Department of Medicine, University of California, Los Angeles, CA 90024, USA.
Bioconjug Chem. 2013 Jan 16;24(1):97-104. doi: 10.1021/bc3005123. Epub 2012 Dec 31.
The chronic administration of recombinant fusion proteins in preclinical animal models may generate an immune response and the formation of antidrug antibodies (ADA). Such ADAs could alter the plasma pharmacokinetics of the fusion protein, and mask any underlying toxicity of the recombinant fusion protein. In the present study, a model IgG-enzyme fusion protein was evaluated with chronic dosing of rhesus monkeys. The IgG domain of the fusion protein is a genetically engineered monoclonal antibody (mAb) against the human insulin receptor (HIR), which is shown to cross-react with the primate insulin receptor. The enzyme domain of the fusion protein is human iduronidase (IDUA), the lysosomal enzyme mutated in Mucopolysaccharidosis Type I (MPSI). MPSI affects the brain, but enzyme replacement therapy is not effective for the brain, because IDUA does not cross the blood-brain barrier (BBB). The HIRMAb domain of the fusion protein acts as a molecular Trojan horse to deliver the IDUA across the BBB. The HIRMAb-IDUA fusion protein was administered to rhesus monkeys with weekly intravenous infusions of 3-30 mg/kg for 6 months, and the pharmacokinetics, immune response, and tissue toxicology were assessed. The pharmacokinetics of plasma clearance of the fusion protein was determined with measurements of plasma IDUA enzyme activity. ADAs formed during the course of the 6 months of treatment, as determined by a sandwich ELISA. However, the plasma clearance of the fusion protein at the start and end of the 6-month study was comparable at all drug doses. Fusion protein administration for 6 months showed no evidence of chronic tissue toxicity. These studies demonstrate that the immune response produced with chronic treatment of primates with an IgG-enzyme fusion protein has no effect on the pharmacokinetics of plasma clearance of the fusion protein.
在临床前动物模型中,慢性给予重组融合蛋白可能会产生免疫反应和抗药物抗体(ADA)的形成。这些 ADA 可能会改变融合蛋白的血浆药代动力学,并掩盖重组融合蛋白的任何潜在毒性。在本研究中,使用恒河猴对模型 IgG-酶融合蛋白进行了评估。融合蛋白的 IgG 结构域是针对人胰岛素受体(HIR)的基因工程单克隆抗体(mAb),它被证明与灵长类动物胰岛素受体发生交叉反应。融合蛋白的酶结构域是人溶酶体酸性麦芽糖酶(IDUA),该酶在黏多糖贮积症 I 型(MPSI)中发生突变。MPSI 会影响大脑,但酶替代疗法对大脑无效,因为 IDUA 不能穿过血脑屏障(BBB)。融合蛋白的 HIRMAb 结构域充当分子木马,将 IDUA 递送到 BBB 中。将 HIRMAb-IDUA 融合蛋白以每周 3-30mg/kg 的剂量静脉输注 6 个月的方式给予恒河猴,并评估药代动力学、免疫反应和组织毒理学。通过测量血浆 IDUA 酶活性来确定融合蛋白的血浆清除率的药代动力学。通过夹心 ELISA 确定在治疗的 6 个月期间形成的 ADA。然而,在 6 个月研究开始和结束时,所有药物剂量的融合蛋白血浆清除率均相当。融合蛋白给药 6 个月未显示出慢性组织毒性的证据。这些研究表明,用 IgG-酶融合蛋白对灵长类动物进行慢性治疗产生的免疫反应对融合蛋白的血浆清除率的药代动力学没有影响。