Study on 20-hydroxyprogesterone: Chiral resolution, content determination and progesterone-like activity

• Published in: The Journal of steroid biochemistry and molecular biology Vol. 244; p. 106592
• Main Authors: Cheng, Yuexuan, Yan, Shujing, Li, Liangyun, Du, Shijie, Zhong, Chunhong, Gao, Xiaoli, Chen, Chunli
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.11.2024
• Subjects: 20-alpha-Dihydroprogesterone - chemistry; 20-alpha-Dihydroprogesterone - metabolism; 20-hydroxyprogesterone; Animals; Chiral resolution; Content determination; Endometrium - metabolism; Female; Humans; Progesterone - blood; Progesterone - metabolism; Progesterone-like activity; Stereoisomerism; PR; Progesterone-like activity; 20-hydroxyprogesterone; P4; IHC; VEGF; HPO; 20-DHP; Content determination; FSH; Chiral resolution; GnRH; LH; HE; RT-qPCR; ELISA
• ISSN: 0960-0760; 1879-1220; 1879-1220
• DOI: 10.1016/j.jsbmb.2024.106592

20-hydroxyprogesterone (20-DHP) is a natural metabolite of progesterone which occurs with two diastereoisomers: 20α-DHP and 20β-DHP. They have drawn attention for their progesterone-like activity since the middle of the 20th century. However, the literature from that era bears witness to a lack of consensus regarding their specific effects. Considered that their stereoisomerism differences, it is essential to investigate their biological activities in vivo separately. In this study, we presented a chemical synthesis technique that yielded highly pure samples of 20α-DHP and 20β-DHP, and performed simultaneous content analysis. Subsequently, we examined and contrasted the progesterone-like properties of 20α-DHP, 20β-DHP, and a 1:1 mixture of 20α-DHP and 20β-DHP. The Morphological observations of the endometrium were conducted via Haematoxylin–eosin staining. Serum hormone levels were measured using enzyme-linked immunosorbent assays. Furthermore, real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were employed to analyse the relevant mRNA and protein expression, respectively. Our comparison revealed that 20α-DHP and P4 share identical progesterone-like actions, while 20β-DHP exhibits partial similarity. The progesterone activity varied when the two were combined in a 1:1 ratio. [Display omitted] •20α-DHP and 20β-DHP with excellent purity could be synthesized through an etherification-reduction-deetherification process.•An HPLC procedure has been developed to determining 20α-DHP and 20β-DHP concentration with remarkable reproducibility, linearity, specificity, and accuracy.•In terms of progesterone-like activity, 20α-DHP exhibits the same level as P4, while 20β-DHP shows partial similar. When the two are mixed in a 1:1 ratio, the resulting progesterone activity varies.