Zhejiang Xianju Pharmaceutical Co., Ltd.

Torreya grandis arils (TGAs), historically underutilized due to processing limitations, have gained value through the efficient extraction of essential oils. However, the post-distillation residue from this process remains rich in non-volatile bioactive compounds. In the present study, the post-distillation TGAs were systematically investigated and led to the identification of 32 diterpenoids, including twenty-three abietane-type 1-23, two pimarane-type 24-25, and seven labdane-type diterpenoids 26-32. Compounds 1-5 were elucidated to possess previously undescribed structures by extensive spectroscopic methods and quantum chemical calculations of NMR coupled with DP4+ probability analysis. Compounds 7, 8, 14, 19, 20, 23, 25, 29, 30, and 31 were obtained for the first time from the genus Torreya. In an in vitro bioassay, compounds 2, 3, 6, 7, 8, 10, 12, 26, 27, and 30 exhibited significant suppression of interleukin-6 (IL-6) and interleukin-1β (IL-1β) gene expression in LPS-induced RAW 264.7 macrophages, demonstrating anti-inflammatory potential.

Steroid drugs play a pivotal role in clinical therapeutics, with androsta-1,4-diene-3,17-dione (ADD) serving as a critical intermediate whose efficient biosynthesis relies on the catalytic activity of 3-ketosteroid-Δ1-dehydrogenase (KstD). Building upon the previously engineered KstD₂^ep variant, this study employed semi-rational design strategies to enhance KstD₂'s high-substrate-loading adaptability. Key residues (V332, L334, G534) were systematically identified through homology modeling and molecular docking, followed by constructing a combinatorial mutant library. Through alanine scanning and iterative screening of single/multiple-site mutations, the optimal mutant KstD₂^ep (V332E/L334T/G534V) demonstrated a 1.1-fold enhancement in catalytic efficiency compared to the KstD₂^ep. Molecular dynamics simulations confirmed significantly enhanced structural stability in the mutant. Whole-cell catalytic optimization revealed expanded operational tolerance to temperature fluctuations, pH variations, and co-solvent exposure. Implementing high-density fermentation coupled with fed-batch substrate supplementation, the process achieved a 1.5-fold increase in substrate conversion efficiency, yielding 117.75 g/L ADD. These advancements position the engineered variant as a high-potential candidate for scalable steroid biotransformation, addressing key barriers to enzymatic stability and process efficiency for enzyme-driven biosynthesis of steroidal pharmaceutical intermediates.

CZ1S injection is a novel, extended-release local anaesthetic formulation of ropivacaine, classified as a type 2.2 new drug, with potential for post-operative analgesia by subcutaneous infiltration and peripheral nerve blockade. This study aimed to validate the superior properties of CZ1S over ropivacaine hydrochloride injection and to evaluate the safety, tolerability, pharmacokinetic (PK) and pharmacodynamic (PD) profiles of a single dose of brachial plexus block with CZ1S in healthy Chinese adults. The tolerability and PK profile of CZ1S were investigated following a unilateral brachial plexus injection in 24 healthy adults. Safety assessments were performed throughout the study and the PD effects of CZ1S injection for unilateral brachial plexus block were also evaluated. The results showed that CZ1S was slowly absorbed after a single injection in healthy subjects, with PK parameters of ropivacaine (including C_max, AUC_t and AUC_∞) increasing with the CZ1S doses. CZ1S demonstrated a prolonged and non-constant release profile compared to a single 75 mg injection of ropivacaine hydrochloride, resulting in a smooth and sustained blood concentration. CZ1S significantly prolonged the duration of sensory and motor blockade, and further analysis revealed a correlation between PK and sensory nerve block. The PK profile of CZ1S was enhanced compared to that of ropivacaine injection, and it was deemed safe and well tolerated in healthy Chinese adults, suggesting its potential application in postoperative analgesia. CLINICAL TRIAL REGISTRATION: Chinadrugtrials.org.cn: CTR20231295 (registration date: 26 April 2023).