NY-ESO-1 Vaccine in Combination With Ipilimumab in Patients With Unresectable or Metastatic Melanoma

Patients were enrolled sequentially, alternating among 3 treatment arms. Study treatment
comprised ipilimumab 3 mg/kg administered intravenously (IV) over 90 minutes every 3 weeks
for 4 doses followed by the NY-ESO-1 vaccine administered subcutaneously (SC). Arm A received
the NY-ESO-1 recombinant protein mixed with polyinosinic-polycytidylic acid-poly-L-lysine
carboxymethylcellulose (Poly-ICLC) and Montanide; Arm B received NY-ESO-1 overlapping long
peptides 4 (OLP4) mixed with Poly-ICLC and Montanide; and Arm C received NY-ESO-1 OLP4 mixed
with Poly-ICLC (without Montanide). The vaccine was administered immediately following the
ipilimumab infusion, and patients were observed for 1 hour following administration. No dose
adjustments or delays were permitted.

Because the study treatment regimens had not been previously investigated in humans, the
first patient in each treatment arm was followed for 28 days and evaluated for any
regimen-limiting toxicity (RLT), defined as any dose-limiting toxicity (DLT) that could not
be attributed solely to either the vaccine or ipilimumab and was therefore considered to be
related to the combination. If an RLT was observed in the first patient, the second patient
was to be evaluated for 28 days before the third patient was enrolled. If at any point ≥ 2
RLTs were observed in a treatment arm, accrual to that arm was to be terminated and the
combination in that arm was to be declared unsafe.

Patients were monitored for safety, immune and tumor response, and immunological changes in
the tumor microenvironment for the duration of study participation, which may have been up to
6 months.

Inclusion Criteria:

1. Patients with unresectable or metastatic melanoma, for whom treatment with ipilimumab
was indicated as per ipilimumab/Yervoy® package insert (applicable for United States
[US] sites) or product information (applicable for Australia site).

2. Radiologically measurable disease by irRC.

3. Tumor expression of NY-ESO-1 or LAGE-1 antigen by immunohistochemistry or reverse
transcriptase-polymerase chain reaction (RT-PCR), or evidence of seropositivity to
NY-ESO-1 or LAGE-1.

4. Willingness to provide at least one pre-and post-vaccination tumor biopsy sample.

5. Expected survival of at least 4 months.

6. At the time of Day 1 of the study, patients must have been at least 3 weeks since
surgery.

7. At the time of Day 1 of the study, patients with brain metastases must have been
asymptomatic and:

- at least 8 weeks without tumor progression after any whole brain radiotherapy;

- at least 4 weeks since craniotomy and resection or stereotactic radiosurgery;

- at least 3 weeks without new brain metastases as evidenced by magnetic resonance
imaging (MRI).

8. Eastern Cooperative Oncology Group performance status of 0 to 2.

9. Laboratory parameters for vital functions must have been in the normal range.
Laboratory abnormalities that were not clinically significant were generally
permitted, except for the following laboratory parameters, which must have been within
the ranges specified:

- hemoglobin: ≥ 10 g/dL;

- neutrophil count: ≥ 1.5 x 10^9/L;

- lymphocyte count: ≥ lower limit of normal (LLN);

- platelet count: ≥ 80 x 10^9/L;

- serum creatinine: ≤ 2 mg/dL;

- serum bilirubin: ≤ 2 x upper limit of normal (ULN);

- aspartate aminotransferase (AST)/alanine aminotransferase (ALT): ≤ 2 x ULN.

10. Had been informed of other treatment options.

11. Age ≥ 18 years.

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

Exclusion Criteria:

1. Any contraindications for ipilimumab/Yervoy® as per package insert (applicable for US
sites) or product information (applicable for Australia site).

2. Prior exposure to NY-ESO-1 vaccine.

3. Active autoimmune disease, symptoms or conditions except for vitiligo, type I
diabetes, treated thyroiditis, asymptomatic laboratory evidence of autoimmune disease
(e.g., +antinuclear antibody [ANA], +rheumatoid factor [RF], antithyroglobulin
antibodies), or mild arthritis requiring no therapy or manageable with nonsteroidal
anti-inflammatory drugs (NSAIDs).

4. Unresolved immune-related adverse events following prior biological therapy.

5. Systemic treatment with high-dose corticosteroids (greater than prednisone 10 mg daily
or equivalent).

6. Treatment with protocol-specified non-permitted concomitant therapies.

7. Metastatic disease to the central nervous system for which other therapeutic options,
including radiotherapy, may have been available.

8. Myocardial infarction, angina, congestive heart failure, cardiomyopathy, stroke or
transient ischemic attack, chest pain or shortness of breath with activity, or other
heart conditions being treated by a doctor.

9. Other malignancy within 3 years prior to entry into the study, except for treated
non-melanoma skin cancer and cervical carcinoma in situ.

10. Known immunodeficiency or human immunodeficiency virus positivity, active Hepatitis B
or active Hepatitis C.

11. History of severe allergic reactions to vaccines or unknown allergens.

12. Other serious illnesses (e.g., serious infections requiring antibiotics, bleeding
disorders).

13. Participation in any other clinical trial involving another investigational agent
within 4 weeks prior to Day 1 of the study.

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

15. Lack of availability for immunological and clinical follow-up assessments.

16. Women who were breast feeding or pregnant as evidenced by positive serum pregnancy
test (minimum sensitivity 25 IU/L or equivalent units of human chorionic gonadotropin
[HCG]) done within 14 days prior to first dosing and urine test within 72 hours prior
to first dosing.

17. Women of childbearing potential not using a medically acceptable means of
contraception for the duration of the study.

18. Any condition that, in the clinical judgment of the treating physician, was likely to
prevent the patient from complying with any aspect of the protocol or that may have
put the patient at unacceptable risk.