title: Toxicity and toxicokinetics of a phosphorothioate oligonucleotide against the c-myc oncogene in cynomolgus monkeys.
abstract: A 2-week toxicity and toxicokinetic study of a 15-mer phosphorothioate oligonucleotide, INX-3280, against the c-myc oncogene was performed in cynomolgus monkeys. As this oligonucleotide readily adopts an aggregate structure, a quadruplex, which may be associated with adverse physiologic effects, this study was performed using INX-3280 that had been converted to its monomeric form. Animals received intravenous (i.v.) infusions of monomeric INX-3280 three times per week for 2 weeks at doses of 3 or 15 mg/kg per administration. The monkeys were examined for clinical signs: changes in hematology, serum chemistry, coagulation, and urinalysis parameters; complement activation; macroscopic findings at necropsy; and histopathologic alterations. In addition, the toxicokinetics of INX-3280 were evaluated, using a validated HPLC assay, after the first and last (sixth) doses. No treatment-related clinical signs of any adverse effects were observed, and there were no test article-related changes in hematology, serum chemistry, or complement activation parameters. The only alteration in clinical pathology parameters was a minor (30%) prolongation of the activated partial thromboplastin time (aPTT), reflecting slight inhibition of the intrinsic coagulation pathway, which was less than that reported with other oligonucleotides given at similar doses. Treatment-related histopathologic alterations consisted of characteristic accumulation of basophilic material in the cytoplasm of tubular epithelial cells in the kidney, resident macrophages in the lymph nodes, and Kupffer cells in the liver. These changes were graded as minimal in all cases. The basophilic material is believed to reflect accumulation of the oligonucleotide or metabolites or both. The pharmacokinetic parameters of INX-3280 were identical on the first and sixth administrations and were similar to those reported for other phosphorothioate oligonucleotides. Maximum concentration (Cmax) values for INX-3280 (101-119 microg/ml) were in excess of the threshold plasma concentrations reported to trigger complement activation by phosphorothioate oligonucleotides. It is concluded that the safety profile of monomeric INX-3280 in cynomolgus monkeys is quite favorable relative to the known effects of other phosphorothioate oligonucleotides, particularly with respect to the blood level-related toxicities of this class of compounds, including complement activation and inhibition of coagulation. This study found no toxicities that were expected to be clinically significant.
authors.affiliation: Inex Pharmaceuticals Corporation, Burnaby, BC, Canada. mwebb@celator.ca
authors.forename: M S
authors.initials: MS
authors.lastname: Webb
authors.forename: N
authors.initials: N
authors.lastname: Tortora
authors.forename: M
authors.initials: M
authors.lastname: Cremese
authors.forename: H
authors.initials: H
authors.lastname: Kozlowska
authors.forename: M
authors.initials: M
authors.lastname: Blaquiere
authors.forename: D V
authors.initials: DV
authors.lastname: Devine
authors.forename: D J
authors.initials: DJ
authors.lastname: Kornbrust
mesh_terms: D000818:Animals; D001483:Base Sequence; D001777:Blood Coagulation / Q000187:drug effects; D003167:Complement Activation / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D005260:Female; D016259:Genes, myc; D066298:In Vitro Techniques; D007684:Kidney Tubules / Q000187:drug effects / Q000473:pathology; D007728:Kupffer Cells / Q000187:drug effects / Q000473:pathology; D008198:Lymph Nodes / Q000187:drug effects / Q000473:pathology; D008252:Macaca fascicularis / Q000235:genetics*; D008297:Male; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*; D009841:Oligonucleotides / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*; D010314:Partial Thromboplastin Time; D012449:Safety; D013873:Thionucleotides / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*
chemical_list: C444099:INX-3280; D020319:Oligodeoxyribonucleotides, Antisense; D009841:Oligonucleotides; D013873:Thionucleotides