Electro-Gene Therapy with IL-12 plasmid in metastatic melanoma
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1 Electro-Gene Therapy with IL-12 plasmid in metastatic melanoma Adil Daud MBBS Clinical Professor of Medicine and Dermatology University of California, San Francisco
2 Disclosures Advisory Boards OncoSec, Amgen Research Funding OncoSec, GSK, Genentech/Roche, Pfizer, Merck, Bristol-Myers Stock Ownership None Employment None Speaker Bureau- None
3 Electrically Mediated Delivery Electrogene Transfer (EGT) DC DNA Surrounds Target Cells Cell Poration, DNA enters cells Cells Reseal & DNA expresses
4
5 % Survival Intra-tumor electroporation with IL-12 Plasmid in B16-F10 Tumors No Treat ment IL-12 Pl asmi d, No El ectroporati on Vector and El ectroporati on IL-12 Pl asmi d and El ectroporati on 3TX D a y s A fter Trea tment Lucas, et al, DNA Cell Biology 22(12):753, 2003.
6 % Survival Intra-tumor electroporation with IL-12 Plasmid in B16.F10 Tumor 100 P-E P+ E- i.t. V+ E+ i.t. P+ E+ i.t. 2 tx P+ E+ i.t. P+ E+ i.t./i.m Two Tumor Model Days after T herapy Lucas, et al, DNA Cell Biology 22(12):753, 2003.
7 Fold increase over Day 0 volume % Survival Treament of B16 Melanoma in Athymic Mice 15 0 P-E P+E-i.t. V+E+ i.t P+E+ i.t P-E- P+E- i.t V+E+ i.t. P+E+ i.t Days after therapy Days after T herapy 1. Lucas ML, Heller L, Coppola D, Heller R. IL-12 plasmid delivery by in vivo electroporation for the successful treatment of established subcutaneous B16.F10 melanoma. Mol. Ther 2002 Jun;5(6):
8
9 Image shown for educational purposes only
10 EP h IL-12 EP h IL-12 EP h IL-12 Excision of any remaining EP lesions and other lesions if indicated Blood Days FNA Blood IL-12 PBMC FNA Blood + Punch IL-12 FNA Blood + Punch IL-12 PBMC Phase I trial of plasmid IL-12 Electroporation in Melanoma First-in-Man trial
11 Table 1 Cohort Patient Age Sex AJCC Stage LDH IL-12 Plasmid Treated Lesions Objective Response Number Site Distant Disease Overall Duration Sites Response (Months) Concentration (mg/ml) Lesion a Volume (mm 3 ) 1 35 M IVA Leg Sub Q, LN PD 2 54 M IVC Trunk Sub Q, LN PD 3 69 M IVC Trunk SubQ PD 4 55 M IVC Trunk Multiple Sites PD 5 66 M IVB Trunk Multiple Sites SD M IVA Trunk, Arm SubQ PD 7 50 M IIIC Trunk, Arm SubQ * > M IIIC Leg SubQ PD 9 80 M IVA Trunk, Arm SubQ CR > M IVA Trunk SubQ PD F IVC Leg SubQ, LN PD M IIIC Trunk - PD M IIIC Arm - PD F IIIC Leg Sub Q CR > M IIIC Arm Sub Q PD M IIIC FV 4 Trunk Sub Q SD F IIIB FV 3 Leg - SD F IIIC FV 4 Leg Sub Q PD M IIIC FV 2 Leg LN PD M IIIB FV 4 Leg - SD M IVA FV 4 Leg SubQ SD M IVA FV 2 Trunk Sub Q PD M IVA FV 4 Leg SubQ, LN SD M IVC FV 3 Neck Liver, Lung PD
12 Expression of IL-12 and IFN- in Tumor Biopsies H I Figure 4: Expression of IL-12 (Panel H) and IFN (Panel I) in post treatment biopsies Daud AI et al., Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 2008 Dec;26(36):
13 Pre-Tx D 256 D 637 Patient 9 Cohort 3 Chest A B C Back D E F Daud AI et al., Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 2008 Dec;26(36):
14 Patient 4, Cohort 5 Day 5 A B Day 513 C D
15 H & E, 200 X Pre-tx Day 22 Day Mo A B C D Pt 9 Pt 10 E F G H Day 22 50X Day X Day 22 CD4,200X Day 22 CD8,200X H & E Immunostains
16 Conclusions from the Phase I Trial of pil-12 EP in Melanoma EP with pil-12 is safe and non toxic No NCI CTC Grade 2 toxicity observed Intra-tumoral pil-12 Electroporation leads to reproducible sustained high level expression of IL-12 and IFN- Very effective local treatment > 90 % Prolonged Systemic Efficacy Distant CR seen in 2 patient/19 (no other Tx) Distant CR in 1/19 (had other treatment) SD and PR in 7/19
17 Ongoing Multicenter Phase II Trial of pil- 12 Electroporation in Melanoma Inclusion In-transit/cut/subQ mets Stage III unresectable or Stage IV Normal blood counts/organ fx No extensive visceral disease >1 tumor Exclusion Prior IL-12 Pacemaker, seizure
18 Blood Immune Response Blood Immune Response Blood Immune Response Blood Immune Response FNA/ Punch Biopsy FNA Biopsy FNA/ Punch Biopsy Days Pre Tumor Eval Tumor Tumor Tumor Tumor Eval Eval Eval Eval pil12/ control EGT pil12/ control EGT pil12/ control EGT Figure 5 Stage IIIB-IV Melanoma with In transit/ cutaneous Metastasis pil mg/ml X 1ml intratumorally with EP, D1,5,8 Patients may be retreated q3 months if no evidence of systemic progression Phase II Trial
19 Plan Statistics 25 patients total 15 patients 1st stage Primary Objective 24 week Distant complete RR Secondary Objectives Local RR, Objective PFS, OS, Exploratory Biomarkers IL-12 and IFN expression TIL and circulating FOXP3 cells
20 Patient Characteristics Patient Age Sex Stage Lesions treated Sites # Cycles BRAF Status M 3 4 Head 1 WT M 3 2 R Leg, R foot, trunk M 3 4 Head, Neck M 4 4 Trunk, Neck 2 WT 2 WT 1 V600E F 4 3 Head 1 V600E M 4 2 Trunk 1 NA F 3 2 Right leg 1 T599I and V600E F 3 3 Left leg 2 V600E M 3 2 Right arm 1 NA M 3 3 Trunk, Right arm 1 V600E F 3 2 Right leg 1 NA F 3 3 Right leg 1 NA M 3 2 Neck 1 NA
21 Treatment-related adverse events (Total = 18) Event Grade Fatigue 2 Anxiety 1 Facial Pain 1 Pruritus 1 Erythema 2 Maculopapular rash 2 Bleeding with treatment 1 Nausea 1 Diarrhea 1 Cough 1 Arthralgia 1 Myalgia 1 Urinary tract pain 1
22 pg/g of tumor IL-12 levels detected within punch biopsies of treated tumors by ELISA after injection of plasmid IL-12 and electroporation Evaluable punch biopsies that were taken before treatment and at day IL-12 levels ELISAs were performed for IL-12 protein levels and scored per gram of tumor tissue Pre-Study Day 11
23 Monitoring of serum autoantibodies using a seromic approach Σ(positive resp) = Antigen Score (0-35) Before pil-12 4 weeks after pil-12 Multiplex miniaturized ELISA format allows simultaneous detection of autoantibodies and high throughput screening of a panel of 35 melanoma-associated antigens.
24 % CD4 CD3 cells % lymphocytes % lymphocytes Preliminary assessment of changes in T cell populations over time CD25 FoxP3 CD8 CD69 CD8 PD Cycle 1 Cycle 2 Cycle 1 Cycle 2 Cycle 1 Cycle 2 Patient Baseline 2. Day Day Day Day 180
25 Suppurative folliculitis at previous site of primary melanoma Day 50 Neutrophilic dermatitis (lt medial ankle) resembling Sweet s syndrome Day Four days after sent us pictures seen on last slide, came to clinic and these were sent to path
26 Self reported photos for rash that popped up o/n, no pain or itching Day 46 E/p scar is lesion 1 E/p scars are lesions 2 and 3 Inferior/anterior to lesion 4 (not pictured)
27 Pre-Study Day 90 PR Day 180 CR for local and systemic c
28 Pre-study txd lesions Day 90 PR (flattening of 1 and 2 and fading of 3 and 4) 1 &
29 Local and Distant Response per Patient Distant Response ORR Patient Day 39 Day 90 Day 120 Day 180 Day 180 Day PR PR Off Study PR CR CR CR SD SD PR PR PR PR CR CR CR Off Study PR Off Study CR CR Off Study CR CR CR CR CR CR CR CR CR PD PR SD Off Study SD Off Study SD PR PD PR PR
30 Durable Response Rate (DRR) 3 Months (Day 90) 6 Months (Day 180) Treated Lesions 68% 45% Treated Patients 69% 38% Of the 13 subjects analyzed at the interim analysis: Of the 31 lesions at Day 39 that were observed to have an ORR (CR or PR), 21 of 31 lesions had a DRR at 3 months. Of the 31 lesions at Day 39 that were observed to have an ORR (CR or PR),14 of 31 lesions had a DRR at 6 months. 9 of 13 patients had a DRR of treated lesions at 3 months. 4 of 13 patients had a DRR of treated lesions at 6 months
31 Disease Evaluation Distant Response (non treated lesions) 2/13 had evaluable distant disease and had a response distantly by D180 4/13 not yet at D180 6/13 have PD 1/13 withdrew from study
32 Conclusions IL-12 plasmid electroporation is associated with a tolerable side effect profile Phase I Widespread local responses and 10% systemic CR Phase II 2/13 Distant Responses 4/13 awaiting D180 eval ~40% durable responses ( 6 month)
33 Acknowledgements Pathology Anna Berry MD Britt-Marie Ljung MD Research Richard Heller Edward Cha MD PHD Larry Fong MD OncoSec Inc Punit Dhillon Avtar Dhillon MD Tu Diep Patients and Families
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