Study of ADI-PEG 20 in Patients With Advanced Melanoma

A 3+3 design was implemented during phase 1, in which 3 to 6 subjects were enrolled
sequentially into the following escalating dose cohorts:

- Cohort 1 (40 IU/m^2)

- Cohort 2 (80 IU/m^2)

- Cohort 3 (160 IU/m^2)

Subjects were monitored for dose-limiting toxicity (DLT) during the first 2 weeks of cycle 1,
with DLT defined as any grade 3 or higher toxicity. The maximum tolerated dose (MTD) was
defined as the cohort in which < 33% of subjects (ie, 0/3 or 1/6 subjects in a cohort)
experienced DLT. In phase 2, the MTD cohort was expanded in up to 25 patients.

Subjects who completed treatment in cycle 1 without DLT were eligible to initiate cycle 2 at
week 10 provided that a computed tomography (CT) scan showed either enlargement of existing
disease without accompanying symptoms OR stable disease or improvement with no unacceptable
toxicity. The same radiologic criteria applied for initiation of subsequent cycles. Subjects
could continue to receive study treatment until disease progression.

Inclusion Criteria:

1. Histologically confirmed malignant melanoma, American Joint Committee on Cancer (AJCC)
stage III (unresectable) or IV. Subjects with uveal and mucosal melanomas were
eligible.

2. Measurable disease using the Response Evaluation Criteria in Solid Tumors (RECIST).

3. Pathology slides reviewed by the Memorial Hospital Department of Pathology or New York
University (NYU) Department of Pathology for confirmation of melanoma diagnosis.

4. Karnofsky performance status of 80% or more.

5. Adequate organ and marrow function, as defined below:

- white blood cell count ≥ 3000/µL

- absolute neutrophil count ≥ 1500/µL

- platelet count ≥ 100,000/µL

- total bilirubin ≤ 2.5 x institutional upper limit of normal (ULN)

- lactate dehydrogenase ≤ 1.5 x institutional ULN

- albumin ≥ 3.0 mg/dL

- creatinine ≤ 2.0 mg/dL

6. Expected survival of at least 3 months.

7. Age ≥ 18 years.

8. Able and willing to give valid written informed consent.

Exclusion Criteria:

1. Receipt of chemotherapy, immunotherapy, or radiotherapy within 3 weeks prior to first
dosing of study agent or lack of recovery from adverse events (AEs) due to agents
administered more than 3 weeks earlier. For nitrosoureas, at least 6 weeks must have
elapsed.

2. Any other malignancy that required concomitant therapy.

3. Any medical condition that could have made it difficult for the subject to complete
the full course of treatments, at the discretion of the Principal Investigator or
co-Principal Investigators.

4. Metastatic disease to the central nervous system, unless treated and stable.

5. Known human immunodeficiency virus (HIV) positivity.

6. Mental impairment that may have compromised the ability to give informed consent and
comply with the requirements of the study.

7. Lack of availability for clinical follow-up assessments.

8. Participation in any other clinical trial involving another investigational agent
within 3 weeks prior to enrollment.

9. Pregnant women or women who were nursing. Women of child-bearing potential and
sexually active men must have used appropriate contraception during the course of this
study. Women of child-bearing potential must not have been pregnant (negative β human
chorionic gonadotropin within 2 weeks of treatment) or nursing during treatment.

10. History of seizure disorder.