effectiveness of human EGF-conjugate in the treatment of breast cancer

人 EGF 结合物治疗乳腺癌的有效性

• 批准号: 14571156
• 负责人: JINNO Hiromitsu
• 金额: $ 1.92万
• 依托单位: KEIO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571156/
• 关键词: Epidermal Growth Factor; Eosinophilic Cationic Protein; Targeting Therapy; Breast Cancer; Endogenous Protein; ECP; EGF; targeting therapy; ターゲッティング療法

Epidermal growth factor receptor (EGFR) overexpression has been found in 30% of breast cancer patients anti is associated with a poor clinical prognosis. Such cases are highly aggressive and resistant to conventional treatment including surgery and chemoendocrine therapies. To selectively kill highly proliferating cancer cells with EGFR overexpression, targeted therapies aiming at EGFR have been developed. However, clinical application of targeted therapies would entail several major problems, because they are usually composed of plant toxin, bacterial toxin, or marine monoclonal antibodies. In an attempt to reduce immunogenicity and toxicity of the targeted therapy, we have developed the conjugate consisting of human endogeneous proteins. Recombinant human eosinophil cationic protein (ECP) was chemically conjugated to recombinant human EGF via a disulfide bond using the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and 2-iminothiolane. The cytotoxic activity of the conjugate was evaluated by MTT assay against EGFR-overexpressing breast cancer cell lines (BT-20, MCF-7) as well as EGFR-negative small cell lung cancer cell line (H69). EGF-ECP conjugate revealed dose-dependent cytotoxicity for EGFR-overexpressing BT-20 breast cancer cells with an IC_<50> of 1.5x10^<-7>M and had no effect on EGFR-negative H69 cells. Recombinant human ECP alone had an IC_<50> of higher than 10^<-5>M. A mixture of flee ECP and EGF showed no more cytotoxic activity than ECP alone. Addition of excess EGF in the medium protected EGFR-overexpressing cells from the cytotoxic activity of the conjugate. The cytotoxic potential of the conjugate was positively correlated with cell's EGFR number. These results suggest that EGF-ECP conjugate may be effective against EGFR-overexpressing breast cancers with less iminunogenicity than conventional targeted therapies.

已发现 30% 的乳腺癌患者表皮生长因子受体 (EGFR) 过度表达与不良临床预后相关。此类病例具有高度侵袭性，并且对包括手术和化学内分泌疗法在内的常规治疗具有抵抗力。为了选择性地杀死 EGFR 过度表达的高度增殖的癌细胞，针对 EGFR 的靶向治疗已被开发出来。然而，靶向治疗的临床应用会带来几个重大问题，因为它们通常由植物毒素、细菌毒素或海洋单克隆抗体组成。为了降低靶向治疗的免疫原性和毒性，我们开发了由人内源性蛋白组成的缀合物。使用异双功能试剂 N-琥珀酰亚胺基 3-(2-吡啶二硫代)丙酸酯 (SPDP) 和 2-亚氨基四氢噻吩，将重组人嗜酸性粒细胞阳离子蛋白 (ECP) 通过二硫键与重组人 EGF 化学缀合。通过 MTT 测定评估缀合物针对 EGFR 过表达乳腺癌细胞系（BT-20、MCF-7）以及 EGFR 阴性小细胞肺癌细胞系（H69）的细胞毒活性。 EGF-ECP缀合物揭示了对EGFR过表达的BT-20乳腺癌细胞的剂量依赖性细胞毒性，IC_ 50 为1.5×10 10 -7 M，并且对EGFR阴性H69细胞没有影响。单独的重组人ECP具有高于10 ^ -5 M的IC 50 。 ECP和EGF的混合物没有表现出比单独的ECP更高的细胞毒性活性。在培养基中添加过量的EGF可以保护EGFR过度表达的细胞免受缀合物的细胞毒活性的影响。缀合物的细胞毒性潜力与细胞的EGFR数量呈正相关。这些结果表明，EGF-ECP 缀合物可能有效对抗 EGFR 过度表达的乳腺癌，且免疫原性低于传统靶向治疗。

项目成果

H.Jinno et al.: "The Grnotoxicity of a Conjugate compoxecl of human EGF and ECP"Aunicancer Res. 22. 4141-4146 (2002)

H.Jinno 等人：“人 EGF 和 ECP 的缀合物复合物的毒性”Aunicancer Res。

H.Jinno et al.: "The cytotoxicity of conjugate composed of human EGF and ECF"Anticancer Res. 22. 4141 (2002)

H.Jinno 等：“人 EGF 和 ECF 缀合物的细胞毒性”抗癌研究。

H.Jinno et al.: "Selective cytotoxicity of human angiogenin conjugated to human EGF against EGFR-overexpressing cancer cells"Proc Am Assoc Cancer Res. 45. 355 (2004)

H.Jinno 等人：“与人 EGF 缀合的人血管生成素对 EGFR 过表达癌细胞的选择性细胞毒性”Proc Am Assoc Cancer Res。

H.Jinno et al.: "The cytotoxicity of a conjugate composed of human epidermal growth factor and eosinophil cationic protein."Anticancer Res. 22. 4141-4146 (2002)

H.Jinno 等人：“由人表皮生长因子和嗜酸性粒细胞阳离子蛋白组成的缀合物的细胞毒性。”Anticancer Res。

H.Jinno et al.: "Selective cytotoxicity of human angiogenin conjugated to human EGF against EGFR-overexpressing cancer cells"Proc Am Assoc Cancer Res. 未定. (2004)

H.Jinno 等人：“与人 EGF 缀合的人血管生成素对 EGFR 过表达癌细胞的选择性细胞毒性”Proc Am Assoc Cancer Res. 待确定。