Preclinical testing of human recombinant von Willebrand factor: ADAMTS13 cleavage capacity in animals as criterion for species suitability.
Von Willebrand factor (VWF) is cleaved by the plasma metalloprotease ADAMTS13 ( A Disintegrin and Metalloproteinase with Thrombo Spondin repeats, number 13) that regulates the hemostatic activity of VWF by limiting its multimeric size in the human system. In vitro and ex vivo studies have shown that human recombinant VWF (rVWF) is virtually resistant to the proteolytic activity of murine ADAMTS13. In contrast, rabbit and cynomolgus ADAMTS13 is able to cleave human rVWF. These findings were consistent with in vivo results showing distinct pharmacological behavior of human rVWF depending on the cleaving capacity of ADAMTS13 present in the species tested. Studies were performed using three mouse strains (ADAMTS13 deficient, C57BL/6J [wild type], VWF deficient), rats, rabbits, and cynomolgus monkeys. All animals were infused once with different doses of human rVWF and, in addition, 14 daily doses were given to rats and cynomolgus monkeys. Exaggerated pharmacological effects were observed in mice, with the ADAMTS13 knockout mouse being the most sensitive strain. Similar findings with decreased incidence and severity were seen in normal C57BL/6J mice and also in VWF-deficient mice, where they were least pronounced. In rats, exaggerated pharmacological effects were observed only after 14 doses. Rabbits and cynomolgus monkeys showed no exaggerated pharmacological effects. These differences between species and between mouse strains suggest that the efficiency of ADAMTS13 to cleave rVWF determines the severity of clinical, laboratory and pathohistological findings. These observations highlight the importance of evaluating species' suitability for the generation of meaningful preclinical data for determining the therapeutic safety margins for human patients. Only animals with a sufficient rVWF cleavage capacity by endogenous ADAMTS13 (rabbits and cynomolgus monkeys) are considered appropriate animal models for preclinical evaluation of the rVWF product.
Muchitsch
Eva-Maria
EM
Baxter BioScience, Vienna, Austria.
Dietrich
Barbara
B
Rottensteiner
Hanspeter
H
Auer
Wilfried
W
Nehrbass
Dirk
D
Gritsch
Herbert
H
Ehrlich
Hartmut J
HJ
Turecek
Peter L
PL
Schwarz
Hans Peter
HP
D051722:ADAM Proteins / Q000378:metabolism*; D000818:Animals; D006801:Humans; D011994:Recombinant Proteins / Q000494:pharmacology; D014841:von Willebrand Factor / Q000494:pharmacology*
D011994:Recombinant Proteins; D014841:von Willebrand Factor; D051722:ADAM Proteins