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

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.

Time frame: From the signing of informed consent until 28 days after the last dose of study treatment, up to 28 weeks

Population: All safety analyses will be conducted on the Safety population. The Safety population is defined as all patients who receive at least one INXN-1001 capsule or, in the event an injection of INXN-2001 is administered before an INXN-1001 capsule is taken, at least one injection of INXN-2001.~The Safety population will be used for the analysis of safety data based on the actual initial dose of INXN-1001 received.

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).

Time frame: Day 2 of Cycle 1, Day 2 of Cycle 2, and Day 2 of Cycle 3.

Population: The analysis was performed on subjects from the Safety Population who had a tumor biopsy sample available for analysis at any of the specified time points. The number of subjects with available data varies by cohort and time point, and the Overall Number of Participants Analyzed for each cohort reflects the unique number of subjects in that cohort who contributed data at any time point.

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.

Time frame: 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).

Population: The PK Population includes all subjects who received at least one dose of INXN-1001 and had sufficient plasma concentration data to derive reliable PK parameters.

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).

Time frame: From the first dose of study treatment for up to 1 year.

Population: Analysis of BOR was performed on all subjects who received at least one dose of study drug and were evaluable for response. Of the 26 treated patients, 15 were evaluable for Best Overall Response per RECIST v1.1. Subjects were excluded if no post-baseline tumor imaging was available. The Phase I 160 mg QD and Phase II 120 mg QOD and 80 mg QOD cohorts had no evaluable subjects at the data cutoff due to early discontinuation or pending imaging. No data were imputed

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.

Time frame: Baseline (Screening) and at Cycle 2, Day 7

Population: A total of 5 of the 26 treated subjects were included in this CTL analysis. Inclusion was based on the availability of paired peripheral blood samples at baseline and Cycle 2, Day 7

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.

Time frame: Baseline (Screening) and at Cycle 2, Day 7.

Population: A total of 5 of the 26 treated subjects were included in this final analysis of peripheral blood Regulatory T-cell levels. Inclusion required availability of paired serum samples from Baseline and Cycle 2, Day 7.

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.

Time frame: Baseline (Screening) and at the Post-Treatment Safety Assessment (approximately 28 days after the end of Cycle 1).

Population: The analysis was performed on participants in the Safety Population who had paired (baseline and post-treatment) tumor biopsy samples suitable for qRT-PCR.~Paired samples were required for inclusion.~The number of participants analyzed per arm/group is lower than the total enrolled population because of limited biopsy availability, insufficient RNA yield, or RNA quality issues.

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

Time frame: Baseline (Screening) and at the Post-Treatment Safety Assessment (28 days after the end of Cycle 1).

Population: The analysis was performed on subjects from the Safety Population who had paired (baseline and post-treatment) tumor biopsy samples available for interferon-gamma (IFN-γ) mRNA analysis. A total of 12 of the 26 treated subjects were included in this final analysis. The number of participants analyzed per cohort reflects those who contributed data at any post-baseline time point, even if data were not available at all scheduled intervals.

The maximum observed plasma concentration (Cmax) of INXN-1001 following oral administration, as determined from plasma sample analysis during the first treatment cycle

Time frame: 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)

Population: The PK Population includes all subjects who received at least one dose of INXN-1001 and had sufficient plasma concentration data to derive reliable PK parameters.

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.

Time frame: From the first dose of study treatment for up to 1 year.

Population: Subjects with at least one post baseline evaluation. Two subjects were enrolled in the 80 mg QOD arm. Both discontinued study treatment early and did not receive post-baseline tumor assessments. One subject completed scheduled visits but had no RECIST-evaluable imaging and was censored at baseline. Therefore, 0 subjects from this arm were included in the PFS analysis. The overall PFS-evaluable population included 17 subjects across all arms.

The time to reach the maximum observed plasma concentration (Tmax) of INXN-1001 following oral administration during the first treatment cycle.

Time frame: 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).

Population: The analysis was performed on the PK Population, which includes all subjects who received at least one dose of INXN-1001 and had sufficient plasma concentration data to derive reliable PK parameters.