Acetohydroxy acid synthase (AHAS) is a key enzyme that catalyzes the synthesis of branched-chain amino acids, which is indispensable for the survival and growth of Mycobacterium tuberculosis (Mtb). Aim to discover new AHAS inhibitors from natural products, here we performed computer assistant target-based screening for MtbAHAS inhibitors using Discovery Studio on TCMSP and SELLECK libraries. Mtb-AHAS structure was first simulated and verified for docking, and 80 compounds with top LIBDOCK and CDDOCK scores were obtained. By experimental verification, four compounds namely Salvianolic acid A, Embelin, Celastrol and Wushanicaritin showed inhibition potency against Mtb-AHAS with IC50 ranging from 805.5 nM-32.36 mu M. The most potential inhibitor Celastrol exhibited bacteriostatic activity for both Mycobacterium smegmatis and Mycobacterium tuberculosis with MIC of 62.5 mu M and 80 mu M, respectively. This study revealed that Celastrol is the potential Mtb-AHAS inhibitor as an anti-tuberculosis lead compound.

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) remains a global public health threat, particularly due to dormant Mtb, which necessitates prolonged drug treatment. Mycobacterium tuberculosis malate synthase (MtbMS) is a key rate-limiting enzyme in the glyoxylate shunt, essential for the survival of dormant Mtb but absent in the host. Using target-based virtual screening and biochemical approaches, we identified novel natural inhibitors of MtbMS. Molecular docking by Schrödinger and subsequent manual selection identified 11 compounds as potential inhibitors. Molecular dynamics (MD) simulations and binding-free energy analysis (MM/GBSA) demonstrated high stability and binding affinity of MtbMS with Nordihydroguaiaretic Acids (NDGA) and Meso-NDGA. NDGA and Meso-NDGA by inhibition experiment exhibited half-maximal inhibitory concentrations (IC50) against MtbMS at 1.10 ± 0.01 μM and 14.29 ± 0.95 μM and by Isothermal Titration Calorimetry (ITC) showed binding constants (Kd) of 5.66 μM and 34.90 μM, respectively. Their minimum inhibitory concentrations (MIC) against Mtb H37Rv were 60.47 μg/mL and 30.24 μg/mL, respectively. In conclusion, natural products NDGA and Meso-NDGA are potent inhibitors of MtbMS and represent promising new scaffolds for combating dormant Mtb.

Purpose: To investigate the synergistic effect of bovine cateslytin-loaded nanoparticles (bCAT-NPs) combined with ultrasound against Candida albicans biofilm and uncover the underlying mechanism. Methods: bCAT-NPs were prepared by the double emulsion method, and toxicity was observed by the hemolysis ratio. The metabolic activity and viable cell biomass, morphology and membrane permeability of C. albicans biofilm were observed. The expression of ALS3 mRNA, the content of reactive oxygen species, was detected. Finally, bCAT structure was analyzed. Results & conclusion: The hemolysis ratio of the bCAT-NPs group was significantly lower than that of the bCAT group. bCAT-NPs combined with ultrasound significantly reduced biofilm metabolic activity, inhibited the formation of hyphae, decreased the expression of ALS3 mRNA and increased the intracellular reactive oxygen species content. In the in vivo experiments, the colony-forming units/ml in the ultrasound+bCAT-NPs group decreased, and a few planktonic fungal cells were observed.

Pharmacovigilance in China has experienced rapid development in the past 30 years. The implementation of Good Pharmacovigilance Practice in China since the end of 2021 heralds a new era of pharmacovigilance affairs, which puts forward higher requirements for the quantity and quality of pharmacovigilance personnel. This study aimed to preliminarily explore the current career situations of pharmacovigilance professionals working in China for pharmaceutical companies. A questionnaire was adapted from research in the USA and Europe with the help of several pharmacovigilance experts. Snowball sampling was used to conduct an exploratory survey to obtain the frequency of basic demographic information, work status, and career expectations of pharmacovigilance professionals working for pharmaceutical companies. The personnel engaged in pharmacovigilance work for pharmaceutical companies were mainly medical or pharmaceutical undergraduates within 3 years of graduation. Their work intensity and pressure were relatively high. The training provided by their universities and enterprises could not well meet their needs to improve their job competence. Although they were optimistic about pharmacovigilance and will not change their career, most of them were planning to change their employers. There was a gap between the demand and supply of pharmacovigilance personnel. Relevant regulatory authorities and industry associations should guide higher education institutions to collaborate with pharmacovigilance specialists to strengthen pharmacovigilance education for medical or pharmaceutical students, on the basis of which pharmacovigilance certification courses and continuing education courses can be developed. Meanwhile, pharmaceutical enterprises should consider reasonably adjusting work intensity and income to avoid a high turnover rate.

Tuberculosis has been the serious disease threatening human health and public safety due to the emergence of MDR and XDR-TB. Mycobacterium tuberculosis peptide deformylase (MtPDF) is a valuable target for antituberculotics. In order to discover new potential inhibitor candidates of MtPDF as leads for antituberculotics, Discovery Studio (DS) 2019 was used to perform molecular docking for virtual screening in silico with the bioactive compound library-I (L1700) against MtPDF. Six compounds with high docking scores and favourable ligand-protein interactions by LibDock and CDOCKER were selected for the evaluation of the inhibition potencies against MtPDF and Mycobacterium smegmatis. GST-6xHis tagged MtPDF was recombinant expressed and purified firstly by Glutathione Sepharose 4B, and secondly by Ni Sepharose 6 FF after the cleavage of human rhinovirus 3C protease. These compounds showed IC50 values from 0.5 mu mol/L to 112 mu mol/L against MtPDF, among which CUDC-101 bearing hydroxamic acid exhibited IC50 of 0.5 mu mol/L on MtPDF and MIC against Mycobacterium smegmatis of 32 mu g/mL, and Ixazomib Citrate with IC50 of 63 mu mol/L and MIC of 16 mu g/mL. CUDC-101 and Ixazomib Citrate are promising as the potential leads for antituberculotics.

Tuberculosis remains a global threat to public health, and dormant Mycobacterium tuberculosis leads to long-term medication that is harmful to the human body. M. tuberculosis isocitrate lyase (MtICL), which is absent in host cells, is a key rate-limiting enzyme of the glyoxylic acid cycle and is essential for the survival of dormant M. tuberculosis. The aim of this study was to evaluate natural compounds as potential MtICL inhibitors through docking and experimental verification. Screening of the TCMSP database library was done using Discovery Studio 2019 for molecular docking and interaction analysis, with the putative inhibitors of MtICL, 3-BP, and IA as reference ligands. Daphnetin (MOL005118), with a docking score of 94.8 and -CDOCKER interaction energy of 56 kcal/mol, was selected and verified on MtICL in vitro and M. smegmatis; daphnetin gave an IC50 of 4.34 μg/mL for the MtICL enzyme and an MIC value of 128 μg/mL against M. smegmatis, showing enhanced potential in comparison with 3-BP and IA. The interactions and essential amino acid residues of the protein were analyzed. In summary, natural daphnetin may be a promising new skeleton for the design of inhibitors of MtICL to combat dormant M. tuberculosis.

基于Discovery Studio软件分子三维结构可视化功能和分子对接模拟模块,设计了山奈酚(kaempferol)与3CL水解酶(3CLpro)的分子对接实验,让药学、中药学和医学相关专业硕士通过分子对接模拟过程,掌握中药中天然药物分子结构特征,了解锁钥原理与构效关系特征,认识酶抑制剂和靶蛋白的作用机制,了解中药现代化和从分子水平进行天然药物研究的新手段.

Ni2+-NTA-boosted magnetic porous silica nanoparticles (Ni@MSN) to serve as ideal support for bi-functional enzyme were fabricated for the first time. The versatility of this support was validated by one-step purification and immobilization of bi-functional enzyme MLG consisting of 3-Quinuclidinone reductase and glucose dehydrogenase, which can simultaneously catalyze both carbonyl reduction and cofactor regeneration, to fabricate an artificial bi-functional nanobiocatalyst (namely, MLG-Ni@MSN). The enzyme loading of 71.7 mg/g support and 92.7% immobilization efficiency were obtained. Moreover, the immobilized MLG showed wider pH and temperature tolerance and greater storage stability than free MLG under the same conditions. The nanosystem was employed as biocatalyst to accomplish the 3-quinuclidinone (70 g/L) to (R)-3-quinuclidinol biotransformation in 100% conversion yield with >99% selectivity within 6 h and simultaneous cofactor regeneration. Furthermore, the immobilized MLG retained up to 80.3% (carbonyl reduction) and 78.0% (cofactor regeneration) of the initial activity after being recycled eight times. In addition, the MLG-Ni@MSN system exhibited almost no enzyme leaching during biotransformation and recycling. Therefore, we have reason to believe that the Ni@MSN support gave great promise for constructing a new biocatalytic nanosystem with multifunctional enzymes to achieve some other complex bioconversions.

目的 分析中药注射剂说明书修订公告及其具体要求,以及据此开展的说明书修订备案情况,为中药注射剂说明书监管工作提供依据.方法 将中药注射剂说明书修订公告和现有中药注射剂数据、说明书修订备案情况对比.统计分析中药注射剂说明书修订范本/要求中警示语、禁忌、不良反应、注意事项各方面具体内容.结果 中药注射剂说明书修订公告数近年增加明显,但批文数量少的中药注射剂品种被公告的比例偏低.公告品种批准文号的整体未修订备案率为23.7％,未备案批准文号约82.8％在2020年面临再注册.30份说明书修订范本/要求中,警示语多数列出过敏性休克,所列不良反应累及的系统-器官数在4～10,肝肾系统损害少有被提及,只有2份的禁忌项涉及中医证候,注意事项中特殊人群和药物相互作用的信息较多,警示语、不良反应、禁忌、注意事项之间的关联度偏低.结论 需持续对中药注射剂特别是批文数量较少的品种进行说明书修订,并通过明确法律责任、说明书社会公开和药品再注册落实企业说明书修订和通知使用的责任.需根据系统的循证研究完善不良反应、警示语、禁忌的修订,可考虑将特殊人群用药和药物相互作用单列,并增强说明书各项目之间的联系.

将思维导图引入医用有机化学的教学实践.以芳香烃为例,从局部到整体,逐层设计并构建了苯及其衍生物的亲电取代反应、化学性质及芳香烃小结等多个层次的思维导图,引导学生对所学知识进行归纳总结和发散思维.并举例说明如何以小组合作方式构建思维导图,培养学生协作能力.形式多样的思维导图可以帮助学生在医用有机化学的学习中理清思路,培养思维习惯和建立思维方法,是一种有效的教学辅助工具.

为了认真贯彻"停课不停学"精神,文章从前期准备、在线课程教学活动的开展、通过监管和反馈保证教学质量三个方面论述了利用在线课堂实现医用有机化学"停课不停学"的实践.

Background: Compounds featuring furan nucleus exhibit diverse biological properties. Lots of furan derivatives have been explored as pharmaceutical compounds. Hence it is of great interest to explore furan derivatives and their precursors as antitumor agents. Objective: A series of novel furan derivatives and their precursors (1-36) were synthesized from α-haloketones and β-dicarbonyl compounds. Methods: The reactions between β-dicarbonyl compounds and α-haloketones under basic conditions produced tricarbonyls or dihydrofurans, which were then condensed into their corresponding furan products. Their potential antiproliferative activity in vitro against two human tumor cell lines-cervical (HeLa) and colorectal (SW620) was evaluated using CCK-8 assay. Compounds 1 and 24 were selected for Western blot analysis. Results: Pronounced anti-proliferative effect in the micromolar level was observed for compounds (1, 4, 17, 20, 21, 24, 27, 31 and 32) in HeLa cells, with their IC50 values ranging from 0.08 to 8.79μM. Additionally, furan compounds (24, 26, 32 and 35) had moderate to potent anti-proliferative activity against the SW620 cell line. Furthermore, the possible targets of these compounds were explored by Western blot analysis. The results indicated that the candidates (compounds 1 and 24) exhibited excellent antiproliferative activity, which may be mediated by promoting the activity of PTEN to suppress PI3K/Akt and Wnt/β-catenin signaling. Conclusion: Most of the furan derivatives and their precursors reported herein exhibited moderate to excellent anti-proliferative activity against HeLa cell line and/or SW620 cell line. Compounds 1 and 24, as well as their analogues may be developed as promising anti-cancer agents.

Baicalin (BCL) is a flavonoid compound with demonstrated potential therapeutic benefits, but the clinical advancements are limited mainly due to its poor aqueous solubility. A guest-host supermolecular inclusion complex of mono-6-O-α-maltosyl-β-cyclodextrin (G2-β-CD) with BCL was developed to improve the aqueous solubility of BCL. The resultant inclusion complex was characterized by DSC, FT-IR, P-XRD, 1HNMR, 2D-ROESY and SEM analysis. Solubility test suggested that G2-β-CD markedly improved the solubility of BCL and showed AL type phase diagrams. The spectral shifts suggested that the phenyl moiety is buried deeper than the chromen moiety and protrudes from the 6-hydroxyl side of G2-β-CD. Molecular docking verified the experimental findings and supported the insertion of benzene ring of BCL into G2-β-CD cavity via hydrophobic interactions and possible hydrogen bonds. The enhanced bioavailability of the inclusion complex was reflected in earlier Tmax, higher Cmax and larger AUC0-∞ than that of BCL after its oral administration. Taken together, these findings suggested that the solid inclusion complex BCL-G2-β-CD may serve as valuable BCL solid dosage form with good solubility and dissolution rate.

Based on the Discovery Studio (DS) software, the receptor/ligand-based pharmacophore model for TGFβR1 inhibitors was designed for undergraduates. This experiment project includes pretreatment of protein, molecular preparation of test sets, construction of pharmacophore model and result analysis. Through the innovative experiment, undergraduates can understand the new means of pharmaceutical research through receptor/ligand-based pharmacophore model process. Furthermore, undergraduates can deeply understand the related knowledge of pharmacophore.

As new drugs for the treatment of malignant tumors, transforming growth factor-beta receptor 1 (TGFβR1) antagonists have attracted wide attention. Based on the crystal structure of TGFβR1-BMS22 complex, the pharmacophore model A02 with two hydrogen bond acceptors (HBAs) and four hydrophobic (HYD) properties was constructed. From the common features of active ligands reported in the literature, pharmacophore model B10 was also generated, which has two aromatic ring centers (RAs) and two HYD properties. The two models have high sensitivity and specificity to the training set, and they are highly consistent in spatial structure. Combining the two pharmacophore models, two novel skeleton structures with potential activity were selected by virtual screening from the DruglikeDiverse, MiniMaybridge, and ZINC Drug-Like databases. Four compounds (YXY01⁻YXY04) with potential anti-TGFβR1 activity were designed based on the new skeleton structures. In combination with Lipinski's rules; absorption, distribution, metabolism, excretion, and toxicity (ADMET); and, toxicological properties predicted in the study, YXY01-03 with the novel skeleton, good drug-like properties, and potential activity were finally discovered and may have higher safety relative to BMS22, which may be valuable for further research.

化学计算软件是药学研究的重要工具之一.对化学计算软件引入药学专业课程教学的必要性进行探讨,并以Gau-ssian和Gaussian View为例,对沙利度胺分子的立体异构体、表观静电势和分子轨道教学进行展示.

Enzymes are biocatalysts. In this study, a novel biocatalyst consisting of magnetic combined cross-linked enzyme aggregates (combi-CLEAs) of 3-quinuclidinone reductase (QNR) and glucose dehydrogenase (GDH) for enantioselective synthesis of (R)-3-quinuclidinol with regeneration of cofactors in situ was developed. The magnetic combi-CLEAs were fabricated with the use of ammonium sulfate as a precipitant and glutaraldehyde as a cross-linker for direct immobilization of QNR and GDH from E. coli BL(21) cell lysates onto amino-functionalized Fe3O4 nanoparticles. The physicochemical properties of the magnetic combi-CLEAs were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and magnetic measurements. Field emission scanning electron microscope (FE-SEM) images revealed a spherical structure with numerous pores which facilitate the movement of the substrates and coenzymes. Moreover, the magnetic combi-CLEAs exhibited improved operational and thermal stability, enhanced catalytic performance for transformation of 3-quinuclidinone (33 g/L) into (R)-3-quinuclidinol in 100% conversion yield and 100% enantiomeric excess (ee) after 3 h of reaction. The activity of the biocatalysts was preserved about 80% after 70 days storage and retained more than 40% of its initial activity after ten cycles. These results demonstrated that the magnetic combi-CLEAs, as cost-effective and environmentally friendly biocatalysts, were suitable for application in synthesis of (R)-3-quinuclidinol essential for the production of solifenacin and aclidinium with better performance than those currently available.

ObjectivesBaicalin (BCL) has potential therapeutic benefits, but its clinical outcomes are restricted mainly because of low water solubility. This study sought to improve the water solubility of BCL by the formation of inclusion complex with -cyclodextrin (-CD).MethodsThe inclusion complex was studied by solubility test, differential scanning calorimeter (DSC), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), H-1 Nuclear magnetic resonance ((HNMR)-H-1) and scanning electron microscopy (SEM). Molecular docking was conducted to verify the experimental findings. The dissolution rate was determined by dialysis membrane method. In vivo absorption studies in rats were conducted and high-performance liquid chromatography (HPLC) was used to analyse the plasma level of BCL after oral administration.Key findingsThe DSC, FTIR, XRD, (HNMR)-H-1 and SEM findings suggested the formation of inclusion complex between BCL and -CD in 1 : 1 stoichiometry. Molecular docking demonstrated the insertion of benzene ring of BCL into -CD cavity by hydrophobic interactions and possible H-bond formation. Moreover, -CD markedly improved the solubility of BCL and displayed A(L)-type phase diagrams. The improvement in dissolution rate of the inclusion complex was reflected in the earlier T-max, higher C-max and larger AUC(0-t) than that of BCL after oral administration.Conclusions-cyclodextrin complex can be used as an effective formulation strategy for development of BCL-loaded delivery system with better therapeutic outcomes.

The geometry of L-Thr-TPPZnI was optimized using density functional theory (DFT) at the B3LYP level of theory with the mixed basis sets:6-31G(d) for C and H, and 6-311++G(2d, p) for Zn, Fe, N and O atoms.Based on the optimized results , the electron transition absorption wavelengths (λ) and rotational strengths (R), and electronic circular dichroism (ECD) spectrum were calculated using time -dependent densi-ty functional theory ( TD-DFT) with the same functional and basis sets.In these calculations , solvent effects of CHCl3 were included using the polarized continuum model ( PCM).Comparing with the experimental date , the calculated absorption peaks exhibit blue shifts and calculated ECD spectrum is in good agreement with experi -mental one.Detailed analyses of the dominant transitions reveal that all the cotton effects in the ECD spectrum are caused by the π→π*charge transfer transition.The circular dichroism band in 353 nm is dominated by the intermolecular charge transfer transition form porphyrin ring to amino acid residue.

采用密度泛函理论B3LYP方法和混合基组(Fe(Ⅱ)采用赝式基组Lan2dz;C、H原子采用6-31G(d);N、O采用6-311++G(2d,p))对卟啉亚铁(TPPFe(Ⅱ))、L-苏氨酸卟啉亚铁(L-Thr-TPPFe(Ⅱ))的电子结构特征进行了研究;为了模拟真实的体内环境和实验条件,理论计算采用了极化连续介质模型(PCM)模拟CHCl3和H2O的溶剂效应.在优化获得稳定构型的基础上,采用Multiwfn软件对获得的波函数进行了拓扑分析和静电势研究,获得了键鞍点电荷密度和分子表观静电势数据.利用VMD软件绘制得到了表现静电势分布图.研究发现:(1)苏氨酸残基中的羰基O原子与Fe(Ⅱ)原子形成配位键,H(7)与N(1)之间存在分子内氢键;以上作用使得卟吩环扭曲变形.(2)苏氨酸残基与Fe(Ⅱ)配位后可减弱O2和Fe(Ⅱ)的配位作用,有利于O2的离去.(3)溶剂效应主要使分子内氢键作用减弱,卟吩环扭曲程度增加;减弱苏氨酸残基与Fe(Ⅱ)的配位作用,增强O2与Fe(Ⅱ)的配位作用.(4) L-Thr-TPPFe(Ⅱ)的表观静电势的负值区域主要集中在O原子、N原子附近和苯环的轴面,分别体现了孤对电子和7电子的贡献;正值区域主要集中在苯环H原子和Fe(Ⅱ)附近区域;中心Fe(Ⅱ)处存在具有较大的表观静电势,容易与O2配位.