A novel AML-selective TRAIL fusion protein that is superior to Gemtuzumab Ozogamicin in terms of in vitro selectivity, activity and stability

• Published in: Leukemia Vol. 23; no. 8; pp. 1389 - 1397
• Main Authors: ten Cate, B, Bremer, E, de Bruyn, M, Bijma, T, Samplonius, D, Schwemmlein, M, Huls, G, Fey, G, Helfrich, W
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-08-2009; Nature Publishing Group
• Subjects: Abl protein; Acute Disease; Acute myeloid leukemia; Aminoglycosides - pharmacology; Antibodies; Antibodies, Monoclonal - pharmacology; Antibodies, Monoclonal, Humanized; Antigens, CD - metabolism; Antigens, Differentiation, Myelomonocytic - metabolism; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Antitumor activity; Apoptosis; Apoptosis - drug effects; BCR protein; Biological and medical sciences; Bystander Effect; Calicheamicin; Cancer Research; Cells, Cultured - drug effects; Chemical compounds; Coupling; Critical Care Medicine; Cytotoxicity; Drug Delivery Systems; Drug Screening Assays, Antitumor; Drug Stability; Enzyme Activation - drug effects; Enzyme inhibitors; Fusion protein; Gemtuzumab ozogamicin; Health aspects; Hematologic and hematopoietic diseases; Hematology; Hepatotoxicity; Humans; Intensive; Internal Medicine; Kinases; Leukemia; Leukemia, Myeloid - pathology; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Leukocytes, Mononuclear - drug effects; Medical sciences; Medicine; Medicine & Public Health; Mitoxantrone; Monocytes; Myelosuppression; Neoplasm Proteins - metabolism; Oncology; original-article; Patients; Pharmacology; Physiological aspects; Protein-tyrosine kinase; Proteins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - pharmacology; Selectivity; Sialic Acid Binding Ig-like Lectin 3; Single-Chain Antibodies; Stability; Subgroups; Toxicity; TRAIL protein; Tumor cells; Tumor Cells, Cultured - drug effects; Tyrosine; Valproic acid; United States; AML; TRAIL; mylotarg; CD33; gemtuzumab ozogamicin; Antineoplastic agent; Immunochemotherapy; Acute myelogenous leukemia; Calicheamicin; Selectivity; Monoclonal antibody; Ozogamicin; Fusion protein; Stability; Hematology; Gemtuzumab; Cytokine; Malignant hemopathy; Immunoconjugate; In vitro; Biological activity; Chemotherapy; Treatment; Tumor necrosis factor; TNF related apoptosis inducing ligand; Cancer
• ISSN: 0887-6924; 1476-5551
• DOI: 10.1038/leu.2009.34

Gemtuzumab ozogamicin (GO, Mylotarg) is a targeted therapeutic agent in which an anti-CD33 antibody is chemically coupled to a highly cytotoxic calicheamicin derivative through a hydrolysable linker. GO has improved the treatment outcome for a subgroup of acute myeloid leukemia (AML) patients, but its use is associated with severe myelosuppression and hepatotoxicity. Here, we report on a novel anti-leukemia agent, designated scFvCD33:sTRAIL, in which an anti-CD33 single chain fragment of variable regions (scFv) antibody fragment is genetically linked to soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL). Normal CD33-positive monocytes were fully resistant to prolonged treatment with scFvCD33:sTRAIL, whereas treatment with GO resulted in substantial cytotoxicity. The activity of scFvCD33:sTRAIL towards AML cells was up to 30-fold higher than GO. The CD33-restricted anti-leukemia activity of scFvCD33:sTRAIL remained stable during prolonged storage at 37 °C, whereas GO showed a rapid increase in CD33-independent cytotoxicity. Moreover, scFvCD33:sTRAIL showed potent anti-leukemia activity towards CD33+ CML cells when treatment was combined with the Bcr-Abl tyrosine kinase inhibitor, Gleevec. Importantly,
ex vivo
treatment of patient-derived CD33+ AML tumor cells with scFvCD33:sTRAIL resulted in potent apoptosis induction that was enhanced by valproic acid, mitoxantrone and 17-(Allylamino)-17-demethoxygeldanamycin (17-AAG). Taken together, scFvCD33:sTRAIL is superior to GO in terms of tumor selectivity, activity and stability, warranting its further development for the treatment of CD33-positive leukemias.