Clinical trial NCT01437605
Conditions: Melanoma
Keywords: Resected Skin Stage IV
Sponsor: H. Lee Moffitt Cancer Center and Research Institute
Start date: 2011-10-11; Completion date: 2018-07-18
Enrollment: 14 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The overall purpose of this research study is to find a better way to treat melanoma.

The goals of this study are:

1. To measure the side effects of and find out how well patients tolerate the recMAGE-A3 + AS15 ASCI (MAGE-A3 ASCI) treatment with or without the Poly IC:LC
2. To see how well the patient's immune system responds to the MAGE-A3 ASCI treatment with or without the Poly IC:LC
3. To measure the rate of return of the patient's tumor after the MAGE-A3 ASCI treatment with or without the Poly IC:LC
4. To measure the rate of return of the patient's tumor in two groups of patients: one group positive for the gene signature, and the other group not positive for the gene signature in their tumor after the MAGE-A3 ASCI treatment with or without the Poly IC:LC.

Detailed description:
In the first year, participants may receive up to 8 injections given in the following order:

1. 5 ASCI injections with or without Poly IC:LC with a 3-week interval between each.
2. 3 ASCI injections with or without Poly IC:LC with a 3-month interval between each.

   During years 2 through 3, participants may receive up to 5 ASCI injections with or without Poly IC:LC given in the following order:
3. During year 2, ASCI injections with or without Poly IC:LC will be given every 3 months for a total of up to 3 injections.
4. During year 3, ASCI injections with or without Poly IC:LC will continue to be given every 3 months for a total of up to 2 more injections.

Arms: arm 1: ASCI injections without Poly IC:LC arm 2: ASCI injections with Poly IC:LC
Interventions: intervention 1: MAGE-A3 ASCI injections without Poly IC:LC intervention 2: MAGE-A3 ASCI injections with Poly IC:LC
Intervention descriptions: intervention 1: MAGE-A3 ASCI injections without Poly IC:LC as outlined in Detailed Description intervention 2: MAGE-A3 ASCI injections with Poly IC:LC as outlined in Detailed Description

Protocol documents:
Prot_SAP_000.pdf: 
CONFIDENTIAL 
Protocol version 7.0  10/28/2015   
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Protocol MCC 16545, IRB #11.05.0007 
TITLE 
 
IND: 14779 
A Randomized Phase II Study to Assess the Safety and 
Immunogenicity of recMAGE-A3+AS15 ASCI with or without 
Poly IC:LC in Patients with Resected MAGE-A3 Positive, 
Stage IV Melanoma  
 
Document type: 
Clinical Trial Protocol  
Development Phase: 
Phase II open randomized trial 
Document status: 
Amendment 
Sponsor: 
Principal Investigator: 
MOFFITT CANCER CENTER 
Nikhil Khushalani, MD 
Date: 
October 28, 2015 
Number of pages: 
90 
Co-Investigators:           
 
                                         
 
                                         
 
                                         
 
                                         
 
                                         
 
                                       
Biostatistics:        
  
 
 
Protocol Management:   
                         
 
Investigational Agents: 
          
Hiltonol (poly IC:LC) provided by Oncovir, Inc. (NSC 301463) 
 
Recombinant MAGE-A3 protein combined with the AS15 
adjuvant (recMAGE-A3 + AS15) ASCI provided by GSK 
Biologicals, SA. (IND 13253) 
 
MOFFITT CANCER CENTER 
12902 MAGNOLIA DRIVE, TAMPA, FL.  33612 
(813) 745-4673 
Moffitt McKinley Outpatient Center 
10920 N. McKinley Drive 
Tampa, FL  33612 


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SYNOPSIS 
 
This is an open, randomized, two-arm, Phase II study in MAGE-A3 positive patients with 
completely resected stage IV melanoma to assess the safety, tolerability, immunogenicity and 
clinical activity (RFS and OS) of recMAGE-A3 + AS15 ASCI in combination with Poly IC:LC  
as adjuvant therapy compared to recMAGE-A3 + AS15 ASCI alone. 
 
The study will be conducted solely at the Moffitt Cancer Center in Tampa, Florida.  
 
Adjuvant treatment for patients with resectable stage IV melanoma:  
 
Many patients will relapse after surgery for stage IV resectable disease. Surgery of stage IV 
melanoma is generally not curative and approximately 90% of the patients will ultimately die of 
metastatic disease by 5 years from diagnosis. There is thus a need for adjuvant therapy to prevent 
disease relapse after surgical resection of isolated metastases. The role of chemotherapy or high 
dose interferon alpha-2b as adjuvant treatment remains unclear with no documented tangible 
benefit in terms of prolonged overall survival.  
 
Non-specific immunotherapy with high-dose interferon has shown activity in disease-free 
survival in resected stage III disease, but its impact on overall survival in that setting, and in 
stage IV resected melanoma is less clear. A recent Phase III trial with pegylated interferon in 
stage III resected disease showed a significant long-term impact of this treatment on relapse-free 
survival, but not on overall survival. Subgroup analyses further showed that patients with 
macroscopic lymph node involvement did not derive a significant clinical benefit from this 
treatment, clouding the outcome for higher risk patients with stage IV resected disease and 
warranting the development of new treatment options for these patients. Active specific 
immunization against tumor antigens is certainly one such possible approach.  
 
The recMAGE-A3 Antigen-Specific Cancer Imunotherapeutics (ASCI) has shown promising 
results in Phase II studies in stage IV melanoma. In a recent Phase II proof-of-concept trial of 
two different formulations of recMAGE-A3 ASCI in melanoma patients with early metastatic 
disease (GSK study 249553/008), a strong and robust immunological response was seen, and the 
safety of the treatments was considered acceptable, raising no specific safety concerns. Objective 
clinical response was observed for 5 of the 75 patients treated and another 7 patients showed 
disease stabilization (over periods varying between 5 and 12 months, some still ongoing). These 
preliminary findings support the development of recMAGE-A3 ASCI as adjuvant therapy for 
stage IV resected melanoma. A double-blind, randomized, placebo-controlled Phase III trial is 
currently evaluating the efficacy of recMAGE-A3 + AS15 ASCI versus placebo in patients with 
resected stage IIIB/C melanoma with macroscopic lymph node involvement (DERMA Study). 
 
Additional support for developing recMAGE-A3 ASCI in the adjuvant melanoma setting may be 
found in a recent Phase II proof-of-concept trial of MAGE-A3 ASCI as adjuvant therapy for 
patients with resectable stage IB and II NSCLC. This study (GSK 249553/004) showed a relative 
reduction of the risk of disease recurrence of 27% in the group receiving the ASCI compared to 
the placebo group. The immunological responses observed in melanoma patients and NSCLC 


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patients in the adjuvant and metastatic settings, respectively, are very similar, so it is reasonable 
to expect that an equivalent tumor response might be seen in the adjuvant melanoma setting.  
 
Taking into account the tumor-specificity of recMAGE-A3, the MAGE-A3 antigen expression 
levels in stage IV melanoma tumors, the acceptable tolerability of the recMAGE-A3 + AS15 
ASCI and the promising results from Phase II proof-of-concept trials (GSK 249553/008 and 
249553/004), this Phase II trial will evaluate the immunogenicity of the recMAGE-A3 + AS15 
ASCI with or without the toll-like receptor (TLR) agonist Poly IC:LC (Hiltonol) as adjuvant 
therapy for MAGEA3-positive patients with resected stage IV melanoma that have been 
rendered free of disease but are at very high risk of relapse.   
 
 
Objectives:  
 
Primary Objectives: 
 
• To evaluate the safety and tolerability of recMAGE-A3 + AS15 ASCI compared to recMAGE-
A3 + AS15 ASCI in combination with Poly IC:LC in patients with resected stage IV 
melanoma. 
 
• To evaluate the immunogenicity of recMAGE-A3 + AS15 ASCI compared to recMAGE-A3 + 
AS15 ASCI in combination with Poly IC:LC as measured by humoral response (antibodies 
serum titers) in patients with resected stage IV melanoma.  
 
 
Secondary Objectives: 
 
• To evaluate the clinical activity of the recMAGE-A3 + AS15 ASCI compared to recMAGE-A3 
+ AS15 ASCI in combination with Poly IC:LC in terms of relapse-free survival (RFS) and 
overall survival (OS) in the overall study population. 
 
• To evaluate the immunogenicity of the recMAGE-A3 + AS15 ASCI compared to recMAGE-
A3 + AS15 ASCI in combination with Poly IC:LC measured by T cell responses against 
MAGE-A3 antigen.  
 
• To assess the gene profile of tumor tissue in the two study arms and evaluate the activity of the 
recMAGE-A3 + AS15 ASCI +/- Poly IC:LC in terms of RFS in the sub-population of patients 
presenting a potentially favorable gene signature (see Section 6.4.4), and in the sub-population 
of patients without this gene signature.  
 
Translational Research Objectives: 
 
• Translational research includes assessment of MAGE-A3 expression of tumor tissue from 
recurrent lesions, and gene profiling of recurrent or new lesions. 
  
 


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Study design: 
 
• Experimental design: Two-arm, open, randomized single center study. 
 
• Treatment allocation: Patients will be randomized (1:1 ratio) to receive either recMAGE-A3 + 
AS15 ASCI or recMAGE-A3 + AS15 ASCI in combination with Poly IC:LC. 
 
• Blinding: None, open label. 
 
• Treatment groups: Two groups, recMAGE-A3 + AS15 ASCI and recMAGE-A3 + AS15 ASCI 
in combination with Poly IC:LC. 
 
• Treatment schedule: The study treatment will consist of an Induction phase and a Maintenance 
phase. During the Induction phase, recMAGE-A3 + AS15 ASCI +/- Poly IC:LC will be 
administered 5 times at 3-week intervals. During the Maintenance phase, recMAGE-A3 + 
AS15 ASCI +/- Poly IC:LC will be administered 8 times at 3-month intervals. A total of 13 
doses of recMAGE-A3 + AS15 ASCI +/- Poly IC:LC will be administered over a period of 27 
months. 
  
• Control: recMAGE-A3 + AS15 ASCI. 
  
• Data collection: Case report form (CRF). 
 
• Duration of the study: The total duration of the study treatment phase (including concluding 
visit) is 30 months per patient. The duration of active follow-up for survival, recurrence and 
serious adverse events (SAEs) related to the study participation will be 5 years from the first 
administration of study treatment.  
 
 
Main inclusion criteria:  
 
• Patients with histologically proven resected stage IV cutaneous or mucosal melanoma rendered 
free of disease. 
 
• The patient’s tumor expresses MAGE-A3, as determined by RT-PCR analysis on formalin-
fixed paraffin-embedded (FFPE) tumor tissue sample obtained during the screening phase. 
 
• ECOG performance status 0 or 1. 
 
• Adequate bone marrow reserve with Hg >10, platelets > 100,000, and WBC >3000 per mm
3. 
 
 
Number of patients:  
 
The study will enroll up to 44 patients to end up with 40 evaluable patients in two cohorts of 20 
patients each.  


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Endpoints: 
 
Primary endpoints: 
 
• Toxicity and tolerability in the two study arms (recMAGE-A3 + AS15 ASCI and recMAGE-
A3 + AS15 ASCI in combination with Poly IC:LC) defined by the adverse events and serious 
adverse events.   
 
• Immunogenicity in the two study arms (recMAGE-A3 + AS15 ASCI and recMAGE-A3 + 
AS15 ASCI in combination with Poly IC:LC) defined by antibodies serum titer (such as anti-
MAGE-A3).   
 
 
Secondary endpoints: 
 
• Immunogenicity as measured by T cell responses directed against MAGE-A3 antigen.  
 
• Relapse-Free Survival (RFS), defined as the time from randomization to the date of first relapse 
of melanoma or of death, whichever comes first. 
 
• Overall Survival (OS), defined as the time from randomization to the date of death. 
 
• A potential correlation between gene expression profile and treatment clinical activity (RFS) in 
both study arms (recMAGE-A3 + AS15 ASCI and recMAGE-A3 + AS15 ASCI in 
combination with Poly IC:LC). 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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TABLE OF CONTENTS                                                                                                      PAGE 
 
SYNOPSIS .....................................................................................................................................2  
LIST OF TABLES ......................................................................................................................10 
LIST OF FIGURES ....................................................................................................................10  
LIST OF APPENDICES ............................................................................................................10 
LIST OF ABBREVIATIONS ....................................................................................................11  
GLOSSARY OF TERMS ..........................................................................................................12  
 
1. INTRODUCTION ..................................................................................................................14  
1.1. Background ............................................................................................................. 14  
1.2. recMAGE-A3 + AS15 Antigen-Specific Cancer Immunotherapeutics ...............14  
1.2.1. Properties of the study investigational products .........................................14  
1.2.2. Preclinical safety and toxicology tests of recMAGE-A3 and    AS15....... 16  
1.2.3. Previous clinical experience with recMAGE-A3 protein ...........................16  
1.2.3.1. First clinical studies with recMAGE-A3 protein .........................16  
1.2.3.2. Phase II proof-of-concept study in NSCLC..................................16  
1.2.3.3. Phase II proof-of-concept study in metastatic melanoma ............17  
1.2.3.4. Summary of the safety data from previous clinical trials with  
 recMAGE-A3 ASCIs....................................................................18 
1.2.3.5. Identification of a gene signature being associated with clinical 
             benefit in response to MAGE-A3 ASCI treatment ......................18 
1.2.4 Rationale for the use of Poly IC:LC in combination with  
         recMAGE-A3 + AS15 ASCI …....................................................………...19 
1.3 Rationale for the study: relapse of patients with resectable stage IV  
      melanoma after surgery ...........................................................................................21  
 
2. OBJECTIVES .........................................................................................................................22  
2.1. Primary objective .....................................................................................................22  
2.2. Secondary objectives ................................................................................................22  
2.3. Translational research objectives ...........................................................................23  
 
3. STUDY DESIGN OVERVIEW .............................................................................................23  
3.1. Study design ..............................................................................................................23 
3.2. Duration of study participation ..............................................................................24  
3.3. Treatment allocation ................................................................................................24  
 
4. STUDY POPULATION .........................................................................................................25  
4.1. Number of patients ..................................................................................................25 
4.2. Inclusion criteria ......................................................................................................25  
4.3. Exclusion criteria .....................................................................................................26  
 
5. INVESTIGATIONAL PRODUCT AND ADMINISTRATION ........................................27  
5.1. Study treatments: recMAGE-A3 + AS15 and Poly IC:LC ..................................27  
5.1.1. recMAGE-A3 + AS15 ASCI ......................................................................27 


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5.1.2. Poly IC:LC (Hiltonol) .................................................................................28   
5.2. Dosage and administration .....................................................................................29  
5.2.1. Contraindications to subsequent treatment administration .........................29  
5.2.1.1. Criteria for postponement of study treatment ..............................30  
5.2.1.2. Criteria for permanent stopping of study treatment and Dose 
Limiting Toxicity ......................................................................................31  
5.3. Treatment assignment and randomization ............................................................32 
5.3.1. Patient identification ...................................................................................32  
5.3.2. Randomization ............................................................................................33  
5.4. Storage ......................................................................................................................34  
5.5. Packaging ..................................................................................................................35  
5.6. Treatment accountability ........................................................................................35  
5.7. Concomitant medication/treatment .......................................................................35  
5.7.1. Permitted medications .................................................................................35  
5.7.2. Prohibited medications or non-drug therapies ............................................35  
5.7.3. Time-window for prophylactic vaccination against infectious diseases .....36  
 
6. STUDY ASSESSMENTS AND PROCEDURES .................................................................37 
6.1. General study aspects ..............................................................................................37  
6.1.1. Attendance for study visits ..........................................................................37  
6.2. Outline of study procedures ....................................................................................37  
6.2.1. Induction phase ...........................................................................................37  
6.2.2. Maintenance phase ......................................................................................39  
6.2.3. Follow-up period .........................................................................................39  
6.3. Detailed description of study visits .........................................................................40  
6.3.1. Screening phase: Procedures from surgical resection until Visit 1  
of the study ................................................................................................40  
6.3.1.1. Visit S1 ........................................................................................40  
6.3.1.2. Visit S2 ........................................................................................41  
6.3.2. Resection procedure ....................................................................................42  
6.3.3. Visit 1: First study treatment administration ..............................................42  
6.3.4. Visit 2: Second study treatment administration ..........................................43  
6.3.5. Visit 3: Third study treatment administration .............................................44  
6.3.6. Visit 4: Fourth study treatment administration ...........................................44  
6.3.7. Visit 5: Fifth study treatment administration ..............................................44  
6.3.8. Visits 6-13: Three-monthly maintenance treatment administrations ..........45  
6.3.9. Visit 14: Concluding examination ..............................................................45  
6.3.10. Post-treatment follow-up ..........................................................................46  
6.3.10.1. Follow-up of patients who have completed the study   
treatment .......................................................................................47  
6.3.10.2. Follow-up of patients who were withdrawn from  
the study treatment ........................................................................47  
6.3.11. Annual contact ..........................................................................................48  
6.3.12. Relapse ......................................................................................................48  
 
6.4. Biologicals sample handling and analysis ..............................................................49  


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6.4.1. Treatment and storage of biological samples ..............................................49  
6.4.2. Laboratory assays and immunological read-outs ........................................49  
6.4.3. MAGE-A3 expression screening of tumor tissue .......................................51  
6.4.4. Gene expression signature associated to treatment benefit of  
resected lesions ........................................................................................ 52  
6.4.4.1. Background ..................................................................................52  
6.4.4.2. Practical aspects ...........................................................................52  
6.4.5. Laboratory assays .......................................................................................53  
6.4.6. Immunological read-outs ............................................................................53 
 
7. TRANSLATIONAL RESEARCH ........................................................................................53  
7.1. MAGE-A3 expression on multiple resected lesions ..............................................54 
7.2. Cellular Immune assays ..........................................................................................54 
7.3. Gene profiling on fresh tissue .................................................................................54  
7.4. Gene profiling research of a relapsed or new tumor ...........................................55  
 
8. ADVERSE EVENTS AND SERIOUS ADVERSE EVENTS .............................................55 
8.1. Definition of an adverse event .................................................................................55  
8.2. Definition of a serious adverse event ......................................................................56 
8.3. Adverse Events of Specific interest ........................................................................58 
8.3.1. Potential immune-mediated diseases .......................................................58 
8.4. Relapse of the disease ...............................................................................................59  
8.5. Clinical laboratory parameters and other abnormal assessments  
       qualifying as adverse events and serious adverse events ......................................59  
8.6. Time period, frequency, and method of detecting adverse events and 
       serious adverse events ..............................................................................................60  
8.7. Evaluating adverse events and serious adverse events .........................................61  
8.7.1. Assessment of intensity ...............................................................................61  
8.7.2. Assessment of causality ..............................................................................62  
8.8. Follow-up of adverse events and serious adverse events and  
       assessment of outcome .............................................................................................63  
8.9. Prompt reporting of serious adverse events ..........................................................63  
8.9.1. Time frames for submitting serious adverse event reports  
           to GSK Bio and Oncovir ...........................................................................63  
8.9.2. Completion and transmission of serious adverse event reports to FDA .....64  
8.10. Regulatory reporting requirements for serious adverse events ........................65 
8.11. Post-study adverse events and serious adverse events ........................................65  
8.12. Pregnancy ...............................................................................................................65 
8.13. Treatment of adverse events .................................................................................66 
 
9. PATIENT COMPLETION AND WITHDRAWAL ............................................................66 
9.1. Patient completion of study treatment ...................................................................66 
9.2. Patient withdrawal ...................................................................................................66 
9.2.1. Patient withdrawal from investigational product ........................................67  
9.2.2. Patient withdrawal from the study ..............................................................67  
9.3. Screen and baseline failures ....................................................................................68  


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10. Statistical Considerations .................................................................................................... 68 
10.1. Primary endpoints .................................................................................................68  
10.1.1. Safety and Tolerability endpoints .............................................................68  
10.1.2. Immunogenicity endpoints ........................................................................68 
10.2. Secondary endpoints ..............................................................................................69 
10.2.1. Secondary clinical activity endpoints .......................................................69 
10.3. Study design/sample size and power/data analyses ............................................69 
10.4. Study populations to be evaluated ........................................................................71  
10.4.1. Total Treated Population (TTP) ................................................................71  
10.4.2. Total Treated Population (TTP) for analysis of efficacy ..........................71  
10.4.3. According to Protocol (ATP) population for analysis of  
            Immunogenicity.........................................................................................72  
10.5. Elimination criteria for analysis ...........................................................................72  
10.6. Derived and transformed data ..............................................................................72  
10.6.1. Group definition ........................................................................................72  
10.6.2. Immunogenicity ........................................................................................73  
10.6.3. Safety ........................................................................................................73 
10.7. Final analyses .........................................................................................................73  
10.7.1. Analysis of demographics and other baseline characteristics ...................73  
10.7.2. Analysis of treatment compliance and treatment modifications................73  
10.7.3. Analysis of efficacy ..................................................................................74  
10.7.3.1. Relapse-free survival (RFS) .......................................................74  
10.7.3.1.1. Relapse-free survival in the overall study  
                  population (Secondary objective) ...............................74  
10.7.3.2. Overall survival (Secondary objective) .....................................74  
10.7.4. Analysis of immunogenicity .....................................................................75  
10.7.5. Analysis of safety ......................................................................................75  
10.7.5.1. Adverse events ...........................................................................75  
10.7.5.2. Serious adverse events ...............................................................75  
10.7.5.3. Deaths ........................................................................................76  
10.7.5.4. Concomitant medications ...........................................................76  
10.8. Gene profile analyses and translational research ...............................................76  
10.8.1. Gene profile ..............................................................................................76  
 
11. CONDUCT OF THE STUDY ..............................................................................................76  
11.1. Ethics and regulatory considerations ...................................................................76  
11.1.1. Institutional Review Board/Independent Ethics Committee (IRB/IEC) ...76  
11.1.2. Informed consent ......................................................................................78  
11.2. Administrative Matters .........................................................................................79  
 
12. REFERENCES ......................................................................................................................80   
  
 
 
 
 


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LIST OF TABLES                                                                                                               PAGE  
 
Table 1. MAGE-A3 expression in various types of cancer ..........................................................15 
Table 2. Dosage and administration..............................................................................................29 
Table 3. Example of postponement of study treatment dose for a maximum of 3 weeks  
              during induction phase ....................................................................................................31  
Table 4. Example of postponement of study treatment dose for more than 3 weeks  
              during induction phase ....................................................................................................31  
Table 5. Timing of study visits and the assessments to be performed (Induction Phase) ............38  
Table 6. Timing of study visits and the assessments to be performed (Maintenance Phase and 
               Concluding Visit) ...........................................................................................................39 
Table 7. Timing of study visits and the assessments to be performed (Follow-up Phase) ..........40  
Table 8. Laboratory assays and immunological read-outs ...........................................................50 
Table 9. List of potential immune-mediated diseases ..................................................................58 
Table 10. Reporting periods for adverse events and serious adverse events ...............................60  
 
LIST OF FIGURES  
   
   
   
   
   
   
   
          
 
Figure 1. Overview of Study design ............................................................................................23  
Figure 2. Overview of Design and Duration of Study .................................................................24  
Figure 3. Timelines for pre-study procedures and obtaining informed consent .........................34  
 
LIST OF APPENDICES     
   
   
   
   
   
 
 
Appendix A. Overview of the Recruitment Plan .......................................................................84  
Appendix B. Laboratory assays .................................................................................................85  
Appendix C. Treatment supplies, packaging and accountability ...............................................86  
Appendix D. TNM classification of malignant melanoma ........................................................88  
Appendix E. Eastern Co-operative Oncology Group (ECOG) Performance Status ..................89  
Appendix F. U.S. National Cancer Institute (NCI) Common Terminology Criteria for  
                       Adverse Events (CTCAE) ....................................................................................90 
 
 
 
 
 
 
 
 
 
 
 
 
 


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LIST OF ABBREVIATIONS 
  
AE     
Adverse event  
AJCC    
American Joint Committee on Cancer  
AS     
Adjuvant System  
ASCI    
Antigen-Specific Cancer Immunotherapeutic  
ATP    
According to protocol  
CI     
Confidence interval  
CRF    
Case Report Form  
CTCAE   
Common Terminology Criteria for Adverse Events  
ECOG    
Eastern Co-operative Oncology Group  
ELISA 
   
Enzyme-linked Immunosorbent assay  
FDA    
Food and Drug Administration, United States  
FFPE    
Formalin-fixed paraffin-embedded  
GCP    
Good Clinical Practice  
GMT    
Geometric Mean Titre  
GSK Bio   GlaxoSmithKline Biologicals SA  
HR     
Hazard Ratio  
IB     
Investigator Brochure  
ICF    
Informed Consent Form  
ICH    
International Conference on Harmonization  
IEC    
Independent Ethics Committee  
IFN    
Interferon  
IND    
Investigational New Drug  
IRB    
Institutional Review Board  
MAGE-A3  
Melanoma AntiGEn-A3  
MCC    
MOFFITT CANCER CENTER 
MedDRA  
Medical Dictionary for Regulatory Activities  
MHC    
Major Histocomptability Complex  
PBMC    
Peripheral blood mononuclear cell  
q   
   
Indicates time interval between administrations of study medication  
RT-PCR   
Reverse Transcriptase Polymerase Chain Reaction  
SAE    
Serious Adverse Event  
SAS®    
Statistical Analysis System 
TNM    
Tumor, nodes and metastases  
 
 
 
 
 
 
 
 
 


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GLOSARY OF TERMS  
 
Adverse event: Any untoward medical occurrence in a patient or clinical investigation patient, 
temporally associated with the use of a medicinal product, whether or not considered related  
to the medicinal product.  
 
An AE can therefore be any unfavorable and unintended sign (including an abnormal laboratory 
finding), symptom, or disease (new or exacerbated) temporally associated with the use of a 
medicinal product. For marketed medicinal products, this also includes failure to produce 
expected benefits (i.e. lack of efficacy), abuse or misuse.  
 
Adequate contraception:  defined as a contraceptive method with failure rate of less than 1% per 
year when used consistently and correctly (when applicable, as mentioned in the product label) 
for example abstinence, combined or progestogen oral contraceptives, injectable progestogen, 
implants of levonorgestrel, oestrogenic vaginal ring, percutaneous contraceptive patches or 
intrauterine device (IUD) or intrauterine system (IUS), vasectomy with documented azoospermia 
of the sole male partner or double barrier method (condom or occlusive cap plus spermicidal 
agent).  
 
Eligible: Qualified for enrolment into the study based upon strict adherence to inclusion/ 
exclusion criteria.  
 
Investigational product: A pharmaceutical form of an active ingredient or placebo being tested or 
used as a reference in a clinical trial, including a product with a marketing authorization when 
used in a way different from the approved form, or when used for an unapproved indication, or 
when used to gain further information about an approved use.  
 
Study Coordinator: An individual medically qualified to assume the responsibilities of the 
sponsor, Moffitt Cancer Center, especially in regards to the ethics, clinical safety of a study and 
the assessment of adverse events.  
 
Patient: Term used throughout the protocol to denote an individual that has been contacted in 
order to participate or participates in the clinical study, either as a recipient of the investigational 
product(s) or as a control.  
 
Patient number: A unique number identifying a patient, assigned to each patient as soon as they 
sign their first informed consent.  
 
ICH defines a protocol amendment as: “A written description of a change(s) to or formal 
clarification of a protocol.”  
 
Protocol administrative change: A protocol administrative change addresses changes to change: 
only logistical or administrative aspects of the study.  
 


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N.B. Any change that falls under the definition of a protocol amendment (e.g. a change that 
affects the safety of patients, scope of the investigation, study design, or scientific integrity of the 
study) MUST be prepared as an amendment to the protocol.  
 
Randomization: Process of random attribution of treatment to patients in order to reduce bias of 
selection  
 
Treatment: Term used throughout the clinical study to denote a set of investigational product(s) 
or marketed product(s) or placebo intended to be administered to a patient, identified by a unique 
number, according to the study randomization or treatment allocation.  
 
Treatment number: A unique number identifying a treatment to a patient, according to the study 
randomization or treatment allocation. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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1.  INTRODUCTION  
 
1.1. 
Background  
 
Cutaneous melanoma is the most aggressive form of all skin cancers. Although it represents only 
4% of all cancers, its incidence is continuing to rise in the world at a rate exceeding all other 
cancers [Jemal, 2007]. Worldwide, it is expected that approximately 132,000 people will be 
diagnosed with melanoma each year and approximately 37,000 people are expected to die of the 
disease annually.  
 
Surgical resection is the treatment of choice for localized melanoma and results in a large 
number of cures with a 90% long term survival rate for stage I disease [Van Akkooi et al 2007]. 
But, patients with resected stage IIB-C (T4N0M0), stage III (T1-4N1-3M0) and stage IV (T1-4, 
N1-3,M1a-c) disease are at high risk of relapse after definitive surgery [Kirkwood, 2001]. The 
risk of recurrence after surgery for the stage IV patients is reported to be 90% at five years. 
[Kirkwood, 2000].  
 
The role of chemotherapy as adjuvant treatment remains unclear with no tangible benefit in 
terms of prolonged overall survival [Nathan, 1995]. High dose interferon remains the most active 
adjuvant agent evaluated to date for high risk stage III, but not stage IV melanoma. It has been 
shown to prolong relapse free survival, while its impact on overall survival in stage III resected 
disease is less clear [Garbe, 2001]. Recently, the results of a Phase III trial showed that pegylated 
interferon (PEG-IFN) in resected stage III melanoma patients had a significant and sustained 
effect on Relapse-Free Survival (RFS) but with no significant effect on Overall Survival 
[Eggermont, 2007]. The significant long-term effect on RFS was only found in melanoma 
patients with microscopic nodal involvement. The trial showed no improvement in the safety 
profile of PEG-IFN as compared to non-pegylated IFN-a [Eggermont, 2007].  
 
Immunity to melanoma appears to be central to disease control. Spontaneous regression has been 
reported in melanoma, suggesting a role for host immunity, indirectly supported by the presence 
of lymphoid infiltrate associated with the primary tumor [Balch, 2001]. It has also been shown 
that patients with melanoma do have tumor rejection antigen recognized by CD4 and CD8 cells 
[Kadison, 2003].  
 
 
 
1.2. recMAGE-A3 + AS15 Antigen-Specific Cancer Immunotherapeutics  
 
1.2.1. Properties of the study investigational products  
 
The primary study investigational product in this trial is an Antigen-Specific Cancer 
Immunotherapeutic (ASCI) comprising the recombinant protein ProtD-MAGE-A3/His 
(abbreviated as recMAGE-A3 in the rest of this document) and the GSK proprietary 
immunological Adjuvant System AS15. The recMAGE-A3 antigen is a 432-amino acid fusion 
protein containing 109 residues of Protein D, a lipoprotein present on the surface of 
Haemophilus influenzae B (ProtD), the MAGE-A3 protein, and a polyhistidine tail (His). 


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MAGE-A3 is a human gene that encodes a tumor-specific antigen (the MAGE-A3 protein) [Van 
Den Eynde, 1997]. The MAGE-A3 gene is silent in all normal human tissues with the exception 
of testis and placenta [De Plaen, 1994; Jungbluth, 2007] and is under epigenetic control 
[Sigalotti, 2002, Jones et al 2005, Baylin et al 2006]. However, the MAGE-A3 antigen is 
believed to be strictly tumor specific, as the cells, where it is expressed (spermatogonia in testis 
and trophoblasts in placenta), do not bear Major Histocompatibility Complex (MHC) molecules 
on their surface and therefore do not present any MAGE-A3 antigen [Boël, 1995; Jungbluth, 
2007].  
 
Because of this high tumor specificity, targeting MAGE-A3 with a cancer immunotherapeutic is 
expected to show a favorable safety profile. The aim of the ASCI approach is to immunize the 
patients against the antigens that are expressed by their tumor cells and thereby to eradicate these 
tumors.  
 
The interest in MAGE-A3 is caused by its expression in various tumor types (Table 1), and its 
expression in both early and more advanced disease stages [Brasseur, 1995; Patard, 1995]. 
Detection of MAGE-A3 expression on tumor samples is easy and can be performed by RT-PCR.  
 
 
 
Tumor 
MAGE-A3 express
ion 
NSCLC 
35-50 % 
Me
lanoma 
65 % 
B
ladder 
62 % 
Esophagus 
47 % 
Head & Neck 
65 % 
Leukem
ia 
29 % 
L
iver 
48 % 
Ovary 
30 % 
Prostate 
18 % 
 
Table 1. MAGE-A3 express
ion 
in var
ious types of cancer. Adapted from Gure 2005 and GSK B
io data. 
 
The expression of MAGE-A3 may also be associated with poor prognosis. Indeed, Gure et al. 
observed that survival of patients with Non-Small Cell Lung Cancer (NSCLC) was shorter when 
MAGE-A3 was highly expressed [Gure, 2005] in comparison to survival of NSCLC patients 
with low MAGE-A3 expression.  
 
To efficiently elicit a strong and enduring immune response upon injection of recMAGEA3 it is 
necessary to combine the MAGE-A3 protein with an immunological Adjuvant System. This 
need was first demonstrated in preclinical studies [Krieg, 2001; Meidenbauer, 2004; Ren, 2004; 
Speiser, 2005], but was soon after demonstrated in humans. In an early clinical trial evaluating 
the administration of recMAGE-A3 either alone or in combination with an immunological 
Adjuvant System, it was shown that combining MAGE-A3 with an immunological Adjuvant 
System was required to efficiently elicit both humoral and cellular immune responses 
[Atanackovic, 2004, Brichard et al 2007].  
 


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The AS15 Adjuvant System consists of the liquid immunological adjuvant AS01B and the 
immunostimulatory nucleotide CpG 7909 (which is co-lyophilized with the recMAGE-A3 
antigen).  
 
Per dose, the liquid immunological adjuvant AS01B contains 50 g of QS21 and 50 g of 
Monophosphoryl lipid A (MPL) [Moore et al, 1999], made up to 500 L with a suspension of 
liposomes. The liposomes are composed of 1000 g of dioleylphosphatidylcholine (DOPC) and 
250 g of cholesterol, in phosphate-buffered saline (PBS).  
 
There is 420 g of immunostimulatory nucleotide CpG 7909 per dose. CpG 7909 is a single-
stranded oligodeoxynucleotiode of 24 bases in length and containing 4 CpG dinucleotides. 
 
For details of the pre-clinical safety and toxicology of the AS15 Adjuvant System and its 
components, please refer to the Investigator’s Brochure. 
 
In this trial, we will determine whether the addition of another adjuvant, poly IC:LC, a Toll-like 
receptor agonist [Pulendran 2004, Seya 2006], will improve the immunogenicity of the 
recMAGE-A3 product with adjuvant AS-15.   
 
 
1.2.2. Preclinical safety and toxicology tests of recMAGE-A3+AS15  
 
For details of preclinical safety and toxicology tests of recMAGE-A3 and AS15, either 
separately or in combination, see the Investigator
's Brochure.  
 
 
1.2.3. Previous clinical experience with recMAGE-A3 protein  
 
1.2.3.1. First clinical studies with recMAGE-A3 protein  
 
Several clinical Phase I/II studies with the recMAGE-A3 protein to be used in this study have 
been performed or are currently in progress. These studies have included mainly melanoma and 
NSCLC patients, and evaluated recMAGE-A3 either alone or combined with various Adjuvant 
Systems. Clinical responses to the treatment with recMAGE-A3 have been observed mainly in 
patients with metastatic melanoma, including partial responses and some complete responses, in 
certain cases of significant long-term duration. An immunological response could be measured in 
the group of patients administered the adjuvanted recMAGE-A3 [Marchand, 2003; Atanackovic, 
2004; Vantomme, 2004; Kruit, 2005]. These early studies also demonstrated that the recMAGE-
A3 administrations are well tolerated.  
 
1.2.3.2. Phase II proof-of-concept study in NSCLC 
 
A study of particular interest for the development of the MAGE-A3 ASCI in the adjuvant 
melanoma setting is a Phase II proof-of-concept study in the NSCLC adjuvant setting.  
 


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This double-blind, randomized, placebo-controlled trial was testing recMAGE-A3 + AS02B as 
adjuvant therapy in 182 MAGE-A3-positive stage IB and II NSCLC patients after complete 
surgical resection of the tumor. The treatment phase of the study is finished, but follow-up is still 
ongoing. The results described here are based on the main analysis performed in November 2006 
after a median follow-up of 28 months.  
 
A total of 1214 ASCI injections have been administered to 122 patients and 60 patients have 
received 561 placebo administrations. The injections were overall well tolerated, with only 3 
Grade 3 events (2 cases of pain in extremity and 1 exacerbation of chronic obstructive 
pulmonary disease (COPD)) considered as possibly related to the ASCI treatment. Only 2 SAEs 
were considered as possibly related to the treatment with the MAGE-A3 ASCI: one injection site 
reaction and one exacerbation of COPD.  
 
The primary objective of the trial was to estimate the clinical activity of the treatment in terms of 
the time-to-recurrence or the disease-free interval. At the time of the main analysis, a Cox 
regression analysis performed to determine the relative improvement of the disease-free interval 
showed a 27% relative reduction in the MAGE-A3 ASCI group compared to the placebo group 
of the risk of cancer recurrence (hazard ratio = 0.73; p = 0.213) [Vansteenkiste, 2007]. 
 
 
1.2.3.3. Phase II proof-of-concept study in metastatic melanoma  
 
The encouraging early clinical data, the high proportion of metastatic melanoma patients with 
MAGE-A3-positive tumors and the important unmet medical need for these patients prompted 
the initiation of a Phase II trial in this disease (GSK study 249553/008), conducted in 
collaboration with the EORTC (EORTC study 16032-18031). In this randomized, open label 
study, the recMAGE-A3 protein is being administered as first-line treatment for metastatic 
disease to patients who have progressive metastatic melanoma with regional or distant skin 
and/or lymph-node lesions (unresectable stage III and stage IV M1a) but without visceral 
disease. The recMAGE-A3 protein is combined with 2 different Adjuvant Systems, AS02B (38 
patients) or AS15 (37 patients). The trial is currently ongoing, with 12 patients still on treatment. 
The results described here are based on interim analyses performed in October 2007 on the total 
cohort of 75 patients.  
 
Almost all adverse events observed were of Grade 1 or 2 and injection site reactions, fever and 
fatigue were by far the most common. In each of the groups, 2 grade 3 events possibly related to 
the treatment were observed: fatigue and muscle pain in the AS15 group, injection site reaction 
and tumor flare in the AS02B group. SAEs were reported for 6 patients in the AS02B arm and 
for 8 patients in the AS15 arm, and all of these except one were assessed to be unrelated to the 
study treatment. For one SAE, a case of disseminated intravascular coagulation occurring in a 
patient in the AS15 arm, the investigator assessed that it could not be excluded that this SAE was 
possibly related to the study treatment.  
 
The clinical activity of the treatments was assessed by the rate of objective clinical response. Of 
the 37 patients enrolled in the AS15 arm, 3 showed a complete response and 1 developed a 
partial response. Two of the complete responses are still ongoing, both with a duration of more 


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18 
than 30 months. One of the 38 patients in the AS02B arm showed a partial response. Another 7 
of the patients in the trial experienced disease stabilization for about 6 months or more.  
 
All the patients developed a specific anti-MAGE-A3 antibody response and some of the tested 
patients also developed a cell-mediated immune response. The observation of the immune 
responses suggests that the ASCI combining recMAGE-A3 with the AS15 Adjuvant System 
induces a more robust immunological response, both humoral and cell-mediated.  
 
The preliminary results of this trial showed an acceptable safety profile for the AS15 Adjuvant 
System and a potential for this ASCI formulation to induce a stronger immune response and lead 
to higher clinical activity. These findings confirm preclinical and clinical findings suggesting 
that AS15 is able to induce stronger immune and anti-tumor responses [Krieg, 2001; 
Meidenbauer, 2004; Ren, 2004; Speiser, 2005]. For this reason, GSK Biologicals has chosen 
recMAGE-A3 combined with AS15 as the preferred ASCI for development in melanoma and 
other cancers. 
 
1.2.3.4. Summary of the safety data from previous clinical trials with recMAGE-A3 ASCI 
  
The immunotherapy with recMAGE-A3 appears to be well tolerated. As of February 2008, more 
than 320 patients have received a total of more than 2,600 doses of recMAGE-A3 combined with 
3 different Adjuvant Systems or alone; approximately 500 doses of the MAGE-A3 + AS15 ASCI 
have been administered to some 60 patients. The AS15 Adjuvant System has also been 
investigated combined with other antigens in the treatment of patients with prostate cancer or 
breast cancer.  
 
Most of the adverse events observed in all these studies were to be anticipated and consist of 
local and systemic symptoms, such as injection site reactions, myalgia and fatigue. These 
adverse events have mainly been of Grade 1 or 2, but systemic Grade 3 events have occasionally 
been reported. Except for fatigue, these Grade 3 symptoms have rarely been assessed to be 
possibly related to the ASCI treatment. Almost all serious adverse events observed in the studies 
were assessed by the investigators as being unrelated to the study treatment.  
 
Please refer to the Investigator’s Brochure for a detailed description of all the clinical studies of 
the recMAGE-A3 protein and the results in terms of safety, clinical activity and immunological 
response.  
 
 
1.2.3.5. Identification of a gene signature being associated with clinical benefit in response 
to MAGE-A3 ASCI treatment 
 
Microarray gene profiling has been shown to be a powerful technique predicting treatment 
response or disease relapse in cancer patients, and a number of large scale clinical trials are 
currently in progress to validate the gene profiles potentially associated with different prognoses 
[Vantomme, 2004; Weigelt, 2005]. Also in the Phase II melanoma trial (GSK 249553/008), an 
attempt was made to identify from a tumor biopsy a gene signature that may predict a favorable 
clinical outcome for patients treated with the recMAGE-A3 + AS15 ASCI [GSK, unpublished]. 


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In this study, tumor samples were tested for gene expression using Affymetrix HU-U133.Plus 
2.0 gene chips covering about 47,000 transcripts. Biopsies from 69 patients were tested. In a first 
step, a supervised comparison was performed in order to find a group of genes able to cluster 11 
patients considered as presenting clinical benefit (complete response, partial response, mixed 
response or stable disease) in one group, and 11 patients with progressive disease in the other 
group. From this analysis, 41 differentially expressed probe sets (34 genes) that could segregate 
the patients with progressive disease and the patients with clinical activity were selected. In a 
second step, these 41 probe sets were used with the gene expression data from 69 patients to 
predict their clinical status in a hierarchical clustering approach. As a result, 2 clusters were 
clearly identified: one associated with patients who have progressive disease, and another one 
associated with clinical benefit. Interestingly, mixed-responders and patients with stable disease 
clustered with the group of objective responses, suggesting an association between the gene 
signature and a favorable clinical outcome following the MAGE-A3 ASCI treatment. 
 
Estimated Kaplan-Meier (KM) curves of the Time to Treatment Failure for the recMAGE-A3 + 
AS15 group of patients from the GSK 249553/008 melanoma trial also suggest that the presence 
of the gene signature correlates with a favorable clinical benefit in response to the recMAGE-A3 
+ AS15 therapy. In the GSK 249553/004 Phase II study of stage IB/II MAGE-A3 positive 
NSCLC patients, the same gene signature was found to be predictive of clinical activity.  In both 
settings (metastatic melanoma and primary NSCLC tumor), approximately 50% of patients 
present with this predictive gene signature. 
 
This set of genes differentially expressed in patients with clinical benefit in response to the 
MAGE-A3 ASCI treatment will in this protocol be referred to as the predictive gene signature. 
Overall, it is considered that selection of NSCLC or melanoma patients with MAGE-A3 positive 
tumors presenting with the predictive gene signature may increase the efficacy of treatment with 
recMAGE-A3 ASCIs by a factor of 2. Based on the gene profile data, GSK Biologicals has 
initiated a Phase II trial to prospectively validate the identified gene signature as a predictive 
marker of the clinical outcome for patients with unresectable stage III or stage IV M1a disease 
treated with the recMAGE-A3 + AS15 ASCI. 
 
 
1.2.4  Rationale for the use of Poly IC:LC in combination with recMAGE-A3 + AS15 ASCI 
 
Polyinosinic-Polycytidylic acid stabilized with poly-L-lysine and carboxymethylcellulose (Poly-
ICLC)
 is a synthetic double-stranded ribonucleic acid (dsRNA) with broad immune-enhancing 
effects.  These effects include induction of interferons (IFNs) and other cytokines, activation of 
cellular immunity (T4 cells, natural killer [NK] cells, and macrophages), an immune adjuvant 
effect, and activation of several nuclear enzyme systems (oligoadenylate synthetase [OAS] and 
the dsRNA dependent p68 protein kinase [PKR]) involved in antiviral and antitumor host 
defenses.  More recently it has been shown to have broad gene regulatory actions as well. 
Poly-IC:LC is a Toll-like receptor 3 agonist (TLT3) has been under clinical investigation since 
the 1970s, including human trials for a wide variety of neoplastic, viral, and neurologic 
conditions [Levy 1975, Levy 1984, Levy 1992; Houston 1976, Carter 1987]. The original US 
IND, held by Dr. Hilton Levy, investigated dose levels of greater than 100 µg/kg of Poly-ICLC.   


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A separate IND, No. 43,984, was opened in 1995 under which 6 clinical studies have thus far 
been conducted in approximately 350 glioma or glioblastoma patients.  Dose regimens used in 
these trials generally consisted of 20-µg/kg Poly-ICLC 3 times each week. The degree of adverse 
effects seen with of Poly-ICLC depends on 3 factors: (1) dose, (2) route of injection, and (3) 
health status of the patient.  Early phase 1 studies were done to determine the maximum tolerated 
dose, under the assumption that this was also the most effective dose.  In these studies of cancer 
patients, it was found that the maximum tolerated dose was about 12 mg/m2 when given 
intravenously in patients who were not terminally ill.  Patients typically developed fevers of 
40°C, myalgias, arthralgias, malaise, and some nausea and vomiting.  Fever was the primary 
dose-limiting factor.  At this dose, the mean serum interferon level was 2000 IU/mL.  While the 
dose of exogenous interferon rarely attains this level, levels of 100 IU/mL after exogenous 
interferon are associated with the same adverse effects as those in high-dose Poly-ICLC.  In most 
of the early cancer trials, doses around 6 mg/m2 of Poly-ICLC IV were generally used.  Mild 
(grade 1), transient (less than 7 days) hepatic enzyme elevations were described in a trial of 100 
µg/kg Poly-ICLC given intravenously in multiple sclerosis patients.  In 3 patients, the elevated 
enzyme levels were prolonged for greater than 7 days, but in all patients, the enzyme levels 
returned to normal after temporary discontinuation of the Poly-ICLC.  One paralyzed multiple 
sclerosis patient in this group suffered a fatal pulmonary embolus that was not judged to be due 
to the drug.  Poly-ICLC has been associated with a coagulopathy in dogs, but not in other species 
(including primates); and there has been no change in the expected incidence of deep venous 
thrombosis, pulmonary embolus, or coagulopathy in multiple sclerosis, AIDS, or malignant 
glioma patients on low-dose intramuscular Poly-ICLC.  It was subsequently shown that a low 
dose of Poly-ICLC was better than a high dose for enhancing immune effects, and that the higher 
dose actually inhibited a number of cell-associated immune functions.  It was also found that 
intramuscular injection brought about much milder side effects. Representative safety data in 
humans at a dose of 20-µg/kg Poly-ICLC 3 times each week have been reported in 2 clinical 
trials:  Protocol NABTC 01-05 entitled: “An open multicenter trial of Poly-ICLC treatment of 
newly diagnosed glioblastoma with Poly-ICLC as an adjuvant to external beam radiation” and 
Protocol NABTC 01-06 entitled: “An open multicenter trial of Poly-ICLC in subjects with 
recurrent anaplastic glioma.”  Treatment with Poly-ICLC in these 2 clinical trials appeared to be 
well tolerated; adverse reactions were usually mild or moderate in nature (grade 1 or 2 by the 
National Cancer Institute Common Terminology Criteria for Adverse Events scale). 
Toll like receptor agonists have been shown to be effective adjuvants and in pre-clinical models 
poly IC:LC has been shown to be an effective booster of immunity when added to a peptide 
vaccine.  Celis and colleagues have shown that in melanoma mouse models that polyIC:LC is a 
potent adjuvant for a peptide vaccine with thre addition of an anti-CD40 antibody, or with the 
addition of other type TLR agonists (Cho and Celis, 2009, Wells et al, 2008)). Studies in 
infectious disease vaccine models have confirmed the synergy of multiple TLR agonists within a 
vaccine strategy (Raman al 2010), Different TLR agonists signal via different pathways, again 
suggesting that combinations of TLR agonists may induce optimal immune responses (Agrawal 
et a, 2003). Based on the compiled data in which different TLR agonists have been shown to 
have immunostimulatory activity, it was felt that the combination of the TLR 3 agonist poly 
IC:LC with ASCI, which contains a CpG ODN TLR9 agonist, was rational. 
Based on the doses used in the above trials, and the side effect profile of the drug, a single 1000 
μg dose of Hiltonol will be used subcutaneously in the same limb as the recMAGE-3 + AS15 
ASCI injections are given.  As a function of surface area, the Human Equivalent Dose of 1000μg 


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of Poly-ICLC corresponds to 5-10μg in the mouse, which is a dose that has marked 
immunomodulatory effects in multiple animal models.   For further details of preclinical safety 
and toxicology tests of Poly IC:LC, see the Investigator
's Brochures.  
 
 
1.3. Rationale for the study: relapse of patients with resectable stage IV melanoma after 
surgery 
 
Surgery of stage IV melanoma is not generally curative and approximately 90% of the patients 
will ultimately die of metastatic disease [Markovic, 2007]. There is thus a need for adjuvant 
therapy to prevent disease relapse after surgical resection of the primary tumor. The role of 
chemotherapy as adjuvant treatment remains unclear with no tangible benefit in terms of 
prolonged overall survival [Nathan, 1995]. Non-specific immunotherapy with high-dose 
interferon has shown activity on disease-free survival, but its impact on overall survival in stage 
III resected melanoma is less clear, and there is simply no track record for interferon in resected 
stage IV melanoma [Garbe, 2001]. Low dose Interferon has been investigated in several studies 
with patients with stage III disease and did not show any significant improvement in Relapse-
Free Survival or Overall Survival. The recent Phase III trial with pegylated interferon showed a 
significant long-term impact of this treatment on relapse-free survival, but not on overall survival 
in resected stage III disease [Eggermont, 2007]. Subgroup analyses in this study further showed 
that patients with macroscopic lymph node involvement did not derive a significant clinical 
benefit from this treatment, warranting the development of new treatment options for these 
patients. Active immunization against tumor antigens is certainly one such possible approach.  
 
The recMAGE-A3 ASCI has shown promising results in Phase II studies: In the recent Phase II 
proof-of-concept trial of two different formulations of the recMAGEA3 ASCI in melanoma 
patients with early metastatic disease (GSK study 249553/008), a strong and robust 
immunological response was seen, and the safety of the treatments was considered acceptable, 
raising no specific safety concerns. Objective clinical response was observed for 5 of the 75 
patients treated and another 7 patients showed disease stabilization (over periods varying 
between 5 and 12 months, some still ongoing). These preliminary findings support the 
development of the recMAGE-A3 ASCI in the adjuvant melanoma setting, particularly in the 
stage IV NED setting in which microscopic residual disease makes relapse of disease likely 
within a year of surgical resection.  
 
Additional support for developing the recMAGE-A3 ASCI in the adjuvant melanoma setting 
may be found in the Phase II proof-of-concept trial of the recMAGE-A3 ASCI as adjuvant 
therapy for patients with resectable stage IB and II NSCLC. This study (GSK 249553/004, see 
Section 1.2.3.2) showed a relative reduction of the risk of disease recurrence of 27% in the group 
receiving the ASCI compared to the placebo group. The immunological responses observed in 
melanoma patients and NSCLC patients in the adjuvant and metastatic settings, respectively, are 
very similar, so it is reasonable to expect that an equivalent tumor response will be seen in the 
adjuvant resected stage IV melanoma setting.  
 
The justification for adding Poly IC:LC as an adjuvant to recMAGE-A3 + AS15 ASCI are data 
suggesting in pre-clinical murine models that the use of two different TLR Agonists added to a 


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vaccine or other immunotherapy approach can augment its anti-tumor activity and 
immunogenicity, as cited in section 1.2 above. Since the AS15, containing CpG709 and MPL 
that are a TLR 9 and TLR3 agonists respectively, is well tolerated when added to a recMAGE-3 
ASCI, and the Poly IC:LC is a TLR3 agonist that is quite well tolerated as well, the addition of 
the different agonists seems sensible and is likely to be safe and well tolerated. 
 
Taking into account the tumor-specificity of MAGE-A3, the MAGE-A3 expression levels in 
stage IV melanomas, the acceptable tolerability of the recMAGE-A3 + AS15 ASCI and 
promising results from Phase II proof-of-concept trials (GSK 249553/008 and 249553/004), it is 
proposed to initiate a randomized Phase II trial to evaluate the immunogenicity, safety and 
tolerability of the recMAGE-A3 + AS15 ASCI with or without TLR 3 agonist Poly IC:LC  in 
patients with stage IV resected melanoma at very high risk of relapse.   
 
 
2. OBJECTIVES 
  
2.1. Primary objectives 
 
The primary objectives of this clinical trial are: 
 
• To evaluate the safety and tolerability of recMAGE-A3 + AS15 ASCI compared to recMAGE-
A3 + AS15 ASCI in combination with Poly IC:LC in patients with resected stage IV 
melanoma. 
 
• To evaluate the immunogenicity of recMAGE-A3 + AS15 ASCI compared to recMAGE-A3 + 
AS15 ASCI in combination with Poly IC:LC as measured by antibodies serum titers (such as 
anti-MAGE-A3) in patients with resected stage IV melanoma.  
 
Refer to Section 10.1 for the definition of the primary endpoints.  
 
 
2.2. Secondary objectives 
   
 
• To evaluate the clinical activity of the recMAGE-A3 + AS15 ASCI compared to recMAGE-A3 
+ AS15 ASCI in combination with Poly IC:LC in terms of relapse-free survival (RFS) and 
overall survival (OS) in the overall study population. 
 
• To evaluate the immunogenicity of the recMAGE-A3 + AS15 ASCI compared to recMAGE-
A3 + AS15 ASCI in combination with Poly IC:LC measured by T cell responses against 
MAGE-A3.  
 
• To assess the gene profile of tumor tissue in the two study arms and evaluate the activity of the 
recMAGE-A3 + AS15 ASCI +/- Poly IC:LC in terms of RFS in the sub-population of patients 
presenting a potentially favorable gene signature (see Section 6.4.4), and in the sub-population 
of patients without this gene signature.  
Refer to section 10.2 for definitions of secondary endpoints.  


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2.3. Translational research objectives  
 
• Translational research will include assessment of MAGE-A3 expression of tumor tissue from 
recurrent lesions and gene profiling of recurrent or new lesions.   
 
3. STUDY DESIGN OVERVIEW 
  
3.1. Study design 
 
This is an open, randomized, two-arm, Phase II study in MAGE-A3 positive patients with 
completely resected stage IV melanoma to assess the safety, tolerability, immunogenicity and 
clinical activity (RFS and OS) of recMAGE-A3 + AS15 ASCI in combination with Poly IC:LC  
as adjuvant therapy compared to recMAGE-A3 + AS15 ASCI alone.  
 
Patients will be randomized (1:1 ratio) to receive either recMAGE-A3 + AS15 ASCI or 
recMAGE-A3 + AS15 ASCI in combination with Poly IC:LC. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 1. Overview of Study Design. 
 
 
 
 




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25 
4. STUDY POPULATION 
  
4.1. Number of patients  
 
According to our current assumptions, up to 44 patients will be enrolled in this trial to end up 
with 40 evaluable patients (considering a possible 10% withdrawal rate from the study) in two 
cohorts of 20 patients each.  
 
Refer to Section 10.3 of the protocol for a detailed description of the assumptions applied for the 
calculation of the sample size.  
 
 
4.2. Inclusion criteria 
 
The following criteria are to be checked at the time of randomization. A patient may only be 
included in the study if ALL of the following criteria are FULFILLED:  
 
1. Written informed consent for the study will be obtained prior to the performance of MAGE-
A3 expression screening on resected tumor tissue or any other protocol-specific procedure. 
 
2. Male or female patient with histologically proven and completely resected stage IV cutaneous 
or mucosal melanoma.  In terms of the AJCC classification [AJCC, 2009, see Appendix D], this 
means that patients with resected M1a-b-c (stage IV) disease may be enrolled.  
 
3. The patient must have been surgically rendered free of disease no more than 12 weeks before 
the randomization.  
 
4. Patient is equal to or greater than 18 years old at the time of signing the informed consent 
form.  
 
5. The patient’s tumor shows expression of the MAGE-A3 gene, as determined by RT-PCR 
analysis on formalin-fixed paraffin-embedded (FFPE) tumor tissue.  In all patients in whom it 
can be obtained, a fresh tumor portion of the resected tumor will be stored in RNA later and 
analyzed for gene profiling (section 6.3.2; section 6.4).  
 
6. The patient has fully recovered from surgery.  
 
7. ECOG performance status of 0 or 1 at the time of randomization.  
 
8. The patient must have adequate bone-marrow reserve, adequate renal function and adequate 
hepatic function as assessed below by standard laboratory criteria.  
 
Absolute neutrophil count 
 1.5 x 109/L  
Platelet count 
 75 x 109/L  
Serum creatinine 
 1.5 times the Upper Limit of Normal (ULN)  
Total bilirubin 
 1.5 times the ULN 


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Transaminase (ALT - AST)  2.5 times the ULN 
 9. If the patient is female, she must be of non-childbearing potential, i.e. have a current tubal 
ligation, hysterectomy, ovariectomy or be post menopausal, or if she is of childbearing potential, 
she must practice adequate contraception for 30 days prior to randomization, have a negative 
pregnancy test and continue such precautions during the entire study treatment period and for 2 
months after completion of the injection series. For further details, refer to Section 6.3.1.2. 
 
10. Men must also agree to use an adequate method of contraception. 
 
11. In the opinion of the investigator, the patient can and will comply with all the requirements 
of the protocol.  
 
 
4.3. Exclusion criteria 
 
The following criteria should be checked at the time of randomization. If any apply, the patient 
must not be included in the study:  
 
1. The patient has an ocular melanoma.  
 
2. The patient has in-transit metastases.  
 
3. The patient has been treated or is scheduled to be treated with an adjuvant anticancer therapy 
after the metastasectomy that qualifies the patient for inclusion in the present trial. 
 
• One prior systemic treatment with an immunomodulator (i.e., interferon, vaccine and/or anti-
CTLA-4) after a previous surgery is permitted, provided that the last dose has been administered 
at least 45 days before randomization in the present trial.  
 
• Previous radiotherapy is permitted, provided that the treatment has been completed before the 
surgery that qualifies the patient for participation in the present trial . 
 
4. The patient requires concomitant chronic treatment (more than 7 consecutive days) with 
systemic corticosteroids or any other immunosuppressive agents. The use of prednisone, or 
equivalent, at a dose of < 0.125 mg/kg/day (absolute maximum 10 mg/day) or topical steroids is 
permitted.  
 
5. Use of any investigational or non-registered product (drug or vaccine) other than the study 
treatment within 30 days preceding the randomization or planned use during the study period.  
 
6. The patient has a history of autoimmune disease such as, but not limited to, multiple sclerosis, 
lupus, and inflammatory bowel disease. Patients with vitiligo are not excluded.  
 
7. The patient has a family history of congenital or hereditary immunodeficiency.  
 


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8. The patient is known to be positive for Human Immunodeficiency Virus (HIV) or has another 
confirmed or suspected immunosuppressive or immunodeficient condition.  
 
9. History of allergic disease or reactions likely to be exacerbated by any component of the 
treatments.  
 
10. The patient has psychiatric or addictive disorders that may compromise his/her ability to give 
informed consent or to comply with the trial procedures.  
 
11. The patient has concurrent severe medical problems, unrelated to the malignancy, that would 
significantly limit full compliance with the study or expose the patient to unacceptable risk.  
12. The patient has previous or concomitant malignancies at other sites, except effectively 
treated non-melanoma skin cancers or carcinoma in situ of the cervix or effectively treated 
malignancy that has been in remission for over 5 years and is highly likely to have been cured.  
 
13. The patient has an uncontrolled bleeding disorder.  
 
14.  For female patients: the patient is pregnant or lactating. 
 
 
 
5. INVESTIGATIONAL PRODUCTS AND ADMINISTRATION  
 
5.1. Study treatment: recMAGE-A3 + AS15 ASCI and Poly IC:LC 
 
The recMAGE-A3 + AS15 ASCI to be used in this study have been developed and manufactured 
by GSK Biologicals; Oncovir manufactures the Poly IC:LC (Hiltonol). 
 
The Quality Control Standards and Requirements for these products are described in separate 
release protocols, and the required approvals have been obtained by manufacturers. 
 
 
5.1.1. recMAGE-A3 + AS15 ASCI  
 
In the study, the recMAGE-A3 + AS15 ASCI will be administered by using a sterile two-vial set 
comprising:  
 
• One vial with the lyophilized preparation containing 300 µg recMAGE-A3 antigen plus 420 µg 
of CpG7909 (a part of the Adjuvant System AS15),  
 
• One vial with liquid adjuvant diluent AS01B (liposomes containing 50 µg of MPL®, 1mg of 
DOPC, 250 µg of cholesterol and 50 µg of QS21 in phosphate-buffered saline), making up the 
remainder of the Adjuvant System AS15. The final recMAGE-A3 + AS15 ASCI for 
administration is obtained by reconstitution of the lyophilized preparation with the adjuvant 
diluent. One recMAGE-A3 + AS15 ASCI dose consists of 0.5 ml.  
 


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28 
 
 
5.1.2. Poly IC:LC (Hiltonol) 
 
Poly IC-LC (Hiltonol) is a synthetic, nuclease resistant, hydrophilic complex of polyinosinic and 
polycytidylic acid, stabilized with poly-L-lysine and carboxymethylcellulose.  Poly-ICLC is an 
opalescent solution in 2-mL vials.  Each 1 mL of Poly-ICLC for injection contains 2-mg poly-IC, 
1.5-mg poly-L-lysine, and 5-mg sodium carboxymethylcellulose in 0.9% sodium chloride 
solution and adjusted to pH 7.6 to 7.8 with sodium hydroxide. 
 
How supplied 
Poly-ICLC is supplied in single-use vials containing 1 mL of a 2 mg/mL opalescent solution.  A 
total of 0.5 ml = 1 mg is withdrawn from the vial using sterile technique and is administered 
deep subcutaneously as supplied.  
 
Each vial poly-ICLC will be labeled with the following information: 
Drug Name 
Concentration 
Storage Conditions 
Lot Number 
Date of Manufacture,  Manufacturer 
Investigational Use Statement 
 
Storage and stability 
Poly-ICLC is stable at room temperature for brief periods (days).  Treatment sites are asked to 
store the drug at approximately 40 F.  Patients should be advised to store their doses in a 
standard refrigerator; when necessary, vials may be kept at room temperature up to 24 hours.  
The vials should not be frozen.   
 
Supplier 
The drug to be used in this study is prepared and packaged under GMP, under contract to 
Oncovir, Inc.  It is then tested for quality, activity and pyrogenicity by Oncovir, Inc. 
 
Drug accountability 
The intent of drug accountability is to assure that supplied agents are only used for patients 
enrolled on an approved trial. According to FDA guidelines the investigator is ultimately 
responsible for all agents shipped in his/her name. FDA regulations require investigators to 
establish a record of the receipt, use, and disposition of all investigational agents. The sponsor of 
investigational trials has the responsibility to assure the FDA that systems for drug accountability 
are being maintained by investigators in their clinical trial program. Investigators may delegate 
responsibility for drug ordering, storage, accountability and preparation to his/her designee. 
 
Drug ordering information:  
 
   
   
 
  
 
 
    
   
 
 




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Protocol version 7.0  10/28/2015   
30 
 
 
 
5.2.1.1. Criteria for postponement of study treatment  
 
If any one of the following events occurs at the time scheduled for administration of the study 
treatment, the patient may be treated at a later date (i.e., the entire program of study visits and 
immunizations is interrupted), within the time window specified below, or withdrawn at the 
discretion of the investigator. The patient must be followed until resolution of the event, as with 
any AE.  
 
• Acute disease at the time of treatment administration. (Acute disease is defined as the 
presence of a moderate or severe illness with or without fever). Study treatment can be 
administered to persons with a minor illness such as diarrhea or mild upper respiratory 
infection. 
 
• Any Grade 2 or more adverse event possibly related to the study treatment and that is 
present at the time of treatment administration. In case of postponement of study treatment 
administration for any of the above reasons, the following rules have to be followed:  
 
• During the induction phase (study treatment administration number 1 to 5), the maximum 
delay for postponement of study treatment administration is 3 weeks.  
 
• During the maintenance phase (study treatment administration number 6 to 13), the 
maximum delay for postponement of study treatment administration is 3 months. When a 
study treatment administration has to be postponed, a visit to administer the missed 
treatment should be planned as soon as possible to catch up with the originally planned 
schedule.  
 
If the administration occurs within the specified delay, the treatment dose to administer 
corresponds to the next one in the initial order of administration. Example: if a patient has 
already received doses 1 and 2 of study treatment but the administration of dose 3 of study 
treatment has to be postponed, the patient will receive dose 3 if (s)he comes for her/his next 
administration a maximum of 3 weeks after the originally planned date of Visit 3 (refer to Table 
3).  
 
If the administration does not occur within the delay stated above, the missed dose will not be 
administered. At the time the patient reintegrates the study, (s)he will receive the dose as planned 
in the initial schedule of administration. Example: if a patient has already received doses 1 and 2 
of study treatment but the administration of dose 3 of study treatment has to be postponed for 
more than 3 weeks, the patient will receive dose 4 if (s)he comes for his/her next administration 
(refer to Table 4).  
 
The next doses will be planned at a time allowing:  
 
• A minimum of 14 days between 2 treatment administrations,  


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Protocol version 7.0  10/28/2015   
31 
 
• To keep up with the schedule as based on the date of first study treatment administration.  
 
 
 
Week  0 1 2 3 4 5 6 7 8 
9 10 11 
12 
In
it
ia
l schedu
le V
is
it  
1    2    
3 
    4    
  
5 
Dose  1    2    
3 
    4    
  
5 
 
  
                    
    
 
Actua
l schedu
le 
due to 
postponement 
V
is
it  
1    2    
3 
 
3    4    
5 
Dose  1    2        
3    4    
5 
  
  
          
  
    
 
  
 
  
 
 
 
 
  
         Adm
in
istrat
ion 
postponed for 
2 weeks 
  2 weeks de
lay for next 
doses 
  
a. A
lso app
l
ies for postponement up to 3 months dur
ing ma
intenance phase. 
Table 3. Examp
le of postponement of study treatment dose for a max
imum of 3 weeks dur
ing Induct
ion Phase
a. 
 
 
 
  
Week  0 1 2 3 4 5 6 7 8 
9 10 11 
12 
In
it
ia
l schedu
le V
is
it  
1    2    
3 
    4    
  
5 
Dose  1    2    
3 
    4    
  
5 
  
  
                  
  
  
  
 
Actua
l schedu
le 
due to 
postponement 
V
is
it  
1    2    
3 
    
  4    
5 
Dose  1    2    
X 
    
  4    
5 
  
 
               
    
    
  
 
 
 
 
 
  
         Adm
in
istrat
ion 
postponed for 
4 weeks 
  
 
   2 week 
de
lay for 
next dose 
  
 
  
a. A
lso app
l
ies for postponement up to 3 months dur
ing ma
intenance phase. 
Table 4. Examp
le of postponement of study treatment dose for more than 3 weeks dur
ing Induct
ion Phase
a. 
 
 
5.2.1.2. Criteria for permanent stopping of study treatment and Dose Limiting Toxicity  
 
If any of the following criteria becomes applicable during the study, the patient will be required 
to discontinue the study treatment: 
 
a. Evidence of disease relapse (see Section 6.3.12 for the definition of relapse).  
 
b. Treatment for melanoma with one of the following:  


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Protocol version 7.0  10/28/2015   
32 
 
• Investigational product or non-registered product other than the study 
recMAGE-A3 + AS15 ASCI +/- Poly IC:LC.  
 
• Other anticancer treatments, including but not limited to chemotherapeutic or 
immunomodulating agents and radiotherapy.  
 
c. Any Grade 2 or more allergic reaction following the administration of study treatment.  
 
d. Any intolerable adverse event defined as any grade 3 or 4 toxicities felt to be treatment 
related; or, any persistent moderate adverse event defined as a grade 2 toxicity that could 
be worsened by subsequent administration of the study treatment, at the investigator’s 
discretion.  
 
e. Clinical signs or symptoms indicative of vasculitis, glomerulonephritis or any 
autoimmune disorder of grade 3 or more, excluding vitiligo or thyroiditis that can be 
treated with replacement therapy; in such cases, appropriate clinical and laboratory 
testing will be performed to identify and characterize that disorder.  
 
f. Appearance of any confirmed or suspected immunosuppressive or immunodeficient 
condition, including human immunodeficiency virus (HIV) infection, or any medical 
condition requiring the administration of immunosuppressive agents or systemic 
corticosteroids.  
 
g. Inability of the patient to complete the study evaluations/visits because of unforeseen 
circumstances.  
 
h. The patient develops other conditions for which, in the investigator’s opinion, it is in  
the patient’s best interest to be withdrawn from the study treatment.  
 
i. The patient requests to be withdrawn from the study. 
 
j. For female patients, pregnancy or decision to become pregnant (see Section 8.12).  
 
Procedures to be followed in any of these situations are described in Section 6.3.10.2.  
 
 
 
5.3. Treatment assignment and randomization  
 
5.3.1. Patient identification  
 
Patient numbers will be assigned sequentially to patients as soon as they sign the informed 
consent form consenting to participate in the study.  
 


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After the patient has signed the informed consent form, the investigator will enter the patient into 
the screening section of the CRF. General demographic details and tumor characteristics will be 
entered in the screening section for each patient for whom a tumor sample is sent for analysis of 
MAGE-A3 expression (GSK Biologicals or contracted lab). When a patient is eligible (i.e., 
meets all the inclusion criteria and does not meet any of the exclusion criteria), the investigator 
or an authorized designee will continue with the actual CRF for this patient.  
 
5.3.2. Randomization  
 
Randomization will take place when it has been ascertained that the patient is fully eligible and 
at most 1 week before administration of the first injection of the study product, which will take 
place a maximum of 12 weeks after surgery.  
 
Randomization of patients and treatment allocation at the investigator site will be performed 
utilizing the Moffitt Cancer Center’s Subject Registration and Randomization Server, a web-
based interactive randomization system (available 24H/day, 7D/week). It will be open-label and 
stratified on the following variables:  
 
1. M Stage of the disease (M1a vs. M1b vs. M1c) 
 
2. Prior treatment with IFN or anti-CTLA-4 (Yes or No) 
 
The list of randomization numbers will be generated by the study statistician and passed on to 
Moffitt Research IT at MCC to be programmed in the automated web-based randomization 
system for this specific trial. 
 
Randomization will be performed a maximum of 1 week before the first study treatment 
administration, which is assigned as Visit 1. Figure 3 presents the timeline for obtainment of 
patients’ informed consent and the procedures to be performed before the beginning of the study.  
 
After having verified that the patient is fully eligible and having obtained the patient’s informed 
consent to participate in the study, the person in charge of the treatment administration will 
access the randomization system on Internet. Upon providing a patient number and the stage, and 
prior treatment with IFN or anti-CTLA-4, the randomization system will use the stratified 
randomization method to determine the treatment number to be used for the patient.  
 
The actual treatment number used for each treatment administration of the patient must be 
recorded in the CRF.  As soon as the targeted number of randomized patients has been reached, 
the enrollment of patients will be frozen. However, to allow patients at the screening phase or 
those who have signed the informed consent form at the time of enrollment hold to be enrolled, 
an over-enrollment of 5% (4 patients) will be allowed.  
 
 
 
 
 


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Protocol version 7.0  10/28/2015   
34 
 
 
 
 
 
 
 
                                     
Screening visit 1 (S1) 
ICF  signed  
 
Surgical resection of  tumor  
        
Screening visit 2 (S2) 
result of MAGE-A3 test known 
check eligibility criteria 
      
Randomization 
 
Study Visit 1 (V1): 
Recheck  eligibility criteria 
First ASCI  +/- Poly IC:LC 
injection 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
 
        
       
 
                                     
  just before/after surgery                    max. 1 week    
                                     
       
                             
 
                                     
                 max. 12 weeks 
            
ICF = informed consent form signature  
                        
 
Figure 3. Timelines for pre-study procedures and obtaining informed consent. 
 
 
 
5.4. Storage  
 
All investigational products must be stored in a safe and locked place with no access by 
unauthorized personnel.  
 
The ASCI treatments will be stored at the defined temperature range (i.e., +2 to +8°C/ 36°F to 
46°F).  
 
The storage temperature of treatments will be monitored daily by means of validated temperature 
monitoring devices and the temperature measurements will be recorded during working days, 
preferably at the same time of the day (e.g., at the beginning of the day). Freezing indication will 
be continuously controlled by an appropriate device placed close to the treatment(s) and recorded 
daily during working days, preferably at the same time of the day (e.g., at the beginning of the 
day).  
 
 
Please refer to the Pharmacy Manual for further details. 
 
 
 


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5.5. Packaging  
 
Please refer to the Pharmacy Manual for further details. 
 
5.6. Treatment accountability  
 
The Sponsor, MCC, is responsible of the Study Drug accountability. 
 
5.7. Concomitant medication/treatment 
 
At first study visit (Visit 1), the investigator should record the chronic medication, i.e., any 
medications taken by the patient from a minimum of 6 months before the surgical resection of 
the tumor.  
 
At each study visit, the investigator should question the patient about any medications taken.  
 
All concomitant medications (including changes in chronic medication but not chronic 
medication itself - see next paragraph), with the exception of vitamins and/or dietary 
supplements, administered at ANY time during the period starting with administration of each 
dose of study treatment and ending 30 days after each dose of study treatment are to be recorded 
with generic name of the medications (trade names are allowed for combination drugs, i.e., 
multi-component drugs), medical indication, total daily dose, route of administration, start and 
end dates of treatment.  
 
 
5.7.1. Permitted medications  
 
Patients should receive medication appropriate to their health condition during the whole study.  
 
During the period starting with administration of each dose of study treatment and ending 30 
days after the administration of each dose, concomitant medications administered for the 
treatment of an AE must be recorded in the CRF with generic name of the medications (trade 
names are allowed for combination drugs only), medical indication (including which AE), total 
daily dose, route of administration, start and end dates of treatment. Similarly, concomitant 
medications administered for the treatment of a SAE, at any time, must be recorded on the SAE 
screens in the CRF, as applicable (Refer to Section 8.2 for the definition of a SAE).  
 
 
5.7.2. Prohibited medications or non-drug therapies  
 
Patients may not receive concomitant treatment during the study with any of the following:  
 


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a. Investigational or non-registered product for melanoma other than the recMAGE-A3 + 
AS15 ASCI ± Poly IC:LC. 
  
b. Other anticancer treatments, including but not exclusively, chemotherapeutic, 
immunomodulating agents and radiotherapy.  
 
c. Chronic use (more than 7 consecutive days) of systemic corticosteroids or any other  
immunosuppressive agents. 
 
The use of prednisone, or equivalent, < 0.125 mg/kg/day (absolute maximum 10 
mg/day), inhaled corticosteroids or topical steroids is permitted.  
 
d. Administration of a vaccine not foreseen by the study protocol during the period within 
the prohibited time as specified in Section 5.8.3.  
 
e. Repetitive administration of immunoglobulins and/or any blood products during the 
study period.  
 
Any of these specifically contraindicated treatments and/or medications administered at any time 
during the study period are to be recorded in the CRF with generic name of the medications 
(trade names are allowed for combination drugs only), medical indication, total daily dose, route 
of administration, start and end dates of treatment.  
 
If any of these is taken, it will require either permanent discontinuation of study treatment (see 
Section 5.2.1.2) or may determine a patient’s evaluability in the according to protocol (ATP) 
analysis (see Section 10.3.1 for definition of study populations to be evaluated).  
 
 
 
5.7.3. Time-window for prophylactic vaccination against infectious diseases  
 
Immunization with any commercial anti-infectious vaccine may be performed during the study. 
However, this may not take place during the period from 7 days before any study treatment 
administration to 7 days after it. Thus, if the study treatment is to be given on a notional Day 0, 
then immunization with any commercial vaccine may be performed on or before Day -8, and on 
or after Day 8.  
 
Any commercial vaccine administered in the period beginning 30 days preceding each dose of 
study treatment and ending one month (minimum 30 days) after each dose of study treatment is 
to be recorded with trade name, route of administration and date(s) of administration.  
 
Any investigational medication or treatment administered throughout the study period (i.e., from 
Day 0 through Visit 13) must be recorded in the CRF. 
 
 
 


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6. STUDY ASSESSMENTS AND PROCEDURES  
 
6.1. General study aspects 
  
6.1.1. Attendance for study visits  
 
It is the investigator’s responsibility to ensure that the intervals between visits/contacts are 
strictly followed.  
 
The timing for study visits can be found in Section 6.2. Permitted deviations from the stipulated 
date of visit (due e.g., to week-ends or public holidays) will be as follows:  
 
Induction phase ± 4 calendar days.  
 
Maintenance and follow-up phases ± 14 calendar days.  
 
In case of deviations exceeding these limits, the sponsor, MCC will decide whether the violation 
is to be regarded as:  
 
• A minor violation (i.e., without consequence),  
 
• A major violation (resulting in exclusion of the patient from the population of patients 
evaluable according to protocol).  
 
When a study treatment has to be postponed (refer to Section 5.2.1.1 for details on postponement 
of study treatment), the next study visit should be planned at a time allowing:  
 
• A minimum of 14 days between 2 treatment administrations,  
 
• To keep up with the schedule as based on the date of first study treatment 
administration (Visit 1).  
 
 
6.2. Outline of study procedures  
 
6.2.1. Induction phase  
 
The first administration of study treatment (Visit 1) will take place a maximum of 12 weeks after 
surgical resection of the tumor. 
 








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Protocol version 7.0  10/28/2015   
41 
tumor tissue for MAGE-A3 expression and expression of a predictive gene list associated to 
treatment. Testing for MAGE-A3 expression will not be completed until informed consent has 
been obtained regardless of when the tumor resection was performed. 
 
 
6.3.1.2 Visit S2 
 
This visit will take place after V1, once the results of MAGE-A3 expression testing are known. 
 
The inclusion and exclusion criteria will be evaluated. Only patients who, in the opinion of the 
investigator, meet all inclusion criteria and none of the exclusion criteria may be included in this 
study. The following will also be performed at visit S2: 
 
Pregnancy test (for women of child-bearing potential)  
 
When considering the enrollment of females of childbearing potential, please take note of the 
following definitions:  
 
• Adequate contraception is defined as a contraceptive method with failure rate of less than 1% 
per year when used consistently and correctly (when applicable, as mentioned in the product 
label) for example abstinence, combined or progestogen oral contraceptives, injectable 
progestogen, implants of levonorgestrel, oestrogenic vaginal ring, percutaneous contraceptive 
patches or intrauterine device (IUD) or intrauterine system (IUS), vasectomy with documented 
azoospermia of the sole male partner or double barrier method (condom or occlusive cap plus 
spermicidal agent).  
 
For azoospermia, “documented” refers to the outcome of the investigator
's or his/her designee’s 
medical examination of the patient or review of the patient
's medical history for study eligibility, 
as obtained via a verbal interview with the patient or from the patient’s medical records.  
 
• Post-menopause: Menopause is the age associated with complete cessation of menstrual cycles, 
menses, and implies the loss of reproductive potential by ovarian failure. A practical definition 
accepts menopause after 12 consecutive months without menses with an appropriate clinical 
profile (for example, at the appropriate age e.g., > 45 years).  
 
• The patient’s medical history (inclusive cancer history) will be recorded.  
 
• SAEs related to study participation or concomitant medications are recorded (see Section 8.2). 
  
• Blood sampling for safety haematological and chemical assays (15 ml, as described in Section 
6.4.2).  
 
• Neck and chest CT scan, complete abdomen CT scan and pelvic CT scan within 4 weeks 
preceding treatment.  
 


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42 
• Brain MRI scan. If such a scan has been performed up to 4 weeks before treatment, this need 
not be repeated. Brain CT may be used instead of brain MRI scan if patients are unable to have 
an MRI, i.e. due to placement of a pacemaker or defibrillator, etc.  
 
• Physical examination  
 
• When the patient is eligible (meets all of the inclusion criteria and none of the exclusion 
criteria), randomization via the internet will take place (for procedure see Section 5.3.2). A 
maximum period of 2 weeks is allowed between randomization and the first administration of the 
study drugs.  
 
 
6.3.2. Resection procedure  
 
After resection, formalin-fixed paraffin-embedded (FFPE) tumor tissue will be tested for 
MAGE-A3 expression. A paraffin-embedded tissue block sample of at least 10 mm³ or 
alternatively 20 unstained slides (19 slides of 10 µm and 1 slide of 5 µm for a total of at least 100 
mm2 tumor tissue) will be provided to GSK Biologicals or a contracted lab. The laboratory will 
then extract the RNA from these samples and assess the MAGE-A3 expression by quantitative 
PCR. If there are several melanoma blocks, it is preferred that 2 of these are sent for 
determination of MAGE-A3 expression (As described above, a FFPE block of at least 10 mm³ is 
preferred). The second block will only be tested, in case the first tested is MAGE-A3-negative. 
The patient will be considered eligible on this criterion, if at least one tested block is MAGE-A3- 
positive. The gene profile analysis performed by GSK Biologicals or a contracted laboratory will 
be performed on RNA extracted from fresh tumor tissue obtained from the biopsy and stored in 
RNA later. Gene expression profiling including the presence or absence of the predictive gene 
signature will be assessed by an appropriate technology such as qRT-PCR, microarray or IHC on 
these samples and correlated with the patient’s clinical data. Additional gene profiling might be 
performed in the screening FFPE tumor samples sent for MAGE-A3 screening. 
 
Confirmation of the pathological diagnosis (melanoma stage IV) will be made by the pathologist 
at MCC using a FFPE section. The extent of sampling is to be entered into the patient
's CRF.  
 
 
6.3.3. Visit 1: First study treatment administration 
 
This visit will take place a maximum of 12 weeks after resection.  
 
The following pre-administration procedures will be performed:  
 
• Physical examination including ECOG performance status and disease assessment.  
• Blood sampling for baseline humoral immunity assays will be taken (10 ml, as described in 
Section 6.4.2). 
• Leukapheresis  
• Female patients will undergo a urine or serum ßHCG pregnancy test (if applicable; see 
Section 6.3.1.2 for definition of female of childbearing potential). For female patients of 


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43 
childbearing potential, pregnancy test results must be negative before the first administration 
of study treatment. For these patients, a pregnancy test will be repeated every 3 months and 
should be performed at any time during the study if the investigator or patient suspects that 
pregnancy has occurred while the patient was being treated on protocol therapy.  
• Recording of concomitant medications (as described in Section 5.8)  
• Recording of potential immune-mediated disease (pIMD).  
• SAEs related to study participation or concomitant medications are recorded (see Section 
8.2) 
 
The leukapheresis will be performed within 1 week of starting treatment . 
 
Study treatment administration and other procedures will be as follows:  
 
• The patient will receive the randomized study treatment (recMAGE-A3+AS15 ASCI +/- 
Poly IC:LC given intramuscularly in the deltoid or preferably in the lateral region of the 
thigh; see Section 5.2) and will thereafter be kept under close observation for at least 30 
minutes, with appropriate medical treatment readily available in case of a rare anaphylactic 
reaction following the injection.  
 
• Any (serious) adverse events that occurred during the study treatment will be recorded.  
 
• For patients who agree, eight green tops of blood totaling 70 mL will be taken on days 1, 3, 
7, 14 and 21 after the first and fifth doses of study drugs for assessment of T cell reactivity. 
Day 21 after the first dose of study drug would be the day of the second injection of study 
drug.  These assays will be performed by 
 
 
 
 
6.3.4. Visit 2: Second study treatment administration  
 
This visit will take place 3 weeks ± 4 calendar days after the day of the first administration (Visit 
1).  
 
The pre-administration procedures for this visit will be:  
 
• Physical examination (as at Visit 1, including ECOG performance status and disease 
assessment).  
• Recording of (serious) adverse events and autoimmune diseases.  
• Recording of pIMD.  
• A pregnancy test should be performed at any time during the study for all women of child-
bearing potential if the investigator or patient suspects that pregnancy has occurred while 
the patient was being treated on protocol therapy.  
 
In addition:  
• Before administration, the criteria for permanent stopping (Section 5.2.1.2) or postponement 
of study treatment (Section 5.2.1.1) will be checked; if any of these is met, the patient will 


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either be withdrawn from study treatment or the present administration of study treatment 
will be postponed. Study treatment will then be administered, provided that this is not 
contra-indicated by the findings of the previous 2 steps.  
 
6.3.5. Visit 3: Third study treatment administration  
 
This visit will take place 6 weeks ± 4 calendar days after the day of the first treatment 
administration (Visit 1). Study procedures will include all those of Visit 2.  
 
In addition:  
 
• Blood sampling for humoral immunity assays will be done (10 ml, as described in Section 
6.4.2). If the second study treatment administration had to be postponed (refer to Section 
5.2.1.1 for criteria for postponement of study treatment), Visit 3 should be planned as close 
as possible to the 6 weeks ± 4 calendar days after Visit 1 schedule, but a minimum of 14 
days after the second treatment administration.  
 
6.3.6. Visit 4: Fourth study treatment administration  
 
This visit will take place 9 weeks ± 4 calendar days after the day of the first administration (Visit 
1). Study procedures will be identical to those of Visit 2.  
 
If the third study treatment administration had to be postponed (refer to Section 5.2.1.1 for 
criteria for postponement of study treatment), Visit 4 should be planned as close as possible to 
the 9 weeks ± 4 days after Visit 1 following the original schedule, but a minimum of 14 days 
after the third treatment administration.  
 
 
6.3.7. Visit 5: Fifth study treatment administration  
 
This visit will take place 12 weeks ± 4 calendar days after the day of the first administration 
(Visit 1). Study procedures will be identical to those of Visit 3. 
 
• For patients who agree, eight green tops of blood totaling 70 mL will be taken on days 1, 3, 
7, 14 and 21 after the first and fifth doses of study drugs for assessment of T cell reactivity. 
Day 21 after the first dose of study drug would be the day of the second injection of study 
drug.  Those assays will be performed by 
 
 
  
In addition:  
 
• A CT scan of the neck, chest and upper abdomen and pelvic CT scans will be performed.  
• Blood sampling for safety haematological and chemical assays (15 ml, as described in 
Section 6.4.2).  
• A pregnancy test will be performed for females of childbearing potential. 6.3.8.  
• Leukapheresis will be repeated. 


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Blood sampling for humoral immunity assays will be done (10 ml, as described in Section 6.4.2). 
 
6.3.8. Visits 6-13: Three-monthly maintenance treatment administrations  
 
These visits will take place at 3-month intervals, i.e., at time-points 6, 9, ..., 27 months after Visit 
1, with a permitted deviation from the scheduled time-point of up to 14 calendar days.  
 
The following procedures will be performed at each of these visits:  
• Physical examination (as at Visit 1, including ECOG performance status and disease 
assessment).  
• A CT scan of the neck, chest and upper abdomen and pelvic CT scan.  
• Before administration of study treatment the criteria for permanent stopping or 
postponement of study treatment (Sections 5.2.1.1 and 5.2.1.2) will be checked; if any of 
these is met, the patient will either be withdrawn from study treatment or the administration 
of study treatment will be postponed.  
• Recording of (serious) adverse events and pIMD.  
• A pregnancy test for females of childbearing potential.  
• Recording of concomitant medications.  
• Administration of the study treatment.  
 
The following procedures will be performed before the administration of study treatment at 
Visits 7, 8 and 10: 
• Blood samples (10 ml, as described in Section 6.4.2) for humoral immunity assays will be 
taken. 
 
The following procedures will be performed once every year after Visit 1 (i.e. at Visits 8 and 
12):  
• Blood sampling (15 ml as described in Section 6.4.2) for haematological and chemical 
safety assays.  
 
If any of the sixth to twelfth study treatment administrations had to be postponed (refer to 
Section 5.2.1.1 for criteria for postponement of study treatment), the subsequent Visit should be 
planned as close as possible to the 3-monthly interval of the original schedule, but with a 
minimum of 14 days between any two treatment administrations.  
 
6.3.9. Visit 14: Concluding or end of study examination  
 
This visit will take place 3 months after Visit 13, i.e., approximately 30 months after the first 
administration of study treatment (Visit 1), with a permitted deviation of up to 14 calendar days. 
 
If a patient on trial relapses, or if the patient withdraws for any reason, including toxicity, they 
will also be asked to return to Moffitt for this end of study visit.  
 
If the thirteenth study treatment administration had to be postponed (refer to Section 5.2.1.1 for 
criteria for postponement of study treatment), the concluding Visit should be scheduled 3 months 
after the actual date of the last administration.  


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The following procedures will be performed:  
• A CT scan of the neck, chest and upper abdomen and pelvic CT scans.  
• Physical examination (as at Visit 1).  
• Recording of (serious) adverse events and pIMD.  
• Blood sampling (15 ml as described in Section 6.4.2) for laboratory safety assays.  
• Blood sampling (10 ml as described in Section 6.4.2) for humoral immunity assays.  
• Blood sampling of 80 ml for cellular immune assays 
 
In addition, these samples could be used for translational research for patients having given their 
specific informed consent to this.  
 
• Pregnancy test for females of childbearing potential.  
• Recording of concomitant medications.  
• Treatment conclusion. 
 
6.3.10 Post-treatment follow-up 
      
  
  
Completed treatment phase?   
  
  
          
      
  
  
YES  
  NO    
  
  
          
      
  
  
  
         
  
  
          
     
Perform Visit 
14a 
  
        
Recurrence 
during 
treatment 
phase? 
          
      
  
  
  
        
          
      
Enter 
Follow-up 
phase b 
  
         
  
  
          
     
  
       
YES  
NO  
        
      
  
  
  
         
  
  
          
     
Relapse 
during 
 
  
    
Perform 
concludin
g visit a 
  
 
Withdrawal 
of consent? 
      
     
  
    
  
 
      
      
  
  
  
    
  
  
          
      YES NO  
  
         
  
  
YES NO       
      
  
  
  
     Annual 
contact up 
to 5 years, 
e.g. by 
phone for 
survival 
  
             
 Annual 
contact 
up to 5 
years, 
e.g. by 
phone 
for 
survival 
  
 
Perform 
Follow-up 
visits up to 
5 years b, d 
  
 
No more 
contact 
  
Perform 
concluding 
visit a 
   
 
  
 
  
 
  
   
 
  
 
  
  
  
   
   
 
  
  
  
         
  
  
          
      
  
 
 
       
  
  
  Perform Follow-up 
visits up to 5 years 
c, d 
  
      
  
 
       
  
  
  
  
      
  
  
  
         
  
  
          
a. For procedures of Visit 14 - concluding visit, refer to Section 6.3.9 . 
b. For 5-year follow-up of patients who have completed the treatment phase, refer to Section 6.3.10.2. 
c. For 5-year follow-up of patients who have not completed the treatment phase, refer to Section 6.3.102. 
d. Five-year follow-up is performed until five years after first administration or until recurrence of disease.  
6.3.10.1. Follow-up of patients who have completed the study treatment  
 


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For patients who have completed the study treatment (up to and including Visit 14), follow-up 
visits will take place every 6 months after the concluding visit (Visit 14) for a maximum of 5 
years after first study treatment administration.  
 
The following procedures will be performed:  
 
• A routine physical examination including assessment of ECOG performance status. 
Determination of disease status, on the basis of the patient
's medical records and all other 
available information. If the patient has relapsed, the investigator will obtain information 
about the recurrence (date, type), as far as this is possible.  
• CT scan of the neck, chest and upper abdomen as well as the pelvis (at Months 36, 48 and 
60, or at any other time if clinically indicated).  
• Recording of pIMD.  
• Recording of SAEs related to study participation and concurrent medications.  
• In addition, one year after the concluding visit, a blood sample (10 ml as described in 
Section 6.4) will be taken for humoral immunity testing.  
 
Follow-up visits will be numbered consecutively from Visit F1 onwards. Findings at these visits 
will be recorded using Moffitt CRFs.  
 
If relapse of disease is established at one of the follow-up visits, no further follow-up visits will 
take place. As for patients who have withdrawn from the study treatment (Section 6.3.10.2) 
because of disease relapse, annual post study contacts, e.g., by telephone, will be instituted and 
continued up to 5 years after the first study treatment administration.  
 
 
6.3.10.2. Follow-up of patients who were withdrawn from the study treatment  
 
Patients will be free to withdraw from the study at any time and for any reason and without 
prejudice to their further treatment. However, all patients who have withdrawn from study 
treatment will be examined at least 30 days after the last administration of study treatment and 
before starting any other anti-cancer treatment. The procedures to be performed at this 
examination are those defined for Visit 14 (see Section 6.3.9). Note that if a CT-scan has been 
performed at the last administration of study treatment this will not be repeated.  
 
In addition, all patients who have withdrawn from study treatment will be followed every 6 
months after the last contact for up to 5 years from the date of first study treatment 
administration to document disease recurrence, survival and safety parameters (until another 
cancer treatment is initiated). The procedures at these examinations are the same as for patients 
who have completed study treatment (See Section 6.3.10.1).  
 
For female patients withdrawing because of pregnancy or decision to become pregnant, in 
addition to the follow-up of the melanoma, there will be a follow-up of the pregnancy as 
described in Section 8.12.  
 
There are two exceptions to this follow-up:  


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a. Patients who have withdrawn from the study because of withdrawal of consent will not 
be contacted again.  
 
b. Patients who were withdrawn from study treatment because of disease relapse or who 
relapsed during the post-treatment study period will not be asked to return for study 
follow-up visits. These patients will be contacted yearly, e.g., by telephone, to follow 
them for survival.  
 
 
6.3.11. Annual contact  
 
The survival of patients withdrawn from the study treatment because of relapse during the 
treatment phase or who relapsed during the follow-up period will be monitored by contacting 
them once a year up to 5 years after the first administration. These contacts will be made by the 
investigator (or a delegate) and may be done for example by telephone.  
 
 
6.3.12. Relapse  
 
Relapse of melanoma will include distant metastases. Any new primary cancer, including second 
primary melanoma, will not be considered as a recurrence and should be reported as a SAE.  
 
All relapses must be documented by photographic or radiological evidence or by biopsy, as 
determined by the investigator. Suspicion of relapse based on physical examination findings only 
or laboratory signs must be confirmed by radiological examination and/or biopsy. Equivocal 
findings on standard radiological imaging should be confirmed by repeated examinations or 
cytology/histology or other imaging techniques.  
 
In the event of unequivocal, objective recurrence (at scheduled or unscheduled visit), the 
investigator will determine and record the recurrence, withdraw the patient from the study 
treatment and perform all procedures as defined for Visit 14 (see Section 6.3.9) before any new 
anti-cancer treatment or procedure to treat the recurrence is initiated.  
 
In case a biopsy of the relapse is performed, it will be useful to test MAGE-A3 expression in 
FFPE recurrent tumor tissue and gene profiling research on the fresh tissue tumor samples stored 
in RNA later and taken during biopsy. The informed consent form will have an optional section 
for patients to consent to allow these assays to be performed by GSK Bio. Instruction for sample 
handling and dispatch are given in the GSK Bio recommendations. 
 
As soon as a planned examination shows objective relapse, the patient will be considered to have 
had a relapse on the day of this planned imaging.  
 
If a planned imaging is equivocal, a confirmatory examination (imaging or biopsy) will be 
planned within 8 weeks after the first equivocal imaging. If this confirmatory examination shows 
an objective relapse, the patient will be considered to have had a recurrence on the day of the 


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first equivocal imaging. On the other hand, if the patient’s condition does not allow for this 
confirmatory examination and another anti-cancer treatment is started, the patient will be 
considered to have had relapse on the day of the first equivocal examination. 
 
If a planned imaging does not show any objective relapse but the patient is symptomatic, 
additional examinations will be conducted as clinically indicated (Chest X-Ray, CT scan, bone 
scan, brain CT scan, brain MRI, biopsy, etc.). If objective relapse is demonstrated, the date of 
this additional imaging or biopsy will be considered as the date of relapse. However, if these 
additional examinations are equivocal or if symptoms persist without any explanation, a 
confirmatory examination will be performed within 8 weeks. If relapse is confirmed or if the 
patient requires immediate treatment with another anticancer therapy, the patient will be 
considered to have had relapse.  
 
If a patient comes to an unplanned visit due to symptoms, additional examinations will be 
conducted as clinically indicated (Chest X-Ray, CT scan, bone scan, brain CT scan, brain MRI, 
biopsy, etc.). If objective relapse is demonstrated, the date of this additional imaging or biopsy 
will be considered as the date of relapse. If these additional examinations are equivocal, a 
confirmatory examination will be performed within 8 weeks as. If relapse is confirmed or if the 
patient requires immediate treatment with another anti-cancer therapy, the patient will be 
considered to have had relapse. However, if these additional examinations are negative but 
symptoms persist without any explanation, and the patient requires immediate treatment with 
another anti-cancer therapy, the patient will be considered to have had relapse on the date of the 
unplanned visit.  
 
 
6.4. Biological sample handling and analysis 
 
6.4.1. Treatment and storage of biological samples  
 
See Appendix B and GSK Bio recommendations.  
 
6.4.2. Laboratory assays and immunological read-outs  
 
Please refer to the GSK recommendations for details of biospecimen management (handling, 
storage and shipment) for specimens shipped to GSK Bio or contracted lab.  
 
Samples will not be labeled with information that directly identifies the patients but will be 
coded with the identification number for the patient (patient number or requisition number).  
 
Collected samples may be used for purposes related to the quality assurance of the laboratory 
tests described in this protocol. This may include the management of the quality of these current 
tests, the maintenance or improvement of these current tests, the development of new test 
methods for the markers described in this protocol, as well as making sure that new tests are 
comparable to previous methods and work reliably.  
 
Table 8 specifies the laboratory assays and immunological read-outs that must be performed. 






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tested block is MAGE-A3- positive. Upon reception of the tumor tissue sample, the MAGEA3 
screening will be done within 5 working days. The cut-off value for the MAGE-A3 expression 
assay is defined as 1% of the positive MAGE-A3 control included in the assay.  
 
If tissue from the primary tumor is also provided to the laboratory, this may be tested to examine 
whether it expresses MAGE-A3 (See Section 7.1).  
 
The gene profile analysis performed by GSK Biologicals or a contracted laboratory will be 
performed on RNA extracted from fresh tumor tissue obtained from the biopsy and stored in 
RNA later. Gene expression profiling including the presence or absence of the predictive gene 
signature will be assessed by an appropriate technology such as qRT-PCR, microarray or IHC on 
these samples and correlated with the patient’s clinical data. Additional gene profiling might be 
performed in the screening FFPE tumor samples sent for MAGE-A3 screening. 
 
 
For details on the handling and shipment of tumor tissue samples, refer to GSK 
recommendations. 
 
 
6.4.4. Gene expression signature associated with treatment benefit of resected lesions  
 
6.4.4.1. Background  
 
Gene expression profiling of the tumor by microarray has been shown in the literature to be 
useful in defining the patient
's prognosis and also response to therapy [Dave, 2004; Weigelt, 
2005; Hu, 2006].  
 
Because it is therefore more beneficial to target a patient population that will specifically benefit 
from treatment, GSK Biologicals has already started evaluating in another clinical trial (GSK 
249553/008) whether patients with MAGE-A3-positive cancer could present a specific gene 
expression signature predictive of their response to recMAGE-A3 + AS15 ASCI treatment. 
Using biopsies of the tumor taken prior to any recMAGE-A3 + AS15 ASCI administration, they 
were able to identify a gene expression signature that identify patients who might benefit from 
treatment with recMAGE-A3 ASCI. The identified, potentially predictive signature defines an 
immune microenvironment in the tumor that is present prior to any therapeutic intervention and 
correlates strongly with the clinical benefit following the MAGE-A3 ASCI administration. Such 
molecular signature could select patients with a higher likelihood of clinical response to MAGE-
A3.  
 
A secondary goal of this study is to evaluate the gene signature as a potential marker of clinical 
benefit according to treatment, recMAGE-A3 + AS15 ± Poly IC:LC  
 
 
6.4.4.2. Practical aspects  
 


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Testing will be performed by GSK Biologicals or contracted lab on RNA extracted from fresh 
tumor tissue obtained during the surgical resection.  
 
The presence or absence of the gene signature will be assessed by quantitative PCR (qPCR) on 
these samples and correlated with the patient’s clinical data.  
 
For handling and shipment of tumor tissue samples, refer to the GSK recommendations.  
 
For gene profile and other biomarkers analysis, refer to Section 10.8.  
 
 
 
 
6.4.5. Laboratory assays  
 
Safety laboratory assays assessing the hematological parameters as well as renal and hepatic 
functions will be performed by the laboratory at the investigational site.  
 
These assays (detailed in Table 8) will be performed before the first administration of the study 
treatment and then each year after Visit 1 until the end of the maintenance phase and at Visit 14 
(Refer to Table 5 and Table 6). These parameters will not be recorded anymore during the 
follow-up phase (Table 7).  
 
If a clinically significant abnormality in any laboratory parameter is detected at one of these 
assessments, it should be followed up as adequate until it has returned to normal or a satisfactory 
explanation has been provided. Refer to Section 8.5 for AE and SAE reporting.  
 
 
6.4.6. Immunological read-outs  
 
Specific antibodies (humoral response) induced by the recMAGE-A3 + AS15 ASCI will be 
measured by ELISA or upgraded technologies, and the results will be provided in EU/ml or other 
appropriate units (Table 8). A patient will be considered as seropositive if the value of the 
antibody titre is superior or equal to the assay cut-off value.  
 
These assays will be performed at GSK Biologicals or by a contracted lab chosen and contracted 
for this task by GSK Biologicals. For handling and shipment of tumor tissue samples, refer to the 
GSK recommendations. 
 
 
7. -TRANSLATIONAL RESEARCH 
  
This section provides details on the research that may be done on samples collected during this 
study. The specified tests will be performed on samples from all patients. Based on the data 
generated during the study and in other studies, not all samples will be processed if there is a 
problem with performing leukaphereses.  


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7.1. MAGE-A3 expression on multiple resected lesions  
 
When multiple lesions are obtained from a patient at the time of screening (primary tumor(s), 
lymph node(s)), it is of interest to investigate how MAGE-A3 is expressed in these different 
lesions. For example, one would like to establish whether recurrent lesions are MAGE-A3 
positive, when the primary lesion is positive. In order to establish this information, the MAGE-
A3 test, as it is applied for patient accrual, would be applied to all material that is collected from 
the patient.  
 
 
7.2. Cellular Immune assays 
  
The aim of a cancer immunotherapeutic is to stimulate the patient’s cellular and humoral 
immune response in order to eliminate the cancer cells. Although such immune responses can be 
detected upon administration of a cancer immunotherapeutic, the direct impact of this response, 
especially the T-cell response, on the clinical activity has not yet been demonstrated; some 
patients who show evidence of tumor regression upon administration of a cancer 
immunotherapeutic have very low T-cells directed against the target antigen of the 
immunotherapeutic [Lurquin, 2005].  
 
 
7.3. Gene profiling on fresh tissue 
 
If fresh tumor tissue stored in RNA later is available at the time of tumor resection, it would be 
interesting to test such samples to exploratory define a gene signature predictive of response to 
therapy.  
 
An adequate amount of RNA will be used to hybridize a human expression chip containing a 
locked gene set (for example, Affymetrix; this commercial expression chip contains a locked set 
of over 47,000 defined gene transcripts annotated from the human genome sequencing project). 
Genes expressed in the resected tumor tissue of the cutaneous melanoma patients will be 
analyzed and correlated with the patients
' clinical data.  
 
For handling and shipment of tumor tissue samples, refer to the GSK recommendations. 
 
For gene profile and other biomarkers analysis, refer to Section 10.8.  
 
 
 
 
 
 
 
 


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7.4. Gene profiling research of a relapse or a new tumor 
 
In the event of tumor relapse or the occurrence of a tumor of new histology in patients that have 
been included in the study and if a biopsy is scheduled by the investigator, the patient will be 
proposed to allow gene profiling on the recurrent or new tumor (including MAGE-A3 expression 
screening of the recurrent tumor).  
 
Details of this research are the same as for gene profiling research of the resected metastasis (see 
Section 6.4.4)  
 
This test will not require tissue sampling additional to the tissue removed during biopsy.  
 
(N.B. a cytological sample obtained by fine-needle aspiration is not sufficient). The patient’s 
decision to participate in this optional research must be documented by his/her initials of a 
check-off section in the informed consent form.  
 
 
 
 
 
8. ADVERSE EVENTS AND SERIOUS ADVERSE EVENTS  
 
The investigators are responsible for the detection and documentation of events meeting the 
criteria and definition of an adverse event (AE) or serious adverse event (SAE) as provided in 
this protocol. During the study, when there is a safety evaluation, the investigator or site staff 
will be responsible for detecting AEs and SAEs, as detailed in this section of the protocol.  
 
Each patient or patient’s guardians will be instructed to contact the investigator immediately 
should the patient manifest any signs or symptoms they perceive as serious.  
 
 
8.1. Definition of an adverse event  
 
An AE is any untoward medical occurrence in a clinical investigation patient, temporally 
associated with the use of a medicinal product, whether or not considered related to the 
medicinal product.  
 
An AE can therefore be any unfavorable and unintended sign (including an abnormal laboratory 
finding), symptom, or disease (new or exacerbated) temporally associated with the use of a 
medicinal product. For marketed medicinal products, this also includes failure to produce 
expected benefits (i.e., lack of efficacy), abuse or misuse.  
 
 
 


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Examples of an AE include:  
 
 Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.  
 New conditions detected or diagnosed after investigational product administration even 
though it may have been present prior to the start of the study.  
 Signs, symptoms, or the clinical sequelae of a suspected interaction.  
 Signs, symptoms, or the clinical sequelae of a suspected overdose of either 
investigational product or a concurrent medications (overdose per se should not be 
reported as an AE/SAE).  
 Signs, symptoms temporally associated with treatment administration.  
 Blood counts or chemistries that were abnormal at baseline and worsen outside the 
patient’s usual range of fluctuations after the start of protocol therapy Examples of an AE 
DO NOT include: Medical or surgical procedure (e.g., endoscopy, appendectomy); the 
condition that leads to the procedure is an AE.  
 Situations where an untoward medical occurrence did not occur (social and/or 
convenience admission to a hospital).  
 Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present or 
detected at the start of the study that do not worsen. 
 The disease/disorder being studied, or expected progression, signs, or symptoms of the 
disease/disorder being studied, unless more severe than expected for the patient’s 
condition.  
 Blood counts or chemistries that were abnormal at baseline and did not worsen outside 
the patient’s usual range of fluctuations after the start of protocol therapy AEs may 
include pre- or post-treatment events that occur as a result of protocol-mandated 
procedures (i.e., invasive procedures, modification of patient’s previous therapeutic 
regimen).  
 
Example of events to be recorded in the medical history section of the CRF:  
 
 Pre-existing conditions or signs and/or symptoms present in a patient prior to the start of 
the study (i.e., prior to the first study treatment administration).  
 
 
8.2. Definition of a serious adverse event  
 
A serious adverse event (SAE) is any untoward medical occurrence that:  
 
a. Results in death,  
b. Is life-threatening,  
 
NOTE: The term ’life-threatening’ in the definition of ’serious’ refers to an event in which  
the patient was at risk of death at the time of the event. It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.  


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c. Requires hospitalization or prolongation of existing hospitalization,  
 
NOTE: In general, hospitalization signifies that the patient has been detained (usually involving 
at least an overnight stay) at the hospital or emergency ward for observation and/or treatment 
that would not have been appropriate in the physician.s office or outpatient setting. 
Complications that occur during hospitalization are AEs. If a complication prolongs 
hospitalization or fulfils any other serious criteria, the event is serious. When in doubt as to 
whether hospitalization occurred or was necessary, the AE should be considered serious.  
 
Hospitalization for elective treatment of a pre-existing condition that did not worsen from 
baseline is not considered an AE.  
 
d. Results in disability/incapacity, or  
 
NOTE: The term disability means a substantial disruption of a person’s ability to conduct normal 
life functions. This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, and 
accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday life functions 
but do not constitute a substantial disruption.  
 
e. Is a congenital anomaly/birth defect in the offspring of a study patient.  
f. Is a Grade 4 adverse event according to the Common Terminology Criteria for Adverse 
Events (CTCAE), Version 4.0 (cf. Appendix F) 
 
Medical or scientific judgment should be exercised in deciding whether reporting is appropriate 
in other situations, such as important medical events that may not be immediately life-
threatening or result in death or hospitalization but may jeopardize the patient or may require 
medical or surgical intervention to prevent one of the other outcomes listed in the above 
definition. These should also be considered serious.  
 
Examples of such events are invasive or malignant cancers, intensive treatment in an emergency 
room or at home for allergic bronchospasm, blood dyscrasias or convulsions that do not result in 
hospitalization.  
 
In this study, Grade 3 autoimmune disorders according to the Common Terminology Criteria for 
Adverse Events (CTCAE), Version 4.0 (cf. Appendix F), will be considered as medically 
significant and will therefore be notified as SAE.  
 
 
 
 
 
 
 


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8.3. Adverse events of specific interest 
 
8.3.1 Potential immune-mediated diseases  
 
Potential immune-mediated diseases (pIMDs) are a subset of AEs that include autoimmune 
diseases and other inflammatory and/or neurologic disorders of interest which may or may not 
have an autoimmune aetiology. AEs that need to be recorded and reported as pIMDs include 
those listed in the table below. However, the investigator will exercise his/her medical and 
scientific judgement in deciding whether other immune-mediated diseases have an autoimmune 
origin (i.e. pathophysiology involving systemic or organ-specific pathogenic autoantibodies) 
and should also be recorded as a pIMD. 
 
Neuroinflammatory disorders   
Musculoskeletal disorders   
Skin disorders  
 
• Cranial nerve disorders, 
including paralyses/paresis (e.g. 
Bell’s palsy), and neuritis (e.g. 
optic neuritis)  
• Multiple sclerosis (including 
variants)  
• Transverse myelitis  
• Guillain-Barré syndrome, 
(including Miller Fisher syndrome 
and other variants)  
• Other demyelinating diseases 
(including acute disseminated 
encephalomyelitis)  
• Myasthenia gravis (including 
Lambert-Eaton myasthenic 
syndrome)  
• Non-infectious encephalitis/ 
encephalomyelitis  
• Neuritis (including peripheral 
neuropathies)  
 
 
• Systemic lupus erythematosus  
• Scleroderma (including, CREST 
syndrome and morphoea)  
• Systemic sclerosis  
• Dermatomyositis  
• Polymyositis  
• Antisynthetase syndrome  
• Rheumatoid arthritis,  
• Juvenile chronic arthritis, 
(including Still’s disease)  
• Polymyalgia rheumatica  
• Reactive arthritis  
• Psoriatic arthropathy  
• Ankylosing spondylitis 
(including undifferentiated 
spondylarthritides)  
• Relapsing polychondritis  
• Mixed connective tissue 
disorder  
 
 
• Psoriasis  
• Vitiligo  
• Raynaud’s phenomenon  
• Erythema nodosum  
• Autoimmune bullous skin 
diseases (including pemphigus, 
pemphigoid and dermatitis 
herpetiformis)  
• Cutaneous lupus 
erythematosus  
• Alopecia areata  
• Lichen planus  
• Sweet’s syndrome  
 
Liver disorders  
Gastrointestinal disorders   
Metabolic diseases  
 
• Autoimmune hepatitis  
• Primary biliary cirrhosis  
• Primary sclerosing cholangitis  
• Autoimmune cholangitis.  
 
 
• Crohn’s disease  
• Ulcerative colitis  
• Ulcerative proctitis  
• Celiac disease  
 
 
• Autoimmune thyroiditis 
(including Hashimoto thyroiditis)  
• Grave's or Basedow’s disease  
• Diabetes mellitus type I  
• Addison’s disease  
 
Vasculitides  
Others  
 
• Large vessels vasculitis including: giant cell 
arteritis such as Takayasu's arteritis and temporal 
arteritis.  
• Medium sized and/or small vessels vasculitis 
including: polyarteritis nodosa, Kawasaki's disease, 
microscopic polyangiitis, Wegener's 
granulomatosis, Churg–Strauss syndrome, 
thromboangiitis obliterans (Buerger’s disease), 
necrotizing vasculitis, allergic granulomatous 
angiitis, Henoch-Schonlein purpura, anti-neutrophil 
cytoplasmic antibody positive vasculitis, Behcet's 
syndrome, leukocytoclastic vasculitis.  
• Vasculitides secondary to other immune mediated 
diseases such as lupus vasculitis and rheumatoid 
vasculitis.  
 
 
• Autoimmune hemolytic anemia  
• Autoimmune thrombocytopenias  
• Antiphospholipid syndrome  
• Pernicious anemia  
• Autoimmune glomerulonephritis (including IgA 
nephropathy, glomerulonephritis rapidly 
progressive, membranous glomerulonephritis, 
membranoproliferative glomerulonephritis, and 
mesangioproliferative glomerulonephritis)  
• Uveitis  
• Autoimmune myocarditis/cardiomyopathy  
• Sarcoidosis  
• Stevens-johnson syndrome  
• Sjögren’s syndrome  
• Idiopathic pulmonary fibrosis  
• Goodpasture syndrome 


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Table 9. List of potential immune-mediated diseases. 
When there is enough evidence to make any of the above diagnoses, the AE must be reported as 
a pIMD. Symptoms, signs or conditions which might (or might not) represent the above 
diagnoses, should be recorded and reported as AEs but not as pIMDs until the final or definitive 
diagnosis has been determined, and alternative diagnoses have been eliminated or shown to be 
less likely. 
 
In order to facilitate the documentation of pIMDs in the CRF, a pIMD standard questionnaire 
and a list of preferred terms (PTs) and PT codes corresponding to the above diagnoses will be 
available.  
 
8.4. Relapse of disease  
 
An event which is part of the natural course of the disease under study (i.e., disease relapse) is 
captured in the study as an efficacy measure; therefore it does not need to be reported as an SAE.  
 
Relapse of the tumor will be recorded in the clinical assessments in the CRF. Death due to 
progressive disease is to be recorded on a specific form in the CRF but not as an SAE. However, 
if the progression of the underlying disease is greater than that which would normally be 
expected for the patient, or if the investigator considers that there was a causal relationship 
between treatment with the recMAGE-A3 + AS15 ASCI +/- Poly IC:LC or protocol 
design/procedures and the disease progression/recurrence, then this must be reported as an SAE.  
 
Any new primary cancer (not related to the cancer under study) must be reported as an SAE.  
 
8.5. Clinical laboratory parameters and other abnormal assessments qualifying as adverse 
events and serious adverse events  
 
All laboratory results (normal and abnormal) will be recorded in the patient’s CRF as described 
in the study procedures (Table 5 and Table 6).  
 
Abnormal laboratory findings (e.g. clinical chemistry, haematology, urinanalysis) or other 
abnormal assessments will be recorded as AEs or SAEs:  
 
 If they occur after the start of protocol therapy and meet the definition of an AE, as 
defined in Section 8.1 or SAE, as defined in Section 8.2. 
 If they are present at baseline and worsen outside the patient’s usual range of fluctuations 
after the start of protocol therapy. 






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8.7.1. Assessment of intensity  
 
Severity of AEs will be assessed according to the International Common Terminology Criteria 
for Adverse Events (CTCAE; version 4.0).  
 
The investigator will make an assessment of intensity for all AEs, including SAEs reported 
during the study. The assessment will be based on the investigator’s clinical judgment. The 
intensity of each AE and SAE recorded in the CRF will be graded according to the table given in 
the CTCAE (version 4.0; cf. Appendix F).  
 
An AE assessed as Grade 3 (severe) should not be confused with a SAE. Grade 3 is a category 
utilized for rating the intensity of an event; and both AEs and SAEs can be assessed as Grade 3. 
An event is defined as ‘serious’ when it meets the definition in Section 8.2.  
 
8.7.2. Assessment of causality 
  
The investigator is obliged to assess the relationship between the investigational product and the 
occurrence of each AE/SAE. The investigator will use clinical judgment to determine the 
relationship. Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors and the temporal relationship of the event to the investigational 
product will be considered and investigated. The investigator will also consult the Investigator 
Brochures and/or Product Information, for marketed products, in considering his/her assessment.  
 
There may be situations when a SAE has occurred and the investigator has minimal information 
to include in the initial report to the FDA. However, it is very important that the investigator 
always makes an assessment of causality for every event prior to submission of the SAE to the 
FDA. The investigator may change his/her opinion of causality in light of follow-up information, 
amending the SAE information accordingly. The causality assessment is one of the criteria used 
when determining regulatory reporting requirements.  
 
In case of concomitant administration of multiple treatments, it may not be possible to determine 
the causal relationship of general AEs to the individual treatments given. The investigator should 
therefore assess whether the AE could be causally related to treatment administration rather than 
to the individual treatments.  
 
Causality of all AEs should be assessed by the investigator using the following question:  
 
Is there a reasonable possibility that the AE may have been caused by the investigational 
product?  
 
NO: The AE is not causally related to administration of the study treatment(s). There are other, 
more likely causes and administration of the study treatment(s) is not suspected to have 
contributed to the AE.  
 
YES: There is a reasonable possibility that the treatment(s) contributed to the AE.  
 


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Non-serious and serious AEs will be evaluated as two distinct events. If an event meets the 
criteria to be determined “serious” (see Section 8.2 for definition of serious adverse event), it 
will be examined by the investigator to the extent to be able to determine ALL contributing 
factors applicable to each serious adverse event.  
 
Other possible contributors include:  
 
• Medical history  
• Other medications  
• Protocol required procedure  
• Other procedure not required by the protocol  
• Erroneous administration  
• Other cause (specify) 
 
 
8.8. Follow-up of adverse events and serious adverse events and assessment of outcome 
 
After the initial AE/SAE report, the investigator is required to proactively follow each patient 
and provide further information to the FDA on the patient’s condition.  
 
All AEs and SAEs documented at a previous visit and designated as not recovered/not resolved 
or recovering/resolving will be reviewed at subsequent visits/contacts.  
 
Investigators will follow-up patients:  
 
 with SAEs or patients withdrawn from the study as a result of an AE, until the event has 
resolved, subsided, stabilized, disappeared, the event is otherwise explained, or the 
patient is lost to follow-up;  
 or, in the case of other non-serious AEs, until they complete the study or they are lost to 
follow-up. 
 
Clinically significant laboratory abnormalities will be followed up until they have returned to 
baseline, or a satisfactory explanation has been provided. Additional information (including but 
not limited to laboratory results) relative to the subsequent course of such an abnormality noted 
for any patient must be made available to the Study Monitor.  
 
The outcome of any non-serious AE occurring within 30 days post-treatment administration (i.e., 
unsolicited AE) or any SAE reported during the entire study will be assessed as:  
 
• Recovered/resolved  
• Not recovered/not resolved  
• Recovering/resolving  
• Recovered with sequelae/resolved with sequelae  
• Fatal (SAEs only).  
 
8.9. Prompt reporting of serious adverse events 


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The paper SAE Report Forms and the facsimile (Fax) system  will be the primary method for 
reporting SAEs during the study period.  
 
 
8.9.1. Time frames for submitting serious adverse event reports to GSK Bio and Oncovir 
 
SAEs (including any grade of pIMD) will be reported promptly to GSK Bio and Oncovir once 
the investigator determines that the event meets the protocol definition of an SAE. The 
investigator or designate will complete and submit relevant information on the SAEs in the paper 
SAE form  WITHIN 24 HOURS OF HIS/HER BECOMING AWARE OF THESE 
EVENTS. Additional or follow-up information relating to the initial SAE report is also to be 
completed and submitted in the paper SAE form within 24 hours of receipt of such information.  
 
GSK Biologicals and Oncovir may request that the investigator perform or arrange for the 
conduct of supplemental measurements and/or evaluations to elucidate as fully as possible the 
nature and/or causality of the AE or SAE.  
 
  
 
  
  
  
 
  
 
24/24 hour and 7/7 day availability  
 
 
8.9.2. Completion and transmission of serious adverse event reports to FDA  
 
Once an investigator becomes aware that a SAE has occurred in a study patient (including a 
screened patient), the investigator or designate will complete and submit the information in the 
paper SAE form within 24 hours as outlined in Section 8.9.1. Submission of safety information to 
FDA will follow 21 CFR 312.32. The SAE form will always be completed as thoroughly as 
possible with all available details of the event and will be submitted by the investigator or 
designate. If the investigator or designate does not have all information regarding an SAE, he/she 
will not wait to receive additional information before notifying FDA of the event and completing 
the SAE forms. The SAE form should be updated when additional information is received and 
forwarded to FDA WITHIN 24 HOURS as outlined in Section 8.9.1. 
 
The investigator will always provide an assessment of causality at the time of the initial report as 
described in Section 8.7.2.  
 
For paper SAE Report Form the investigator or designate will report relevant information on 
SAEs to FDA within the 24 hours as outlined in Section 8.9.1. The SAE Report Form will 


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always be completed as thoroughly as possible with all available details of the event, signed by 
the investigator or designate, and forwarded to FDA within the designated time frames. If the 
investigator or designate does not have all information regarding an SAE, he/she will not wait to 
receive additional information before notifying FDA of the event and completing the form.  
When additional information is received on a SAE after freezing of the patient
's CRF, new or 
updated information is to be recorded on the paper SAE Report Form, with all changes signed 
and dated by the investigator. The updated SAE Report Form should be resent to FDA WITHIN 
24 HOURS of receipt of the follow-up information as outlined in Section 8.9.1.  
 
 
8.10. Regulatory reporting requirements for serious adverse events  
 
The investigator, as the sponsor’s representative will promptly report all SAEs to FDA in 
accordance with the procedures detailed in Section 8.9 and to GSK on a quarterly basis. The 
sponsor, MCC has a legal responsibility to promptly notify, as appropriate, both the local 
regulatory authority and other regulatory agencies about the safety of a product under clinical 
investigation. Prompt notification of SAEs by the investigator is essential so that legal 
obligations and ethical responsibilities towards the safety of other patients are met.  
 
The investigator, or responsible person according to local requirements, will comply with the 
applicable local regulatory requirements related to the reporting of SAEs to the IRB/IEC and, if 
required, to the applicable government authority.  
 
An investigator who receives an investigator safety report describing a SAE or other specific 
safety information (e.g., summary or listing of SAEs) from GSK Biologicals or Oncovir will file 
it with the respective Investigator Brochure or other appropriate study documentation and will 
notify the IRB or IEC, if appropriate according to local requirements.  
 
 
8.11. Post-study adverse events and serious adverse events 
  
A post-study AE/SAE is defined as any event that occurs outside of the AE/SAE detection 
period defined in Section 8.6. Investigators are not obligated to actively seek AEs or SAEs in 
former study participants.  
 
However, if the investigator learns of any SAE, including a death, at any time after a patient has 
been discharged from the study, and (s)he considers the event reasonably related to the 
investigational product, the investigator will promptly notify the Study Contact for Reporting 
SAEs.  
 
 
8.12. Pregnancy  
 
Patients who become pregnant during the study treatment phase must not receive additional 
doses of study treatment but may continue other study procedures at the discretion of the 
investigator.  


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The investigator or designate will collect pregnancy information on any patient who becomes 
pregnant while participating in this study, i.e., from signature of the first informed consent till the 
end of the 5-year follow-up period, or till recurrence/death if occurring before the end of the 5-
year follow-up period. The investigator or designate will record pregnancy information on the 
Pregnancy Report Form and submit it to FDA, GSK Biologicals and Oncovir within 24 hours of 
learning of a patient
's pregnancy. The patient will be followed to determine the outcome of the 
pregnancy. At the end of the pregnancy, whether that be full-term or prematurely, information on 
the status of the mother and child will be forwarded to GSK Biologicals and Oncovir. Generally, 
follow-up will be no longer than six to eight weeks following the estimated delivery date.  
While pregnancy itself is not considered an AE or SAE, any pregnancy complication or elective 
termination of a pregnancy for medical reasons will be recorded as an AE or a SAE, as described 
in Sections 8.1 and 8.2, and will be followed as described in Section 8.6.  
 
A spontaneous abortion is always considered to be a SAE and will be reported as described in 
Section 8.9. Furthermore, any SAE occurring as a result of a post-study pregnancy AND 
considered reasonably related in time to receipt of the investigational product by the investigator, 
will be reported to FDA as described in Section 8.9. While the investigator is not obliged to 
actively seek this information from former study participants, he/she may learn of a pregnancy 
through spontaneous reporting. 
 
Information on pregnancies identified during the screening phase prior to treatment 
administration does not need to be collected; this information need not be communicated to 
safety.  
 
 
8.13. Treatment of adverse events  
 
Treatment of any adverse event is at the sole discretion of the investigator and according to 
current good medical practice. Any medications administered for the treatment of an AE should 
be recorded in the patient’s CRF. Refer to Section 8.8.  
 
 
9. PATIENT COMPLETION AND WITHDRAWAL  
 
9.1. Patient completion of study treatment 
  
A patient will be considered to have completed the study treatment when he/she has been under 
treatment until Month 27 and turns up for the concluding examination 30 months after the first 
administration of the study treatment.  
 
Administration of the investigational treatment (recMAGE-A3 + AS15 ASCI +/- Poly IC:LC) 
should continue until completion, documented disease recurrence, withdrawal due to an 
unacceptable toxicity, or withdrawal for other reasons (see Section 9.2).  
 


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Once the patient has been withdrawn from the investigational treatment, the reason must be 
documented in the patient’s medical records and CRF.  
 
9.2. Patient withdrawal  
 
Patients who are withdrawn because of AEs must be clearly distinguished from patients who are 
withdrawn for other reasons. Investigators will follow patients who are withdrawn as a result of a 
SAE/AE until resolution of the event (see Section 8.8).  
 
Once a patient has been withdrawn, the reason must be documented in the patient’s medical 
records and CRF.  
 
Withdrawn patients will not be replaced.  
 
 
9.2.1. Patient withdrawal from investigational product  
 
A patient will be considered as being withdrawn from the investigational product when he/she 
does not receive any further planned dose after the date of withdrawal.  
 
The investigator will document in the CRF whether the decision to discontinue further treatment 
was made by the patient or the investigator and which of the following possible reasons was 
responsible for withdrawal:  
 
• (Serious) adverse event (including intercurrent illness, unacceptable toxicity) 
• Disease recurrence 
• Protocol violation 
• Other (specify) 
 
In case of withdrawal from the investigational product, the concluding visit will be performed as 
described in Section 6.3.9 and the patient will enter the follow-up phase with follow-up visits up 
to five years after first study dose. If the withdrawal from the study treatment is due to disease 
recurrence, the patient should be asked to come for the concluding visit but will not be asked to 
return for the six-monthly follow-up visits. The investigator will contact these patient once a 
year, e.g., by telephone, until 5 years after the first administration of the study treatment.  
 
If the patient refuses or cannot undergo these study procedures, (s)he will be considered as 
withdrawn from the study (Section 9.2.2).  
 
 
 
9.2.2. Patient withdrawal from the study  
 
A patient will be considered as being withdrawn from the study when (s)he does not undergo any 
further planned study activity after the date of withdrawal.  
 


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A patient may voluntarily discontinue participation in this study at any time. The investigator 
may also, at his/her discretion, discontinue the patient from participating in this study at any 
time. In addition, if the sponsor decides to discontinue the study, no further study procedures 
(including administration of the study treatment) will occur.  
 
Information relative to the withdrawal will be documented on the Study Conclusion page of the 
CRF. The investigator will document whether the decision to withdraw from the study was made 
by the patient or the investigator and which of the following possible reasons was responsible for 
withdrawal:  
 
• Death (any cause),  
• (Serious) adverse event (including intercurrent illness, unacceptable toxicity),  
• Disease recurrence,  
• Consent withdrawal, not due to an adverse event,  
• Lost to follow-up,  
• Other (specify).  
 
The investigator will contact these patients once a year, e.g., by telephone, until 5 years after the 
first administration of the study treatment.  
 
 
9.3. Screen and baseline failures  
 
A patient is considered to be a screen/baseline failure if the patient signs the first ICF but 
withdraws before receiving study treatment. 
 
All patients having given consent to undergo MAGE-A3 expression screening and gene 
expression signature analysis upon resection of their tumor will be referenced into a screening 
section of the CRF.  
 
 
10. STATISTICAL CONSIDERATIONS 
 
10.1. Primary endpoints 
 
Safety and Toxicity of the two regimens: adverse events and serious adverse events.  
 
10.1.1. Safety and tolerability endpoints  
 
Occurrence of adverse events up to 30 days after each study dose, including abnormal 
haematological and biochemical parameters.  
 
Occurrence of serious adverse events and autoimmunity during the whole study duration (up to 
30 days after the last administration of study treatment). For methods of analysis, see Section 
10.7.5.  
 


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10.1.2. Immunogenicity endpoints  
 
The primary laboratory endpoint is an assessment of the immunogenicity of the two regimens.  
Serum antibodies (such as Anti-MAGE-A3) seropositivity status (a seropositive patient is a 
patient whose titre is greater than or equal to the cut-off value) will be the primary immune 
endpoints assessed. Seropositivity will be assessed at baseline, after 2, 4, 6, 7 and 9 
administrations, post-treatment (i.e., at concluding visit) and one year after concluding visit (i.e., 
at follow-up visit 2). Cellular immune assays will also be determined as a secondary endpoint. 
For methods of analysis, see Section 10.7.4. 
 
Refer to Section 10.3 for details on the statistical analyses.  
 
 
10.2. Secondary endpoints 
  
10.2.1. Secondary clinical activity endpoints  
 
Relapse-Free Survival (RFS), defined as the time from randomization to either the date of first 
recurrence of the disease or the date of death (whatever the cause), whichever occurs first.  
 
Therefore, any death occurring without prior documentation of tumor relapse will be considered 
as an event (and will not be censored in the statistical analysis) as this approach is less prone to 
introduce bias. If no event has occurred by the time of the analysis, then the time to event will be 
censored at the date of the last assessment of the patient in question.  
 
Any new primary cancer at another site, including second primary melanoma, will not be 
considered as a relapse and should be reported as a SAE.  
 
Overall Survival (OS) defined as the interval from randomization to the date of death, 
irrespective of the cause of death; patients still alive will be censored at the date of the last 
assessment.  
 
10.3. Study design/sample size and power/data analyses 
 
Study Design 
 
This is a 1:1 randomized, two-arm, open-label, Phase II, single-center study to evaluate the 
safety, tolerability and immunogenicity of recMAGE-A3 + AS15 ASCI with Poly IC:LC as 
adjuvant therapy compared to recMAGE-A3 + AS15 ASCI alone in MAGE-A3 positive patients 
with completely resected stage IV melanoma.  Secondary objectives include: to describe the 
efficacy in terms of Relapse-Free Survival (RFS) and Overall Survival (OS) of the two arms; to 
collect preliminary data on the T cell immune response to the ASCI in the overall study 
population as well as subsets of patients of interest (e.g., arms and gene signature groups), and to 
evaluate the gene signature in all patients when possible. The study treatment will consist of an 
induction phase and a maintenance phase. During the induction phase, immunization with or 


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without Poly IC:LC will be administered 5 times at 3-week intervals. During the maintenance 
phase, immunization with recMAGE-A3 + AS15  with or without Poly IC:LC will be 
administered 8 times at 3-month intervals. The 13 immunizations with or without Poly IC:LC 
will be administered over a period of 27 months. The total duration of the study treatment phase 
(including concluding visit) is 30 months per patient. The duration of active follow-up for 
survival, recurrence and serious adverse events (SAEs) related to the study participation and 
concurrent GSK medications will be 5 years from the first administration of study treatment. 
  
 
 
 
Sample Size/Power Calculations and Accrual 
 
The sample size for this trial is calculated based on the primary immune endpoint of a serologic 
response to treatment. In a GSK trial using recMAGE-A3 + AS15 ASCI in a similar patient 
population, the total anti-MAGE-A3 IgG antibody measured at approximately 6 months at the 
first vaccine of the maintenance phase for the ASCI arm without Poly IC:LC had a geometric 
mean of about 7,000 and a standard deviation of no more than 0.28 on the log scale. It is 
anticipated that the ASCI arm with Poly IC:LC will further increase such anti-MAGE-A3 IgG 
antibody measurement at 6 months to about 10,500 for a 50% enhancement of the effect. 
Therefore, 20 patients per arm will yield 62% power for such detection of the anti-MAGE-A3 
IgG antibody difference between the two arms at a one-sided significance level of 0.05. Power 
calculations for this trial will be only for the immune primary endpoint, since little toxicity is 
expected from either of the treatment arms on which to base a power calculation. Table 1 lists the 
change in power if the standard deviation deviates from the assumed value of 0.28 when 
everything else is the same. We plan to enroll 20 patients per arm for a total of 40 patients. To 
account for an approximate 10% loss-to-follow-up rate in this study, up to 44 patients may be 
enrolled. 
 
The change in power if the standard deviation deviates from the assumed value of 0.28: 
 
Standard 
Deviation 
0.28  
0.26  
0.24  
0.22  
0.20 
Power  
0.62  
0.68  
0.74  
0.80  
0.86 
 
A median RFS of about 8 months was observed for similar patients treated without adjuvant 
therapy. A median RFS of 12 months is expected of patients treated with recMAGE-A3 + AS15 
ASCI alone in resected stage IV melanoma. Adding Poly IC:LC to this ASCI regimen might 
double the median RFS to 24 months. The power calculation shows that 20 patients per arm 
would have a power of about 61% for detecting such difference in RFS between the two arms 
with a one-sided type error rate of 0.05, assuming an accrual rate of 2.5 patients per month and a 
3-year follow-up after the completion of accrual.   
 
Assuming an accrual rate of 2.5 patients per month, it would take up to18 months to enroll 44 
patients to this trial. 
 


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Data Analyses Plan 
 
Demographic and disease characteristics as well as other baseline variables for all patients 
randomized will be tabulated and analyzed using the standard descriptive statistics such as 
frequency and percentage tables for discrete variables and mean, median, SD, range, and 
interquartiles for continuous variables. These data will be reported by arm and may be further 
stratified by subgroups, e.g., those patients with favorable/unfavorable gene signature, of 
interest. 
 
The total anti-MAGE-A3 IgG antibody measured at various time points over the course of 
treatment will be descriptively reported by arm with the usual summary statistics as well as with 
plots and confidence intervals (CIs) to discern any time change pattern of interest. Comparisons 
between the two arms will be on the log scale using the two sample t test. If the normal 
distribution assumption is violated for log (total anti-MAGE-A3 IgG antibody), then non-
parametric Wilcoxon test and/or other transformations such as ln and square root will be 
considered as well.  
 
In order to determine whether recMAGE-A3 + AS15 ASCI +/- poly IC:LC is able to induce a 
specific T-cell response to the MAGE-A3 protein and to evaluate the T-cell response in both 
treatment groups, the percentage of T-cells simultaneously producing IFNγ and IFNα will be 
assessed after 5, and after 9 recMAGE-A3 + AS15 ASCI +/- poly IC:LC injections.   
 
The cutoff value for a positive response is defined as 2 standard deviations over the mean 
percentage of double positive events in control wells. Anti-MAGE-A3 T-cell response is 
considered as positive when there are at least 3 more positive wells at any point after 
immunization than at baseline. This variable will be reported using both point estimates and 95% 
CIs for various subgroups of patients, e.g., arms or patients with favorable/unfavorable gene 
signature. Antibodies (such as Anti-MAGE-A3) seropositivity status, defined for a patient whose 
anti-body titer is greater or equal to the cutoff value, will be analyzed similarly.  
 
RFS and OS will be analyzed using the Kaplan-Meier product-limit method as well as the Cox 
proportional hazards regression models. The Log-rank test for comparing RFS between the two 
arms will be performed and reported. Confidence interval for the median and survival rates at 
different time points will be constructed if needed. 
 
The descriptions and grading scales found in the CTEP Active Version of the NCI Common 
Terminology Criteria for Adverse Events (CTCAE v.4) will be utilized for AE reporting.  The 
CTEP Active Version of the CTCAE is identified and located on the CTEP website at: 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm.  All appropriate 
treatment areas should have access to a copy of the CTEP Active Version of CTCAE. Toxicity 
data will be summarized using the usual toxicity table by category and worst grade for the two 
ASCI regimens.  
 
 
10.4. Study populations to be evaluated  


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10.4.1. Total Treated Population (TTP) 
 
The Total Treated Population will include all the enrolled patients that have received at least one 
dose of the study product (recMAGE-A3 + AS15 ASCI +/- poly IC:LC ).  
 
The total treated population will be used for the analysis of clinical activity. 
  
 
10.4.2. Total Treated Population (TTP) for analysis of efficacy  
 
The intent-to-treat approach will define the Total Treated Population to be used for data analysis 
of this trial. That means all evaluable patients (i.e. those meeting the eligibility criteria) enrolled 
in this trial who have received at least one dose of the study treatment, i.e., recMAGE-A3 + 
AS15 ASCI with or without Poly IC:LC, will be included in the final data analyses. Patients who 
are lost to follow-up or drop out of study prior to their scheduled evaluation time due to any 
reason will be treated as non-responders for response determination and censored for time-to-
event type of endpoints at the time of their last assessment/follow-up if no relevant event has 
occurred by then. By-arm analysis will be presented as the main analysis.   
 
 
10.4.3. According to Protocol (ATP) population for analysis of clinical activity and 
immunogenicity  
 
A secondary analysis that includes only all eligible patients may be conducted in addition to the 
primary analysis described above at the discretion of the trial PI and study biostatistician. Other 
subanalyses may also be considered on the basis of a subset of patients, excluding those for 
whom major protocol deviations have been identified (e.g., early discontinuation of treatment, 
major protocol violations, etc.).  However, these subanalyses may not serve as the basis for 
drawing conclusions concerning treatment efficacy, and the reasons for excluding patients from 
the analysis should be clearly reported. Missing data will not be imputed.  A p-value of <0.05 
will be considered significant. No multiplicity adjustment will be made in this study.  
 
10.5. Elimination criteria for analysis  
 
As defined in Section 5.8.2, certain medications are prohibited during this study. Among those 
medications, the following will not require withdrawal of the patient from the study but may 
determine the patient’s evaluability in the according-to-protocol (ATP) analysis.  
 
 Chronic administration of immunosuppressants or other immune-modifying drugs during 
the study period. The use of prednisone, or equivalent, < 0.125 mg/kg/day (absolute 
maximum 10 mg/day), inhaled corticosteroids or topical steroids is permitted.  
 Administration of a vaccine not foreseen by the study protocol during the prohibited time 
period as specified in Section 5.8.3.  
 Repetitive administration of immunoglobulins and/or any blood products during the study 
period.  


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Patients to be excluded from the ATP populations will be identified during a review of data. 
 
 
10.6. Derived and transformed data  
 
10.6.1. Group definition  
 
The patients will be analyzed according to the treatment group assigned by randomization.  
 
 
 
 
10.6.2. Immunogenicity  
 
The cut-off value of the ELISA assay is defined by the laboratory before the analysis and is 
described in Section 6.4.6.  
 A seropositive patient is a patient whose titre is greater than or equal to the cut-off value. 
The seropositivity rate is defined as the proportion of seropositive patients.  
 Antibody response is defined as antibody concentrations ≥ the cut-off value among 
patients with concentrations < the cut-off value before treatment initiation and as 
antibody concentration at least twice the patient’s own baseline value among patients 
with concentrations ≥ the cut-off value before treatment initiation.  
 The Geometric Mean Titre (GMT) is calculated by taking the anti-logarithm of the mean 
of the log concentration transformations. Antibody titres below the cut-off value of the 
assay will be given an arbitrary value of half the cut-off value for the purpose of this 
calculation.  
 For a given patient and a given immunogenicity measurement, results of missing or non-
evaluable measurements will not be replaced. Therefore, an analysis will exclude patients 
with missing or non-evaluable measurements.  
 
10.6.3. Safety  
 
For the analysis of adverse events, such as serious adverse events or adverse events by primary 
MedDRA term, all patients treated will be considered. Patients who did not report an event will 
be considered as patients without the event.  
 
 
10.7. Final analyses  
 
All statistical analyses will be performed using SAS, R and/or StatXact statistical software.  
10.7.1. Analysis of demographics and other baseline characteristics  
 
Demographic characteristics (age, gender, etc.) and other baseline characteristics such as 
MAGE-A3 expression of the tumor specimen, tumor classification, previous surgery etc. for all 
patients randomized will be tabulated and analyzed by appropriate descriptive statistics:  


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 Frequency tables will be generated for categorical variables such as gender; 
 Mean, median, SD, range, and interquartiles will be provided for continuous data such as 
age. These data will be tabulated by treatment group, and the tables will be repeated for 
the different analysis populations: Total Treated Population, ATP cohort for \clinical 
activity and ATP cohort for immunogenicity.  
 
10.7.2. Analysis of treatment compliance and treatment modifications  
 
The total number of doses administered and the length of follow-up will be summarized.  
 
For the Total Treated population cohort, overall compliance will be calculated for each dose as 
the percentage of patients who received the dose as per protocol (see Section 10.4) among all 
patients still on treatment at the target day of dose administration. Compliance will be considered 
unknown if it cannot be calculated because of missing data. Dose delays recorded on the CRF 
will not be taken into account in the measure of compliance.  
 
The number of patients discontinuing treatment with the investigational product, as well as the 
reason for discontinuation, will be summarized and listed.  
 
These data will be tabulated by treatment group for the total treated population.  
 
10.7.3. Analysis of clinical activity  
 
10.7.3.1. Relapse-free survival (RFS) 
 
10.7.3.1.1. Relapse-free survival in the overall study population (secondary objective) 
 
The analysis of RFS will be performed on the overall study Total Treated population. The 
primary analysis will be based on the adjusted Cox regression model.  
 
Estimates of HR and their 95% CI will be obtained and the Wald test will be used to compare the 
groups.  
 
Events of relapse of disease will be assigned to the visit at which they are detected (see Relapse 
Algorithm in Section 6.3.12), ignoring previously missed evaluation visits. 
  
Or rather, interval-censored time-to-event data analysis methods such as Turbull’s (1976) and 
Finkinstein’s (1985) could be used.) 
 
This primary analysis will be complemented by the following sensitivity analyses:  
 
 an analysis of RFS based on the adjusted Cox regression model (see above) performed on 
the Total treated population, taking the date of first missed evaluation visit before 
detection of recurrence as date of event for patients not compliant with the scheduled 
evaluations; 


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 an unadjusted analysis of RFS based on the Logrank test, performed on the Total treated 
population. For the purpose of this sensitivity analysis, estimates of HR and their 95% 
CIs will be obtained by an unadjusted Cox model. More details will be given in the 
Report Analysis Plan (RAP).  
 
 
10.7.3.2. Overall survival (Secondary objective)  
 
The analysis of Overall survival will include the analysis of Overall survival in the overall study 
population, the analysis of Overall survival in the population of patients presenting the 
potentially favorable gene signature and the analysis of Overall survival in the population not 
presenting this gene signature. These analyses will be performed using the same methods as 
those described for the primary analysis of RFS (see Section 10.7.3.1.1).  
 
10.7.4. Analysis of immunogenicity  
 
The analysis of immunogenicity will be performed on the ATP population, cf. Section 10.4.3.  
 
The immunogenicity induced by the recMAGE-A3 + AS15 +/- poly IC:LC ASCI will be 
assessed through calculation of the geometric mean titres (with 95% CI), the proportion of 
seropositive patients and antibody response in the two treatment groups. This will be done for 
antibodies at baseline, after 2, 4, 6, 7, and 9 dose administrations and post-treatment (i.e., at the 
concluding examination at Visit 14, 3 months after the injection of dose 13, cf. Section 6.3).  
 
Antibody titres may also be investigated using reverse cumulative curves.  
 
A possible correlation between the immunological response (such as antibody response to 
MAGE-A3) and RFS may be explored using a time-dependent Cox model.  
 
The immunogenicity will also be evaluated in the gene signature-positive and gene signature 
negative populations, separately.  
 
 
10.7.5. Analysis of safety  
 
The safety analyses will be based on the Total Treated population.  
 
The duration of treatment and total number of doses of investigational product administered will 
be summarized by treatment group.  
 
10.7.5.1. Adverse events  
 
Adverse events (AEs), including abnormal laboratory values, will be graded according to the 
CTCAE, Version 4.0 and coded to the preferred term (PT) level using the MedDRA dictionary.  
 


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The percentage of patients with adverse event(s) within 30 days of each recMAGE-A3 + AS15 
ASCI dose +/- Poly IC:LC will be reported by treatment group with exact 95% confidence 
intervals, as per MedDRA terminology, by Preferred Term (PT) and System Organ Class (SOC), 
if appropriate. The same tabulation will be presented for grade 3, grade 4, related, grade 3 related 
and grade 4 related events, if appropriate.  
 
 
10.7.5.2. Serious adverse events  
 
Serious adverse events occurring at any time after study initiation will be summarized by 
treatment group and described in detail by patient narratives. 
 
 
10.7.5.3. Deaths  
 
Time to death will be analyzed as described in Section 10.7.3. Causes of death (e.g., 
hematological toxicity, non-hematological toxicity, disease under study, or other) will be 
summarized by treatment group.  
 
 
10.7.5.4. Concomitant medications  
 
For the analysis of the use of concomitant medications, the Total Treated population will be 
considered. Patients, for whom no use of concomitant medications has been reported, will be 
considered as patients without any use of concomitant medications.  
 
 
10.8. Gene profile analyses and translational research 
  
10.8.1. Gene profile  
 
The gene profile will be evaluated by an interaction test between the treatment status 
(recMAGEA3+AS15 versus recMAGEA3+AS15 in combination with Poly IC:LC) and the gene 
signature status (positive versus negative) in a Cox regression analysis using the secondary 
clinical endpoint (RFS) as dependent variable in an exploratory hypothesis driven analysis.  
 
 
 
11. CONDUCT OF THE STUDY  
 
11.1. Ethics and regulatory considerations 
 
The study will be conducted according to Good Clinical Practice (GCP), the Declaration of 
Helsinki (US 21 CFR Part 50—Protection of Human Patients, and Part 56— Institutional Review 
Boards) and local rules and regulations of the country.  
 


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Submission of the protocol and any protocol amendments to regulatory agencies will occur in 
accordance with local regulatory requirements at the Moffitt Cancer Center.  
 
 
11.1.1. Institutional Review Board/Independent (IRB/IEC)  
The IRB/IEC must be constituted according to the local laws/customs of each participating 
country. The ICH Harmonized Tripartite Guideline for Good Clinical Practice recommends that 
the IRB/IEC should include:  
 
a. At least five members.  
b. At least one member whose primary area of interest is in a non-scientific area.  
c. At least one member who is independent of the institution/ study site.  
 
Only those IRB/IEC members who are independent of the investigator and the sponsor of the 
study should vote or provide opinion on a study-related matter.  
 
A list of IRB/IEC members, their professions and other qualifications should be obtained by the 
Study Monitor.  
 
This protocol and any other documents that the IRB/IEC may need to fulfil its responsibilities, 
including patient recruitment procedures and information about payments and compensation 
available to patients, will be submitted to the IRB/IEC by the Study Monitor. Written and dated 
unconditional approval from the IRB/IEC of the protocol and amendment (if any and applicable), 
written informed consent form, consent form updates (if any), patient recruitment procedure(s) 
(e.g. advertisements), and any other written information to be provided to patients must be in the 
possession of the investigator and sponsor, MCC, before commencement of the study. This 
approval must refer to the study by study title and number with exact protocol version and date, 
and should identify the documents reviewed and state the date of review. Relevant GSK 
Biologicals and Oncovir’s data such as the Investigator Brochures will be supplied by the Study 
Monitor to the appropriate IRB/IEC for review and approval of the protocol. Verification of the 
unconditional approval of the IRB/IEC will be transmitted by the Study Monitor prior to 
shipment of treatment supplies and CRFs to the site. 
 
No deviations from, or changes to, the protocol should be initiated without prior written sponsor 
and IRB/IEC approval of an appropriate amendment, except when necessary to eliminate 
immediate hazards to the patients or where permitted by all applicable regulatory requirements 
or when the change(s) involves only logistic or administrative aspects of the study (e.g., change 
of monitor(s), telephone number(s)). Administrative changes and amendments not submitted for 
approval are submitted to the IRB/IEC for information only. However, written verification that 
such documents were submitted should be obtained.  
 
The IRB/IEC must be informed by the Sponsor of:  
 
 all subsequent protocol amendments, informed consent changes or revisions of other 
documents originally submitted for review,  
 serious and/or unexpected adverse events occurring during the study, where required,  


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 all subsequent protocol administrative changes (for information, except for US studies),  
 new information that may affect adversely the safety of the patients or the conduct of the 
study,  
 an annual update and/or request for re-approval, where required,  
 when the study has been completed, where required.  
 
If a trial is prematurely terminated or suspended for reasons including, but not limited to, safety 
or ethical issues or severe non-compliance, the sponsor will promptly inform the regulatory 
authorities of the termination or suspension and the reason(s) for the termination or suspension. 
If required by applicable regulations, the investigator must inform the IEC/IRB promptly and 
provide the reason for the suspension or termination.  
 
11.1.2. Informed consent  
 
In obtaining and documenting informed consent, the investigator should comply with the 
applicable regulatory requirement(s), and should adhere to GCP and to the ethical principles that 
have their origin in the Declaration of Helsinki. Prior to the beginning of the trial, the 
investigator should have the IRB/IEC’s written approval of the written informed consent form 
and any other written information to be provided to the patients.  
 
Informed consent will be obtained in accordance with 21 CFR 50.25.  
 
Freely given informed consent should be obtained from every patient prior to clinical trial 
participation.  
 
Information should be given in both oral and written form whenever possible and as deemed 
appropriate by the IRB/IEC. 
 
An investigator or designate will describe the protocol to potential patients face to face. The 
Informed Consent Form may be read to the patients, but, in any event, the investigator or 
designate shall give the patients ample opportunity to inquire about details of the study and ask 
any questions before dating and signing the Informed Consent Form.  
 
While informed consent information can be presented to groups at an initial information session, 
each patient must be given the opportunity to individually pose questions to the investigator or 
designate prior to the patient dating and signing the Informed Consent Form.  
 
The Informed Consent Form must be in a language fully comprehensible to the prospective 
patients. Informed consent shall be documented by the use of a written consent form approved by 
the IRB/IEC and signed and dated by the patients and by the person who conducted the informed 
consent discussion. The signature confirms that the consent is based on information that has been 
understood. All illiterate individuals will have the study and the Informed Consent Form 
explained to them point by point by the interviewer in the presence of an impartial witness. The 
witness will personally sign and date the consent form. Orally witnessed consent will replace 
written consent only in countries where the local custom is contrary or if the patient’s incapacity 
precludes this and provided that the local legal obligations are fulfilled.  


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Each patient
's signed informed consent form must be kept on file by the investigator for possible 
inspection by Regulatory Authorities. The patients should receive a copy of the signed and dated 
written informed consent form and any other written information provided to the patients and 
should receive copies of any signed and dated consent form updates. Any amendments to the 
written information will be provided to the patients.  
 
Both the informed consent discussion and the written informed consent form and any other 
written information to be provided to the patients should include explanations of the following:  
 
a. That the trial involves research.  
b. The purpose of the trial.  
c. The trial treatments and the probability for random assignment to each treatment.  
d. The trial procedures to be followed, including all invasive procedures.  
e. The patient’s responsibilities.  
f. Those aspects of the trial that are experimental.  
g. The reasonably foreseeable risks or inconvenience to the patients and, when 
applicable, to an embryo, fetus or nursing infant.  
h. The reasonable expected benefits. When there is no intended clinical benefit to 
patients, the patients should be made aware of this.  
i. The alternative procedures or courses of treatment that may be available to patients and 
their important potential benefits and risks. 
j. The compensation and/or treatment available to patients in the event of trial-related 
injury.  
k. The anticipated prorated payment, if any, to patients or patients’ guardians for 
participating in the trial.  
l. The anticipated expenses, if any, to patients for participating in the trial.  
m. That the patients’ participation in the trial is voluntary and patients may refuse to 
participate or withdraw from the trial, at any time, without penalty or loss of benefits to 
which patients are otherwise entitled.  
n. That the monitor(s), the auditor(s), the IRB/IEC, and the regulatory authority(ies) will 
be granted direct access to the patient’s original medical records for verification of 
clinical trial procedures and/or data, without violating the confidentiality of patients, to 
the extent permitted by the applicable laws and regulations and that, by signing a written 
informed consent, the patient authorize such access.  
o. That records identifying patients will be kept confidential and, to the extent permitted 
by the applicable laws and/or regulations, will not be made publicly available. If the 
results of the trial are published, patients’ identity will remain confidential.  
p. That the patients will be informed in a timely manner if information becomes available 
that may be relevant to the patients’ willingness to continue participating in the trial.  
q. The person(s) to contact for further information regarding the trial and the rights of 
trial patients, and who to contact in the event of trial-related injury.  
r. The foreseeable circumstances and/or reasons under which a patient’s participation in  
the trial may be terminated.  
s. The expected duration of a patient’s participation in the trial.  
t. The approximate number of patients involved in the trial.  


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The Sponsor has the final responsibility for the final presentation of the Informed Consent Form, 
respecting the mandatory requirements of local regulations. The consent form generated by the 
sponsor, MCC, must be approved (along with the protocol, and any other necessary 
documentation) by the IRB/IEC.  
 
 
11.2. Administrative Matters 
  
To comply with Good Clinical Practice important administrative obligations relating to 
investigator responsibilities, monitoring, archiving data, audits, confidentiality and publications 
must be fulfilled.  
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Lurquin C, Lethe B, De Plaen E et al. Contrasting frequencies of antitumor and anti-vaccine T 
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Marchand M, Punt CJ, Aamdal S et al. Immunisation of metastatic cancer patients with MAGE-3 
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Raman, V.S., Bhatia, A., Picone, A., Whittle, J, et al  Applying TLR synergy in immunotherapy: 
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Speiser DE, Lienard D, Rufer N, Rubio-Godoy V, Rimoldi D et al.Rapid and strong human 
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Vansteenkiste J, Zielinski M, Linder A, et al.: Final results of a multi-center, double-blind, 
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Appendix A. Overview of the Recruitment Plan  
 
The recruitment of patients will include the following essential aspects:  
 
 There will be one study centre at the Moffitt Cancer Center, which is the sponsor 
 Enrollment will cease when the target of 40 evaluable patients (up to 44 patients 
recruited) is reached.  
 An anticipated average of 2-3 patients per month will be recruited.  
 Recruitment is anticipated to commence in March 2011and is expected to be complete by 
February 2013.  
 Recruitment tracking will be provided by the centralized randomization procedure.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Appendix B. Laboratory assays  
 
1. Laboratory assays part of the study protocol: Laboratory assays part of the study protocol are 
summarized in Table 8. Refer to Section 6.4 for details of the mandatory assays and to Section 7 
for the optional translational research assays.  
 
2. Handling and shipment of biological samples collected by the Investigator: For the samples to 
be analyzed by GSK Bio materials to be used will be agreed between the Sponsor and GSK Bio. 
GSK Bio will provide to the Sponsor some recommendations for the collection, storage and 
shipments of samples to be analyzed by GSK Bio or contracted lab (refer to Table 8). However, 
when GSK Biologicals does not recommend material for collecting and storing clinical samples, 
then appropriate materials from the investigator’s site are to be used.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Appendix C. Treatment supplies, packaging and accountability  
 
1. Treatment and/or other supplies  
 
GSK Biologicals will supply the following study treatments, sufficient number of doses to 
administer to all patients as described in the present protocol.  
 
recMAGE-A3 + AS15 ASCI in two vials  
 
Oncovir will ship the Poly IC:LC, in a sufficient number of doses to administer to all patients as 
described in the present protocol.  
  
2. Treatment packaging  
The treatments will be packed in labelled boxes. The box label will contain, as a minimum, the 
following information: study number, treatment number, lot number, instructions for treatment 
administration and any other relevant regulatory requirements.  
 
3. Treatment shipment from GSK Biologicals Rixensart to the investigational site: on reception 
of the shipment, its content, quality and maintenance of the cold-chain must be checked.  
 
The supplies receipt documents must then be returned to:  
 
  
  
  
  
 
 
4. Treatment accountability  
Treatment accountability is responsibility of the Sponsor, MCC. At all times, the figures on the 
treatment supplied by GSK Bio and Oncovir, used and remaining treatment doses should match.  
Used and unused treatment vials should be destroyed at the study site using locally approved 
biosafety procedures and documentation and a copy of the certificate of destruction must be sent 
to GSK Bio and Oncovir. If no adequate biosafety procedures are available at the study site, the 
used and unused treatment vials are to be returned to an appropriate site for destruction.  
 


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5. Transfers of clinical treatments or products from GSK Bio to the study sites: Storage 
temperatures must be maintained during transport and deviations must be reported to GSK Bio 
for guidance.  
 
 
 
 
 
 
 
Appendix D. TNM classification of malignant melanoma  
The TNM staging system characterizes melanoma with respect to several factors. The current 
version of the staging system used has been published in the Seventh Edition of the AJCC 
Cancer Staging Manual (AJCC 2010, Balch CM et al, Final Version of 2009 AJCC Melanoma 
Staging and Classification. J Clin Oncol 2009)  
 
http://www.mmmp.org/MMMP/import.mmmp?page=tnm_stagingmmmp 
 
Melanoma TNM Classification 
T Classification Thickness 
Ulceration Status 
T1 
≤ 1.0 mm 
a:  without ulceration and mitotic 
rate <1/mm2 
b:  with ulceration or mitotic rate 
1 or greater 
T2 
1.01 – 2.0 mm 
a:  without ulceration 
b:  with ulceration 
T3 
2.01 – 4.0 mm 
a:  without ulceration 
b:  with ulceration 
T4 
> 4.0 mm 
a:  without ulceration 
b:  with ulceration 
 
 
 
N Classification # of Metastatic Nodes  
Nodal Metastatic Mass 
N1 
1 node 
a:  micrometastasis* 
b:  macrometastasis† 
N2 
2 – 3  nodes 
a:  micrometastasis* 
b:  macrometastasis† 
c: in transit met(s)/satellite(s) 
without metastatic nodes 
N3 
4 or more metastatic nodes, 
or matted nodes, or  
in transit 
met(s)/satellites(s) with 
metastatic node(s) 
 
 
 
 
M Classification Site 
Serum Lactate Dehydrogenase 
M1a 
Distant skin, subcutaneous Normal 


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or nodal mets 
M1b 
Lung metastases  
Normal 
M1c 
All other visceral 
metastases 
Normal 
Any distant metastatsis  
Elevated 
*  Micrometastases are diagnosed after sentinel or elective lymphadenectomy. 
†  Macrometastases are defined as clinically detectable nodal metastases confirmed by therapeutic 
lymphadenectomy or when nodal metastasis exhibits gross extracapsular extension. 
 
Stage Groupings for Cutaneous Melanoma  
  
Clinical Staging  
Pathologic Staging 
  
T  
N  
M  
T  
N  
M 
0 
Tis  N0  
M0  
Tis  
N0  
M0 
IA 
T1a  N0  
M0  
T1a  N0  
M0 
IB 
T1b 
T2a 
N0 
N0 
M0 
M0 
T1b 
T2a 
N0 
N0 
M0 
M0 
IIA T2b 
T3a 
N0 
N0 
M0 
M0 
T2b 
T3a 
N0 
N0 
M0 
M0 
IIB T3b 
T4a 
N0 
N0 
M0 
M0 
T3b 
T4a 
N0 
N0 
M0 
M0 
IIC T4b 
N0  
M0  
T4b  N0  
M0 
III‡ Any T N1 
N2 
N3 
M0 
 
III
A 
 
T1-4a 
T1-4a 
N1a 
N2a 
M0 
M0 
III
B 
 
T1-4b 
T1-4b 
T1-4a 
T1-4a 
T1-
4a/b 
N1a 
N2a 
N1b 
N2b 
N2c 
M0 
M0 
M0 
M0 
M0 
III
C 
 
T1-4b 
T1-4b 
Any T 
N1b 
N2b 
N3 
M0 
M0 
M0 
IV Any T Any N Any M1 Any T Any N Any M1 
 
*  Clinical staging includes microstaging of the primary melanoma and clinical/radiologic evaluation for 
metastases.  By convention, it should be used after complete excision of the primary melanoma with clinical 
assessment for regional and distant metastases. 
†  Pathologic staging includes microstaging of the primary melanoma and pathologic information about the 
regional lymph nodes after partial or complete lymphadenectomy.  Pathology stage 0 or stage 1A patients are 
the exception; they do not require pathologic evaluation of their lymph nodes. 
‡  There are no stage III or IV subgroups for clinical staging. 
 
 
 
 


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Appendix E. Eastern Co-operative Oncology Group (ECOG) Performance Status  
 
 
Grade Performance 
0 
Fully active, able to carry on all pre-disease performance without restriction 
1 
Restricted in physically strenuous activity but ambulatory and able to carry out work of 
a light or sedentary nature, e.g. light house work, office work 
2 
Ambulatory and capable of all selfcare, but unable to carry out any work activities. Up 
and about more than 50% of waking hours 
3 
Capable of only limited selfcare, confined to bed or chair more than 50% of waking 
hours 
4 
Completely disabled. Cannot carry on any selfcare.  Totally confined to bed or chair 
5 
Dead 
 
 
As published in Am. J. Clin. Oncol.:  Oken, M.M., Creech, R.H., Tormey, D.C., Horton, J., 
Davis, T.E., McFadden, E.T., Carbone, P.P.: Toxicity And Response Criteria Of The Eastern 
Cooperative Oncology Group. Am J Clin Oncol 5:649-655, 1982. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Appendix F.   U.S. National cancer Institute (NCI) Common Terminology Criteria for 
Adverse Events (CTCAE)  
 
The severity of adverse events will be assessed by reference to the Common Terminology 
Criteria for Adverse Events (CTCAE), version 4.0 published May 28, 2009 (v4.03: June 14, 
2010). A copy can be downloaded from the internet website: 
 
http://evs.nci.nih.gov/ftp1/CTCAE/CTCAE_4.03_2010-06-14_QuickReference_5x7.pdf