Enzyme specific activation of benzoquinone ansamycin prodrugs using HuCC49ΔCH2-β-galactosidase conjugates
Enzyme specific activation of benzoquinone ansamycin prodrugs using HuCC49ΔCH2-β-galactosidase conjugates
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DOI：
10.1021/jm060647f
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发表时间：
2006-10-19
影响因子：
7.3
通讯作者：
Sun, Duxin
中科院分区：
文献类型：
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作者：
Fang, Lanyan;Battisti, Robert F.;Sun, Duxin
为了激活用于癌症治疗的前药，使用抗 TAG-72 抗体 (HuCC49 Delta CH2) 来递送激活酶（β-半乳糖苷酶），以特异性激活针对结肠癌的格尔德霉素前药 (17-AG-C2-Gal)。格尔德霉素前药 17-AG-C2-Gal 通过将半乳糖胺衍生物与格尔德霉素在 C-17 位偶联来合成。使用两种不同程序（Affinity 和 Autodock）进行分子对接表明前药（17-AG-C2-Gal）无法与 Hsp90 结合；然而，产物 (17-AG-C2) 被 β-半乳糖苷酶缀合物酶促裂解，以与格尔德霉素和 17-AG 类似的方式与 Hsp90 结合。计算对接结果在四唑[3-(4,5-二甲基噻唑-2-基)]-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑测定和质谱的实验测试中得到进一步证实。 HuCC49 Delta CH2 与 β-半乳糖苷酶化学缀合。该抗体-酶缀合物能够靶向肿瘤抗原TAG-72，并保持良好的酶活性来激活17-AG-C2-Gal前药。所释放的活性药物 17-AG-C2 被证明可诱导高达 70% 的 AKT 降解，与前药相比，抗癌活性增强超过 25 倍。
To activate prodrugs for cancer treatment, an anti-TAG-72 antibody (HuCC49 Delta CH2) was used for delivery of an activation enzyme (beta-galactosidase) to specifically activate a geldanamycin prodrug (17-AG-C2-Gal) against colon cancer. The geldanamycin prodrug 17-AG-C2-Gal was synthesized by coupling a galactose-amine derivative with geldanamycin at the C-17 position. Molecular docking with two different programs (Affinity and Autodock) showed that the prodrug (17-AG-C2-Gal) was unable to bind to Hsp90; however, the product (17-AG-C2), enzymatically cleaved by beta-galactosidase conjugate, bound to Hsp90 in a similar way as geldanamycin and 17-AG. The computational docking results were further confirmed in experimental testing by the tetrazolium [3-(4,5-dimethythiazol-2-yl)]-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay and mass spectrometry. HuCC49 Delta CH2 was chemically conjugated to beta-galactosidase. The antibody-enzyme conjugate was able to target tumor antigen TAG-72 with the well preserved enzymatic activity to activate 17-AG-C2-Gal prodrug. The released active drug 17-AG-C2 was demonstrated to induce up to 70% AKT degradation and enhance anticancer activity by more than 25-fold compared to the prodrug.