title: The in vivo and in vitro metabolic profile of 99mTc-NC100668, a new tracer for imaging venous thromboembolism: identification and biodistribution of the principal radiolabeled metabolite.
abstract: (99m)Tc-NC100668 [Acetyl-Asn-Gln-Glu-Gln-Val-Ser-Pro-Tyr(3-iodo)-Thr-Leu-Leu-Lys-Gly-NC100194] is a radiopharmaceutical imaging agent being developed to aid the diagnosis of thromboembolism. The stability profile of (99m)Tc-NC100668 was investigated by high-performance liquid chromatography (HPLC) after in vitro exposure to blood and plasma obtained from rat and human, as well as to urine and bile obtained from rat. The metabolic profile of (99m)Tc-NC100668 exposed to human and rat hepatic S9 (a liver homogenate-rich cytochrome P450) was also studied. The profile of (99m)Tc-labeled species in plasma, urine, and bile was investigated following i.v. administration of (99m)Tc-NC100668 to rat. The major species observed in vitro and in vivo consisted of the (99m)Tc-chelator (NC100194) [N,N-Bis(N-(1,1-dimethyl-2-(hydroxylimino-)propyl)aminoethyl)aminoethylamine] attached to the C-terminal amino acid residue and referred to as (99m)Tc-complex of Gly-NC100194. The identity of the major metabolite was confirmed by cochromatography with an authentic standard and the genuine metabolite using a second HPLC method. The minor metabolites were sodium pertechnetate ((99m)Tc) and (99m)Tc-NC100194. In addition, a small number of other species were transiently observed in vitro; they were not investigated further. The biodistribution of the major metabolite was studied in male Wistar rats. The affinity of the major metabolite toward plasma clot was established using a plasma clot-forming assay. A minor uptake of (99m)Tc-complex of Gly-NC100194 in the plasma clot and a rapid removal from the body were noted. In conclusion, the metabolites of (99m)Tc-NC100668 are not anticipated to have a negative impact on the ability of the test substance to image blood clots.
authors.affiliation: Research and Development, GE Healthcare Bio-Sciences, The Grove Centre, Little Chalfont, Buckinghamshire HP7 9LL, UK. davidedwards@ge.com
authors.forename: David
authors.initials: D
authors.lastname: Edwards
authors.forename: Mark
authors.initials: M
authors.lastname: Battle
authors.forename: Rochelle
authors.initials: R
authors.lastname: Lear
authors.forename: Gill
authors.initials: G
authors.lastname: Farrar
authors.forename: D Jon
authors.initials: DJ
authors.lastname: Barnett
authors.forename: Vanessa
authors.initials: V
authors.lastname: Godden
authors.forename: Catherine
authors.initials: C
authors.lastname: Coombes
authors.forename: Alexandra
authors.initials: A
authors.lastname: Oliveira
authors.forename: Håkan
authors.initials: H
authors.lastname: Ahlström
mesh_terms: D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004355:Drug Stability; D006801:Humans; D006868:Hydrolysis; D036341:Intercellular Signaling Peptides and Proteins; D008099:Liver / Q000378:metabolism; D008297:Male; D009842:Oligopeptides / Q000737:chemistry / Q000493:pharmacokinetics*; D015609:Organotechnetium Compounds / Q000737:chemistry / Q000493:pharmacokinetics*; D010455:Peptides / Q000737:chemistry / Q000493:pharmacokinetics*; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000737:chemistry / Q000493:pharmacokinetics*; D051381:Rats; D017208:Rats, Wistar; D013927:Thrombosis / Q000000981:diagnostic imaging / Q000378:metabolism*; D014018:Tissue Distribution; D014357:Trypsin
chemical_list: C512909:99mTc-NC100668; D036341:Intercellular Signaling Peptides and Proteins; D009842:Oligopeptides; D015609:Organotechnetium Compounds; D010455:Peptides; D019275:Radiopharmaceuticals; C507860:acetyl-Asn-Gln-Glu-Gln-Val-Ser-Pro-Tyr(3-iodo)-Thr-Leu-Leu-Lys-Gly-NC100194; D014357:Trypsin