The invention of the intellectualization cytokine using a synthetic biological response modifier.

本研究的目的是进一步探讨功能性高分子修饰剂偶联在生物活性蛋白临床应用中的作用，并通过体内偶联提高肿瘤坏死因子α (tnf - α)的治疗效果。我们合成了二乙烯基醚-马来酸酐共聚物（DIVEMA）偶联的tnf - α，作为一种合成的生物反应调节剂，具有固有的抗肿瘤活性。divema - tnf - α的合成可以通过添加2,3-二甲基马来酸酐（DMMAn）来控制，当pH改变时，DMMAn与氨基结合或分离。DMMAn合成的divema - tnf - α(+)的体外比活性几乎没有下降。然而，divema - tnf - α(-)在没有被DMMAn阻断的情况下偶联，其比活性明显降低。在肉瘤-180实体瘤小鼠体内静脉注射divema - tnf - α(+)后24小时，与天然tnf - α相比，divema - tnf - α(+)对肿瘤产生了显著的出血性坏死作用。此外，divema - inf - α(+)在每只小鼠仅100日本参考单位的剂量下显示出显著的抗肿瘤作用，大约是天然tnf - α的100倍，并且可以诱导所有5只患有甲基苯二甲实体瘤的小鼠完全消退，而没有任何明显的副作用。由于DIVEMA单独或tnf - α与DIVEMA的混合物在静脉给药时均不能引起抗肿瘤活性，因此tnf - α抗肿瘤效力的增加可能是由与DIVEMA的共价偶联引起的。divema - tnf - α在低剂量下显示出显著的抗肿瘤作用。由于体内注射的tnf - α剂量极低，体内内禀tnf - α浓度不受影响。因此，体内的细胞因子网络不会被破坏。这些结果表明DIVEMA是一种有效的结合tnf - α的聚合物修饰剂，可以提高其抗肿瘤活性。

The purpose of this study is to further explore the usefulness of conjugation with functional polymeric modifiers for clinical application of bioactive proteins and to increase the therapeutic efficacy of tumor necrosis factor alpha (TNF-alpha) by conjugation in vivo. We synthesized TNF-alpha conjugated with the copolymer of divinyl ether and maleic anhydride (DIVEMA), which has intrinsic antitumor activity as a synthetic biological response modifier. The synthesis of DIVEMA-TNF-alpha could be controlled by the addition of 2,3-dimethylmaleic anhydride (DMMAn), which binds to or separates from amino groups when the pH is changed. The specific activity of DIVEMA-TNF-alpha (+) synthesized with DMMAn was hardly decreased in vitro. However, DIVEMA-TNF-alpha (-), which is conjugated without blocking by DMMAn, had a markedly diminished specific activity. DIVEMA-TNF-alpha (+) caused a dramatic hemorrhagic necrotic effect on the tumor when compared to native TNF-alpha 24 h after i.v. injection into mice bearing Sarcoma-180 solid tumors. In addition, DIVEMA-INF-alpha (+) at a dose of only 100 Japan reference units per mouse revealed a dramatic antitumor effect that is approximately 100 times greater than native TNF-alpha and that could induce complete regression in all five mice bearing Meth-A solid tumors without any apparent side effects. Because neither DIVEMA alone nor a mixture of TNF-alpha and DIVEMA caused antitumor activity with i.v. administration, the increase in antitumor potency of TNF-alpha may be caused by the covalent conjugation with DIVEMA.DIVEMA-TNF-alpha at low dose revealed dramatic antitumor potency. Because TNF-alpha injected in vivo is extremely low-dose, concentration of intrinsic TNF-alpha in vivo is not influenced. Therefore, the cytokine network in vivo is not destroyed. These results suggest that DIVEMA is a useful polymeric modifier for conjugation of TNF-alpha to increase its antitumor activity.