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Eight muteins of recombinant human tumor necrosis factor-alpha (rhTNF; 1SSSRTP...29RR...155L), in which 29Arg was replaced by another amino acid, were prepared and their anti-tumor effects in BALB/c mice bearing Meth A fibrosarcoma were evaluated. The therapeutic indices, which mark the extent of the therapeutically effective dose, of V29 (29Arg-->Val) and D29 (-->Asp) were 3.5 and 3.2, respectively, whereas that of rhTNF was 1.4. Clearly, the therapeutically effective range of these muteins was extended along with a decrease in lethal toxicity. V29 did not produce hypotension in the rat system, but D29 did. In addition, V29 showed potent anti-tumor activity (Tumor Volume Inhibition Rate = 81% on day 15 after implantation) in 3 consecutive injection schedules despite the decreases in toxicity compared with rhTNF. The relative receptor binding constant was determined using HEp-2 cells (expressing mainly 55-kDa-TNF receptor; p55R) and HL60 cells (expressing mainly 75-kDa-TNF receptor; p75R), and revealed that the reduced toxicity of V29 in mice was due to the reduced binding to p55R (34% of rhTNF). On the other hand, the ratio of the constants HEp-2/HL60 of V29 was 11 in comparison with the value of 1.0 for rhTNF, suggesting that this mutein binds preferentially to p55R. The biological activities in human cell lines (HEp-2 and HL60 cells) correlated well with the binding activities to each receptor in vitro. Therefore, the much lower toxicity and the potent anti-tumor activity of this mutein suggest that V29 merits further investigation in pre-clinical and clinical trials.
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K Miyata, and
H Shikama, and
T Kawagoe, and
K Nishimura, and
K Takeda, and
N Sakae, and
M Kato
April 1991,
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K Takeda, and
N Sakae, and
M Kato
July 2001,
Acta pharmacologica Sinica,
K Kuroda, and
K Miyata, and
H Shikama, and
T Kawagoe, and
K Nishimura, and
K Takeda, and
N Sakae, and
M Kato
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M Kato
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K Kuroda, and
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K Kuroda, and
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M Kato
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K Kuroda, and
K Miyata, and
H Shikama, and
T Kawagoe, and
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N Sakae, and
M Kato
May 1991,
Cancer research,
K Kuroda, and
K Miyata, and
H Shikama, and
T Kawagoe, and
K Nishimura, and
K Takeda, and
N Sakae, and
M Kato
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K Kuroda, and
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M Kato
June 1989,
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K Kuroda, and
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M Kato
November 2005,
Acta pharmacologica Sinica,
