Daruwati Isti, Gwiharto Abednego Kristande, Kurniawan Ahmad, Mahendra Isa, Achmad Tri Hanggono, Syaifudin Mukh, Muchtaridi Muchtaridi
Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, Indonesia.
Center for Applied Nuclear Research and Technology, Nuclear Energy Research Organization, National Research and Innovation Agency (BRIN), Indonesia.
Heliyon. 2021 Nov 18;7(11):e08438. doi: 10.1016/j.heliyon.2021.e08438. eCollection 2021 Nov.
Estradiol is a steroid hormone that works as an agonist estrogen receptor (ER). This compound is widely used as a ligand and bind specifically to the ERα. Radioligand binding assay is an in vitro method for drug development from natural products by synthesizing estradiol through radiolabeling using the radioiodination method. Synthesis of I-estradiol was perfomed by direct method using chloramine T as an oxidizer and by indirect labeling using I-histamine. The purity of chemical was determined by thin-layer chromatography and paper electrophoresis, as well as its stability for 30 days of storage in refrigerator, freezer and room temperature. The cellular uptake test of the radioligands from both methods was carried out with MCF7 and T-47D cell lines at 60 min. The results exhibited that I-estradiol was succesfully obtained with radiochemical purity greater than 95% and more stable in the refrigerator until 21 days than freezer and room temperature. I-estradiol and I-his-estradiol were internalized higher in T-47D cells than MCF7 cells (44.34 ± 5.93% vs. 17.27 ± 1.71% and 45.34 ± 6.42% vs. 4.92 ± 1.59%, respectively). Furthermore, the radioligands can be used to binding assay in determining the agonist or antagonist to ER of new drugs development.
雌二醇是一种类固醇激素，作为雌激素受体（ER）激动剂发挥作用。这种化合物被广泛用作配体，并特异性结合雌激素受体α（ERα）。放射性配体结合测定是一种体外方法，用于通过使用放射性碘化法进行放射性标记来合成雌二醇，从而从天然产物开发药物。I-雌二醇的合成通过使用氯胺T作为氧化剂的直接法以及使用I-组胺的间接标记法进行。通过薄层色谱法和纸电泳法测定化学纯度，以及其在冰箱、冷冻柜和室温下储存30天的稳定性。在60分钟时，使用MCF7和T-47D细胞系对两种方法得到的放射性配体进行细胞摄取试验。结果表明，成功获得了放射性化学纯度大于95%的I-雌二醇，并且在冰箱中比在冷冻柜和室温下更稳定，可保存21天。I-雌二醇和I-组氨酸雌二醇在T-47D细胞中的内化高于MCF7细胞（分别为44.34±5.93%对17.27±1.71%和45.34±6.42%对4.92±1.59%）。此外，放射性配体可用于结合测定，以确定新药开发中对雌激素受体的激动剂或拮抗剂。
