Synthesis of 20α/β-hydroxyprogesterone-trimethyl lock-triglyceride-mimetic prodrugs and its bioavailability study in rats

• Published in: Bioorganic & medicinal chemistry Vol. 128; p. 118285
• Main Authors: Li, Liangyun, Yan, Shujing, Cheng, Yuexuan, Aisan, Muyasaer, Du, Shijie, Zhong, Chunhong, Chen, Chunli, Gao, Xiaoli
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.10.2025
• Subjects: 20-Hydroxyprogesterone; Animals; Biological Availability; Female; Hydroxyprogesterones - chemical synthesis; Hydroxyprogesterones - chemistry; Hydroxyprogesterones - pharmacokinetics; Liquid chromatography-tandem mass spectrometry; Molecular Structure; Prodrugs - administration & dosage; Prodrugs - chemical synthesis; Prodrugs - chemistry; Prodrugs - pharmacokinetics; Rats; Rats, Sprague-Dawley; Relative bioavailability; Tandem Mass Spectrometry; Triglyceride-mimetic prodrug; Triglycerides - chemistry; Trimethyl lock; Liquid chromatography-tandem mass spectrometry; Relative bioavailability; Trimethyl lock; 20-Hydroxyprogesterone; Triglyceride-mimetic prodrug
• ISSN: 0968-0896; 1464-3391; 1464-3391
• DOI: 10.1016/j.bmc.2025.118285

[Display omitted] •20α-DHP and 20β-DHP are bioactive progesterone diastereoisomers with natural progestin potential.•Four single-configured 20-DHP-TML-triglyceride prodrugs were synthesized and characterized.•A sensitive LC-MS/MS method quantified 20α/β-DHP in rat plasma.•TML release mechanism boosts 20-DHP bioavailability.•β-Configuration prodrugs with linker side chains show optimal bioavailability. 20α-hydroxyprogesterone (20α-DHP) and 20β-hydroxyprogesterone (20β-DHP), the C-20 hydroxylated metabolites of progesterone, constitute a pair of diastereoisomers of 20-hydroxyprogesterone (20-DHP) that demonstrate progesterone-like biological activities. This study employed the triglyceride (TG) −mimetic prodrug strategy and the trimethyl lock (TML) lactonization mechanism to chemically synthesize four 20-DHP-TML-TG-mimetic prodrugs (20α-DHP-TML-C5-TG, 20β-DHP-TML-C5-TG, 20α-DHP-TML-C5βMe-TG, and 20β-DHP-TML-C5βMe-TG) by coupling 20α-DHP or 20β-DHP with the TG backbone through two different TML-containing linkers. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established to determine the concentrations of 20α-DHP and 20β-DHP in rat plasma. Subsequently, pharmacokinetic experiments were conducted to investigate the pharmacokinetic parameters of 20α-DHP, 20β-DHP, and the four prodrugs in ovariectomized female rats after oral gavage. The results showed that compared with oral gavage of 20α-DHP, the relative bioavailability of 20α-DHP-TML-C5-TG and 20α-DHP-TML-C5βMe-TG increased by 9.6 and 21.8 times, respectively. Compared with oral gavage of 20β-DHP, the relative bioavailability of 20β-DHP-TML-C5-TG and 20β-DHP-TML-C5βMe-TG increased by 19.7 and 33.5 times, respectively. This enhancement may be attributed to the methyl side chain of the linker, which prolongs the prodrug’ retention time in the gastrointestinal tract, maintains a degree stability in vivo, and the TML structure in the linker main chain, which trigger rapid release of the parent drug once the prodrug enters the systemic circulation, thereby increasing the blood concentration. Compared with prodrugs of α configuration, prodrugs of β configuration have higher bioavailability, which may be due to their higher gastrointestinal stability or easier recognition by specific esterases in plasma. It can be concluded that TG-mimetic prodrugs with a TML structure and a methyl side chain in the linker can effectively improve the oral bioavailability of the parent drug, while the diastereomers of TG-mimetic prodrugs may have different in vivo processes.