
• 
The treatment subject informed consent forms contain all elements required for consent. 
In addition, the Principal Investigator, or their designee will obtain oral consent and will 
be available to answer all patient questions.  
10 PHARMACEUTICAL INFORMATION:  
10.1 ECI301 
10.1.1 Source: 
Effector Cell Institute.   
10.1.2 Toxicity 
ECI301 has not been delivered to humans.  Single dosing studies of IV (bolus) administration of 
ECI301 in 5-week-old male ICR mice at a dose of 0 (PBS; non-treated [NT], n=2), 1, 10, or 300 
mg/kg ECI301 (n=3/dose) were completed.  All mice were fasted prior to dosing and were 
observed for 4 hours post dose for signs of general condition and mortality. Body weight 
measurements were taken on Days -1, 0, 1, and 2. Animals were sacrificed 2 days after dosing. 
Parameters evaluated included: hematology for albumin/globulin ratios, blood urea nitrogen, 
creatinine, lactic dehydrogenase, aspartate aminotransaminase, alanine aminotransaminase, and 
inorganic phosphorus of the isolated plasma. Urine was analyzed for glucose, urobilinogen, and 
pH. Histological assessment of brain, lung, liver, kidney, thymus, intestine, and colon was 
conducted. No animals died following administration of single doses of ECI301 up to 300 
mg/kg. There were no significant treatment-related differences in clinical signs, body weights, 
plasma examination, histology, or organ weights in any dose group. Protein levels in urine 
showed minor elevations in the 10-mg/kg and 300-mg/kg groups. There were no other observed 
changes. These results indicate that a single dose of ECI301 of up to 300 mg/kg did not cause 
any overt toxicity during observations for 2 days post-treatment. It was concluded that a single 
dose of ECI301, up to 300 mg/kg, was well tolerated, and therefore the NOAEL was 300 mg/kg 
for this study. [28] 
A single dose study was conducted in accordance with GLP to evaluate the acute toxicity of IV 
administration of ECI301 in rats. A single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.5, or 
5 mg/kg ECI301 was administered IV (bolus) to Crl:CD® (SD) IGS BR rats (5/sex/dose). All 
rats were observed daily for 14 days after dosing for general clinical condition, signs of toxicity, 
and mortality. All animals were subjected to necropsy and macroscopic investigation. An 
additional group of animals was administered a single dose of 0.05, 0.5, or 5 mg/kg ECI301 IV 
for toxicokinetic analysis (6/sex/dose). Animals were bled at 5, 15, and 30 minutes, and 1, 2, and 


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4 hours postdose for toxicokinetic analysis. There were no treatment-related deaths for this 
study. There were no significant treatment-related clinical signs or effects on body weight. Organ 
weights were not affected, and there were no treatment-related macroscopic findings. The 
NOAEL was determined to be greater than the 5-mg/kg ECI301 used in this study. The 
toxicokinetic results indicated that the rate and extent of systemic exposure of rats to ECI301 
appeared to be characterized by nonlinear (dose-dependent) kinetics over the dose range. 
Increasing the dose of ECI301 above 0.05 mg/kg was considered likely to result in a higher 
systemic exposure than would be predicted from a linear relationship, which is consistent with 
the possibility of a capacity-limited process for ECI301 elimination. There were no differences 
between the sexes for systemic exposure. It was concluded that a single dose of ECI301 at 5 
mg/kg did not cause any toxicological effects. [28] 
Repeated dosing studies of ECI301 in 5- to 7-week-old ICR mice were conducted as a non-GLP 
investigation to evaluate the potential toxicity of repeat IV administration. ECI301 was 
administered IV at 0 (PBS; 2/dose), 1, 10, or 100 mg/kg/day (3/dose) for two 5-day dosing 
cycles, split by a 3-day off-dose period. An additional group receiving bovine serum albumin 
(BSA; 100 mg/kg) was added to serve as an additional control group for using a different 
protein. Animals were observed for 4 hours post-dose on Day 3 and Day 13 for signs of general 
condition and mortality. Animals were autopsied 24 hours following the last dose. Parameters 
evaluated included daily body weights, urinalysis, and hematology. Organ weights and histology 
of brain, lung, liver, kidney, thymus gland, and colon tissue were conducted. There was no 
treatment-related mortality. There were no treatment-related effects on body weights or any of 
the hematological parameters measured. The protein in urine increased slightly in 1 of 3 animals 
in the 10-mg/kg group and 1 of 3 animals in the 100-mg/kg group. At 100 mg/kg ECI301, there 
was an increase in lung and spleen weights. Histology of the kidney showed an increase in the 
adhesion of glomerular basement membrane and renal glomeruli, and disruption of the 
glomerulus structure in all animals in the group receiving ECI301 at 100 mg/kg, and 1 of 3 
animals in the 10-mg/kg group. It is unclear if these results are due to administration of ECI301 
or to immune complex deposition. In spleen samples from both BSA and ECI301 (100 mg/kg), 
the boundary between the red and white pulp was unclear. There was structural damage to the 
white pulp, and the central artery was unclear. There were no treatment-related changes in any of 
the other organs or tissues. This experiment demonstrated that repeated doses of ECI301, to a 
maximum total dose of 1000 mg/kg (1000 times the effective dosage), had no critical effect on 
the kidney, but abnormal tissue images and urine protein elevations were observed that indicated 
some renal damage. Further study is required to determine if this damage is attributable to 
ECI301 or another protein. Data from this non-GLP study supported further study of repeat 
doses over 2 to 4 weeks.[28] 
A separate study was conducted in accordance with GLP to evaluate the systemic toxicity and 
toxicokinetics of IV administration of ECI301 alone or in combination with docetaxel in rats for 
three 5-day dosing cycles. A single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.15, or 1.5 
mg/kg/day ECI301, 12.5 mg/kg docetaxel, or a 0.05-/12.5- mg/kg/day combination of ECI301 
and docetaxel, was administered IV (bolus) to Crl: CD® (SD) IGS BR rats daily for 5 days. 
After 51 days, 10 animals/sex/dose were sacrificed and an additional 5 animals/sex/dose were 
sacrificed following a 10-day recovery period. All rats were observed daily after dosing for 
general clinical condition, signs of toxicity, and mortality. Additionally, food consumption, 
ophthalmology, hematology, blood chemistry, urinalysis, anti-ECI301 antibody assessments, 
peripheral blood immunophenotyping, and organ weights were analyzed. All animals were 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 43 of 60 
subject to necropsy with macroscopic and microscopic pathology. An additional group of 
animals for each group (6/sex/dose) except control (3/sex/dose) was bled for 
toxicokineticanalysis of ECI301 and docetaxel plasma levels. Animals were bled at 5, 15, and 30 
minutes, and 1, 2, and 4 hours post dose for toxicokinetic analysis. There were no deaths in this 
study attributable to ECI301 treatment alone. Six animals died following docetaxel treatment. 
There were no treatment-related clinical signs or any effect upon bodyweight and food 
consumption. Similarly, there were no treatment-related ophthalmoscopic findings or any effect 
upon the composition of the urine. The hematological examination indicated a slight increase of 
lymphocyte count after the first treatment cycle, resulting in an increased total leukocyte count, 
in males given 0.15 or 1.5 mg/kg, with similar trends also occurring at all doses in females after 
Cycle 3. This was not evident in the males after Cycle 3 or in the females at the end of the 
recovery period. Biochemical changes in the blood plasma were confined to a reduction of 
glucose concentrations after Cycles 1 and 3 in males given 1.5 mg/kg and after Cycle 3 in males 
given 0.15 mg/kg and a small increase of plasma urea after Cycle 3 in males given 1.5 mg/kg. 
An analysis of organ weights after the completion of the third dosing cycle indicated a small 
reduction of spleen and thymus weight in males given 0.05 mg/kg, which was attributed to 
treatment since this dose was at the maximum biological effect level for ECI301. There were no 
treatment-related macroscopic or histopathological findings. The plasma samples taken from the 
control animals were below the limit of quantification in all cases, demonstrating that there was 
no quantifiable contamination with ECI301 in these animals.[28] 
The toxicokinetic investigations indicated that the extent of systemic exposure of rats to ECI301 
appeared to be characterized by non-linear (dose-dependent) kinetics over the dose range 0.05 to 
1.5 mg/kg/day on Days 1 and 47. Increasing the dose of ECI301 above 0.05 mg/kg/day resulted 
in a higher systemic exposure than would be predicted from a linear relationship, indicative of 
limited elimination capacity. In general, there were no differences in the systemic exposure of 
males and females to ECI301 and there was no accumulation after repeated doses. One female 
given 0.05 mg/kg ECI301 showed a positive anti-ECI301 antibody response. All other animals 
showed negative responses. Immunophenotyping after completion of Cycle 3 indicated, in 
animals given 1.5 mg/kg, an increased percentage of CD4+ T cells and the males showed a 
corresponding decrease in the percentage of CD8+ T cells, although the total numbers for all 
lymphocyte subsets studied were not affected. The numbers of NK lymphocytes were reduced in 
females given 1.5 mg/kg.  Following recovery, males given 1.5 mg/kg still showed a reduced 
percentage of CD4+ lymphocytes and an increased percentage of CD8+ lymphocytes.  It was 
concluded that IV (bolus) administration of ECI301 to CD rats at doses up to 1.5 mg/kg during 
three 5-day cycles of treatment, each separated by 16 days, was well tolerated, was not 
immunogenic, and caused no toxicologically significant findings.  Docetaxel treatment 2 days 
after each cycle in which animals were given ECI301 at 0.05 mg/kg (a pharmacologically active 
dose) at 12.5 mg/kg in the first cycle in both sexes and the second cycle in males, subsequently 
reduced to 6.25 mg/kg for the remaining cycles, caused a range of findings that were attributable 
to its anti-mitotic activity. Compared with animals given docetaxel on the same study days, but 
with no ECI301 treatment, all differences in the response to docetaxel were attributed to normal 
biological variation in response to treatment and, consequently, there was no evidence that 
ECI301 modulated the effects of docetaxel treatment.[28] 
A separate study was conducted to evaluate the potential for systemic toxicity of 3 different 
doses of ECI301, when administered IV to common marmosets for a 5-day dosing period with an 
18-day observation period, to confirm doses to be used in the main toxicity study. Common 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 44 of 60 
marmosets (Callithrix jacchus) received 0.05, 0.15, or 1.5 mg/kg ECI301 (1/sex/group) IV bolus, 
once a day, for 5 days. Animals were observed for 18 days for clinical condition, injection-site 
reactions, and body weight. Hematology, blood chemistries, organ weight, and macroscopic 
pathology investigations were undertaken. There were no treatment-related deaths during this 
study. There were no treatment-related clinical signs of toxicity, and body weights were 
unaffected. There was no effect of treatment on organ weights or macropathology. All animals 
had reduced hematocrit, hemoglobin concentration, and/or erythrocyte count on Day 9 and/or 
Day 22, often accompanied by increased reticulocyte count. Mean cell volume was increased in 
the male receiving 0.05 mg/kg, the female receiving 0.5 mg/kg, and both animals receiving 1.5 
mg/kg. Mean cell hemoglobin concentration was decreased in the male receiving 0.05 mg/kg and 
the female receiving 1.5 mg/kg. Anisocytosis was reported on the blood film for all females. 
There was no strong trend with dosage, and it is possible that these changes may have been 
caused by repeated blood sampling. However, with the absence of control animals on this study, 
the possibility that these changes were due to treatment with ECI301 cannot be excluded. An 
increase in monocyte count occurred on Day 9 in the female marmosets receiving 0.05 mg/kg or 
0.15 mg/kg. These values normalized by Day 22. Minor blood chemistry changes occurred on 
Day 9 and Day 23, but these changes were small and were not dose related in degree. All animals 
showed reduced alkaline phosphatase activity on Day 23. Creatinine concentrations were 
reduced on Day 23 in the males given 0.05 mg/kg and in animals given 0.15 mg/kg or 1.5 mg/kg. 
Total triglyceride concentrations were high on Day 9 and Day 23 for all animals except males 
given 1.5 mg/kg. In the animals given 0.05 mg/kg and the females given 0.15 mg/kg, there was a 
reduction of total cholesterol level. All animals showed an increase of sodium concentration on 
Day 9 and, at 0.15 and 15 mg/kg, this was associated with increased chloride concentration. 
Potassium concentrations were low on Day 9 in all animals, and in the males, this persisted to 
Day 23. Total protein concentrations were low on Day 23 in animals given 0.05 mg/kg or 0.15 
mg/kg, but not in those given 1.5 mg/kg, and most showed an increase in the albumin to globulin 
ratio. It was concluded that the IV administration of ECI301 to common marmosets for a single 
5-day dosing period was well tolerated up to doses of 1.5 mg/kg. There were no toxicologically 
significant findings, and therefore it was concluded that 1.5 mg/kg was a suitable dose for future 
studies in marmosets. [28] 
Study EFC0010 was conducted to evaluate the systemic toxicity and toxicokinetics of IV 
administration of ECI301 to Callithrix jacchus, the common marmoset for three 5-day cycles. A 
single dose of 0 (0.01% polysorbate 80 in PBS), 0.05, 0.15, or 1.5 mg/kg ECI301 was 
administered IV (bolus) to marmosets (3/sex/dose). Radiation with chemotherapy is standard of 
care regimen for treatment of cancer. Docetaxel is one of the common chemotherapy drugs; 
however, significant AEs such as suppression of bone marrow function are associated with the 
administration of docetaxel. In phase 1 clinical study, ECI301 is expected to be administered to 
cancer patients in combination with radiation treatment. However, it may be necessary for 
docetaxel to be administered to patients in addition to ECI301. Therefore, toxicity data of 
ECI301 in combination with docetaxel were found to be necessary. The design of the study was 
as indicated below and also included 2 groups that were dosed with docetaxel, one in 
combination with ECI301. All animals were observed daily throughout the study period for 
general clinical condition, injection-site observations, body weight, ophthalmology, 
electrocardiography, blood pressure, hematology, blood chemistry, urinalysis, peripheral blood 
immunophenotyping, organ weights, macroscopic and microscopic pathology, and mortality. An 
additional group of animals for each group except control (2/sex/dose) were bled for 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 45 of 60 
toxicokinetic analysis. Animals were bled at 5, 15, and 30 minutes, and 1, 2, and 4 hours 
postdose for toxicokinetic analysis of plasma concentrations of ECI301 or docetaxel, and anti-
ECI301antibody assessment. Two animals were sacrificed prematurely during this study. Neither 
death was attributed to ECI301 treatment, but 1 of them was determined to be possibly due to 
docetaxel treatment. There were no clinical signs that were attributable to treatment, and there 
was no effect upon bodyweight. One male receiving 0.15 mg/kg was killed on Day 59 due to 
gradual decline in its general clinical condition. The macroscopic and histopathological 
examinations did not identify any finding that would account for the ill health of this animal and, 
consequently, this death was not attributed to treatment. There were no treatment-related 
ophthalmoscopic findings and no effect upon the electrophysiology of the heart or blood 
pressure. There were no hematological findings or changes in blood plasma or urinary 
composition that were attributable to treatment. There were no toxicologically significant organ 
weight differences after 3 cycles of dosing with ECI301 and no macroscopic or histopathological 
findings that were due to treatment.[28] 
The plasma samples taken from the control animals were below the limit of quantification in all 
cases, demonstrating that there was no quantifiable contamination with ECI301 in these animals. 
Mean plasma concentrations (Cmax) and ECI301 concentration-time curves estimated up to 4 
hours post dose (AUC4) on Day 1 and 47 are also shown. The toxicokinetic investigations 
indicated that the extent of systemic exposure of marmosets to ECI301 appeared to be 
characterized by non-linear (dose-dependent) kinetics over the dose range 0.05 to 1.5 mg/kg on 
Days 1 and 47. Increasing the dose of ECI301 above 0.15 mg/kg was considered likely to result 
in a higher systemic exposure than would be predicted from a linear relationship, which is 
consistent with the possibility of a capacity limited process for the elimination of ECI301. In 
general, there were no differences in the systemic exposure of male and female marmosets to 
ECI301, and there was no accumulation after repeated doses. All animals showed negative anti-
ECI301 antibody responses. Immunophenotyping after completion of Cycle 3 indicated no 
significant change at any dose.[28] 
The toxicokinetic investigations indicated that the co-administration of ECI301 with docetaxel 
generally appeared to have little effect on the systemic exposure of male and female marmosets 
to docetaxel. The co-administration of ECI301 with docetaxel did not elicit an anti-ECI301 
antibody response in any animal. Peripheral blood immunophenotyping indicated a reduction in 
the number of CD3+ T lymphocytes in males and females given docetaxel alone and a reduction 
in CD20+ lymphocytes in the males. These males also showed an increase in the percentage of 
CD3+ lymphocytes and a corresponding decrease in the percentage of CD20+ B lymphocytes. 
This T/B cell ratio change was not seen in the females. A reduction in the number and percentage 
of CD14+ monocytes was also seen in the males, but not in the females. It was concluded that IV 
(bolus) administration of ECI301 to common marmosets at doses up to 1.5 mg/kg during three 5-
day cycles of treatment, each separated by 16 days, was well tolerated, was not immunogenic, 
and caused no toxicologically significant findings. Docetaxel treatment at 3 mg/kg, 2 days after 
each cycle in which animals were given ECI301 at 0.05 mg/kg (a pharmacologically active 
dose), caused several findings that were attributable to its anti-mitotic activity. There was no 
evidence in this study that ECI301 modulated adversely the effect of docetaxel treatment, but 
ECI301 coadministration did appear protective with regard to the effect upon lymphocyte 
populations in the peripheral blood.[28] 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 46 of 60 
10.1.3 Formulation and preparation 
The ECI301 drug product is presented as a sterile solution for injection, 1.3 mL in a 6 mL glass 
vial, with a closure consisting of a rubber stopper and crimp cap.  It was made according to 
current Good Manufacturing Practice (cGMP) for ECI by Toyobo, Japan.  The excipients 
(inactive ingredients) in the ECI301 drug product are of compendial grade and are commonly 
used in parenteral dosage forms.  The quantitative composition of the ECI301 drug product is 
shown in Table 4, including the pharmacopoeial reference for each component, where 
applicable. 
TABLE 4 
Quantitative Composition of the ECI301 Drug Product 
Component 
Amount (per mL) 
Pharmacopeial/Quality 
Standard 
Drug substance: 
ECI301  
1.00  mg  
In-house 
Inactive Ingredients: 
 
 
Sodium hydrogen phosphate 
hydrate 
2.90 mg 
JP, USP 
Potassium dihydrogen phosphate 
0.20 mg  
NF 
Sodium chloride 
8.00 mg  
JP, USP 
Potassium chloride 
0.20 mg 
JP, USP 
Polysorbate 80 
0.10 mg  
JP, USP 
Water for injection 
q.s. 
JP, USP 
10.1.4 Stability and storage  
The ECI301 drug product is stored frozen at -20°C until use, and may not be re-frozen. It is 
stable at -20°C for up to 36 months.  ECI301 is stable at room temperature for at least two hours. 
10.1.5  Administration procedures 
The concentration of ECI301 in the drug product is 1000 µg/mL.  Dose escalation will be 
achieved by extracting the correspondingly larger volumes from multiple vials as required, 
according to the following table (Table 5).  ECI301 will be administered without dilution (Table 
5).  The study drug will be filtered with 0.20 or 0.22-micron pore size in the pharmacy and then 
administered as a bolus injection via an in-dwelling peripheral intravenous line (PICC line 
preferred) at a rate of 1 mL/second.  The catheter will immediately be flushed with 20 mL 0.9% 
NaCl at 1 mL/sec  The dose will be delivered from the pharmacy in a single syringe; 
accordingly, doses requiring volumes greater than 10 mL will be delivered in larger syringes. 
TABLE 5 
Volume of Drug Product Required for the ECI301 Dose Levels 
ECI301 
Dose 
(µg/kg) 
ECI301 
Dose 
(µL/kg) 
Volume 
Required in µL 
Final Volume 
to be Injected 
25 
25 
W  x 25 
W x 25 µL 
(no dilution) 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 47 of 60 
ECI301 
Dose 
(µg/kg) 
ECI301 
Dose 
(µL/kg) 
Volume 
Required in µL 
Final Volume 
to be Injected 
50 
50 
W x 50 
 W x 50 µL 
(no dilution) 
82.5 
82.5 
W x 82.5 
 W x 82.5 µL 
(no dilution) 
125 
125 
W x 125 
W x 125 µL 
 (no dilution) 
175 
175 
W x 175 
W x 175 µL 
 (no dilution) 
 
 
W = weight of patient in Kgs 
10.1.6  Incompatibilities 
No incompatibilities or drug interactions have been described.  
 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 48 of 60 
11 REFERENCES 
1. 
Friedman, E.J., Immune modulation by ionizing radiation and its implications for cancer 
immunotherapy. Curr Pharm Des, 2002. 8(19): p. 1765-80. 
2. 
Sharp, H.J., et al., Synergistic antitumor activity of immune strategies combined with 
radiation. Front Biosci, 2007. 12: p. 4900-10. 
3. 
Maurer, M. and E. von Stebut, Macrophage inflammatory protein-1. Int J Biochem Cell 
Biol, 2004. 36(10): p. 1882-6. 
4. 
Rollins, B.J., Chemokines. Blood, 1997. 90(3): p. 909-28. 
5. 
Wolpe, S.D. and A. Cerami, Macrophage inflammatory proteins 1 and 2: members of a 
novel superfamily of cytokines. Faseb J, 1989. 3(14): p. 2565-73. 
6. 
Wazer, D.E. and B.L. Willett, Clinical considerations in palliative treatment of 
metastatic prostate carcinoma. Int J Radiat Oncol Biol Phys, 1987. 13(1): p. 145-6. 
7. 
Wu, J.S., et al., Meta-analysis of dose-fractionation radiotherapy trials for the palliation 
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8. 
Muranski, P., et al., Increased intensity lymphodepletion and adoptive immunotherapy--
how far can we go? Nat Clin Pract Oncol, 2006. 3(12): p. 668-81. 
9. 
Paulos, C.M., et al., Microbial translocation augments the function of adoptively 
transferred self/tumor-specific CD8+ T cells via TLR4 signaling. J Clin Invest, 2007. 
117(8): p. 2197-204. 
10. 
Klebanoff, C.A., et al., Sinks, suppressors and antigen presenters: how lymphodepletion 
enhances T cell-mediated tumor immunotherapy. Trends Immunol, 2005. 26(2): p. 111-7. 
11. 
Roses, R.E., et al., Radiation therapy and Toll-like receptor signaling: implications for 
the treatment of cancer. Oncogene, 2008. 27(2): p. 200-7. 
12. 
Hauser, S.H., et al., Radiation-enhanced expression of major histocompatibility complex 
class I antigen H-2Db in B16 melanoma cells. Cancer Res, 1993. 53(8): p. 1952-5. 
13. 
Klein, B., et al., The effect of irradiation on expression of HLA class I antigens in human 
brain tumors in culture. J Neurosurg, 1994. 80(6): p. 1074-7. 
14. 
Hallahan, D.E., et al., X-ray-induced P-selectin localization to the lumen of tumor blood 
vessels. Cancer Res, 1998. 58(22): p. 5216-20. 
15. 
Gulley, J.L., et al., Combining a recombinant cancer vaccine with standard definitive 
radiotherapy in patients with localized prostate cancer. Clin Cancer Res, 2005. 11(9): p. 
3353-62. 
16. 
Dewan, M.Z., et al., Fractionated but not single-dose radiotherapy induces an immune-
mediated abscopal effect when combined with anti-CTLA-4 antibody. Clin Cancer Res, 
2009. 15(17): p. 5379-88. 
17. 
Demaria, S., et al., Immune-mediated inhibition of metastases after treatment with local 
radiation and CTLA-4 blockade in a mouse model of breast cancer. Clin Cancer Res, 
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18. 
Brody, J.D., et al., In situ vaccination with a TLR9 agonist induces systemic lymphoma 
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19. 
Yoneyama, H., et al., Regulation by chemokines of circulating dendritic cell precursors, 
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20. 
Zhang, Y., et al., Mobilization of dendritic cell precursors into the circulation by 
administration of MIP-1alpha in mice. J Natl Cancer Inst, 2004. 96(3): p. 201-9. 


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21. 
Teitz-Tennenbaum, S., et al., Radiotherapy potentiates the therapeutic efficacy of 
intratumoral dendritic cell administration. Cancer Res, 2003. 63(23): p. 8466-75. 
22. 
Shiraishi, K., et al., Enhancement of antitumor radiation efficacy and consistent induction 
of the abscopal effect in mice by ECI301, an active variant of macrophage inflammatory 
protein-1alpha. Clin Cancer Res, 2008. 14(4): p. 1159-66. 
23. 
Broxmeyer, H.E., et al., Myeloid progenitor cell proliferation and mobilization effects of 
BB10010, a genetically engineered variant of human macrophage inflammatory protein-
1alpha, in a phase I clinical trial in patients with relapsed/refractory breast cancer. 
Blood Cells Mol Dis, 1998. 24(1): p. 14-30. 
24. 
Clemons, M.J., et al., A randomized phase-II study of BB-10010 (macrophage 
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fluorouracil, adriamycin, and cyclophosphamide chemotherapy. Blood, 1998. 92(5): p. 
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25. 
Hunter, M.G., et al., BB-10010: an active variant of human macrophage inflammatory 
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8. 
26. 
Marshall, E., et al., Clinical effects of human macrophage inflammatory protein-1 alpha 
MIP-1 alpha (LD78) administration to humans: a phase I study in cancer patients and 
normal healthy volunteers with the genetically engineered variant, BB-10010. Eur J 
Cancer, 1998. 34(7): p. 1023-9. 
27. 
Bernstein, S.H., et al., A randomized phase II study of BB-10010: a variant of human 
macrophage inflammatory protein-1alpha for patients receiving high-dose etoposide and 
cyclophosphamide for malignant lymphoma and breast cancer. Br J Haematol, 1997. 
99(4): p. 888-95. 
28. 
ECI, I., ECI301 Investigator Brochure. 2010. 
29. 
Kaech, S.M., E.J. Wherry, and R. Ahmed, Effector and memory T-cell differentiation: 
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Thompson, E.D., et al., Tumor masses support naive T cell infiltration, activation, and 
differentiation into effectors. J Exp Med. 207(8): p. 1791-804. 
 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 50 of 60 
12 APPENDICES 
12.1 APPENDIX I :  ACUTE RTOG TOXICITY SCORING 
 
[ 0 ] 
[ 1 ] 
[ 2 ] 
[ 3 ] 
[ 4 ] 
SKIN 
No 
change 
over 
baseline 
Follicular, faint or 
dull erythema/ 
epilation/dry 
desquamation/ 
decreased 
sweating 
Tender or bright 
erythema, patchy 
moist 
desquamation/ 
moderate edema 
Confluent, moist 
desquamatiom other 
than skin folds, pitting 
edema 
Ulceration, hemorrhage, 
necrosis 
MUCOUS 
MEMBRANE 
No 
change 
over 
baseline 
Injection/ may 
experience mild 
pain not requiring 
analgesic 
Patchy mucositis 
which may 
produce an 
inflammatory 
serosanguinitis 
discharge/ may 
experience 
moderate pain 
requiring 
analgesia 
Confluent fibrinous 
mucositis/ may include 
severe pain requiring 
narcotic 
Ulceration, hemorrhage or 
necrosis 
EYE 
No 
change 
Mild 
conjunctivitis 
with or without 
scleral injection/ 
increased tearing 
Moderate 
conjunctivitis 
with or without 
keratitis requiring 
steroids &/or 
antibiotics/ dry 
eye requiring 
artificial tears/ 
iritis with 
photophobia 
Severe keratitis with 
corneal ulceration/ 
objective decrease in 
visual acuity or in visual 
fields/ acute glaucoma/ 
panopthalmitis 
Loss of vision (unilateral 
or bilateral) 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 51 of 60 
EAR 
No 
change 
over 
baseline 
Mild external 
otitis with 
erythema, pruritis, 
secondary to dry 
desquamation not 
requiring 
medication. 
Audiogram 
unchanged from 
baseline 
Moderate external 
otitis requiring 
topical 
medication/ 
serious otitis 
medius/ 
hypoacusis on 
testing only 
Severe external otitis 
with dischange or moist 
desquamation/ 
symptomatic 
hypoacusis/tinnitus, not 
drug related 
Deafness 
SALIVARY 
GLAND 
No 
change 
over 
baseline 
Mild mouth 
dryness/ slightly 
thickened saliva/ 
may have slightly 
altered taste such 
as metallic taste/ 
these changes not 
reflected in 
alteration in 
baseline feeding 
behavior, such as 
increased use of 
liquids with meals 
Moderate to 
complete dryness/ 
thick, sticky 
saliva/ markedly 
altered taste 
------- 
Acute salivary gland 
necrosis 
PHARYNX & 
ESOPHAGUS 
No 
change 
over 
baseline 
Mild dysphagia or 
odynophagia/ 
may require 
topical anesthetic 
or non-narcotic 
analgesics/ may 
require soft diet 
Moderate 
dysphagia or 
odynophagia/ 
may require 
narcotic 
analgesics/ may 
require puree or 
liquid diet 
Severe dysphagia or 
odynophagia with 
dehydration or weight 
loss(>15% from pre-
treatment baseline) 
requiring N-G feeding 
tube, I.V. fluids or 
hyperalimentation 
Complete obstruction, 
ulceration, perforation, 
fistula 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 52 of 60 
LARYNX 
No 
change 
over 
baseline 
Mild or 
intermittent 
hoarseness/cough 
not requiring 
antitussive/ 
erythema of 
mucosa 
Persistent 
hoarseness but 
able to vocalize/ 
referred ear pain, 
sore throat, 
patchy fibrinous 
exudate or mild 
arytenoid edema 
not requiring 
narcotic/ cough 
requiring 
antitussive 
Whispered speech, 
throat pain or referred 
ear pain requiring 
narcotic/ confluent 
fibrinous exudate, 
marked arytenoid edema
Marked dyspnea, stridor 
or hemoptysis with 
tracheostomy or 
intubation necessary 
UPPER G.I. 
No 
change 
Anorexia with 
<=5% weight loss 
from pretreatment 
baseline/ nausea 
not requiring 
antiemetics/ 
abdominal 
discomfort not 
requiring 
parasympatholytic 
drugs or 
analgesics 
Anorexia with 
<=15% weight 
loss from 
pretreatment 
baseline/nausea 
&/ or vomiting 
requiring 
antiemetics/ 
abdominal pain 
requiring 
analgesics 
Anorexia with >15% 
weight loss from 
pretreatment baseline or 
requiring N-G tube or 
parenteral support. 
Nausea &/or vomiting 
requiring tube or 
parenteral 
support/abdominal pain, 
severe despite 
medication/hematemesis
or melena/ abdominal 
distention (flat plate 
radiograph demonstrates 
distended bowel loops 
Ileus, subacute or acute 
obstruction, performation, 
GI bleeding requiring 
transfusion/abdominal 
pain requiring tube 
decompression or bowel 
diversion 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 53 of 60 
LOWER G.I. 
INCLUDING 
PELVIS 
No 
change 
Increased 
frequency or 
change in quality 
of bowel habits 
not requiring 
medication/ rectal 
discomfort not 
requiring 
analgesics 
Diarrhea 
requiring 
parasympatholytic 
drugs (e.g., 
Lomotil)/ mucous 
discharge not 
necessitating 
sanitary pads/ 
rectal or 
abdominal pain 
requiring 
analgesics 
Diarrhea requiring 
parenteral support/ 
severe mucous or blood 
discharge necessitating 
sanitary pags/abdominal 
distention (flat plate 
radiograph demonstrates 
distended bowel loops) 
Acute or subacute 
obstruction, fistula or 
perforation; GI bleeding 
requiring transfusion; 
abdominal pain or 
tenesmus requiring tube 
decompression or bowel 
diversion 
LUNG 
No 
change 
Mild symptoms 
of dry cough or 
dyspnea on 
exertion 
Persistent cough 
requiring narcotic, 
antitussive agents/ 
dyspnea with 
minimal effort but 
not at rest 
Severe cough 
unresponsive to narcotic 
antitussive agent or 
dyspnea at rest/ clinical 
or radiologic evidence 
of acute pneumonitis/ 
intermittent oxygen or 
steroids may be required 
Severe respiratory 
insufficiency/ continuous 
oxygen or assisted 
ventilation 
GENITOURINARY
No 
change 
Frequency of 
urination or 
nocturia twice 
pretreatment 
habit/ dysuria, 
urgency not 
requiring 
medication 
Frequency of 
urination or 
nocturia which is 
less frequent than 
every hour. 
Dysuria, urgency, 
bladder spasm 
requiring local 
anesthetic (e.g., 
Pyridium) 
Frequency with urgency 
and nocturia hourly or 
more frequently/ 
dysuria, pelvis pain or 
bladder spasm requiring 
regular, frequent 
narcotic/gross hematuria 
with/ without clot 
passage 
Hematuria requiring 
transfusion/ acute bladder 
obstruction not secondary 
to clot passage, ulceration 
or necrosis 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 54 of 60 
HEART 
No 
change 
over 
baseline 
Asymptomatic 
but objective 
evidence of EKG 
changes or 
pericardial 
abnormalities 
without evidence 
of other heart 
disease 
Symptomatic with 
EKG changes and 
radiologic 
findings of 
congestive heart 
failure or 
pericardial 
disease/ no 
specific treatment 
required 
Congestive heart failure, 
angina pectoris, 
pericardial disease 
responding to therapy 
Congestive heart failure, 
angina pectoris, 
pericardial disease, 
arrhythmias not 
responsive to non-surgical 
measures 
CNS 
No 
change 
Fully functional 
status (i.e., able to 
work) with minor 
neurologic 
findings, no 
medication 
needed 
Neurologic 
findings present 
sufficient to 
require home 
case/ nursing 
assistance may be 
required/ 
medications 
including 
steroids/anti-
seizure agents 
may be required 
Neurologic findings 
requiring hospitalization 
for initial management 
Serious neurologic 
impairment which 
includes paralysis, coma 
or seizures>3 per week 
despite 
medication/hospitalization
required 
HEMATOLOGIC 
WBC (X 1000) 
>=4.0 
3.0 - <4.0 
2.0 - <3.0 
1.0 - <2.0 
<1.0 
PLATELETS (X 
1000) 
>=100
75 - <100 
50 - <75 
25 - <50 
<25 or spontaneous 
bleeding 
NEUTROPHILS 
>=1.9 
1.5 - <1.9 
1.0 - <1.5 
0.5 - <1.0 
<0.5 or sepsis 
HEMOGLOBIN 
(GM %) 
>11 
11-9.5 
<9.5 - 7.5 
<7.5 - 5.0 
------- 
HEMATOCRIT 
(%) 
>=32 
28 - <32 
<28 
Packed cell transfusion 
required 
------- 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 55 of 60 
 
 
 
GUIDELINES: The acute morbidity criteria are used to score/grade toxicity from radiation therapy. The critera are relevant 
from day 1, the commencement of therapy, through day 90. Therafter, the EORTC/RTOG Criteria of Late Effects are to be 
utilized. 
 
The evaluator must attempt to discriminate between disease- and treatment-related signs and symptoms. 
 
An accurate baseline evaluation prior to commencement of therapy is necessary. 
 
All toxicities Grade 3, 4 or 5* must be verified by the Principal Investigator. 
 
*ANY TOXICITY WHICH CAUSED DEATH IS GRADED 5. 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 56 of 60 
12.2 APPENDIX II:  LATE RTOG TOXICITY SCORING 
 
ORGAN TISSUE 
0 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
SKIN 
None 
Slight atrophy 
Pigmentation 
change 
Some hair loss 
Patch atrophy; Moderate 
telangiectasia; Total hair 
loss 
Marked atrophy; Gross 
telangiectasia 
Ulceration 
SUBCUTANEOUS
TISSUE 
None 
Slight 
induration 
(fibrosia) and 
loss of 
subcutaneous fat 
Moderate fibrosis but 
asymptomatic 
Slight field contracture 
<10% linear reduction 
Severe induration and loss of 
subcutaneous tissue 
Field contracture 
>10% linear measurement 
Necrosis 
MUCOUS 
MEMBRANE 
None 
Slight atrophy 
and dryness 
Moderate atrophy and 
telangiectasia 
Little mucous 
Marked atrophy with complete 
dryness 
Severe telangiectasia 
Ulceration 
SALIVARY 
GLANDS 
None 
Slight dryness 
of mouth 
Good response 
on stimulation 
Moderate dryness of 
mouth 
Poor response on 
stimulation 
Complete dryness of mouth 
No response on stimulation 
Fibrosis 
SPINAL CORD 
None 
Mild 
L'Hermitte's 
syndrome 
Severe L'Hermitte's 
syndrome 
Objective neurological findings 
at or below cord level treated 
Mono, para 
quadraplegia 
BRAIN 
None 
Mild headache 
Slight lethargy 
Moderate headache 
Great lethargy 
Severe headaches 
Severe CNS dysfunction 
(partial loss of power or 
dyskinesia) 
Seizures or 
paralysis 
Coma 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 57 of 60 
ORGAN TISSUE 
0 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
EYE 
None 
Asymptomatic 
cataract 
Minor corneal 
ulceration or 
keratitis 
Symptomatic cataract 
Moderate corneal 
ulceration 
Minor retinopathy or 
glaucoma 
Severe keratitis 
Severe retinopathy or 
detachment 
Severe glaucoma 
Panopthalmitis/ 
Blindness 
LARYNX 
None 
Hoarseness 
Slight arytenoid 
edema 
Moderate arytenoid 
edema 
Chondritis 
Severe edema 
Severe chondritis 
Necrosis 
LUNG 
None 
Asymptomatic 
or mild 
symptoms (dry 
cough) 
Slight 
radiographic 
appearances 
Moderate symptomatic 
fibrosis or pneumonitis 
(severe cough) 
Low grade fever 
Patchy radiographic 
appearances 
Severe symptomatic fibrosis or 
pneumonitis 
Dense radiographic changes 
Severe 
respiratory 
insufficiency/ 
Continuous O2/ 
Assisted 
ventilation 
HEART 
None 
Asymptomatic 
or mild 
symptoms 
Transient T 
wave inversion 
& ST changes 
Sinus 
tachycardia 
>110 (at rest) 
Moderate angina on 
effort 
Mild pericarditis 
Normal heart size 
Persistent abnormal T 
wave and ST changes 
Low ORS 
Severe angina 
Pericardial effusion 
Constrictive pericarditis 
Moderate heart failure 
Cardiac enlargement 
EKG abnormalities 
Tamponade/ 
Severe heart 
failure/ Severe 
constrictive 
pericarditis 
ESOPHAGUS 
None 
Mild fibrosis 
Slight difficulty 
in swallowing 
solids 
No pain on 
swallowing 
Unable to take solid food 
normally 
Swallowing semi-solid 
food 
Dilatation may be 
indicated 
Severe fibrosis 
Able to swallow only liquids 
May have pain on swallowing 
Dilation required 
Necrosis/ 
Perforation 
Fistula 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 58 of 60 
ORGAN TISSUE 
0 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
SMALL/LARGE 
INTESTINE 
None 
Mild diarrhea 
Mild cramping 
Bowel 
movement 5 
times daily 
Slight rectal 
discharge or 
bleeding 
Moderate diarrhea and 
colic 
Bowel movement >5 
times daily 
Excessive rectal mucus 
or intermittent bleeding 
Obstruction or bleeding 
requiring surgery 
Necrosis/ 
Perforation 
Fistula 
LIVER 
None 
Mild lassitude 
Nausea, 
dyspepsia 
Slightly 
abnormal liver 
function 
Moderate symptoms 
Some abnormal liver 
function tests 
Serum albumin normal 
Disabling hepatitic 
insufficiency 
Liver function tests grossly 
abnormal 
Low albumin 
Edema or ascites 
Necrosis/ 
Hepatic coma 
or 
encephalopathy
KIDNEY 
None 
Transient 
albuminuria 
No hypertension 
Mild 
impairment of 
renal function 
Urea 25-35 
mg% 
Creatinine 1.5-
2.0 mg% 
Creatinine 
clearance >75% 
Persistent moderate 
albuminuria (2+) 
Mild hypertension 
No related anemia 
Moderate impairment of 
renal function 
Urea>36-60 mg% 
Creatinine clearance (50-
74%) 
Severe albuminuria 
Severe hypertension 
Persistent anemia (<10g%) 
Severe renal failure 
Urea >60 mg% 
Creatinine >4.0 mg% 
Creatinine clearance <50% 
Malignant 
hypertension 
Uremic 
coma/Urea 
>100% 


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 59 of 60 
ORGAN TISSUE 
0 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
BLADDER 
None 
Slight epithelial 
atrophy 
Minor 
telangiectasia 
(microscopic 
hematuria) 
Moderate frequency 
Generalized 
telangiectasia 
Intermittent macroscopic 
hematuria 
Severe frequency and dysuria 
Severe generalized 
telangiectasia (often with 
petechiae) 
Frequent hematuria 
Reduction in bladder capacity 
(<150 cc) 
Necrosis/ 
Contracted 
bladder 
(capacity <100 
cc) 
Severe 
hemorrhagic 
cystitis 
BONE 
None 
Asymptomatic 
No growth 
retardation 
Reduced bone 
density 
Moderate pain or 
tenderness 
Growth retardation 
Irregular bone sclerosis 
Severe pain or tenderness 
Complete arrest of bone growth 
Dense bone sclerosis 
Necrosis/ 
Spontaneous 
fracture 
JOINT 
None 
Mild joint 
stiffness 
Slight limitation 
of movement 
Moderate stiffness 
Intermittent or moderate 
joint pain 
Moderate limitation of 
movement 
Severe joint stiffness 
Pain with severe limitation of 
movement 
Necrosis/ 
Complete 
fixation 
 
 
 
 
GUIDELINES: The late morbidity criteria are used to score/grade toxicity from radiation therapy. The critera are relevant from 
day 90 after completion of therapy.  
 
The evaluator must attempt to discriminate between disease- and treatment-related signs and symptoms. 
 
All toxicities Grade 3, 4 or 5* must be verified by the Principal Investigator. 
 
*ANY TOXICITY WHICH CAUSED DEATH IS GRADED 5.


Abbreviated Title: Phase 1 ECI301 with radiation 
Version Date: November 1, 2011 
Page 60 of 60 
12.3 APPENDIX III.  EXCLUDED IMMUNOSUPRESSIVE MEDICATIONS 
Adalimumab (HUMIRA) 
Azathioprine 
Basiliximab (Simulect) 
Certolizumab pegol (Cimzia) 
Cyclophosphamide 
Cyclosporin 
Daclizumab (Zenapax) 
Dactinomycin 
Etanercept (Enbrel) 
Golimumab (Simponi) 
Ifliximab (Remicade) 
Methotrexate 
Mercaptopurine 
Muromonab-CD3 
Mycophenolate 
Rituxan 
Sirolimus 
Tacrolimus 
Oral or intravenously delivered glucocorticoids 
 
 

ICF_001.pdf: 
 
 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
               •  Adult Patient or     •  Parent, for Minor Patient 
 
 
INSTITUTE: 
National Cancer Institute 
 
 
STUDY NUMBER: 
11-C-0248 
PRINCIPAL INVESTIGATOR:  Deborah E. Citrin, M.D. 
 
 
STUDY TITLE: 
Phase I Trail of ECI301 in Combination with Radiation in Patients with Advanced or 
Metastatic 
 
 
Initial Review Approved by the IRB on 07/11/11 
Amendment Approved by the IRB on 11/15/11 (B)  
 
 
 
Date Posted to Web: 12/16/11 
Standard 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY 
 
•   Adult Patient or    •   Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent (1) 
 
 
INTRODUCTION 
We invite you to take part in a research study at the National Institutes of Health (NIH). 
First, we want you to know that: 
Taking part in NIH research is entirely voluntary.  
You may choose not to take part, or you may withdraw from the study at any time.  In either case, you will not 
lose any benefits to which you are otherwise entitled.  However, to receive care at the NIH, you must be taking 
part in a study or be under evaluation for study participation. 
You may receive no benefit from taking part.  The research may give us knowledge that may help people in the 
future. 
Second, some people have personal, religious or ethical beliefs that may limit the kinds of medical or research treatments 
they would want to receive (such as blood transfusions).  If you have such beliefs, please discuss them with your NIH 
doctors or research team before you agree to the study. 
Now we will describe this research study.  Before you decide to take part, please take as much time as you need to ask 
any questions and discuss this study with anyone at NIH, or with family, friends or your personal physician or other 
health professional.   
Why is this study being done?  
In this study we will be giving an experimental anti-cancer drug, called ECI301, with radiation therapy to patients with 
metastatic or locally advanced cancer.  
ECI301 is an experimental drug that has not yet been studied in humans and is not approved by the U.S. Food and Drug 
Administration (FDA).  Radiation therapy is one way to treat body sites where cancer has spread. In this study, radiation 
therapy will be given to one or two areas of cancer that are causing pain or might cause an emergency if left untreated.  
The radiation therapy is not experimental and it is given over two weeks in the standard way that it would be given to 
those not on a study.  The difference between this study and standard treatment is that the experimental drug, ECI301 
will be given for two weeks during radiation therapy and for an additional week after radiation therapy.  ECI301 is man-
made and is similar to a protein residing inside the human body.  ECI301 affects a type of white blood cells (effector 
cells). It is possible that ECI301 will make your cancer more sensitive to radiation therapy. It is also possible that ECI301 
will help your body’s immune system recognize and destroy the cancer at sites that have not been treated with radiation 
therapy.  


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  2  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
The purpose of this study is to evaluate the potential benefits and side effects of ECI301 when given with radiation 
therapy.  This will help us learn the best dose of ECI301 to give with radiation therapy. In addition, this study will also 
help us learn how long ECI301 remains in the human body and if ECI301 can change cancers outside of the area treated 
with radiation therapy. 
Why are you being asked to take part in this study? 
You are being asked to participate because you have either metastatic or locally advanced cancer, and your doctor has 
determined that you would benefit from radiation therapy.   
How many people will take part in this study?  
A total of up to 30 people will take part in this study. 
Description of Research Study 
What will happen if you take part in this research study?   
Before you begin the study 
All research studies have specific criteria for entry to allow for valid interpretation of the study results and safety of 
participants, known as eligibility criteria.  Before you begin this study you will need to have the following exams and tests 
to make sure you are eligible for this study.  The exams and tests are part of regular cancer care and may be done even 
if you do not join the study.  It is possible that if you recently had some of these tests, they may not need to be repeated.   
• History and physical examination, including vital signs (height, weight, blood pressure, heart rate, temperature, 
breathing rate). 
• Standard blood tests to measure: your liver and kidney function, white blood cells, red blood cells and platelets, 
your blood’s ability to clot, your blood sugar and blood electrolytes.  If you are a woman who is able to get 
pregnant, you will also be tested for pregnancy. 
• Detailed measurement of your kidney function.  This will require you to collect your urine for 24 hours. 
• Scans, CT scan, ultrasound, MRI and X-rays of your tumors to measure your disease within 3 weeks of your first 
dose of ECI301.   
• HIV Testing:  As part of this study, we will test you for infection with the human immunodeficiency virus (HIV), 
the virus that causes AIDS.  If you are infected with HIV you will not be able to participate in this study.  We will 
tell you what the results mean, how to find care, how to avoid infecting others, how we report newly diagnosed 
HIV infection, and the importance of informing your partners at possible risk because of your HIV infection. 
• Hepatitis B and C screening. 
• Electrocardiogram to measure your heart function 
• Confirmation of diagnosis and stage of disease. 
If you meet the eligibility criteria for this study you will be offered the option to participate. 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  3  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
 
During the study 
Everyone enrolled in the study will receive the same dose of radiation therapy (3 Gy per daily fraction) Monday through 
Friday, to a total dose of 30 Gy delivered over two weeks. On this study, one or two sites may be treated. In addition to 
the radiation therapy, a total of 5 groups of 3-6 patients each will receive increasing doses of ECI301, until we find the 
highest dose that can be given safely.  This is called “dose escalation”.  Ask your doctor what dose of ECI301 you will be 
receiving. 
Before starting treatment, you will have the option of consenting to a biopsy of the site with that will receive radiation.   
During 2, the ECI301 infusion will be given to you in the outpatient clinic or the inpatient unit about one to two hours 
after your radiation treatment Monday through Friday. Your radiation treatment will be completed at the end of week 2.   
At week 3, you will have the option of consenting to a second biopsy of the site that received radiation and a biopsy of 
another site of cancer that did not receive radiation. 
Study Chart 
 
Day 
What to do and what will happen to you 
Screening/ 
Baseline  
Get routine blood tests; urine tests will be done, including the collection of urine for 24 hours. If you 
are female you will be tested for pregnancy. 
Provide a history of how you feel and undergo a physical examination by the research team’s Health 
Care Provider. 
Research blood and a tumor biopsy (optional). 
EKG, CT Scan, MRI, x-rays and ultrasound may be done. 
Week 1 
(Monday-Friday) 
Provide a history of how you feel and undergo a physical examination by the research team’s Health 
Care Provider.  
Research blood samples will be taken (Monday). 
Receive radiation to the sites that your doctor has chosen (Monday-Friday). 
Report any side effects to the research team. 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  4  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
Week 2 
(Monday-Friday) 
Provide a history of how you feel and undergo a physical examination by the research team’s Health 
Care Provider.  
Get routine blood tests; urine test will be done (Monday and Friday). 
Research blood samples will be taken (Monday and Friday). 
A blood sample will be taken to test the amount of ECI301 in your blood (Monday, Tuesday, Thursday 
and Friday).  On the Monday and Thursday , blood samples will be drawn at 0, 15 and 30 minutes, 
and 1, 2, 4, 6, 8, and 24 hours after ECI 301 is givenand just before the final dose is given  
EKG (Monday and Friday) 
Receive radiation to the sites that your doctor has chosen (Monday-Friday). 
Receive ECI301 in the vein (IV) in the day hospital approximately 1 to 2 hours after completion of 
radiation (Monday-Friday). 
Report any side effects to the research team. 
Week 3 
Follow-up 
Return to the NIH clinic to see your doctor.  Provide a history of how you feel and have a physical 
exam by your Health Care Provider. 
Get routine blood tests; urine test will be done. 
Research blood and a tumor biopsy (optional). 
 
Week 4 
Follow-up 
Return to the NIH clinic to see your doctor.  Provide a history of how you feel and have a physical 
exam by your Health Care Provider. 
Get routine blood tests; urine test will be done. 
Research blood and a tumor biopsy (optional).  
Week 6  
Follow-up 
Return to the NIH clinic to see your doctor.  Provide a history of how you feel and have a physical 
exam by your Health Care Provider. 
Get routine blood tests.  
Collect urine for 24 hours. 
CT Scan, MRI, x-rays and ultrasound may be done. 
Week 12  
Follow-up 
Return to the NIH clinic to see your doctor.  Provide a history of how you feel and have a physical 
exam by your Health Care Provider. 
Get routine blood tests. 
CT Scan, MRI, x-rays and ultrasound may be done. 
6 months 
Follow-up 
Return to the NIH clinic to see your doctor.  Provide a history of how you feel and have a physical 
exam by your Health Care Provider. 
Get routine blood tests. 
 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  5  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
Birth Control 
If you are a woman who is breast feeding or pregnant, you may not take part in the study because we don’t know how 
this medicine would affect your baby or your unborn child. If you are a woman who can become pregnant, or are the 
partner of a woman who can become pregnant, you will need to practice an effective form of birth control before starting 
study treatment, during study treatment, and for 6 months after you finish study treatment.  If you think that you or your 
partner is pregnant, you should tell your study doctor or nurse at once.   
Effective forms of birth control include: 
• 
abstinence  
• 
intrauterine device (IUD) 
• 
hormonal [birth control pills, injections, or implants] 
• 
tubal ligation 
• 
vasectomy 
Alternative Approaches or Treatments 
What other choices do I have if I do not take part in this study?  
Instead of being in this study, you have these options: 
• 
Getting treatment or care for your cancer without being in a study; 
• 
Taking part in another study; 
• 
Getting comfort care, also called palliative care.  This type of care helps reduce pain, tiredness, appetite problems 
and other problems caused by the cancer.  It does not treat the cancer directly.  Instead, it tries to improve how 
you feel.  Comfort care tries to keep you as active and comfortable as possible. 
Please talk to your doctor about these and other options. 
Risks or Discomforts of Participation 
What side effects or risks can I expect from being in this study?  
ECI301 
You may have side effects while you are taking the study drugs.  Everyone taking part in the study will be watched 
carefully for any side effects.  However, doctors don’t know all the side effects that may happen, so it is important to 
report any changes that you notice, even if your study team does not ask specifically about them.  Side effects may be 
mild or severe.  Your study team will give you medicines to help lessen side effects.  Many side effects go away with 
those medicines and others may go away soon after you stop the study drugs.  In some cases, side effects can be 
serious, long lasting, or may never go away.  There are no known long-lasting side effects from ECI301.  If new 
information about side effects becomes available, we will make you aware of this new information.   
This is the first time that the drug ECI301 has been given to humans.  This means that ECI301 has never been tested in 
humans before this study.  The side effects of ECI301 infusions in man are unknown.  Studies in animals suggest that 
ECI301 should be safe at the doses we will use in this protocol. 
It is possible that ECI301 will make your cancer more sensitive to radiation therapy. It is also possible that ECI301 will 
help your body’s immune system recognize and destroy the cancer at sites that have not been treated with radiation 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  6  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
therapy. This could result in longer lasting relief of your symptoms or a decrease in the size of your tumor.  It is also 
possible that ECI301 could increase the effects of radiation therapy on normal tissues that are in the treatment area 
which could result in inflammation (redness or swelling).   
The expected discomforts of infusions of ECI301 are the same as those to be expected from other infusions. These 
include pain and discomfort from a vein being punctured. 
Radiation Therapy 
Some additional side effects may occur with radiation therapy. These side effects are listed below.  There may be 
additional side effects that depend on which parts of your body are being treated.  Your doctor will discuss these with 
you.  It is important to let your doctor know if you develop these side effects because changes may need to be made to 
your treatment.  The short-term side effects of radiation tend to go away a few weeks after treatment is completed. 
 
Likely 
Less Likely 
• 
Skin irritation in the area radiated  
• 
Fatigue 
• 
Hair loss in the area treated with radiation 
• 
Decrease in blood counts 
• 
Pain in the irradiated area  
Late Side Effects of Radiation 
Some side effects of radiation can happen many months or years after treatment.  It is impossible to predict who will 
develop long-term side effects.  Your doctor will discuss these risks with you.   
Potential Benefits of Participation 
Are there benefits to taking part in this study?  
The aim of this study is to see if this experimental treatment will cause your cancer to shrink.  We hope that you will get 
personal medical benefit from taking part in this study, but we cannot be certain.  These potential benefits could include 
shrinking your cancer or decreasing your symptoms, such as pain, that are caused by the cancer.  Because there is not 
much information about ECI301’s effect on your cancer, we do not know if you will benefit from taking part in this study, 
although the knowledge gained from this study may help others in the future who have cancer.   
Research Subject’s Rights 
What are the costs of taking part in this study?  
If you choose to take part in the study, the following will apply, in keeping with the NIH policy: 
• 
You will receive study treatment at no charge to you. This may include surgery, medicines, 
laboratory testing, x-rays or scans done at the Clinical Center, National Institutes of Health 
(NIH), or arranged for you by the research team to be done outside the Clinical Center, NIH if 
the study related treatment is not available at the NIH.  
• 
There are limited funds available to cover the cost of some tests and procedures performed 
outside the Clinical Center, NIH.  You may have to pay for these costs even if they are not 
covered by your insurance company. 
• 
Medicines that are not part of the study treatment will not be provided or paid for by the Clinical 
Center, NIH. 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  7  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
• 
Once you have completed taking part in the study, medical care will no longer be provided by 
the Clinical Center, NIH. 
Stopping Therapy  
Your doctor may decide to stop your therapy for the following reasons: 
• 
if he/she believes that it is in your best interest 
• 
if your disease gets worse during treatment 
• 
if you have side effects from the treatment that your doctor thinks are too severe 
• 
if new information shows that another treatment would be better for you 
If this happens, you will be informed of the reason therapy is being stopped. 
You can choose to stop taking part in the study at any time. However, if you decide to stop taking part in the study, we 
would like you to talk to both the study doctor and your regular doctor first. 
 
Conflict of Interest 
The National Institutes of Health (NIH) reviews NIH staff researchers at least yearly for conflicts of interest.  The 
following link contains details on this process http://ethics.od.nih.gov/procedures/COI-Protocol-Review-Guide.pdf.  You 
may ask your research team for a copy of the Protocol Review Guide or for more information.  Members of the research 
team who do not work for NIH are expected to follow these guidelines but they do not need to report their personal 
finances to the NIH. 
 
Members of the research team working on this study may have up to $15,000 of stock in the companies that make 
products used in this study.  This is allowed under federal rules and is not a conflict of interest.  
 
 
Optional Biopsies 
A core tumor biopsy uses a hollow needle to extract tissue from the tumor. The most common side effect of a core tumor 
biopsy is pain. Other rare complications include: bleeding, which in rare cases may be severe enough to require a 
transfusion; puncturing a nearby organ; or very rarely (less than 1%) death. 
Optional core tumor biopsies will be taken before treatment and again one week after finishing radiation.  These biopsies 
will be taken from the target lesion (the site that is being radiated) before treatment and from both the target lesion and 
a separate un-irradiated lesion (a site that was not irradiated) at the one week follow up time point.  Biopsies will be 
performed in collaboration with interventional radiology.  Standard techniques will be used for needle biopsies which may 
include CT and/or ultrasound-guided biopsy. In some cases, such biopsies may be done quickly and helped by the use of 
navigation tools, such as a laser guide or GPS-electromagnetic tracking of the needle or ultrasound, to determine which 
exact angle the biopsy needle will be inserted. 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  8  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
 
The biopsies to be performed are exclusively for research purposes and will not benefit you.  It might help other people in 
the future.  Even if you sign "yes" to have the biopsy you can change your mind at any time.  Please read each sentence 
below and think about your choice. After reading each sentence, circle and initial the answer that is right for you. The 
decision to participate in this part of the research is optional, and no matter what you decide to do, it will not affect your 
care.  
 
I agree to have the tumor biopsies for the research tests in this study.  
 
Yes              No               Initials_________ 
 
Optional Studies 
We would like to keep some of the tissue and blood that are collected for future research.  These specimens will be 
identified by a number and not your name.  Your specimens will be used for research purposes only and will not benefit 
you.  It is also possible that the stored specimens may never be used.  Results of research done on your specimens will 
not be available to you or your doctor.  It might help people who have cancer and other diseases in the future.   
If you decide now that your tissue and blood can be kept for research, you can change your mind at any time. Just 
contact us and let us know that you do not want us to use your tissue, blood, and urine.  Then any tissue and blood that 
remain will be destroyed.  
Please read each sentence below and think about your choice. After reading each sentence, circle and initial the answer 
that is right for you.  No matter what you decide to do, it will not affect your care. 
1. My tissue and blood specimens may be kept for use in research to learn about, prevent, or treat cancer.  
 
Yes              No               Initials_________ 
2. My specimens may be kept for use in research to learn about, prevent or treat other health problems (for example: 
diabetes, Alzheimer's disease, or heart disease).  
 
Yes              No               Initials_________ 
3. Someone may contact me in the future to ask permission to use my specimen(s) in new research not included in this 
consent.  
 
Yes              No               Initials_________ 
 


 
  
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
•  Adult Patient or    •  Parent, for Minor Patient 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  9  of  10  pages 
 
 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY (Continuation Sheet) 
 
•  Adult Patient or   •  Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent 
 
 
OTHER PERTINENT INFORMATION 
1. 
Confidentiality.  When results of an NIH research study are reported in medical journals or at scientific 
meetings, the people who take part are not named and identified.  In most cases, the NIH will not release any 
information about your research involvement without your written permission.  However, if you sign a release of 
information form, for example, for an insurance company, the NIH will give the insurance company information from your 
medical record.  This information might affect (either favorably or unfavorably) the willingness of the insurance company 
to sell you insurance. 
The Federal Privacy Act protects the confidentiality of your NIH medical records.  However, you should know that the Act 
allows release of some information from your medical record without your permission, for example, if it is required by the 
Food and Drug Administration (FDA), members of Congress, law enforcement officials, or authorized hospital 
accreditation organizations. 
2. 
Policy Regarding Research-Related Injuries.  The Clinical Center will provide short-term medical care for 
any injury resulting from your participation in research here.  In general, no long-term medical care or financial 
compensation for research-related injuries will be provided by the National Institutes of Health, the Clinical Center, or the 
Federal Government.  However, you have the right to pursue legal remedy if you believe that your injury justifies such 
action. 
3. 
Payments.  The amount of payment to research volunteers is guided by the National Institutes of Health 
policies.  In general, patients are not paid for taking part in research studies at the National Institutes of Health.  
Reimbursement of travel and subsistence will be offered consistent with NIH guidelines. 
4. 
Problems or Questions.  If you have any problems or questions about this study, or about your rights as a 
research participant, or about any research-related injury, contact the Principal Investigator, Deborah Citrin, M.D., 
Building 10, Room CRC B2-3500, Telephone:  301-496-5457. You may also call the Clinical Center Patient Representative 
at (301) 496-2626. If you have any questions about the use of your tissue for future research studies, you may also 
contact the Office of the Clinical Director, Telephone:  301-496-4251.  
5. Consent Document.  Please keep a copy of this document in case you want to read it again. 


 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER: 
11-C-0248 
CONTINUATION:  page  10  of  10  pages 
 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
 
 
 
COMPLETE APPROPRIATE ITEM(S) BELOW: 
 
A. Adult Patient’s Consent 
B. Parent’s Permission for Minor Patient. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby consent to take 
part in this study. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby give permission 
for my child to take part in this study.   
(Attach NIH 2514-2, Minor’s Assent, if applicable.) 
 
 
 
  
 
 
  
 
Signature of Adult Patient/Legal Representative 
Date 
 
 
 
 
 
Print Name 
Signature of Parent(s)/Guardian 
Date 
 
 
 
 
 
Print Name 
C. Child’s Verbal Assent (If Applicable) 
The information in the above consent was described to my child and my child agrees to participate in the study. 
 
 
 
 
  
 
 
 
 
 
Signature of Parent(s)/Guardian 
Date 
Print Name 
THIS CONSENT DOCUMENT HAS BEEN APPROVED FOR USE  
FROM JULY 11, 2011 THROUGH JULY 10, 2012.  
 
 
 
 
  
 
 
  
 
Signature of Investigator 
Date 
 
 
 
 
 
Print Name 
Signature of Witness 
Date 
 
 
 
 
 
Print Name