A novel recombinant tumor necrosis factor-alpha mutant with increased anti-tumor activity and lower toxicity.
We prepared a novel recombinant tumor necrosis factor-alpha (TNF) mutant (mutant 471), in which 7 N-terminal amino-acids were deleted and Pro8Ser9Asp10 was replaced by ArgLysArg, and compared its biological activity with that of wild-type recombinant TNF. Mutant 471 had a 7-fold higher anti-tumor activity against murine L-M cells in vitro, and a higher binding activity to TNF receptors on L-M cells, than wild-type TNF. Furthermore, mutant 471 showed a higher anti-tumor effect on murine Meth A-HM tumors transplanted into BALB/c mice, with complete regression of the tumors being observed in the animals. The possible cachectin activity of mutant 471 was almost the same as that of wild-type TNF. The acute lethal toxicity of mutant 471 in beta-D-galactosamine-sensitized C3H/HeJ mice was 18 times lower than that of wild-type TNF. These results suggest that mutant 471 might be a more promising anti-cancer agent than wild-type TNF.
Nakamura
S
S
Biotechnology Research Laboratories, Teijin Limited, Tokyo, Japan.
Kato
A
A
Masegi
T
T
Fukuoka
M
M
Kitai
K
K
Ogawa
H
H
Ichikawa
Y
Y
Maeda
M
M
Watanabe
N
N
Kohgo
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D005354:Fibrosarcoma / Q000188:drug therapy* / Q000473:pathology; D007739:L Cells / Q000166:cytology / Q000187:drug effects; D008071:Lipoprotein Lipase / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011956:Receptors, Cell Surface / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins / Q000494:pharmacology / Q000627:therapeutic use* / Q000633:toxicity; D012513:Sarcoma, Experimental / Q000188:drug therapy* / Q000473:pathology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use* / Q000633:toxicity
D011956:Receptors, Cell Surface; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D008071:Lipoprotein Lipase