❖ GL-ONC1 treatments are well tolerated, with transient overnight flu-like symptoms. Daily i.v. hydration during treatment relieved symptoms and prevented dehydration. ❖ Mechanisms of Action are demonstrated: ➢ Direct lysis: Virus colonized and replicated in the tumor, killing of tumor cells in ascites, and reduced circulating tumor cells. ➢ Immunotherapy: Virus-induced immune activation with enhanced tumor infiltration of CD8+ T cells and generation of tumor-specific T- cell response were observed. ❖ Clinical significant disease control (including objective response) and extended PFS were documented at both dose levels. ❖ Phase 2 trial (VIRO-15) in ROC pts with adequate nutritional & immune status is currently enrolling at Cohort 1 dose level. #5577 Phase 1b Study of Oncolytic Vaccinia Virus GL-ONC1 in Recurrent Ovarian Cancer RESULTS ❖ Primary Analysis of adverse events ❖ Secondary Anti-tumor response by RECIST1.1 & survival (PFS/OS) ❖ Translational Evaluate virus-encoded transgene expression, tumor biomarkers, circulating tumor cells (CTCs), TILs in tumor biopsy, tumor-specific T-cell response in peripheral blood, cytology in ascites, immunohistochemistry of PD-L1 expression in tumor biopsies pre- and post-GL-ONC1 Tx. OBJECTIVES CONCLUSIONS Acknowledgement: All staff at Genelux for support; Prof. Lisa Butterfield lab at Univ. of Pittsburg for ELISPOT analysis. Funding for this research was provided by Genelux Corporation, San Diego, California, USA June 1-5, 2018; Chicago, IL, USA; Corresponding author: [email protected] Patient Characteristics Eleven heavily pretreated end-stage ROC pts were enrolled: ❖ Characteristics related to prior platinum Tx: Platinum-resistant (n=9; 82%), Platinum-refractory (n=1; 9%), Intermediate platinum-sensitive (n=1; 9%) ❖ # of prior lines therapy: 3-4 (n=3; 27%), ≥ 5 (n=8; 73%) ❖ ECOG 0 (n=7; 64%) or 1 (n=4; 36%) ❖ With ascites/pleural effusion at baseline (n=9; 82%) ❖ Progressive disease (PD) at baseline (n=10; 91%) ❖ Cohort 1: 3 × 10 9 pfu (n=6); Cohort 2: 1 × 10 10 pfu (n=5) Protocol #: GL-ONC1-015; ClinicalTrials.gov Identifier: NCT02759588; Copyright © 2018 Genelux Corp. All rights reserved. ABSTRACT Safety Tumor Infiltrating Lymphocytes (TILs) Anti-Tumor Response & Disease Control Observed Distal Anti-tumor Effects from i.pe. Route of Delivery Case Report (Ch1: #15A-05): OR & Tumor-specific T-cell Response Clinical trial design considerations: ➢ I.pe. route of drug delivery is relevant to ovarian cancer (OC) ➢ OCs are immunogenic → VACV is excellent adjuvant for tumor antigen presentation ➢ High tumor-infiltrating lymphocytes (TILs) favors survival → Oncolytic VACV stimulates TILs ➢ Oncolytic VACV may overcome chemo- and/or radiation-resistance ➢ For patients with chemo resistant ovarian cancer that would otherwise consider palliative care or use of drugs with poor Response Rate ❖ Disease Control Rate (DCR = OR + SD≥15 wks) = 55 % in 6/11 evaluable pts (4 in Ch1, 2 in Ch2). ❖ Extended PFS of 23, 35, 59 (with confirmed PR) & 71 wks observed in 4 pts (3 in Ch1, 1 in Ch2). ❖ More than doubling of PFS compared to the last chemotherapy regimen was recognized in 4/11 (36%) pts (2 in Ch1, 2 in Ch2). ❖ Heavily treated w/ 9 prior regimens of chemo; no Tumor-specific T-cell response at baseline ❖ Documented Objective Response (OR) from GL-ONC1 Tx after Failure of Last Chemotherapy ❖ Favorable & long-lasting Tumor-specific T-cell Response (TSTcR) by ELISPOT analysis Exemplary IHC analysis - pt.#15A-06, with PFS of 71 weeks: Significant infiltration of CD8+ cytotoxic T cells into tumors after virus treatment indicates activation of immunosurveillance Clinically Significant Results Immune Modulation peritoneal fluid Prior to 1 st dose W2D17 W2D12 pleural fluid W2D9 W2D15 W2D17 Robust lymphocyte count increase (immune activation) Direct lysis of cancer cells & immune activation in different cavities Tumor cell clusters present Tumor cell clusters absent Decrease of Circulating Tumor Cells (CTCs) may indicate systemic effect against metastases Baseline 5-mon post Tx Objective response (OR) of metastatic lesions pancreatic met. liver met. No any other Tx in the meantime 0 5 10 15 20 25 30 35 40 45 CTC COUNT PRE- & POST TX baseline post Tx Flu-like symptoms in general; lasting a few hours overnight post each Tx No DLT; MTD not reached No discontinuation due to treatment-related AEs Summary: GL-ONC1 Tx is well tolerated. baseline Week 36 CD3 CD8 Baseline 2.94 0.58 Week 36 5.06 3.54 0 1 2 3 4 5 6 Cell Counts Per 80x Field Immune Cell Counts in Tumor biopsies p<0.001 CD3+ CD8+ p<0.001 Acute inflammatory responses and PD-L1 upregulation in tumors by oncolytic virus GL-ONC1 can sensitize tumors to PD-1/PD-L1 blockade. Adverse Events # of Pts (n=11) Grades 1 & 2 AEs (occurred in ≥ 3 patients) Chills 7 (63.6%) Nausea 7 (63.6%) Fever 6 (54.5%) Abdominal distention 4 (36.4%) Abdominal pain 4 (36.4%) Vomiting 3 (27.3%) Grade 3 AEs (occurred in the same patient) Nausea 1 (9.1%) Vomiting 1 (9.1%) No Grade 4 AEs Robert W Holloway 1 , James E Kendrick 1 , Amanda J Stephens 1 , Jessica A Kennard 1 , Jeremy Burt 2 , Jane LeBlanc 3 , Karen Sellers 3 , Jamie Smith 3 , and Susan Coakley 3 1 Gynecologic Oncology Program, Florida Hospital Cancer Institute, Orlando, Florida; 2 Department of Radiologic Services, Florida Hospital, Orlando, Florida; 3 Office of Clinical Research, Florida Hospital Cancer Institute, Orlando, Florida 0 10 20 30 40 50 60 70 80 90 100 Swim Lane Plot of Survival & Tumor Response by RECIST 1.1 PR SD PD Alive WEEKS 1 Year Week 24 15A-05 15A-06 15A-09 15A-04 15A-15 15A-02 15A-01 15A-10 15A-03 15A-13 15A-07 Cohort 1 Cohort 2 (Data cut off: April 17, 2018) Prophylactic hydrations given daily to avoid dehydration, and to reduce symptoms Background : Immunotherapy can trigger immune activation including tumor-infiltrating CD8+ T cells, leading to antitumor response and survival benefits. Immunotherapeutic GL-ONC1 (modified vaccinia virus (VACV)) causes oncolysis, immune activation and durable anti-cancer memory. Methods : Intraperitoneal (i.pe.) infusion of GL-ONC1 monotherapy was given at high repeated doses in patients (pts) with platinum refractory/resistant disease. Primary endpoint : adverse events; Secondary endpoints: anti-tumor response by RECIST1.1 & survival. Eleven heavily pretreated pts with end-stage recurrent ovarian cancer (ROC) were enrolled: 3-4 prior lines (n=3), ≥ 5 lines (n=8), ECOG 0 (n=7) or 1 (n=4), ascites/pleural effusion (n=9) & progressive disease (PD) at baseline (n=10). There were two dose cohorts: 3 × 10 9 (Cohort 1: n=6) or 1 × 10 10 (Cohort 2: n=5) plaque forming units/day on 2 consecutive days. Results : (1) Adverse reactions included Grade 1-2 chills (n=7), nausea (7), fever (6), abdominal pain/distention (4), & vomiting (3). There were no differences in toxicity for the two dose levels. (2) GL-ONC1 colonized and replicated in the tumor, as indicated by a virus-encoded glucuronidase (GusA) assay. (3) Clearance of tumor cells in ascites with induction of lymphocyte infiltration was shown in 5 pts with ascites. (4) Reduction of circulating tumor cells (CTC) was identified in 6/8 (75%) pts who had baseline CTC, ranging 1-42 per 7.5 mL blood. (5) Enhanced infiltration of CD8+ T cells into tumor tissue was demonstrated by repeat biopsy. (6) A tumor-specific T cell response was absent at baseline but confirmed at Week-30 in patient with objective response (OR) by IFN-γ ELISPOT assay. (7) Disease Control Rate (DCR = OR + stable disease (SD) ≥ 15 weeks) was 6/11 (55%). (8) Extended progression-free survival (PFS) of 23, 35, 59 (with confirmed PR) & 71 weeks were observed in 4 pts, respectively. (9) More than doubling of PFS compared to the last chemotherapy regimen was recognized in 4/11 (36%) pts. Conclusions : Promising safety data, anti-tumor activity, and immune activation mechanisms were documented in this Ph1b trial, and a Ph2 trial (VIRO-15) is currently enrolling. Future studies combining GL-ONC1 and other immune therapies and/or chemotherapy are under consideration. Biomarkers 1.0E-04 1.0E-03 1.0E-02 1.0E-01 1.0E+00 1.0E+01 1.0E+02 0 2 4 6 8 10 12 14 GusA activity in blood [units/mL] Days LOD: limit of detection Virus-encoded glucuronidase LOD -70 -60 -50 -40 -30 -20 -10 0 10 20 Best change of CA-125 from baseline (%) 9/11 (82%) pts experienced CA-125 reduction Cohort 1 Cohort 2 Tumor shrinkage observed by RECIST1.1 -40 -30 -20 -10 0 10 20 30 40 Best change of SLD from baseline (%) Partial response Progressive disease Individual target lesion change Target lesion SLD change overtime ➢ 17/38 (45 %) individual target lesions had size reduction ➢ In 4 pts with SLD reduction, all individual target lesions had reduction in size -70 -60 -50 -40 -30 -20 -10 0 10 20 30 40 50 60 70 80 90 100 13 15 7 7 15 10 3 15 7 3 7 6 9 10 3 7 10 15 13 13 2 13 10 4 4 4 13 4 2 2 1 5 1 5 4 5 5 5 Best change of individual target lesion size from baseline (%) Patient ID # Progressive disease Partial response -40 -30 -20 -10 0 10 20 30 40 0 5 10 15 20 25 30 35 40 45 Overtime change of SLD from baseline (%) WEEKS PD SD PR Cohort 1 Cohort 2 Strong PD-L1 staining at the tumor-stromal interface Baseline GL-ONC1 Upregulates Immunomodulatory Target Proteins, such as PD-L1 20 days post Tx SLD: sum of longest diameters 120 140 160 180 200 220 240 260 280 300 SLD (mm) SLD of target lesions over time pemetrexed GL-ONC1 Tx + 90% PD - 34% PR Wks Last Chemotherapy: Pemetrexed 3/30-5/18/2016 Progressive disease (PD) (CT scans on 6/30/2016 & 8/30/2016) GL-ONC1 Tx Partial response (confirmed on Week 24 & 36; durable) Stable disease @ Week 48 (-27% compared to baseline) Confirmed TSTcR No TSTcR 0 18 36 -24