NOVEL MUTEIN OF TUMOR-NECROSIS-FACTOR-ALPHA (F4614) WITH REDUCED HYPOTENSIVE EFFECT

To eliminate systemic toxicity, including the hypotension associated w ith human tumor necrosis factor cy (TNF-(alpha), we constructed mutant proteins (muteins) by mean of genetic engineering. A novel mutein, F4 614, containing mutations of (5)Thr --> Gly and (6)Pro --> Asp, which resulted in the introduction of cell-adhesive Arg-Gly-Asp and (29)Arg --> Val, had remarkably reduced hypotensive effects and lower lethalit y, We present evidence that the Arg --> Val mutation at position 29 is largely responsible for the reduced hypotensive effect. This effect o f F3614 was thought to be closely correlated with its low inducibility of nitric oxide and prostaglandin E(2) in vivo. In addition, the ther apeutically effective dose of F46P4 to MethA fibrosarcoma-transplanted mice was increased compared with that of TNF-alpha. Indicating a wide therapeutic index. These results indicated that F4614 has several adv antages as a systemic therapeutic drug in the treatment of cancer.