This is a Phase I, open-label, randomized study of NY-ESO-l protein with immune adjuvants CpG 7909 and Montanide ISA-51 VG in patients with tumors that often express NY-ESO-1.

The vaccinations was to be administered subcutaneously every 3 weeks for 4 doses.

Patients with any malignancy that is known to frequently express NY-ESO-1 were eligible, regardless of whether antigen expression in the autologous tumor could be demonstrated or not by either PCR or immunohistochemistry.

The primary objective of the study was to define safety.

Secondarily, the study was to evaluate whether patients developed a specific immunologic response to the NY-ESO-1 protein. Blood samples were to be obtained at baseline, prior to each vaccination, one week after each vaccination, and at the last study visit for the assessment of NY-ESO-1-specific CD4+ and CD8+ T cells. Cytokine secretion by NY-ESO-1-specific CD8+ and CD4+ T cells, as a measure of T cell activation, was to be determined by FACS analysis. In addition, humoral immunity was to be determined by the presence of NY-ESO-1-specific antibodies which were to be assessed in all patients by ELISA.

Disease status was to be assessed at baseline and 2-4 weeks after the fourth vaccination in patients with evaluable (measurable and non-measurable) disease.

Subjects were to receive an investigational (research) cancer vaccine every 3 weeks for a total of 4 treatments. It was given by injection underneath the skin in an extremity (leg or arm). A vaccine is a compound designed to strengthen the immune system (the cells and substances that protect the body from infection and foreign matter) to fight an illness such as infections or cancer. This vaccine is called NY-ESO-1 protein. NY-ESO-1 protein (an antigen, which is a compound that is recognized by the immune system) is found in many cancers. Proteins such as NY-ESO-1 and their fragments are the targets the immune system needs to recognize cancer cells. If the immune system can recognize these antigens (foreign substances) it may be able to kill the cells that carry them. NY-ESO-1 can be found at different stages of cancers, and is likely to be expressed (shown) at some point in the lifecycle of these types of cancer (that are eligible for this study). Therefore this study tries to boost (strengthen) the immune system toward NY-ESO-1 protein regardless of whether it is found in your tumor or not.

Since we do not know whether different doses of the NY-ESO-1 protein may result in varying degrees of immune stimulation, we will be randomizing (that is, at the flip of a coin or, in other words, by chance). You may receive either the lower dose of NY-ESO-1 protein (100 µg) or the higher dose (400 µg).

The NY-ESO-1 protein vaccine was to be mixed with 2 substances, called adjuvants (the full names are: CPG 7909 and Montanide ISA-51 VG). Adjuvants are substances to increase the vaccine's ability to stimulate the immune system. By adding two adjuvants to the vaccine, it is hoped that the boosting of the immune system will be especially effective.

It is important to understand, that vaccines are only experimental (investigational, research) for the treatment of cancer. They are not approved by the Food and Drug Administration (FDA) as treatment. Therefore they are only offered in clinical trials.

All tests and treatments were performed as an outpatient.

Inclusion Criteria:

1. Histological diagnosis of hepatocellular carcinoma, bladder cancer, breast cancer, non-small lung cancer (NSCLC), melanoma, sarcoma, prostate cancer, esophageal cancer, or ovarian cancer, independent of NY-ESO-1 expression in a tumor biopsy.

   or

   Histological diagnosis of other types of cancers, provided NY-ESO-1 or LAGE-1 expression can be shown in a tumor biopsy.

2. At least 4 weeks since surgery prior to first dosing of study agent.

3. Laboratory values within the following limits:

   • Hemoglobin ≥ 11.0 g/dL
   • Neutrophil count ≥ 1.5 x l0^9/L
   • Lymphocyte count ≥ lower limit of institutional normal
   • Platelet count ≥ 80 x l0^9/L
   • Serum creatinine ≤ 2.0 mg/dL
   • Serum bilirubin ≤ 2 x upper limit of institutional normal
   • AST/ALT ≤ 2 x upper limit of institutional normal

4. Patients must have a Karnofsky performance status of ≥70%.

5. Life expectancy ≥ 6 months.

6. Age ≥ 18 years.

7. Able and willing to give witnessed, written informed consent for participation in the trial.

Exclusion Criteria:

1. Clinically significant heart disease (i.e. NYHA class 3 congestive heart failure; myocardial infarction within the past six months; unstable angina; coronary angioplasty within the past 6 months; uncontrolled atrial or ventricular cardiac arrhythmias).
2. Other serious illnesses, eg, serious infections requiring antibiotics, bleeding disorders.
3. Previous bone marrow or stem cell transplant.
4. History of immunodeficiency disease or autoimmune disease except vitiligo.
5. Metastatic disease to the central nervous system, unless treated and stable.
6. Other malignancy within 3 years prior to entry into the study, except for treated early-stage melanoma or non-melanoma skin cancer, or cervical carcinoma in situ.
7. Known HIV, Hepatitis B or Hepatitis C positivity.
8. Chemotherapy, radiation therapy or immunotherapy within 4 weeks prior to first dose of study agent (6 weeks for nitrosoureas).
9. Concomitant treatment with steroids. Topical or inhalational steroids are permitted.
10. Participation in any other clinical trial involving another investigational agent within 4 weeks prior to first dose of study agent.
11. Pregnancy or lactation.
12. Women of childbearing potential not using a medically acceptable means of contraception.
13. Psychiatric or addictive disorders that may compromise the ability to give informed consent.
14. Lack of availability of the patient for immunological and clinical follow-up assessment.

All subjects who entered the study and received the NY-ESO-1 protein vaccine.

up to 14 weeks

All AEs occurring during the study were documented in the source records and on the respective AE case report form (CRF) pages, regardless of the assumption of a causal relationship. All events that occurred after the first administration of study drug up to the end of week 14 were to be documented. The severity of an AE was assessed according to the NCI CTCAE scale (Version 3.0).