Basic study on targeted sonodynamic therapy of cancer using photosensitizers and antibodies against tumor antigens.

为了提高光动力治疗癌症的疗效，我们尝试利用超声和抗肿瘤抗体共轭光敏剂开发一种新的靶向声动力治疗方法。为此，我们首先制备了由光敏剂ATX-70和抗癌胚胎抗原（CEA）小鼠单克隆抗体（mAb）组成的免疫偶联物，使用EDC和磺胺- nhs。该偶联物（F39/ATX-70）对体外超声照射肿瘤细胞的细胞毒性有cea特异性增强作用。此外，F39/ATX-70静脉注射后，超声照射对裸鼠肿瘤细胞生长有明显抑制作用。接下来，我们尝试使用人单抗而不是小鼠单抗，以减少应用于患者时可能出现的副作用。用家蚕制备的重组MK-1 （Ep-CAM）蛋白免疫具有人免疫球蛋白基因的KM小鼠，将其脾细胞与小鼠骨髓瘤细胞融合。经过筛选和克隆，获得了几种能分泌针对MK-1的人IgM单抗的杂交瘤克隆。我们目前正在使用不同的免疫方法制备igg级hmab。我们还获得了一种新的、改进的光敏剂DCPH，并试图建立一种合适的方法将其偶联到单抗上。

To improve the efficacy of the photodynamic therapy of cancer, we tried to develop a new targeted sonodynamic therapy using ultrasound and anti-tumor antibody-conjugated photosensitizers. For this, we first prepared immunoconjugates consisting of the photosensitizer ATX-70 and anti-carcinoembryonic antigen (CEA) mouse monoclonal antibody (mAb), using EDC and sulfo-NHS. This conjugate (F39/ATX-70) showed CEA-specific enhancing effects on the cytotoxicity induced by ultrasound irradiation against tumor cells in vitro. Furthermore, the growth of tumor cells in nude mice was greatly inhibited by ultrasound irradiation when F39/ATX-70 was i.v. injected.Next we tried to use human mAb, instead of mouse mAb, to reduce possible side effects when applied to patients. KM mice, which have the human immunoglobulin genes, were immunized with recombinant MK-1 (Ep-CAM) protein prepared in silkworms, and their spleen cells were fused with mouse myeloma cells. After selection and cloning, several hybridoma clones were obtained that secrete human IgM mAbs against MK-1. We are currently preparing IgG-class HmAbs using different immunization methods. We also obtained a new, improved photosensitizer, DCPH, and are trying to establish a suitable method to conjugate it to mAbs.