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

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.