title: Toxicology and pharmacokinetics of DT388-GM-CSF, a fusion toxin consisting of a truncated diphtheria toxin (DT388) linked to human granulocyte-macrophage colony-stimulating factor (GM-CSF) in C57BL/6 mice.
abstract: Because the majority of acute myeloid leukemia (AML) blasts express the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor, we are developing a fusion toxin consisting of a truncated diphtheria toxin (DT388) linked to human GM-CSF for multi-drug resistant AML. Our goal was to determine the toxicity and pharmacokinetics of DT388-GM-CSF in C57BL/6 mice. Because human GM-CSF does not cross-react with the mouse GM-CSF receptor, the toxicity observed should be nonspecific toxicity of DT388. We injected C57BL/6 mice i.p. with 0.1, 0.5, 1.0, 1.5, 1.75, 2.0, 3.5, 5.0, or 10 micrograms/day of DT388-GM-CSF for 5 consecutive days. For pharmacokinetics, blood samples were drawn at 20, 40, 60, 120, and 180 min after i.p. administration of 81 micrograms/kg of DT388-GM-CSF. In mice, the LD10 of DT388-GM-CSF is between 84.4 (1.5) and 104.4 (1.75) micrograms/kg (microgram) when administered for 5 consecutive days. All mice receiving > or = 201 micrograms/kg (3.5 micrograms) for 5 consecutive days died. Histopathologic examination of morbid animals showed only renal toxicity with acute proximal tubular necrosis. DT388-GM-CSF is stable in vivo based on nonreducing SDS-PAGE gel of plasma samples of 125I-labeled DT388-GM-CSF injected i.p.. The peak concentration of DT388-GM-CSF was 3.3 x 10(-8) M at 40 min and exhibited a t1/2 of 24 min. Based on its half-life, DT388-GM-CSF concentrations in the plasma are above the concentration inhibiting 50% protein synthesis and inducing apoptosis in 50% of HL-60 cells (AML cell line) for 5.2 h. Only four of 17 mice developed a weak immune response (0.9-160 ng/mL) 3 weeks after treatment. DT388-GM-CSF exhibits a short t1/2, but concentrations exceed those required in vitro to inhibit AML cell lines.
authors.affiliation: College of Pharmacy, Medical University of South Carolina, Charleston 29425, USA.
authors.forename: P D
authors.initials: PD
authors.lastname: Hall
authors.forename: R J
authors.initials: RJ
authors.lastname: Kreitman
authors.forename: M C
authors.initials: MC
authors.lastname: Willingham
authors.forename: A E
authors.initials: AE
authors.lastname: Frankel
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000187:drug effects; D004167:Diphtheria Toxin / Q000494:pharmacology* / Q000633:toxicity*; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology* / Q000633:toxicity*; D018922:HL-60 Cells; D006207:Half-Life; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D000922:Immunotoxins / Q000493:pharmacokinetics* / Q000633:toxicity*; D007668:Kidney / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D011500:Protein Synthesis Inhibitors / Q000493:pharmacokinetics* / Q000633:toxicity*
chemical_list: D004167:Diphtheria Toxin; D007074:Immunoglobulin G; D000922:Immunotoxins; D011500:Protein Synthesis Inhibitors; C112695:dt(388) granulocyte-macrophage colony-stimulating factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor