RNA-based therapeutics represent a pivotal modality in precision medicine, and their clinical advancement hinges on the development of highly efffcient and safe delivery systems. Given their superior safety, substantial drug-loading capacity, and scalability for industrial production, non-viral delivery systems have become predominant in the ffeld. This review systematically summarizes the approved RNA drugs and analyzes their core delivery strategies in detail. Furthermore, it discusses the key bottlenecks limiting the clinical translation of non-viral platforms, including low delivery efficiency, inefficient endosomal escape, short systemic circulation time, and insufficient extrahepatic targeting capabilities, as well as existing mitigating strategies, with the aim of promoting more effective clinical transformation of RNA therapeutics.

Epigenetics refers to the study of heritable changes in gene expression that occur without alterations to the DNA base sequence, primarily through mechanisms such as DNA methylation, histone modification, and non-coding RNA. Traditional genotoxicity testing focuses mainly on damage to the DNA sequence itself, making it difffcult to identify potential toxicity risks that do not involve sequence changes but may lead to long-term or transgenerational phenotypic effects. As research into epigenetic regulatory mechanism advances, the importance of epigenetics as a novel target for toxicity assessment has become increasingly evident. This article systematically reviews recently developed methods for detecting epigenetic alterations, including techniques for analyzing DNA methylation, histone post-translational modiffcation, and non-coding RNA. It compares the principles, advantages, and limitations of these methods and discusses their applicability across different application scenarios, providing a reference for selecting appropriate detection strategies in drug safety evaluation and related toxicological studies.

KSQ-4279(1), the ffrst ubiquitin-speciffc protease 1(USP1) inhibitor to enter phase Ⅰ clinical trials, is being evaluated for the treatment of multiple cancers, including ovarian and triple-negative breast cancer. Due to its novel and unique structure, 1 has emerged as a key lead compound for the design and scaffold modiffcation of this class of inhibitors. Systematic modifications and structure-activity relationship analyses of the four structural units of 1 have revealed the functional characteristics of each fragment. In recent years, structural modiffcations of 1 have yielded numerous “me-too” candidates. This review summarizes the modiffcation strategies and structure-activity relationship studies of 1, aiming to provide valuable insights and references for future research in this ffeld.

In order to effectively control the quality of bempedoic acid(1), seven related substances were synthesized, namely 2,2-dimethyl-7-(p-tolylthio)heptanoic acid(related substance A), (8R/S)-15-ethoxy-8-hydroxy-2,2,14,14-tetramethyl-15-oxopentadecanoic acid(related substance B), 8-oxo-2,2,14,14-tetramethylpentadecanedioic acid(related substance C), 8,14-dihydroxy-2,2,20,20-tetramethylhenicosanedioic acid(related substance D), 2,2,8,8-tetramethylnonanedioic acid(related substance E), (7R/S,8R/S)-8-hydroxy-2,2,14,14-tetramethyl-7-(p-tolylthio)-pentadecanedioic acid(related substance F) and (8R/S)- 8,15-dihydroxy-2,2,14,14-tetramethylpentadecanoic acid(related substance G). Among them, the related substances A and F and their synthetic routes were reported for the ffrst time. The seven related substances were synthesized via short routes under mild reaction conditions, with purities of no less than 95％ . Their structures were conffrmed by 1 H NMR,13 C NMR and HRMS. This paper could provide some references for the quality control of 1.

A new synthetic process of bilastine(1) has been reported. Commercially available 1-(2-ethoxyethyl)-2-(piperidin-4-yl)-1H-benzo[d]imidazole(2) was reacted with 2-chloroethan-1-ol to afford 2-[1-(2-hydroxyethyl)piperidin-4-yl]-1-(2-ethoxyethyl)-1H-benzo[d]imidazole(3). Compound 3 was transformed into 2-[1-(2-bromoethyl)piperidin-4-yl]-1-(2-ethoxyethyl)-1H-benzo[d]imidazole(4) by bromination reaction in the presence of N-bromosuccinimide/triphenylphosphine system. Bilastine methyl ester hydrochloride(6) was obtained through Nigishi cross-coupling reaction from 4 and methyl 2-(4-bromophenyl)-2-methylpropanoate(5), followed by saliffcation with hydrochloric acid. Finally, the target compound 1 was obtained through hydrolysis reaction of 6 and recrystallization from methanol. The total yield was 70.7％ with a purity of 99.7％ . The novel synthetic process has mild reaction conditions and simple operation, which makes it suitable for industrial production. The synthetic method for 6 from 4 and 5 was reported for the ffrst time.

A novel synthetic method of 10-aza-isoergoline scaffold compound, named 6,6a,7,8,9,10-hexahydro-4H-pyrazino[1,2-a]pyrrolo[4,3,2-de]quinoline(1), was reported in this paper. Commercially available 4-aminoindole(10) as starting material was subjected to N-dialkylated cyclization with bis(2-chloroethyl)amine hydrochloride(11) to get 4-piperazin-1-yl-1H-indole(12), followed by telescoped acylation with CbzCl in one-pot to provide benzyl 4-(1H-indol-4-yl)piperazine-1-carboxylate(5), which via Vilsmeier-Haack reaction in the presence of DMF/POCl3 to give benzyl 4-(3-formyl-1H-indol-4-yl)piperazine-1-carboxylate(6), and then sulfonylated with p-toluenesulfonyl chloride to furnish benzyl 4-(3-formyl-1-p-toluenesulfonyl-1H-indol-4-yl)piperazine-1-carboxylate(7). Compound 7 was condensed with p-tosylhydrazide to prepare benzyl 4-[1-tosyl-3-[(2-tosylhydrazineylidene)methyl]-1H-indol-4-yl]piperazine-1-carboxylate(8), then immediately telescoped intramolecular cyclization via C-H insertion of Carbene in one-pot to prepare benzyl 4-tosyl-6,6a,7,8,9,10-hexahydro-8H-pyrazino[1,2-a]pyrrolo[4,3,2-de]quinoline-8-carboxylate(9). After that, compound 9 deprotected via base and recrystallized from ethanol to afford 1 with an HPLC purity of 99.57％ and an overall yield of 8.1％ (based on 10). The improved route avoided special equipment and precious metal catalyst, further improved the process stability and product quality, so this process was extraordinarily suitable for lab-scale preparation and already provided a sufffcient hectogram-scale of qualiffed products.

In this study, a mouse model of ulcerative colitis(UC) was induced using dextran sulfate sodium(DSS), and the pharmacological activities of the supernatant fraction from the ethanol precipitation of the aqueous extract of Luohua Zizhu granules were verified, including reducing the disease activity index(DAI), alleviating colon shortening, relieving colonic pathological lesions, and decreasing the levels of inffammatory factors in serum. Subsequently, an ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method was employed to analyze the chemical constituents and the absorbed prototype constituents in serum of the supernatant. A total of 61 chemical constituents and 11 prototype constituents absorbed into serum were identiffed. This study provides a scientiffc basis for clarifying the pharmacodynamic material basis of the supernatant fraction from the ethanol precipitation of the aqueous extract of Luohua Zizhu granules and offers data support for further research on the mechanism of Luohua Zizhu granules in the treatment of UC.

This study investigated the bed collapse issue during the ffuidized bed granulation of Codonopsis Radix aqueous extract and evaluated the anti-collapse effects of dextrin, maltodextrin, and HPMC to optimize the granulation process parameters. The dynamic surface tension of the systems containing different excipients in the aqueous extract of Codonopsis Radix, glass transition temperature(Tg) and surface elemental composition of the prepared particles were measured to investigate their differences in anti-collapse mechanisms. The results indicated that dextrin and maltodextrin primarily prevented bed collapse by increasing Tg, thereby enhancing the material’s resistance to sticking. In contrast, HPMC exhibited superior anti-collapse performance by signiffcantly reducing the dynamic surface tension and preferentially accumulating on the droplet surface to form a protective barrier. A Box-Behnken design was employed to optimize the inlet air temperature, liquid feed rate, and atomization pressure. The mathematical models were established for particle yield, Hausner ratio, and span. A robust design space was created to determine the optimal range of above process parameters, ensuring the stability of the granulation process and the consistent quality of the particles. This research provides a theoretical basis for the industrial production of Codonopsis Radix formulations.

Taking Xiao’er Qixingcha prescription, Xiaochaihu prescription and Fenghan Ganmao prescription as models, composite particles were prepared from the aqueous extracts of each prescription via co-spray drying technique with hydroxypropyl-β-cyclodextrin(HP-β-CD) as a modifier. The properties of the spray-dried feed solution, the structure properties, surface free energy and flowability of the obtained composite particles, as well as the compactibility and disintegration time of the tablets were comprehensively characterized. The results showed that compared with the corresponding original extract powders, the yields of the composite particles with HP-β-CD were increased by 4.4％ - 11.7％, and their morphology were smoother and more regular. The particle size of the composite particles increased by 17.0％ - 25.0％ , and the cohesion index, caking strength, dispersive component, and surface free energy decreased by 8.3％ - 35.6％ , 12.7％ - 74.1％ , 9.9％ - 15.5％ , and 2.4％ - 4.0％ , respectively. The ffowability and compactibility of the composite particles were significantly improved, with the angle of repose, Carr index, Hausner ratio decreased by 3.5％ - 5.3％ , 4.6％ - 9.3％ , and 2.5％ - 5.1％ , respectively. Meanwhile, the tensile strength of the tablets compressed from the composite particles increased by 13.6％ - 45.7％, and the disintegration time decreased by 14.6％ - 16.2％. This study demonstrates that co-spray drying with HP-β-CD can signiffcantly improve the structure properties, reduce the surface free energy and enhance ffowability of the traditional Chinese medicine powders, and shorten the disintegration time of the tablets. This research can provide some references for the development of the co-processing modiffcation technology of traditional Chinese medicine powders and direct powder compaction technology.

In this study, using porcine bile after bilirubin extraction as the starting material, hyodeoxycholic acid was prepared via whole-cell hydrolysis, magnesium salt formation, and subsequent puriffcation. Orthogonal experiments were employed to optimize the key process parameters of whole-cell hydrolysis, magnesium salt precipitation, and ethyl acetate reffning. The optimal conditions were determined as follows: hydrolysis was performed at 37 ℃ and pH 7.0 with 1.0％ whole-cell bacterial suspension; magnesium salt was prepared at 55 ℃ and pH 9.0 – 10.0 with 1.5％ magnesium sulfate; purification was carried out by refluxing with 8-fold mass of ethyl acetate at pH 6.0 for 1 h. By the optimized process, the extraction yield of hyodeoxycholic acid reached up to 90 ％ , with a melting point of 195 ℃ . These results indicate that this process is stable, controllable, and suitable for industrial production.

A UPLC-MS/MS method was established for the simultaneous determination of four potential mutagenic impurities in vonoprazan fumarate tablets, namely 1-[5-(2-fluorophenyl)-1-(pyridine-3-sulfonyl)-1H-pyrrol-3-yl]-Nmethylnitrosamine, 3-[[2-(2-fluorophenyl)-4-[(methylamino)methyl]-1H-pyrrol-1-yl]sulfonyl]pyridine-1-oxide, N-[[5- (2-fluorophenyl)-1-(pyridine-3-sulfonyl)-1H-pyrrol-3-yl]methylene]methylamine oxide, and N-[[5-(2-fluorophenyl)- 1-(pyridine-3-sulfonyl)-1H-pyrrol-3-yl]methyl]-N-methylhydroxylamine. A Waters ACQUITY UPLC Peptide BEH C18 column(2.1 mm×150 mm, 1.7 μm) was used with 0.1％ formic acid solution as mobile phase A and acetonitrile as mobile phase B for gradient elution. An electrospray ionization source was employed in the positive ion mode with multiple reaction monitoring. The results demonstrated that the four impurities showed good linearity in the concentration ranges of 0.13 – 3.89, 1.94 – 58.07, 1.77 – 53.25 and 1.83 – 54.99 ng/mL, respectively. The LODs were determined to be 0.08, 0.08, 0.04 and 0.37 ng/mL, and the average recoveries were 97.9％ , 96.2％ , 102.6％ and 96.6％ , respectively. The established method is accurate, sensitive and convenient, and can provide a reference for the preparation process development and quality control of vonoprazan fumarate raw material and tablets.

An LC-MS method for the determination of five potential genotoxic impurities in tedizolid phosphate was established. The ACQUITY UPLC BEH C18 column(2.1 mm×100 mm, 1.8 μm) was used with electrospray ionization and multiple reaction monitoring mode. The analysis was performed using gradient elution with 0.1％ formic acid aqueous solution as mobile phase A and acetonitrile as mobile phase B. Three potential genotoxic impurities and two nitrosamines showed good linearity(r>0.990) in the ranges of 3.0 - 155.6 and 14.4 - 742.5 ng/mL, respectively. The average recoveries of all impurities were 87.9 ％ - 104.1 ％ . The established method is simple and accurate, which can be used for the determination of potential genotoxic impurities in tedizolid phosphate.

This study investigated the effects of packaging containers and sterile oral applicators on the oral trivalent reassortant live-attenuated rotavirus vaccines, and evaluated the applicability of the containers. Three batches of the vaccine were fflled and sealed in neutral borosilicate glass vials with halogenated butyl rubber stoppers, then stored at (25±2)℃ for 6 months and at 2 - 8 ℃ for 18 months, followed by routine tests, migration tests, adsorption tests and safety studies. The results showed that the appearance, ffll volume, abnormal toxicity, sterility test and identiffcation test of the samples all met the qualification criteria; the osmolality and pH value remained stable; the migration amount of elements increased but was below the safety limit; the particulate matter amounts was lower than the standard in Chinese Pharmacopoeial 2025 Edition. The virus titer of the vaccines decreased after storage at (25±2)℃ for 3 months and at 2 - 8℃ for 12 months. A simulated administration test was conducted by connecting a vaccine with a sterile oral applicator, and the results indicated that there were no signiffcant changes in the quality indicators of the vaccines within 1 h(P>0.05). This study demonstrated that the impacts of the selected packaging containers and applicators on the vaccine quality were acceptable, with good compatibility between them and the vaccines.

An ion chromatography method was developed for the simultaneous determination of glucose, mannose and fructose in erythritol. The analysis was performed on a CarboPac PA20 guard column(3.0 mm×30 mm) and a CarboPac PA20 analytical column(3.0 mm×150 mm), using gradient elution with water and 100 mmol/L sodium hydroxide solution as the eluent. The column temperature was set at 30 ℃ with a flow rate of 0.4 mL/min, and the detection was carried out by an integrated pulsed amperometric detector. Glucose, mannose and fructose showed good linearity in the range of 0.05 - 0.50 μg/mL, with average recoveries of 84.3％ - 101.5％. The results of 13 batches of samples demonstrated that the residues of the three reducing sugars were generally well controlled in quality in erythritol, though slight differences were observed among samples from different sources. For amine-containing pharmaceutical preparations, the detection and control of reducing sugar impurities should be emphasized when erythritol is used as an excipient.