pharmaceutics, Mustansiriyah University, Baghdad, Baghdad Governorate, 10011, Iraq.
pharmaceutics, Al-Farahidi University, Baghdad, Baghdad Governorate, 10011, Iraq.
F1000Res. 2024 Mar 11;12:1197. doi: 10.12688/f1000research.140284.2. eCollection 2023.
Conventional chemotherapy results in severe toxic side effects due to affecting normal and cancer cells. The conjugation of chemotherapy with mAb will improve the chemotherapy selectivity towards cancer cells and at the same time will potentiate immune system to detect and kill cancer cells. The aim of the study was to prepare atezolizumab-pemetrexed conjugate using two types of linkers (linker conjugated with -NH2 of lysine amino acid in the mAb).
This study utilizes (for the first time) the mAb atezolizumab (AtZ) to prepare a new, selective conjugate carrier for pemetrexed (PMX) by using gamma amino butyric acid (GABA) as linker for the first time in comparison to the commonly used linker polyethylene glycol (PEG) using carbodiimide (EDC) / N-hydroxysulfosuccinimide (Sulfo-NHS) zero length cross linker. Stepwise evaluation for PMX-linkers linkage as well as mAb conjugates was evaluated by FTIR, HNMR, DSC, LC-MS, gel-electrophoresis as well as the anticancer activity against lung cells A549.
The work revealed that two molecules of GABA combined with PMX, which in turn conjugated with an average ratio of 4:1 with mAb, while one molecule of PEG combined with PMX, which in turn conjugated with mAb in the same average ratio. The IC for the prepared PMX-GABA-AtZ conjugate was 0.048 µM, which was much lower than PMX alone, antibody AtZ alone as well as PMX-PEG-AtZ conjugate in a dose and time dependent manner.
The potential use of such conjugate that selectively directed to the overexpressed lung cells antigen in a low dose leading to reduction of serious side effects of PMX and the cost of therapeutically AtZ mAb used.
传统化疗会因影响正常细胞和癌细胞而导致严重的毒副作用。将化疗与单克隆抗体结合使用将提高化疗对癌细胞的选择性，同时增强免疫系统检测和杀死癌细胞的能力。本研究的目的是制备阿特珠单抗-培美曲塞缀合物，使用两种类型的连接子（连接子与单克隆抗体赖氨酸氨基酸的-NH2 共轭）。
本研究首次利用单克隆抗体阿特珠单抗（AtZ），通过使用γ-氨基丁酸（GABA）作为连接子，与常用的连接子聚乙二醇（PEG）相比，通过碳二亚胺（EDC）/N-羟基磺基琥珀酰亚胺（Sulfo-NHS）零长度交联剂，首次为培美曲塞（PMX）制备新型选择性缀合载体。通过傅里叶变换红外光谱（FTIR）、HNMR、差示扫描量热法（DSC）、液质联用（LC-MS）、凝胶电泳以及对肺癌细胞 A549 的抗癌活性，对 PMX-连接子连接以及单克隆抗体缀合物进行逐步评估。
结果表明，有两个 GABA 分子与 PMX 结合，然后与单克隆抗体以平均 4:1 的比例结合，而一个 PEG 分子与 PMX 结合，然后以相同的平均比例与单克隆抗体结合。所制备的 PMX-GABA-AtZ 缀合物的 IC50 为 0.048 µM，远低于 PMX 单独、抗体 AtZ 单独以及 PMX-PEG-AtZ 缀合物，呈剂量和时间依赖性。
这种缀合物具有潜在的用途，可以选择性地针对过度表达的肺细胞抗原，以低剂量使用，从而减少 PMX 的严重副作用和治疗性使用 AtZ 单克隆抗体的成本。
