Genotoxicity studies on HM10760A, recombinant human erythropoietin conjugated to globin fragment.
HM10760A is a recombinant human erythropoietin chemically conjugated to the N-terminus of human immunoglobulin Fc fragment through a polyethylene glycol linker. HM10760A was shown to have a relatively long half-life, compared with unconjugated recombinant erythropoietin. In this study, the genotoxicity of HM10760A was investigated by using a test battery of three different methods. In the Ames assay, five strains (TA100, TA1535, TA98, TA1537, and Escherichia coli WP2 uvrA) were tested at six concentrations of 3.13, 6.25, 12.5, 25, 50, and 100microg/plate. HM10760A did not increase the number of revertant colonies in any tester strains with and without metabolic activation by rat-liver S9 mix. Subsequently, in vitro chromosomal aberration test, using Chinese hamster lung cells, were conducted at the concentrations of 25, 50, and 100microg/mL. HM10760A did not induce chromosomal aberrations either in the short-period (6 hours) test with or without rat-liver S9 mix or in the continuous-treatment (24 hours) test. In the in vivo bone marrow micronucleus assay using the male ICR (imprinting control region) mouse, HM10760A was subcutaneously administered twice at 24-hour intervals at doses of 0, 150, 300, and 600microg/kg. HM10760A produced a slight, but statistically significant, increase in the frequency of micronucleated polychromatic erythrocytes at 600microg/kg. However, no biological significance was assumed, because this value was within the historical control range. From these findings obtained from the genotoxicity assays performed in this study, it appears unlikely that HM10760A acts as a genotoxic agent in vitro and in vivo.
Kwon
Se Chang
SC
Research Center, Hanmi Pharm Co., Hwaseong, Korea.
Lee
Gwan Sun
GS
Han
Jae Yong
JY
Lee
Michael
M
Lee
Young-Mi
YM
Koh
Woo Suk
WS
Kim
Ji-Young
JY
D000818:Animals; D001711:Biotransformation; D001854:Bone Marrow Cells / Q000187:drug effects; D002478:Cells, Cultured; D002869:Chromosome Aberrations / Q000187:drug effects; D006224:Cricetinae; D004921:Erythropoietin / Q000737:chemistry / Q000633:toxicity*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D005260:Female; D006397:Hematinics / Q000737:chemistry / Q000633:toxicity*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry; D008168:Lung / Q000187:drug effects; D008297:Male; D051379:Mice; D048629:Micronuclei, Chromosome-Defective / Q000139:chemically induced; D015162:Micronucleus Tests; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D016296:Mutagenesis / Q000187:drug effects*; D009153:Mutagens / Q000737:chemistry / Q000633:toxicity*; D011092:Polyethylene Glycols / Q000737:chemistry; D051381:Rats; D011994:Recombinant Proteins; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics
D006397:Hematinics; D007141:Immunoglobulin Fc Fragments; D009153:Mutagens; D011994:Recombinant Proteins; D004921:Erythropoietin; D011092:Polyethylene Glycols