CL-PTL-130 IND 014205 Confidential and proprietary information of Gradalis, Inc. It is the user’s responsibility to verify adequate training before executing a document. Do not make unauthorized copies. COVER PAGE OFFICIAL PROTOCOL TITLE: A Multi -Center Phase III, Randomized, Open -Label Trial of Vigil (b i-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy) in combination with Irinotecan and Temozolomide as a Second -Line Regimen for Ewing’s Sarcoma PROTOCOL NUMBER: CL-PTL-130 IND NUMBER: 014205 NCT NUMBER: NCT03495921 PROTOCOL DATE: Amendment 2 dated December 5, 2018 IRB APPROVAL DATE: December 14, 2018 CONFIDENTIAL Information and data included in this protocol contain trade secrets and privileged or confidential information which is the property of Gradalis, Inc. No person is authorized to make it public without written permission of Gradalis, Inc. These restrictions on disclosure will apply equally to all future information supplied to you, which is indicated as privileged or confidential. This material may be disclosed to and used by your staff and associates as may be necessary to conduct this cl inical study. PROTOCOL TITLE: A Multi -Center Phase III, Randomized, Open -Label Trial of Vigil (bi-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy) in combination with Irinotecan and Temozolomide as a Second-Line Regimen for Ewing’s Sarcoma IND NUMBER: 014205 PROTOCOL NUMBER: CL-PTL-130 STUDY AGENT(S): Vigil bi-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy SPONSOR: Gradalis, Inc. 2545 Golden Bear Drive, Suite 110 Carrollton, TX 75006 Phone: (214) 442-8124 Fax: (214) 442-8101 PROTOCOL DATE: Amendment 2 dated December 5, 2018 Amendment 1 dated June 20, 2018 Initial Protocol dated September 20, 2017 Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 2 of 74 Confidential and proprietary information of Gradalis, Inc. Effective: 05-DEC-2018 It is the user’s responsibility to verify adequate training before executing a document. Do not make unauthorized copies. INVESTIGATOR PROTOCOL SIGNATURE PAGE PROTOCOL TITLE: A Multi -Center Phase III, Randomized, Open -Label Trial of Vigil (bi-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy) in combination with Irinotecan and Temozolomide as a Second-Line Regimen for Ewing’s Sarcoma PROTOCOL NUMBER: CL-PTL-130 STUDY AGENT(S): Vigil bi-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy SPONSOR: Gradalis, Inc. 2545 Golden Bear Drive, Suite 110 Carrollton, TX 75006 Phone: (214) 442-8124 Fax: (214) 442-8101 PROTOCOL DATE: Amendment 2 dated December 5, 2018 Amendment 1 dated June 20, 2018 Initial Protocol dated September 20, 2017 I have read and understand the contents of the indicated clinical protocol and will adhere to the trial requirements as presented, including all statements regarding confidentiality. In addition, should I choose to participate as an investigator, I and my sub-investigator(s) agree to conduct the study as outlined herein, in accordance with Good Clinical Practices (GCPs), the Declaration of Helsinki , in compliance with the obligations and requirements of clinical investigators and all other requirements listed in Title 21 Code of Federal Regulations (CFR) Part 312. Name of Investigator (please print) Signature of Investigator Date Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 3 of 74 Confidential and proprietary information of Gradalis, Inc. Effective: 05-DEC-2018 It is the user’s responsibility to verify adequate training before executing a document. Do not make unauthorized copies. TABLE OF CONTENTS INVESTIGATOR PROTOCOL SIGNATURE PAGE .................................................................... 2 TABLE OF CONTENTS ............................................................................................................... 3 ABBREVIATIONS ........................................................................................................................ 6 SYNOPSIS .................................................................................................................................... 8 STUDY SCHEMA ....................................................................................................................... 15 STUDY PROCESS FLOW .......................................................................................................... 16 1.0 INTRODUCTION .......................................................................................................... 17 1.1 Summary ................................................................................................................... 17 1.2 Relapsed and Refractory Ewing’s sarcoma .............................................................. 17 1.3 Salvage chemotherapy in Ewing’s sarcoma ............................................................. 18 1.4 Immunotherapeutic considerations in Ewing’s sarcoma ........................................... 20 2.0 VIGIL ............................................................................................................................ 22 2.1 Vigil Phase II Trial: Ovarian Cancer .......................................................................... 26 2.2 Vigil Phase I Trial: Ewing’s sarcoma Subset ............................................................. 29 2.3 Efficacy Evidence of Vigil in Ewing’s sarcoma .......................................................... 30 2.4 Vigil Phase II Trial: Ewing’s sarcoma (CL-PTL-121, Part 1) ..................................... 32 2.5 Vigil Phase II Trial: Ewing’s sarcoma (CL-PTL-121, Part 2) ..................................... 34 3.0 STUDY RATIONALE ................................................................................................... 38 4.0 OBJECTIVES ............................................................................................................... 39 4.1 Primary objective ...................................................................................................... 39 4.2 Secondary objective(s) ............................................................................................. 39 5.0 STUDY DESIGN ........................................................................................................... 40 6.0 STUDY POPULATION ................................................................................................. 41 6.1 Sample Size .............................................................................................................. 41 6.2 Tissue Procurement Inclusion Cri teria ...................................................................... 41 6.3 Tissue Procurement Exclusion Criteria ..................................................................... 41 6.4 Study Enrollment Inclusion Criteria ........................................................................... 42 6.5 Study Enrollment Exclusion Criteria .......................................................................... 43 7.0 WITHDRAWAL ............................................................................................................ 43 7.1 Discontinuation from Study Treatment ...................................................................... 43 7.2 Treatment Delay ....................................................................................................... 44 8.0 INVESTIGATIONAL PLAN .......................................................................................... 44 8.1 Subject Screening and Registration .......................................................................... 44 8.2 Tumor Procurement .................................................................................................. 45 8.3 Vigil Immunotherapy Manufacturing ......................................................................... 45 Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 4 of 74 Confidential and proprietary information of Gradalis, Inc. 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Do not make unauthorized copies. 8.4 Study Treatment Administration ................................................................................ 45 8.4.1 Schedule, Dose and Administration ................................................................... 46 8.5 Concomitant Medications and Supportive Care ........................................................ 48 8.6 Contraception ............................................................................................................ 49 8.7 Toxicity ...................................................................................................................... 49 9.0 SCHEDULE OF ASSESSME NTS ................................................................................ 49 10.0 CROSS -OVER ............................................................................................................. 53 10.1 Inclusion Criteria for Cross -Over ............................................................................... 54 10.2 Exclusion Criteria for Cross -Over ............................................................................. 54 11.0 CONDUCT OF THE STUDY ........................................................................................ 55 11.1 Ethics and Regulatory Considerations ...................................................................... 55 11.2 IRB ............................................................................................................................ 55 11.3 Written Informed Consent ......................................................................................... 55 11.4 Confidentiality of Records ......................................................................................... 56 11.5 Modification of Protocol ............................................................................................. 56 11.6 Protocol Questions and Deviations ........................................................................... 56 11.7 Data Safety Monitoring Board ................................................................................... 57 12.0 EVALUATION OF TUMORS ........................................................................................ 57 12.1 Tumor Measurements and Response (RECIST 1.1) ................................................ 57 13.0 DRUG INFORMATION ................................................................................................. 57 13.1 Vigil Investigational Product ...................................................................................... 57 14.0 ADVERSE EVENTS ..................................................................................................... 59 14.1 Adverse Event and Serious Adverse Event Definitions ............................................ 59 14.2 Attribution of Causality .............................................................................................. 60 14.3 Vigil Expected Side Effects ....................................................................................... 60 14.4 Recording of an Adverse Event ................................................................................ 60 14.5 Ser ious Adverse Event Reporting ............................................................................. 61 15.0 PATIENT COMPLETION A ND WITHDRAWAL .......................................................... 61 15.1 Indication for Taking Patients Off Study .................................................................... 61 16.0 STATISTICAL CONSIDER ATIONS ............................................................................. 62 16.1 Sample Size Justification .......................................................................................... 62 16.2 Definition of Progression Free Survival (PFS) .......................................................... 62 16.3 Analysis Populations ................................................................................................. 63 16.4 Efficacy Analyses ...................................................................................................... 63 16.4.1 Primary Efficacy Analysis .................................................................................. 63 16.4.2 Secondary Efficacy Analyses ............................................................................ 63 Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 5 of 74 Confidential and proprietary information of Gradalis, Inc. 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Do not make unauthorized copies. 16.4.3 Sensitivity Analyses of the Primary Endpoint..................................................... 64 16.4.4 Sensitivity Analysis of Overall Survival .............................................................. 64 16.4.5 Subgroup Analyses of the Primary (PFS) and Key Secondary Endpoints (OS, ORR) 64 16.5 Safety Analyses ........................................................................................................ 65 16.6 Interim Analysis ......................................................................................................... 65 17.0 DOCUMENTATION ...................................................................................................... 65 17.1 Case Report Form (CRF) Proc edures ...................................................................... 66 APPENDIX A KARNOFSKY AND LANSKY PERFORMA NCE SCALES ................................. 67 APPENDIX B SCHEDULE OF ASSESSMENTS ....................................................................... 68 REFERENCES ........................................................................................................................... 69 Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 6 of 74 Confidential and proprietary information of Gradalis, Inc. 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ABBREVIATIONS Abbreviation Term AE Adverse event ALT Alanine transaminase (also referred to as SGPT) ANC Absolute neutrophil count AST Aspartate transaminase (also referred to as SGOT) BUN Blood urea nitrogen CBC Complete blood count CD Cluster of differentiation CO 2 Total carbon dioxide CR Complete response CRF Case report form CT Cancer testis CT Computerized Tomography CTCAE Common Toxicity Criteria for Adverse Events ctDNA Circulating tumor DNA DFS Disease -Free Survival DSMB Data Safety Monitoring Board DTIC dacarbazine EDC Electronic Data Capture EFS Event Free Survival ELISPOT Enzyme -Linked ImmunoSorbent Spot ERG ETS-Related Gene ESFT Ewing’s Sarcoma Family of Tumors ESPS Ewing’s Sarcoma Prognostic Scores ETS E26 transformation -specific or E-twenty -six FANG bi-shRNAfurin and GMCSF Augmented Autologous Tumor Cell Immunotherapy FISH Fluorescent in situ hybridization FLI-1 Friend Leukemia Virus Integration 1 GMCSF Granulocyte Macrophage -Colony Stimulating Factor GVAX GMCSF Secreting autologous or allogenic tumor vaccine HLA Human Leukocyte Antigen IDMC Independent Data Monitoring Committee IE Ifosfamide and etoposide IEC Independent Ethics Committee IFN Interferon Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 7 of 74 Confidential and proprietary information of Gradalis, Inc. 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Abbreviation Term IRB Institutional Review Board ITT Intent to Treat KPS Karnofsky Performance Score LDH Lactate dehydrogenase LS Lansky Performance Score MRI Magnetic resonance imaging NCI National Cancer Institute NED No evidence of disease NGS Next Generation Sequencing NK Natural Killer NKT Natural Killer T cell(s) ORR Objective Response Rate OS Overall Survival PBMC Peripheral Blood Mononuclear Cells PCR Polymerase Chain Reaction PD-L1 Programmed death-ligand 1 PFS Progression Free Survival PI Principal Investigator PR Partial response PS Performance Status RECIST Response Evaluation Criteria in Solid Tumors RFS Recurrence Free Survival RT-PCR Real Time Polymerase chain reaction TGFβ Transforming growth factor -β TIL Tumor Infiltrating Lymphocytes TTP Time to Progression TTR Time to Recurrence VAC Vincristine, Actinomycin, and Cyclophosphamide Ph III Ewing’s Sarcoma Vigil ± Irinotecan +Temozolomide IND 014205 CL-PTL-130 Version: 03 Page 8 of 74 Confidential and proprietary information of Gradalis, Inc. Effective: 05-DEC-2018 It is the user’s responsibility to verify adequate training before executing a document.