Rattanapisit Kaewta, Suwanchaikasem Pipob, Bulaon Christine Joy I, Guo Shiying, Phoolcharoen Waranyoo
Baiya Phytopharm Co., Ltd., Bangkok, Thailand.
GemPharmatech Co., Ltd, Nanjing, China.
PLoS One. 2025 Jan 14;20(1):e0316790. doi: 10.1371/journal.pone.0316790. eCollection 2025.
Anti-programmed cell death 1 (PD-1) monoclonal antibodies (mAbs) have proven to be effective in treating various cancers, including colorectal, lung, and melanoma. Despite their clinical success, some patients develop resistance to mAbs, requiring co-treatments with radio- or chemotherapy. Interleukin-15 (IL-15) is an immunostimulatory cytokine that promotes immune cell production and proliferation. It has been combined with mAbs and other immunotherapies to improve efficacy and reduce side effects. Fusion of anti-PD-1 mAb and IL-15 streamlines drug administration and management. In this study, we developed a prototype by conjugating the IL-15 receptor subunit alpha (IL-15Rα) and IL-15 complex to the C-terminus of anti-PD-1 Pembrolizumab (Pembrolizumab-IL-15Rα-IL15) using plant molecular farming for production. LC-MS revealed the presence of plant N-glycans (GnGnXF, GnXF and Man9GlcNAc2) on the molecule, which may affect receptor-binding avidity. However, ELISA demonstrated comparable binding efficacy of Pembrolizumab-IL-15Rα-IL15 to human PD-1 protein as commercial Pembrolizumab. In a mouse anti-cancer study, Pembrolizumab-IL-15Rα-IL15 (3 mg kg-1) exhibited slightly improved tumor-growth inhibition, reducing tumor size by 94% compared to commercial Pembrolizumab (5 mg kg-1) with an 83% reduction, regardless of statistically significant difference. In conclusion, Pembrolizumab-IL-15Rα-IL-15 was successfully produced using plant molecular farming and shows promise in addressing mAb drug resistance and enhancing the immunomodulatory effects of IL-15 payload.
抗程序性细胞死亡蛋白1（PD-1）单克隆抗体（mAb）已被证明在治疗包括结直肠癌、肺癌和黑色素瘤在内的各种癌症方面有效。尽管它们在临床上取得了成功，但一些患者会对mAb产生耐药性，需要联合放疗或化疗进行治疗。白细胞介素-15（IL-15）是一种免疫刺激细胞因子，可促进免疫细胞的产生和增殖。它已与mAb和其他免疫疗法联合使用，以提高疗效并减少副作用。抗PD-1 mAb与IL-15的融合简化了药物给药和管理。在本研究中，我们通过使用植物分子农业生产，将IL-15受体亚基α（IL-15Rα）和IL-15复合物偶联到抗PD-1帕博利珠单抗（帕博利珠单抗-IL-15Rα-IL15）的C末端，开发了一种原型。液相色谱-质谱联用（LC-MS）显示该分子上存在植物N-聚糖（GnGnXF、GnXF和Man9GlcNAc2），这可能会影响受体结合亲和力。然而，酶联免疫吸附测定（ELISA）表明，帕博利珠单抗-IL-15Rα-IL15与人PD-1蛋白的结合效力与市售帕博利珠单抗相当。在一项小鼠抗癌研究中，帕博利珠单抗-IL-15Rα-IL15（3 mg kg-1）表现出略好的肿瘤生长抑制效果，与市售帕博利珠单抗（5 mg kg-1）使肿瘤大小减少83%相比，其使肿瘤大小减少了94%，尽管差异无统计学意义。总之，使用植物分子农业成功生产了帕博利珠单抗-IL-15Rα-IL-15，它在解决mAb耐药性和增强IL-15有效载荷的免疫调节作用方面显示出前景。
