Malignant Melanoma
Conditions
Keywords
Melanoma, Vaccine, Peptide, Montanide
Brief summary
The purposes of this study are to describe the immune response to individual peptides after immunization with a combination of 8 peptides and CpG 7909 or Montanide ISA51; to determine the safety of the vaccines and; to document the tumor response in patients receiving the vaccines.
Detailed description
Patients received six sequential immunizations with 8 peptides presented by HLA-A2 and mixed with either CpG 7909 or Montanide ISA51, at 2-week intervals. The 8 peptides were injected at 8 distinct injection sites. These peptides are the following: MAGE-1.A2, MAGE-3.A2, MAGE-4.A2, MAGE-10.A2, MAGE-C2.A2 (ALKD), and NA17.A2 (20% intradermally and 80% subcutaneously); NY-ESO-1.A2 and Tyrosinase.A2 (100% subcutaneously). 300 µg of each peptide (except MAGE-10.A2 150 µg) was mixed with 4 mg CpG 7909 (Cohort 1) or 0.5ml of Montanide ISA51 (Cohort 2). In Cohort 2, the Tyrosinase.A2 was administered without Montanide ISA51. Tumor staging was performed before inclusion and at week 13. Peripheral Blood Lymphocytes (PBL) collections were performed before starting the treatment, and at weeks 3, 7 and 13. They provided the T lymphocytes for the immunological analysis. At week 13, the PCR results of the pre-immune tumor biopsy must be available. Additional cycles of immunization, ONLY with the peptides expressed by the tumor, mixed with Montanide ISA51, will be proposed to patients without tumor progression requiring another treatment. A second cycle of 3 injections at 6-week intervals will be started at week 17, followed by a third cycle of 12 injections at 3-month intervals, starting at month 11. At any time, progression of the disease necessitating any treatment not allowed during the study will result in withdrawal. The immune response may well be a limiting factor to the therapeutic efficacy of the vaccine. If this is the case, it then becomes crucial to understand why some patients develop a cytotoxic t-lymphocyte (CTL) response against the vaccine, while the majority of them do not. One possible explanation for the low frequency of clinical responses is that each injection of a single peptide has a low probability to provide the adequate stimulus to activate very rare CTL precursors. This probability should be increased if several peptides known to be undoubtedly associated with tumor regressions were used together to immunize patients.
Interventions
BIOLOGICAL8 HLA-A2-restricted peptides and Montanide ISA51
BIOLOGICAL8 HLA-A2-restricted peptides and CpG 7909
Sponsors
Ludwig Institute for Cancer Research
Study design
Allocation
NON_RANDOMIZED
Intervention model
SEQUENTIAL
Primary purpose
TREATMENT
Masking
NONE
Intervention model description
The protocol planned the inclusion of 2 cohorts, each of them including 14 patients, with metastatic cutaneous melanoma, with at least one detectable metastasis (grades AJCC 2002 III N2b to N3, grades IV M1a, M1b, and M1c without elevation of the LDH rate and without central nervous system toxicity).
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
No
Inclusion criteria
1. Histologically proven cutaneous melanoma, or clear cell sarcoma, which is considered as a subtype of melanoma. 2. Melanoma must be at one of the following AJCC 2002 stages: * Regional metastatic disease (any T; N2b, N2c or N3; M0). * Distant metastatic disease (any T; any N; M1a, M1b or M1c), except brain or leptomeningeal localizations, and except elevated LDH. 3. Patients must be HLA-A2. 4. A pre-immune tumor biopsy must be kept frozen for post-study PCR analysis. 5. Presence of at least one measurable or non-measurable tumor lesion. 6. Expected survival of at least 3 months. 7. Karnofsky performance scale ≥70 or WHO performance status of 0 or 1. 8. Within the last 4 weeks prior to study day 1, vital laboratory parameters should be within normal range, except for the following laboratory parameters, which must be within the ranges specified: Lab Parameter Range * Hemoglobin ≥ 10 g/dl or ≥ 6,25 mmol/l * Granulocytes ≥ 1,500/µl * Lymphocytes ≥ 700/µl * Platelets ≥ 100,000/µl * Serum creatinine ≤ 2.0 mg/dl or ≤ 177 μmol/l * Serum bilirubin ≤ 2.0 mg/dl or ≤ 34.2 μmol/l * ASAT and ALAT ≤ 2 x the normal upper limits * LDH ≤ the normal upper limit. 9. Viral tests: * HIV (human immunodeficiency virus): negative antibodies. * HBV (hepatitis B virus): negative antigens; antibodies may be positive. * HCV (hepatitis C virus): negative antibodies. 10. Age ≥ 18 years. 11. Able and willing to give valid written informed consent.
Exclusion criteria
1. Previous treatment with more than one regimen of systemic chemotherapy, combined or not with non-specific immunotherapy such as interferon alpha or interleukins. Chemoimmunotherapy or radiotherapy must be stopped within the preceding 4 weeks (6 weeks for nitrosoureas and mitomycin C). 2. Clinically significant heart disease (NYHA Class III or IV) 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. 3. Active immunodeficiency or autoimmune disease. Vitiligo was not an exclusion criterion. 4. Other serious acute or chronic illnesses, e.g. active infections requiring antibiotics, bleeding disorders, or other conditions requiring concurrent medications not allowed during this study. 5. Other malignancy within 3 years prior to entry into the study, except for treated non-melanoma skin cancer and cervical carcinoma in situ. 6. Lack of availability for immunological and clinical follow-up assessments. 7. Participation in any other clinical trial involving another investigational agent within 4 weeks prior to enrollment. 8. Pregnancy or breastfeeding. 9. Women of childbearing potential: Refusal or inability to use effective means of contraception.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51. | Week 13 | Peripheral blood lymphocytes (PBL) were collected prior to the first dose of vaccine and after the completion of the six vaccinations in Week 13. Specific CTL directed against the 8 vaccine antigens ( NA17.A2, MAGE-1.A2, MAGE-3.A2, MAGE-4.A2, MAGE-10.A2, MAGE-C2.A2 and Tyrosinase.A2) was assessed by using re-stimulation in vitro, followed by staining with the corresponding tetramer (MLPC/tetramer). A patient was considered to have a positive CTL response when the post-vaccine CTL response against at least one of the vaccine antigens was ten times higher than the corresponding pre-treatment value. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Number of Patients With Dose Limiting Toxicities (DLT). | up to Week 13 | Toxicity was evaluated according to the National Cancer Institute (CTC Scale Version 3.0, published December 12, 2003). Dose limiting toxicity (DLT) is defined as: * Any Grade 3 hematological or non-hematological toxicity other than skin or flu-like symptoms * Any Grade 4 toxicity To be dose-limiting, an adverse event must be definitely, probably, or possibly related to the administration of the investigational agent. |
| Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Week 13 | Tumor response was assessed according to the Response Evaluation Criteria in Solid Tumors (RECIST). Tumor measurements were taken at screening and at the end of Cycle 1 in Week 13. Per RECIST, target lesions are categorized as follows: Complete Response (CR): Disappearance of all target lesions; Partial Response (PR): ≥ 30% decrease in the sum of the longest diameter of target lesions; Progressive Disease (PD): ≥ 20% increase in the sum of the longest diameter of target lesions; Stable Disease (SD): small changes that do not meet above criteria. |
| Number of Patients With Cytotoxic T-lymphocyte (CTL) Responses and Tumor Expression of the Corresponding Genes. | Week 13 | Gene expression was determined by RT-PCR on a pre-treatment tumor sample. The correlation of the induction of a CTL response against a defined antigen to the prior expression of the gene coding for this antigen by the tumor removed before vaccination was assessed. |
Countries
Belgium
Participant flow
Recruitment details
The study was to recruit 28 patients; 14 in each group. The first subject was treated on January 18, 2005. The study was prematurely closed in November 2007 as 6 of the 8 peptides had expired.
Participants by arm
| Arm | Count |
|---|---|
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 Patients with metastatic cutaneous melanoma, with at least one detectable metastasis (grades AJCC 2002 III N2b to N3, grades IV M1a, M1b, and M1c without elevation of the LDH rate and without central nervous system toxicity) received six sequential immunizations with 8 peptides presented by HLA-A2 and mixed with CpG 7909, at 2-week intervals. The 8 peptides were to be injected at 8 distinct injection sites. These peptides are the following: MAGE-1.A2, MAGE-3.A2, MAGE-4.A2, MAGE-10.A2, MAGE-C2.A2 (ALKD), and NA17.A2 (20% intradermally and 80% subcutaneously); NY-ESO-1.A2 and Tyrosinase.A2 (100% subcutaneously). | 16 |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 Patients with metastatic cutaneous melanoma, with at least one detectable metastasis (grades AJCC 2002 III N2b to N3, grades IV M1a, M1b, and M1c without elevation of the LDH rate and without central nervous system toxicity) received six sequential immunizations with 8 peptides presented by HLA-A2 and mixed with Montanide ISA51, at 2-week intervals. The 8 peptides were to be injected at 8 distinct injection sites. These peptides are the following: MAGE-1.A2, MAGE-3.A2, MAGE-4.A2, MAGE-10.A2, MAGE-C2.A2 (ALKD), and NA17.A2 (20% intradermally and 80% subcutaneously); NY-ESO-1.A2 and Tyrosinase.A2 (100% subcutaneously). The Tyrosinase.A2 was administered without Montanide ISA51. | 7 |
| Total | 23 |
Withdrawals & dropouts
| Period | Reason | FG000 | FG001 |
|---|---|---|---|
| Overall Study | premature withdrawal | 2 | 3 |
Baseline characteristics
| Characteristic | Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Total |
|---|---|---|---|
| Age, Customized ≥ to 18 years | 16 Participants | 7 Participants | 23 Participants |
| Region of Enrollment Belgium | 16 participants | 7 participants | 23 participants |
| Sex: Female, Male Female | 9 Participants | 5 Participants | 14 Participants |
| Sex: Female, Male Male | 7 Participants | 2 Participants | 9 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | 0 / 16 | 0 / 7 |
| other Total, other adverse events | 16 / 16 | 7 / 7 |
| serious Total, serious adverse events | 7 / 16 | 3 / 7 |
Outcome results
Primary
Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51.
Peripheral blood lymphocytes (PBL) were collected prior to the first dose of vaccine and after the completion of the six vaccinations in Week 13. Specific CTL directed against the 8 vaccine antigens ( NA17.A2, MAGE-1.A2, MAGE-3.A2, MAGE-4.A2, MAGE-10.A2, MAGE-C2.A2 and Tyrosinase.A2) was assessed by using re-stimulation in vitro, followed by staining with the corresponding tetramer (MLPC/tetramer). A patient was considered to have a positive CTL response when the post-vaccine CTL response against at least one of the vaccine antigens was ten times higher than the corresponding pre-treatment value.
Time frame: Week 13
Population: Patients who received at least six vaccinations and had samples taken at Week 13.
| Arm | Measure | Category | Value (COUNT_OF_PARTICIPANTS) |
|---|---|---|---|
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51. | Patients with no CTL Response | 8 Participants |
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51. | Patients with CTL Response | 6 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51. | Patients with no CTL Response | 2 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Response to Individual Peptides After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51. | Patients with CTL Response | 1 Participants |
Secondary
Number of Patients With Cytotoxic T-lymphocyte (CTL) Responses and Tumor Expression of the Corresponding Genes.
Gene expression was determined by RT-PCR on a pre-treatment tumor sample. The correlation of the induction of a CTL response against a defined antigen to the prior expression of the gene coding for this antigen by the tumor removed before vaccination was assessed.
Time frame: Week 13
Population: Patients who had pretreatment tumor samples analyzed for gene expression, received at least six immunizations and had CTL responses at Week 13.
| Arm | Measure | Value (COUNT_OF_PARTICIPANTS) |
|---|---|---|
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Responses and Tumor Expression of the Corresponding Genes. | 3 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Cytotoxic T-lymphocyte (CTL) Responses and Tumor Expression of the Corresponding Genes. | 1 Participants |
Secondary
Number of Patients With Dose Limiting Toxicities (DLT).
Toxicity was evaluated according to the National Cancer Institute (CTC Scale Version 3.0, published December 12, 2003). Dose limiting toxicity (DLT) is defined as: * Any Grade 3 hematological or non-hematological toxicity other than skin or flu-like symptoms * Any Grade 4 toxicity To be dose-limiting, an adverse event must be definitely, probably, or possibly related to the administration of the investigational agent.
Time frame: up to Week 13
Population: All patients who received at least one immunization.
| Arm | Measure | Value (COUNT_OF_PARTICIPANTS) |
|---|---|---|
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Dose Limiting Toxicities (DLT). | 2 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Dose Limiting Toxicities (DLT). | 0 Participants |
Secondary
Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST.
Tumor response was assessed according to the Response Evaluation Criteria in Solid Tumors (RECIST). Tumor measurements were taken at screening and at the end of Cycle 1 in Week 13. Per RECIST, target lesions are categorized as follows: Complete Response (CR): Disappearance of all target lesions; Partial Response (PR): ≥ 30% decrease in the sum of the longest diameter of target lesions; Progressive Disease (PD): ≥ 20% increase in the sum of the longest diameter of target lesions; Stable Disease (SD): small changes that do not meet above criteria.
Time frame: Week 13
Population: Patients who received at least six immunizations and had tumor response measured at Week 13.
| Arm | Measure | Category | Value (COUNT_OF_PARTICIPANTS) |
|---|---|---|---|
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Partial Response (PR) | 0 Participants |
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Progressive Disease (PD) | 10 Participants |
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Stable Disease (SD) | 4 Participants |
| Cohort 1: 8 HLA-A2-restricted Peptides and CpG 7909 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Complete Response (CR) | 0 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Progressive Disease (PD) | 4 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Stable Disease (SD) | 0 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Partial Response (PR) | 0 Participants |
| Cohort 2: 8 HLA-A2-Restricted Peptides and Montanide ISA51 | Number of Patients With Tumor Responses After Immunization With a Combination of 8 Peptides and CpG 7909 or Montanide ISA51 as Measured by RECIST. | Complete Response (CR) | 0 Participants |