A phase I trial of intraperitoneal GEN-1, an IL-12 plasmid formulated with PEG-PEI-cholesterol lipopolymer, administered with pegylated liposomal doxorubicin in patients with recurrent or persistent epithelial ovarian, fallopian tube or primary peritoneal cancers: An NRG Oncology/Gynecologic Oncology Group study

Published:August 10, 2017DOI:


      • Recurrent EOC patients were treated with GEN-1 and pegylated liposomal doxorubicin.
      • Three had partial responses and five had stable disease of the 16 patients.
      • IL-12 expression and downstream immunological effect were found.
      • Maximum tolerated dose of GEN-1 was not achieved.



      The study's purpose was to assess safety and efficacy of escalating doses of weekly GEN-1 with pegylated liposomal doxorubicin (PLD) in patients with recurrent or persistent epithelial ovarian, fallopian tube or primary peritoneal cancers (EOC).


      Patients had persistent or recurrent platinum-resistant EOC. The trial was a standard 3 + 3 phase I dose escalation design with patients receiving intravenous PLD 40 mg/m2 (dose level 1 and 2) or 50 mg/m2 (dose level 3) every 28 days and intraperitoneal GEN-1 at 24 mg/m2 (dose level 1) or 36 mg/m2 (dose level 2 and 3) on days 1, 8, 15, and 22 of a 28 day cycle. Cycles were repeated every 28 days until disease progression. Patients were monitored for toxicity, clinical efficacy, and evidence of systemic and intraperitoneal immunologic effect.


      Sixteen evaluable patients received a median of 4 cycles (range 1–8). No dose limiting toxicities were found. The adverse side effects were 4 grade 3 anemia, 2 grade 3 abdominal pain, 7 grade 3 neutropenia, and 2 grade 4 neutropenia. A clinical benefit of 57.1% (PR = 21.4%; SD = 35.7%) was found in the 14 patients with measurable disease. The highest number of partial responses (28.6%) and stable disease (57.1%) were found at dose level 3. The maximum tolerated dose was not reached. Increases in IL-12, IFN-γ, and TNF-α levels were found in peritoneal fluid following GEN-1 treatment.


      GEN-1 in combination with PLD has encouraging clinical benefit and biological activity in recurrent or persistent EOC and warrants further investigation with escalating doses of GEN-1.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Gynecologic Oncology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Howlader N.
        • Noone A.M.
        • Krapcho M.
        • Garshell J.
        • Miller D.
        • Altekruse S.F.
        • et al.
        SEER Cancer Statistics Review.
        National Cancer Institute, Bethesda, MD1975–2012 (based on November 2014 SEER data submission, posted to the SEER web site, April 2015)
        • Whitworth J.M.
        • Alvarez R.D.
        Evaluating the role of IL-12 based therapies in ovarian cancer: a review of the literature.
        Expert. Opin. Biol. Ther. 2011; 11: 751-762
        • Duda D.G.
        • Sunamura M.
        • Lozonschi L.
        • Kodoma T.
        • Egawa S.I.
        • Matsumoto G.
        • et al.
        Direct in vitro evidence and in vivo analysis of the anti-angiogenesis effects of interleukin 12.
        Cancer Res. 2000; 60: 111-116
        • Lenzi R.
        • Edwards R.
        • June C.
        • Seiden M.V.
        • Garcia M.E.
        • Rosenblum M.
        • et al.
        Phase II study of intraperitoneal recombinant interleukin-12 (rhIL-12) in patients with peritoneal carcinomatosis (residual disease <1 cm) associated with ovarian cancer or primary peritoneal carcinoma.
        J. Transl. Med. 2007; 5: 66
        • Fewell J.G.
        • Matar M.
        • Slobodkin G.
        • Han S.O.
        • Rice J.
        • Hovanes B.
        • et al.
        Synthesis and application of biocompatible non-viral gene delivery system for immunogene therapy of cancer.
        J. Control. Release. 2005; 109: 288-298
        • Fewell J.G.
        • Rice J.
        • Matar M.
        • Brunhoeber E.
        • Pence C.
        • Slobodkin G.
        • et al.
        Use of interleukin-12 gene nanocomplexes as a general treatment against disseminated peritoneal malignancies.
        Proc Am Assoc Cancer Res. 2007; 48: 3304
        • Fewell J.G.
        • Matar M.M.
        • Rice J.S.
        • Brunhoeber E.
        • Slobodkin G.
        • Pence C.
        • et al.
        Treatment of disseminated ovarian cancer using nonviral interleukin-12 gene therapy delivered intraperitoneally.
        J. Gene Med. 2009; 11: 718-728
        • Anwer K.
        • Barnes M.N.
        • Fewell J.
        • Lewis D.H.
        • Alvarez R.D.
        Phase-I clinical trial of IL-12 plasmid/lipopolymer complexes for the treatment of recurrent ovarian cancer.
        Gene Ther. 2010; 17: 360-369
        • Anwer K.
        • Kelly F.J.
        • Chu C.
        • Fewell J.G.
        • Lewis D.
        • Chu J.G.
        • et al.
        Phase I trial of a formulated IL-12 plasmid in combination with carboplatin and docetaxel chemotherapy in the treatment of platinum-sensitive recurrent ovarian cancer.
        Gynecol. Oncol. 2013; 131: 169-173
        • Markman M.
        • Gordan A.N.
        • McGuire W.P.
        • Muggia F.M.
        Liposomal anthracycline treatment for ovarian cancer.
        Semin. Oncol. 2004; 31: 91-105
        • Gordon A.N.
        • Granai C.O.
        • Rose P.G.
        • Hainsworth J.
        • Lopez A.
        • Weismann C.
        • et al.
        Phase II study of liposomal doxorubicin in platinum- and paclitaxel-refractory epithelial ovarian cancer.
        J. Clin. Oncol. 2000; 18: 3093-3100
        • Colombo N.
        • Kutarska E.
        • Dimopoulos M.
        • Bae D.S.
        • Rzepka-Gorska I.
        • Bidzinski M.
        • et al.
        Randomized, open-label phase III study comparing patupilone (EPO906) with pegylated liposomal doxorubicin in platinum-refractory or -resistant patients with recurrent epithelial ovarian, primary fallopian tube, or primary peritoneal cancer.
        J. Clin. Oncol. 2012; 30: 3841-3847
        • Kepp O.
        • Galluzzi L.
        • Martins I.
        • Schlemmer F.
        • Adjemian S.
        • Michaud M.
        • et al.
        Molecular determinants of immunogenic cell death elicited by anticancer chemotherapy.
        Cancer Metastasis Rev. 2011; 30: 61-69
        • Eisenhauer E.A.
        • Therasse P.
        • Bogaerts J.
        • Schwartz L.H.
        • Sargent D.
        • Ford R.
        • et al.
        New response evaluation criteria in solid tumours: revised RECIST guideline (version 1.1).
        Eur. J. Cancer. 2009; 45: 228-247
        • Le Tourneau C.
        • Lee J.J.
        • Siu L.L.
        Dose escalation methods in phase I cancer clinical trials.
        J. Natl. Cancer Inst. 2009; 101: 708-720
        • Deprimo S.E.
        • Bello C.L.
        • Smeraglia J.
        • Baum C.M.
        • Spinella D.
        • Rini B.I.
        • et al.
        Circulating protein biomarkers of pharmacodynamics activity of sunitinib in patients with metastatic renal carcinoma: modulation of VEGF and VEGF-related proteins.
        J. Transl. Med. 2007; 5: 32-38
        • Alidzanovic L.
        • Starlinger P.
        • Schauer D.
        • Maier T.
        • Feldman A.
        • Buchberger E.
        • et al.
        The VEGF rise in blood of bevacizumab patients is not based on tumor escape but a host-blockade of VEGF clearance.
        Oncotarget. 2016; 7: 57197-57212
        • Keunen O.
        • Johansson M.
        • Oudin A.
        • Snazey M.
        • Rahim S.A.
        • Fack F.
        • et al.
        Anti-VEGF treatment reduces blood supply and increases tumor cell invasion in glioblastoma.
        Proc. Natl. Acad. Sci. U. S. A. 2011; 108: 3749-3754
        • Stefnani M.O.
        • Wu F.T.
        • mac Gabhann F.
        • Popel A.S.
        Increase of plasma VEGF after intravenous administration of bevacizumab is predicted by a pharmacokinetic model.
        Cancer Res. 2010; 70: 9886-9894
        • Alvarez R.D.
        • Sill M.W.
        • Davidson S.A.
        • Muller C.Y.
        • Bender D.P.
        • Sill M.W.
        • et al.
        A phase II trial of intraperitoneal EGEN-001, an IL-12 plasmid formulated with PEG-PEI-cholesterol lipopolymer in the treatment of persistent or recurrent epithelial ovarian, fallopian tube or primary peritoneal cancer: a Gynecologic Oncology Group study.
        Gynecol. Oncol. 2014; 133: 433-438
        • Zhu S.
        • Waguespack M.
        • Barker S.A.
        • Li S.
        Doxorubicin directs the accumulation of interleukin-12 induced IFN gamma into tumors for enhancing STAT1 dependent anti-tumor effect.
        Clin. Cancer Res. 2007; 13: 4252-4260
        • Zagozdon R.
        • Golab J.
        • Stoklosa T.
        • Giermasz A.
        • Nowicka D.
        • Feleszko W.
        • et al.
        Effective chemo-immunomodulatory of L1210 leukemia in vivo using interleukin-12 combined with doxorubicin but not with cyclophosphamide, paclitaxel or cisplatin.
        Int. J. Cancer. 1998; 77: 720-727
        • Alagkiozidis I.
        • Facciabene A.
        • Carpenito C.
        • Benecia F.
        • Jonak Z.
        • Adams S.
        • et al.
        Increased immunogenicity of surviving tumor cells enables cooperation between liposomal doxorubicin and IL-18.
        J. Transl. Med. 2009; 7: 104
        • Teicher B.A.
        • Ara G.
        • Buxton D.
        • Leonard J.
        • Schaub R.G.
        Optimal scheduling of interleukin 12 and chemotherapy in the murine MB-49 bladder carcinoma and B16 melanoma.
        Clin. Cancer Res. 1997; 3: 1661-1667
        • Torrerro M.N.
        • Xia X.
        • Henk W.
        • Yu S.
        • Li S.
        Stat1 deficiency in the host enhances interleukin-12-mediated tumor regression.
        Cancer Res. 2006; 66: 4461-4467
        • Ujhazy P.
        • Zaleskis G.
        • Mihich E.
        Doxorubicin induces specific immune functions and cytokine expression in the peritoneal cells.
        Cancer Immunol. Immunother. 2003; 52: 463-472
        • Orsini F.
        • Pavelic Z.
        • Mihich E.
        Increased primary cell mediated immunity in culture subsequent to Adriamycin or daunorubicin treatment of spleen donor mice.
        Cancer Res. 1997; 37: 1719-1726
        • Maccubbin D.L.
        • Wing K.R.
        • Mace K.F.
        • Ho R.L.X.
        • Ehrke M.J.
        • Mihich E.
        Adriamycin-induced modulation of host defenses in tumor-bearing mice.
        Cancer Res. 1992; 52: 3572-3576
        • Mace K.
        • Mayhew E.
        • Mihich E.
        • Ehrke M.J.
        Alterations in murine host defense functions by Adriamycin or liposome-encapsulated Adriamycin.
        Cancer Res. 1988; 48: 130-136
        • Alizadeh D.
        • Trad M.
        • Hanke N.T.
        • Larmonier C.B.
        • Janikashvili N.
        • Bonnotte B.
        • et al.
        Doxorubicin eliminates myeloid-derived suppressor cells and enhances the efficacy of adoptive T-cell transfer in breast cancer.
        Cancer Res. 2014; : 74104-74118
        • Mutch D.G.
        • Orlando M.
        • Goss T.
        • Teneriello M.G.
        • Gordon A.N.
        • McMeekin S.D.
        • et al.
        Randomized phase III trial of gemcitabine compared with pegylated liposomal doxorubicin in patients with platinum-resistant ovarian cancer.
        J. Clin. Oncol. 2007; 25: 2811-2818
        • Gordon A.N.
        • Fleagle J.T.
        • Guthrie D.
        • Parkin D.E.
        • Gore M.E.
        • Lacave A.J.
        Recurrent epithelial ovarian carcinoma: a randomized phase III study of pegylated liposomal doxorubicin versus topotecan.
        J. Clin. Oncol. 2001; 19: 3312-3322
        • Gollob J.A.
        • Mier J.W.
        • Veenstra K.
        • McDermott D.F.
        • Clancy D.
        • Clancy M.
        • et al.
        Phase I trial of twice-weekly intravenous IL-12 in patients with metastatic renal cancer or malignant melanoma: ability to maintain IFN-gamma induction is associated with clinical response.
        Clin. Cancer Res. 2000; 6: 1678-1692
        • Gollob J.A.
        • Veenstra K.G.
        • Jyonouchi H.
        • Kelly A.M.
        • Ferrieri P.
        • Panka D.J.
        • et al.
        Impairment of STAT activation by IL-12 in a patient with atypical mycobacterial and staphylococcal infections.
        J. Immunol. 2000; 65: 4120-4126
        • Lippitz B.E.
        Cytokine patterns in patients with cancer: a systematic review.
        Lancet Oncol. 2013; 14: e218-228
        • Yuzhalin A.E.
        • Kutikhin A.G.
        Interleukin-12: clinical usage and molecular markers of cancer susceptibility.
        Growth Factors. 2012; 30: 176-191
        • Thaker P.H.
        • Leath C.A.
        • Bradley W.H.
        • Moore K.N.
        • Borys N.
        • Anwer K.
        • et al.
        Phase I study of the safety and biological activity of intraperitoneal IL-12 plasmid formulated with PEG-PEI-cholesterol lipopolymer administered in combination with standard neoadjuvant chemotherapy in patients with newly diagnosed ovarian cancer.
        J. Clin. Oncol. 2016; 34TPS5605