A pilot Phase I study combining peptide-based vaccination and NGR-hTNF vessel targeting therapy in metastatic melanoma

Abstract

Administration of NGR-TNF, a tumor vessel-targeting and tumor necrosis factor α TNFα) peptide conjugate, with immunotherapy has been shown to inhibit tumor growth in mice. Thus, we planned a Phase I pilot clinical trial to assess safety, immune and clinical response of this combination treatment for advanced melanoma. NA17.A2 and MAGE-3.A1 peptides were used as vaccine. HLA-A*0201 or HLA-A*01 metastatic melanoma patients received human NGR-hTNF i.v. alternating with s.c. weekly injections of either of the peptides emulsified in Montanide. The T-cell response was assessed ex-vivo using peripheral blood mononuclear cells (PBMCs) before, during and after therapy. The serum level of chromogranin A (CgA), soluble TNF receptors (sTNFR1/2), vascular endothelial growth factor (VEGF), and MIP-1β and MCP-1 chemokines, was determined. In 3 subjects, pre- and post-treatment tumor lesions were examined by immunohistochemistry. Clinically, chills were observed in 4 patients during NGR-hTNF infusion and erythema at vaccination site was seen in 7 patients. T-cell response against the vaccine or against other melanoma-associated antigens was detectable after treatment in 6 out of 7 tested patients. Low level or reduction of CgA and sTNFR and increase of MIP-1β and MCP-1 were found in patients sera. In the lesions examined the immune infiltrate was scanty but macrophage number increased in post-therapy lesions. From a clinical standpoint, a long term survival (>4 months) was found in 6 out of 8 evaluable patients (4, 4, 7, 11, 23+, 25+, 25+, 29+ months). The combination of NGR-hTNF and vaccine in metastatic melanoma patients was well tolerated, often associated with an ex-vivo T cell response and long-term overall survival. These findings warrant confirmation in a larger group of patients.

Keywords:

• anti-vascular target therapy
• combination therapy
• inflammatory cytokines
• melanoma
• peptide-based vaccines
• T cells

Abbreviations:

• APC, antigen presenting cell
• CgA, chromogranin A
• CT, cancer/testis
• DFS, disease-free survival
• MAA, melanoma-associated antigens
• MCP-1, macrophage chemoattractant protein 1
• MIP-1β, macrophage inflammatory protein 1β; OS, overall survival
• PD, progression of disease
• PFS, progression-free survival
• PBMC, peripheral blood mononuclear cell
• RR, response rate
• sTNFR, soluble tumor necrosis factor receptor
• TNFα, tumor necrosis factor α

Disclosure of Potential Conflicts of Interest

A Corti is the inventor of NGR-TNF (patent).

Acknowledgment

The procedures of this study were approved by the Internal Ethics Committee of San Raffaele Hospital on February 4, 2010 (code N. 49/932) and by the Italian Agency for Pharmacological Agents on April 19 and May 18, 2010 respectively and are in accordance with the Helsinki Declaration of 1975. The final authorization to start the protocol was released by the Ethics Committee of the San Raffaele Hospital on June 7, 2010. EUDRACT N. 2009-017818-67 Received.

Funding

This spontaneous protocol was financially supported by a Grant of the Alliance against Cancer (Italian Ministry of Health, Rome) to G.P.

Supplemental Material

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