Acute and repeated dose (28 days) toxicity studies in rats and dogs of recombinant batroxobin, a snake venom thrombin-like enzyme expressed from Pichia pastoris.
Recombinant batroxobin is a thrombin-like enzyme of Bothrops atrox moojeni venom. To evaluate its toxicological effect, it was highly expressed in Pichia pastorisand successfully purified to homogeneity from culture broth supernatant following Good Manufacturing Practice (GMP). The maximum tolerated dose of the recombinant batroxobin was examined in Sprague-Dawley (SD) rat and Beagle dogs following Good Laboratory Practice (GLP) regulations. The approximate lethal dose of recombinant batroxobin was 10 National Institute of Health (NIH) u/kg in male and female rats. Slight test substance-related effects were clearly in male and female dogs at more than 10 NIH u/kg. The maximum tolerated dose (MTD) was considered to be greater than 30 NIH u/kg in dogs. To investigate the repeated dose toxicity of batroxobin, the test item was intravenously administered to groups of SD rat and Beagle dog every day for 4 weeks. We observed that all animals survived the duration of the study without any effects on their mortality. There were no effects in both rats and dogs regarding their clinical signs, body weight, food consumption, ophthalmological examination, urinalysis, hematology, clinical chemistry, organ weightand gross post mortem examinations. The no adverse effect level (NOAEL) of recombinant batroxobin for both males and females is considered to be greater than 2.5 NIH u/kgin rats and 1 NIH u/kg in dogs, respectively. No toxic effects were noted in target organs. In conclusion, these results show a favorable preclinical profile and may contribute clinical development of recombinant batroxobin.
Thrombosis and Vascular Biochemistry Lab., Dept of Biotechnology, College of Life Science, CHA University, CHA Bio-Complex, 335 Pangyo-ro, Seongnam-si 463-400, South Korea.
Ok Hwan
OH
Kim
Thrombosis and Vascular Biochemistry Lab., Dept of Biotechnology, College of Life Science, CHA University, CHA Bio-Complex, 335 Pangyo-ro, Seongnam-si 463-400, South Korea.
Kil-Sang
KS
Cho
R&D Center, NC-BIT Co., Ltd., B-904 Korea Bio Park, Seongnam-si 463-400, South Korea.
Young
Y
Seomun
R&D Center, NC-BIT Co., Ltd., B-904 Korea Bio Park, Seongnam-si 463-400, South Korea.
Jong-Tak
JT
Kim
Thrombosis and Vascular Biochemistry Lab., Dept of Biotechnology, College of Life Science, CHA University, CHA Bio-Complex, 335 Pangyo-ro, Seongnam-si 463-400, South Korea. Electronic address: hoe@cha.ac.kr.
Kwang-Hoe
KH
Chung
D000818:Animals; D012103:Batroxobin / Q000633:toxicity*; D001835:Body Weight; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D005260:Female; D005285:Fermentation; D007928:Lethal Dose 50; D008297:Male; D019055:No-Observed-Adverse-Effect Level; D009929:Organ Size / Q000187:drug effects; D010843:Pichia / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000633:toxicity; D012910:Snake Venoms / Q000737:chemistry*; D013917:Thrombin; D023382:Toxicity Tests, Acute
D011994:Recombinant Proteins; D012910:Snake Venoms; D012103:Batroxobin; D013917:Thrombin
Mass production; Recombinant batroxobin; Repeated dose toxicity; Single dose toxicity; Snake venom thrombin-like enzyme