Treatment schema.

Subjects underwent a series of pretreatment evaluations and met all the eligibility criteria before enrolled in the study. Subjects received a single intraprostatic injection of the Ad5-yCD/mutTKSR39rep-hIL-12 adenovirus at one of five dose levels (see Table 1) on day 1. Two days later (day 3), subjects received a one-week (7 day) course of 5-FC (150 mg/kg/day) + vGCV (1,800 mg/day) prodrug therapy. Toxicity assessments occurred twice a week during the first two weeks and then at every scheduled follow-up visit. The primary endpoint was toxicity through day 30.</p

An estimation of peripheral blood mononuclear cells (PBMCs) for all cohorts.

An estimation of peripheral blood mononuclear cells (PBMCs) for all cohorts.</p

Change in patient PSA in response to gene therapy.

PSA doubling time (PSADT) for all the patients were estimated before gene therapy (PSADT.PRE) and after adenoviral injections (PSADT.POST). Mean, median, minimum, and maximum PSADT are tabulated (A; years). Median and IQR of pre- and post-gene therapy PSADT is plotted in B. Statistical mean, median, minimum, and maximum PSADT for patients that were injected with low-dose of adenovirus (cohorts #1–3) and from patients from high-dose (cohorts #4–5) were estimated and plotted in C. A plot of mean, median, minimum, maximum and IQR for both the groups is shown to the right (D). Absolute PSA count (pg/mL) for patient #12 (E) and #13 (F) over time is plotted. Red dotted line indicates the day of Ad5-IL-12 adenoviral injection. Blue arrows indicate date of androgen suppression therapy (AST).</p

Patient baseline characteristics and adenoviral injection.

Each cohort had 3 patients that were injected with an increasing Ad5-IL-12 dose.</p

Changes in serum IL-12, IFNγ, and CXCL10 levels in each cohort.

Patient blood was drawn within 30 days of the Ad5-IL-12 adenoviral injection or on the day of adenovirus injection prior to the procedure (day 0) and on the days as described in the clinical protocol (up to day 24). Cytokines were measured by ELISA based on cytokine specific standard curves (IL-12 0–500 pg/mL; CXCL10 and IFNγ 0–1,000 pg/mL). The peak cytokine value for each patient was estimated (pg/mL; as presented in Table 3). A mathematical mean value for each cytokine for all the patients within a cohort was estimated and plotted using MS Excel. The mean serum IL-12 (A), IFNγ (B), and CXCL10 (C) with errors (standard error of the mean; SEM) is shown for each cohort along with goodness of fit (R2) and significance (p). P values from Jonckheere-Terpstra test is also provided (p*).</p

PSA through primary toxicity endpoint (day 30).

In a phase I dose escalation and safety study (NCT02555397), a replication-competent oncolytic adenovirus expressing yCD, TK and hIL-12 (Ad5-yCD/mutTKSR39rep-hIL-12) was administered in 15 subjects with localized recurrent prostate cancer (T1c-T2) at increasing doses (1 × 1010, to 1 × 1012 viral particles) followed by 7-day treatment of 5-fluorocytosine (5-FC) and valganciclovir (vGCV). The primary endpoint was toxicity through day 30 while the secondary and exploratory endpoints were quantitation of IL-12, IFNγ, CXCL10 and peripheral blood mononuclear cells (PBMC). The study maximum tolerated dose (MTD) was not reached indicating 1012 viral particles was safe. Total 115 adverse events were observed, most of which (92%) were grade 1/2 that did not require any treatment. Adenoviral DNA was detected only in two patients. Increase in IL-12, IFNγ, and CXCL10 was observed in 57%, 93%, and 79% patients, respectively. Serum cytokines demonstrated viral dose dependency, especially apparent in the highest-dose cohorts. PBMC analysis revealed immune system activation after gene therapy in cohort 5. The PSA doubling time (PSADT) pre and post treatment has a median of 1.55 years vs 1.18 years. This trial confirmed that replication-competent Ad5-IL-12 adenovirus (Ad5-yCD/mutTKSR39rep-hIL-12) was well tolerated when administered locally to prostate tumors.</div

Request for planned change(s).

In a phase I dose escalation and safety study (NCT02555397), a replication-competent oncolytic adenovirus expressing yCD, TK and hIL-12 (Ad5-yCD/mutTKSR39rep-hIL-12) was administered in 15 subjects with localized recurrent prostate cancer (T1c-T2) at increasing doses (1 × 1010, to 1 × 1012 viral particles) followed by 7-day treatment of 5-fluorocytosine (5-FC) and valganciclovir (vGCV). The primary endpoint was toxicity through day 30 while the secondary and exploratory endpoints were quantitation of IL-12, IFNγ, CXCL10 and peripheral blood mononuclear cells (PBMC). The study maximum tolerated dose (MTD) was not reached indicating 1012 viral particles was safe. Total 115 adverse events were observed, most of which (92%) were grade 1/2 that did not require any treatment. Adenoviral DNA was detected only in two patients. Increase in IL-12, IFNγ, and CXCL10 was observed in 57%, 93%, and 79% patients, respectively. Serum cytokines demonstrated viral dose dependency, especially apparent in the highest-dose cohorts. PBMC analysis revealed immune system activation after gene therapy in cohort 5. The PSA doubling time (PSADT) pre and post treatment has a median of 1.55 years vs 1.18 years. This trial confirmed that replication-competent Ad5-IL-12 adenovirus (Ad5-yCD/mutTKSR39rep-hIL-12) was well tolerated when administered locally to prostate tumors.</div

Peak cytokine day and concentration.

In a phase I dose escalation and safety study (NCT02555397), a replication-competent oncolytic adenovirus expressing yCD, TK and hIL-12 (Ad5-yCD/mutTKSR39rep-hIL-12) was administered in 15 subjects with localized recurrent prostate cancer (T1c-T2) at increasing doses (1 × 1010, to 1 × 1012 viral particles) followed by 7-day treatment of 5-fluorocytosine (5-FC) and valganciclovir (vGCV). The primary endpoint was toxicity through day 30 while the secondary and exploratory endpoints were quantitation of IL-12, IFNγ, CXCL10 and peripheral blood mononuclear cells (PBMC). The study maximum tolerated dose (MTD) was not reached indicating 1012 viral particles was safe. Total 115 adverse events were observed, most of which (92%) were grade 1/2 that did not require any treatment. Adenoviral DNA was detected only in two patients. Increase in IL-12, IFNγ, and CXCL10 was observed in 57%, 93%, and 79% patients, respectively. Serum cytokines demonstrated viral dose dependency, especially apparent in the highest-dose cohorts. PBMC analysis revealed immune system activation after gene therapy in cohort 5. The PSA doubling time (PSADT) pre and post treatment has a median of 1.55 years vs 1.18 years. This trial confirmed that replication-competent Ad5-IL-12 adenovirus (Ad5-yCD/mutTKSR39rep-hIL-12) was well tolerated when administered locally to prostate tumors.</div

Changes in serum IL-12, IFNγ, and CXCL10 in subjects # 12 and 13.

Cytokines were measured by ELISA based on cytokine specific standard curves (IL-12 0–500 pg/mL; CXCL10 and IFNγ 0–1,000 pg/mL). Panels A-C show serum IL-12, IFNγ and CXCL10 for patient #12 while panels D-F are for patient #13.</p