Janssen Biopharmaceuticals (formerly Biotechnology Center of Excellence) R&D of J&J Spring House Pennsylvania 19477.
Discovery Sciences Janssen Pharmaceutical R&D of J&J Spring House Pennsylvania 19477.
Pharmacol Res Perspect. 2016 Apr 25;4(3):e00218. doi: 10.1002/prp2.218. eCollection 2016 Jun.
Although much speculation has surrounded intestinally expressed FcRn as a means for systemic uptake of orally administered immunoglobulin G (IgG), this has not been validated in translational models beyond neonates or in FcRn-expressing cells in vitro. Recently, IgG1 intestinal infusion acutely in anesthetized cynomolgus resulted in detectable serum monoclonal antibody (mAb) levels. In this study, we show that IgG2 has greater protease resistance to intestinal enzymes in vitro and mice in vivo, due to protease resistance in the hinge region. An IgG2 mAb engineered for FcRn binding, was optimally formulated, lyophilized, and loaded into enteric-coated capsules for oral dosing in cynomolgus. Small intestinal pH 7.5 was selected for enteric delivery based on gastrointestinal pH profiling of cynomolgus by operator-assisted IntelliCap System(®). Milling of the lyophilized IgG2 M428L FcRn-binding variant after formulation in 10 mmol/L histidine, pH 5.7, 8.5% sucrose, 0.04% PS80 did not alter the physicochemical properties nor the molecular integrity compared to the batch released in PBS. Size 3 hard gel capsules (23.2 mg IgG2 M428L ~3 mg/kg) were coated with hydroxypropyl methylcellulose acetate succinate for rapid dissolution at pH 7.5 in small intestine and FcRn binding of encapsulated mAb confirmed. Initial capsule dosing by endoscopic delivery into the small intestine achieved 0.2 + 0.1 ng/mL (n = 5) peak at 24 h. Weekly oral capsule dosing for 6 weeks achieved levels of 0.4 + 0.2 ng/mL and, despite increasing the dose and frequency, remained below 1 ng/mL. In conclusion, lyophilized milled mAb retains FcRn binding and molecular integrity for small intestinal delivery. The low systemic exposure has demonstrated the limitations of intestinal FcRn in non-human primates and the unfeasibility of employing this for therapeutic levels of mAb. Local mAb delivery with limited systemic exposure may be sufficient as a therapeutic for intestinal diseases.
尽管人们对肠道表达的 FcRn 作为口服给予的免疫球蛋白 G(IgG)全身摄取的一种手段进行了大量推测,但在新生儿以外的转化模型或体外表达 FcRn 的细胞中尚未得到验证。最近,在麻醉恒河猴中急性肠内输注 IgG1 导致可检测到的血清单克隆抗体(mAb)水平。在这项研究中,我们表明 IgG2 在体外和体内具有更大的对肠道酶的蛋白酶抗性,这是由于铰链区的蛋白酶抗性。一种为 FcRn 结合而设计的 IgG2 mAb 被优化配方、冻干,并装入肠溶胶囊中用于恒河猴的口服给药。根据恒河猴胃肠道 pH 分析,选择 pH 7.5 的小肠用于肠溶递送,该分析由操作员辅助的 IntelliCap System(®)进行。在 10 mmol/L 组氨酸、pH 5.7、8.5%蔗糖、0.04% PS80 中进行配方后,冻干的 IgG2 M428L FcRn 结合变体的研磨不会改变物理化学性质或分子完整性与在 PBS 中释放的批次相比。大小为 3 号硬胶囊(23.2 mg IgG2 M428L~3 mg/kg)用羟丙基甲基纤维素乙酸琥珀酸酯包衣,以在小肠中快速溶解,pH 7.5,并确认包封的 mAb 的 FcRn 结合。通过内窥镜将胶囊直接递送到小肠进行初始胶囊给药,在 24 小时时达到 0.2+0.1ng/mL(n=5)的峰值。每周口服胶囊给药 6 周可达到 0.4+0.2ng/mL 的水平,尽管增加了剂量和频率,但仍低于 1ng/mL。总之,冻干研磨的 mAb 保留了用于小肠递送的 FcRn 结合和分子完整性。低全身暴露表明肠道 FcRn 在非人类灵长类动物中的局限性,并且不可行的采用这种方法来实现 mAb 的治疗水平。具有有限全身暴露的局部 mAb 递送可能足以作为肠道疾病的治疗方法。