Safety Study of Adenovirus Vector Engineered to Express hIL-12 in Combination With Activator Ligand to Treat Melanoma

A Phase I/II, Open Label Study of Ad-RTS-hIL-12, an Adenovirus Vector Engineered to Express hIL-12, in Combination With an Oral Activator Ligand, in Subjects With Unresectable Stage III or IV Melanoma

This research study involves two investigational drugs, an Activator Ligand (INXN-1001) in combination with an Adenovirus Vector Engineered to Express hIL-12 (INXN-2001). IL-12 is a protein that may improve the body's natural response to disease by enhancing the ability of the immune system to kill tumor cells and may interfere with blood flow to the tumor.

The main purpose of this study is to evaluate the safety and tolerability of tumor injections of INXN-2001 given in combination with different doses of INXN-1001.

Study Overview

• Status: Completed
• Conditions: Melanoma
• Intervention / Treatment: Biological: INXN-2001; Drug: INXN-1001

Detailed Description

Single-arm, open label, Phase I/II dose escalation study of intratumoral injections INXN-2001 and oral INXN-1001 in subjects with unresectable Stage III or IV melanoma.

Four sequential dose escalation cohorts of INXN-1001 in combination with a fixed dose of INXN-2001 are planned. Subject enrollment and dose escalation will proceed according to a standard 3+3 design.

Approximately 15 additional subjects will be enrolled as an expansion cohort at a single dose level at or below the MTD.

• Safety and tolerability will be assessed by the incidence and severity of adverse events.
• The antitumor activity of study treatment will be assessed according to RECIST v1.1 guidelines. Additional assessment of anti-tumor activity will be explored based on total measurable tumor burden.
• Immunological and biological markers of response will include examinations of tumor biopsy samples, cytokine levels, peripheral blood mononuclear cells (PBMC) and antibody response to INXN-2001.

• Study Type: Interventional
• Enrollment (Actual): 26
• Phase: Phase 2; Phase 1

Contacts and Locations

Study Locations

• United States
  • California
    • Los Angeles, California, United States, 90404
      • The Angeles Clinic
  • Illinois
    • Park Ridge, Illinois, United States, 60068
      • Oncology Specialists
  • Indiana
    • Goshen, Indiana, United States, 46526
      • Indiana University Health Goshen Center for Cancer Care
  • Kentucky
    • Louisville, Kentucky, United States, 40202
      • James Graham Brown Cancer Center
  • Missouri
    • St Louis, Missouri, United States, 63110
      • Washington University
  • New Jersey
    • Morristown, New Jersey, United States, 07960
      • Atlantic Melanoma Center
  • Pennsylvania
    • Easton, Pennsylvania, United States, 15232
      • St. Lukes
  • Texas
    • Dallas, Texas, United States, 75201
      • Mary Crowley Cancer Research Center
  • Vermont
    • Burlington, Vermont, United States, 05401
      • Fletcher Allen Health

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• Males or females of all races ≥ 18 years of age, who have provided written informed consent prior to completing any study specific procedure.
• Unresectable Stage III or Stage IV melanoma arising from any site other than ocular melanoma.
• A minimum of 2 accessible nonvisceral lesions (shortest diameter ≥1 cm) or palpable tumor-involved lymph nodes (shortest diameter ≥1.5 cm).
• ECOG performance status of 0 or 1 (Appendix 1).
• Adequate bone marrow, liver, and renal function.
• An expected survival of at least approximately 6 months.
• Male and female subjects must agree to use a highly reliable method of birth control (expected failure rate less than 5% per year) from the screening visit through 28 days after the last dose of study drug.

Exclusion Criteria:

• Any prior anti-cancer therapy or investigational agent within 28 days prior to the first dose of study drug. (NOTE: For the expansion cohort ONLY, if subjects received ipilimumab, a 90-day washout period since last dose of ipilimumab is required. If subjects received other immunomodulating therapies (eg, anti-PD1 antibodies), the medical monitor should be contacted and an evaluation will be made.)
• Clinically significant infection requiring systemic antibacterial, antifungal, or antiviral therapy within 2 weeks of the first dose of study drug.
• History of HIV infection.
• Active autoimmune disease requiring steroids (>10 mg prednisone or comparable) or other immunosuppressive therapy (e.g., methotrexate, etc.).
• Documented symptomatic brain metastases. Screening for brain lesions by CT or MRI is not required for all potential subjects; however, if there are any neurological signs or symptoms consistent with brain metastases, then a brain CT or MRI should be performed as clinically indicated.
• Any medications that induce, inhibit or are substrates of CYP450 3A4 within 7 days prior to the first dose of study drug.
• Prior history of hematopoietic stem cell transplant or organ allograft.
• Other concurrent clinically active malignant disease, with the exception of other cancers of the skin.
• Females who are nursing or pregnant.
• Subjects who have a history of hypersensitivity that may relate to any component of the study drugs, e.g. to benzoic acid since INXN-1001 contains two benzene rings.
• Unstable or clinically significant concurrent medical condition that would, in the opinion of the investigator, jeopardize the safety of a subject and/or their compliance with the protocol.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

Experimental: INXN-1001 in combination with INXN-2001; Intratumoral injections of INXN-2001 (Ad-RTS-hIL-12) at a constant dose in combination with inter-cohort escalating doses of INXN-1001 (activator ligand).

Biological: INXN-2001; approximately 1.0 x 1012 viral particles (vp) per injection; one intratumoral injection of INXN-2001 per study cycle; maximum of 6 study cycles; Other Names: Ad-RTS-hIL-12; Drug: INXN-1001; 4 dose cohorts (5mg/day, 20mg/day, 100mg/day, and 160mg/day); 7 oral daily doses of INXN-1001 per study cycle; maximum of 6 study cycles; 1 Expansion cohort at a single dose level at or below MTD (160mg/day); Other Names: Activator Ligand

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of Participants With Treatment-Emergent Adverse Events	Evaluation will be based on the incidence, intensity, and type of Adverse Events (AEs). Clinically significant changes in the subjects' physical examinations, vital signs, and ECG evaluations, and clinical manifestations relevant to abnormal laboratory values will be captured as AEs.	From the signing of informed consent until 28 days after the last dose of study treatment, up to 28 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Area Under the Plasma Concentration-Time Curve (AUC) of INXN-1001 (Veledimex) in Cycle 1	The area under the plasma concentration versus time curve from time 0 to the last measurable concentration (AUC0-t) for INXN-1001, calculated during the first treatment cycle.	Samples were collected in Cycle 1 on Day 1 (pre-dose and 0.5, 1, 2, 4, and 6 hours post-dose), Day 2 (pre-dose), Day 7 (pre-dose and 1-2 and 4-6 hours post-dose), and Day 8 (24 hours after the Day 7 dose).
Progression-Free Survival (PFS)	Progression-Free Survival (PFS) was defined as the time in days from the first dose of study treatment to the first assessment of disease progression or death from any cause, whichever occurred first. Subjects who withdrew from the study were censored at the date of their last non-progressive disease assessment. The final analysis was performed using Kaplan-Meier methodology to determine the median PFS.	From the first dose of study treatment for up to 1 year.
Change From Baseline in Tumor IL-12 Messenger RNA (mRNA) Expression Change in IL-12 mRNA Expression Level (Arbitrary Units)	Tumor biopsy samples were analyzed for IL-12 mRNA expression using quantitative real-time polymerase chain reaction (qRT-PCR). Expression levels were normalized to the housekeeping gene ACTB (β-actin) and quantified using the ΔΔCt method. For each participant, the median ΔΔCt-based fold-change from baseline to post-treatment was calculated. These individual participant-level medians were then averaged to derive the mean of medians (measure type) across all participants within each arm/group. The reported values therefore reflect the mean of medians in arbitrary units (AU), representing relative IL-12 mRNA expression normalized to ACTB- calculated as 2^(-ΔΔCt) fold-change relative to baseline Not all subjects in each arm contributed evaluable IL-12 mRNA expression data due to biopsy availability and RNA quality.	Baseline (Screening) and at the Post-Treatment Safety Assessment (approximately 28 days after the end of Cycle 1).
Maximum Plasma Concentration (Cmax) of INXN-1001 (Veledimex) in Cycle 1	The maximum observed plasma concentration (Cmax) of INXN-1001 following oral administration, as determined from plasma sample analysis during the first treatment cycle	Samples were collected in Cycle 1 on Day 1 (pre-dose and 0.5, 1, 2, 4, and 6 hours post-dose), Day 2 (pre-dose), Day 7 (pre-dose and 1-2 and 4-6 hours post-dose), and Day 8 (24 hours after the Day 7 dose)
Change From Baseline in Peripheral Blood Cytotoxic T-Lymphocytes (CTLs)	The percentage of Cytotoxic T-Lymphocytes (CTLs) within the CD8+ T-cell population was measured from serum samples by flow cytometry to assess changes in immune cell populations.	Baseline (Screening) and at Cycle 2, Day 7
Change From Baseline in Peripheral Blood Regulatory T-cells (Tregs)	The percentage of Regulatory T-cells (Tregs) in peripheral blood lymphocytes was measured from serum samples by flow cytometry to assess changes in immune cell populations.	Baseline (Screening) and at Cycle 2, Day 7.
Change From Baseline in Tumor Interferon-gamma (IFN-γ) Messenger RNA (mRNA) Expression	Tumor biopsy samples were analyzed for IFN-γ mRNA expression using quantitative real-time polymerase chain reaction (qRT-PCR). This measure reports the fold change in expression from baseline. Due to very low or undetectable baseline IFN-γ expression in many tumor biopsy samples, the fold change values calculated by qRT-PCR using the delta-delta Ct method may result in large numeric values. All values reported reflect fold change from baseline and were calculated per the qRT-PCR assay standard procedures	Baseline (Screening) and at the Post-Treatment Safety Assessment (28 days after the end of Cycle 1).
Ad-RTS-hIL-12 Vector Copy Numbers in Tumor Biopsy Samples	To evaluate the presence of the viral vector in the tumor, biopsy samples were assessed for vector copy numbers using a quantitative polymerase chain reaction (qPCR) assay. The results are reported as the number of vector copies per 100 nanograms (ng) of deoxyribonucleic acid (DNA).	Day 2 of Cycle 1, Day 2 of Cycle 2, and Day 2 of Cycle 3.
Best Overall Response (BOR) by RECIST 1.1 Criteria	Best Overall Response (BOR) was a pre-specified secondary endpoint assessed according to Response Evaluation Criteria In Solid Tumors (RECIST) v1.1. BOR is the best response recorded from the start of treatment until disease progression. Responses include Complete Response (CR), Partial Response (PR), Stable Disease (SD), and Progressive Disease (PD).	From the first dose of study treatment for up to 1 year.
Time to Maximum Plasma Concentration (Tmax) of INXN-1001 (Veledimex) in Cycle 1	The time to reach the maximum observed plasma concentration (Tmax) of INXN-1001 following oral administration during the first treatment cycle.	Samples were collected in Cycle 1 on Day 1 (pre-dose and 0.5, 1, 2, 4, and 6 hours post-dose), Day 2 (pre-dose), Day 7 (pre-dose and 1-2 and 4-6 hours post-dose), and Day 8 (24 hours after the Day 7 dose).

Collaborators and Investigators

• Sponsor: Alaunos Therapeutics
• Investigators: Study Director: Jaymes Holland, Alaunos Therapeutics

Study record dates

Study Major Dates

• Study Start: August 1, 2011
• Primary Completion (Actual): September 1, 2014
• Study Completion (Actual): September 1, 2014

Study Registration Dates

• First Submitted: July 14, 2011
• First Submitted That Met QC Criteria: July 18, 2011
• First Posted (Estimated): July 19, 2011

Study Record Updates

• Last Update Posted (Estimated): October 29, 2025
• Last Update Submitted That Met QC Criteria: October 15, 2025
• Last Verified: October 1, 2025

More Information

Terms related to this study

• Keywords: Melanoma; Unresectable
• Additional Relevant MeSH Terms: Neoplasms by Site; Neoplasms; Neoplasms by Histologic Type; Skin Diseases; Neuroectodermal Tumors; Neoplasms, Germ Cell and Embryonal; Neoplasms, Nerve Tissue; Neuroendocrine Tumors; Nevi and Melanomas; Skin Neoplasms; Skin and Connective Tissue Diseases; Melanoma
• Other Study ID Numbers: ADA1001; ATI001-101 (Other Identifier: ZIOPHARM)