DIGOXIN-SPECIFIC HUMAN ANTIBODIES FROM TRANSGENIC MICE

该项目的长期目标是开发人单克隆抗体， 针对强心苷、地高辛和 洋地黄毒苷，其可用作治疗剂治疗毒性 过量服用这些药物。产生的抗体将是相当大的 因为这两种药物是最常用的处方药之一， 在美国，他们的利益之间的安全边际非常狭窄， 及其毒性作用。目前，只有Fab片段的制备， 可以获得异种绵羊抗体，但它们的使用是有限的 因为它们的非人类来源和可变的结合特性。在 我们计划利用GenPharm International最近的 利用转基因技术培育出一种小鼠品系， 产生人IgM和IgG免疫球蛋白。这些老鼠已经被证明 对初次和二次免疫的免疫应答 应答 因此，我们将使用地高辛-和洋地黄毒苷-卵清蛋白 引发人IgG抗洋地黄生成的缀合物 抗体的标准杂交瘤技术将用于生产，鉴定 并克隆分泌高亲和力、人抗药物 克隆抗体它们的结合亲和力和特异性 单克隆抗体将被充分表征， 作为临床上有用的药物进行II期开发。 拟议的商业应用：强心苷是最 在美国常用的处方药。据估计，13%的 65岁以上的人口接受治疗， 因明显心力衰竭而入院的患者 这些药物的毒性过量。虽然大多数这些患者没有 需要给予羊抗地高辛Fab片段， 据估计，目前美国每年约有2,500名患者接受这种治疗。 治疗人类单克隆抗体不仅应该取代目前的 产品，而是更安全和更广泛使用的临床试剂。

The long-term goal of this project is to develop human monoclonal antibodies directed against the cardiac glycosides, digoxin and digitoxin,that can be used as therapeutic agents in the treatment of toxic overdoses of these drugs. The generated antibodies would be of considerable value because these two drugs are among the most commonly prescribed drugs in the US, yet have a very narrow margin of safety between their beneficial and their toxic effects. Currently only an Fab fragment preparation of heterogeneous sheep antibodies is available and their use is limited because of their nonhuman origin and variable binding characteristics. In this proposal, we plan to utilize GenPharm International's recent achievements in using transgene technology to develop a mice strain that produces both human IgM and IgG immunoglobulins. These mice have been shown to respond to immunizations with both primary and secondary immune responses. Therefore we will use digoxin- and digitoxin-ovalbumin conjugates to provoke the generation of human IgG anti-digitalis antibodies. Standard hybridoma technology will be used to produce, identify and clone hybridoma cells secreting high affinity, human anti-drug monoclonal antibodies. The binding affinities and specificities of these monclonal antibodies will be fully characterized and those most suitable for Phase II development as clinically useful agents will be identified. PROPOSED COMMERCIAL APPLICATION: The cardiac glycosides are among the most commonly prescribed drugs in the US. It is estimated that 13% of our population over 65 receive treatment with them and that 4-6% of all patients admitted to the hospital for apparent heart failure are suffering from toxic overdoses of these drugs.While most of these patients do not require administration of the sheep anti-digoxin Fab fragments, it is estimated that currently about 2,500 patients/year in the US undergo such treatment. A human monoclonal antibody should not only replace the present product but be a much safer and more widely used clinical agent.