阿霉素与转铁蛋白受体单抗偶联方法探讨─以葡聚糖DextranT＿10为偶联剂

CONJUGATION OF ADRIAMYCIN WITH MONOCLONAL ANTI-TRANSFERRIN RE-CEPTOR ANTBODY VIA A DEXTRAN T10 BRIDGE

以７５％、５０％、３０％和２５％氧化程度的葡聚糖为偶联剂，制备阿霉素与转铁蛋白受体单抗的偶联物Ｃ７５、Ｃ５０、Ｃ３０和Ｃ２５，并对其各方面参数作了比较，结果表明，其偶联度分别为１６２、１０３、７２与５２Ｍｏｌｅｓ／每个抗体分子，即葡聚糖的氧化程度越高，其偶联度越高。然而，以１３１Ｉ－ＳｐＡ为二抗的细胞结合分析结果表明，Ｃ７５、Ｃ５０均丧失大部分的抗体活性。３Ｈ－ＴｄＲ掺入细胞抑制实验也表明，Ｃ７５与Ｃ５０丧失大部分药物活性，而Ｃ３０均保持大部分的抗体活性与药物活性，认为３０％氧化葡聚糖为最佳偶联方案。实验还表明，Ｃ３０对体外靶细胞的杀伤具有高度的选择性。而且，其分药物活性的丧失，可以通过延长与靶细胞的孵育时间而得到补偿。号。就Ａｄｒ－Ｄｅｘ－ＷｕＴ９偶联物而言，同样孵育４ｈ，对肝癌细胞和正常肝细胞却有不同和杀伤率，（ｐ＜０．０１，见表中的▲号）。说明偶联物Ｃ３０对肝癌细胞的杀伤作用具有高度的选择性。与此相反，孵育４ｈ，Ａｄｒ不但对肝癌细胞，而且对正常肝细胞也产生明显的细胞毒作用（ｐ＞０．０５，见表中的△△号），说明游离的Ａｄｒ无明显的选择性。而且Ａｄｒ与ＷｕＴ９偶联之后，所产生的部分药物活性的丧失（约丧失２?

By using 75%,50%,30%and 25% oxidized Dextran T10 as a bridge,the conjugates(c75,c50,c30 and c25) of Adriamycin with monoclonal anti-transferrin receptor(TFR)antibody wuT9were prepared respectively and compared with each other in various parameters.Resultsindicated that the ratios of drug to antibody in conjugates are 162,103,72 and 50 Moles/Mole antibody,i.e. the higher degrees of drug substitution are associated with the higheroxidation of Dextran. However, the findings derived from cell binding analysis and 3H-TdRincorporation test showed that c75 and c50 lose almost all the immunological as well aspharmacological activities, whereas c30 significantly retained the both. This result indicated that30% oxidized Dextran was best bridge for conjugation. In addition,it was suggested that c30has high preference for killing target cells in vitro culture,and some loss of pharmacologicalactivity of the conjugate may be compensated for by prologing incubation time with targetcells.The principle of conjugation with Dextran T 10 as a bridge was also discussed in this article.