The discovery of drug targets is crucial for understanding the mechanism of drug action and facilitating the development of novel drug therapeutics. Microscale thermophoresis (MST), renowned for its minimal sample consumption, rapid detection speed, high sensitivity, and the absence of the need for protein immobilization, has emerged as a pivotal tool in drug target research. This article outlines the strategies for identifying drug targets, with a focus on elucidating the principles of MST and its application in drug target discovery. Furthermore, it summarizes the promising prospects of MST in this field. The applications of MST are found in both forward and reverse strategies for target discovery, enabling the identification of target proteins for compounds, single herbal ingredients, and compound prescriptions, as well as facilitating drug screening. Additionally, the integration of MST with other molecular interaction techniques, such as surface plasmon resonance (SPR), provides a potent arsenal for studying the interactions between drugs and their targets. Despite certain limitations, MST holds immense potential for application in the realms of novel drug development and drug mechanism research.

Pain management remains as a global public health issue, with 20% of the adult population worldwide impacted and many patients still suffer from unrelieved or undertreated pain. Classic opioids exert antinociceptive effects by activating μ opioid receptor (MOR), but they are associated with many side effects (including respiratory depression, addiction, and constipation) which have aggravated the ongoing and escalating opioid crisis in worldwide. Therefore, there is an urgent need for the development of novel analgesics free of MOR-mediated unwanted adverse effects. Multi-target therapeutics target two or more related targets in a disease network system simultaneously, which can potentially provide greater efficacy and improve treatment outcomes compared with single-target therapies. Due to the key role of MOR in regulating pain signal transmission, MOR-based multi-target ligands have been accepted as one of the mainstream strategies for designing new potent analgesics without side effects. In this review, recent advances in multitarget-directed drugs toward MOR are summarized, which may provide references for the development of potent and safer analgesics.

OBJECTIVE To design, synthesize and evaluate a series of 3', 4'-ethylendioxy chalcone derivatives as monoamine oxidase B (MAO-B) inhibitors, and summarize the structure-activity relationship (SAR). METHODS The targeted compounds were synthesized via Claisen-Schmidt condensation reaction starting from 6-acetyl-1, 4-benzodioxan and corresponding benzaldehydes. The inhibition of these compounds on human MAO-B (hMAO-B) was determined, and the inhibiting selectivity, dynamics and reversibility were investigated as well. The binding mode between active compounds and hMAO-B was revealed by molecular docking study. Additionally, the inhibitory effect of active compounds on the proliferation of BV2 cell line was determined by MTT assay. RESULTS Sixteen targeted compounds were successfully prepared. Most compounds showed good inhibitory effects on hMAO-B. Representative compounds 9 and 13 exhibited IC₅₀ values of 0.021 and 0.042 μmol·L^-1, respectively, which showed high inhibiting selectivity towards hMAO-B. Both compounds acted as competitive and reversible hMAO-B inhibitors. The main interactions between active compounds and hMAO-B were hydrophobic interaction and hydrogen bond. The most active compound 9 exhibited low cytotoxicity in BV2 cells. CONCLUSION This class of 3', 4'-ethylendioxy chalcone derivatives represent potential novel inhibitors of hMAO-B, and compound 9 could be further investigated as a potent lead for future studies.

OBJECTIVE To accurately distinguish deer-derived medicinal materials from four different position: Cervi Cornu Pantotrichum, Cervi Cornu, Cervi Cornu Degelatinatum and Deer hide. METHODS The extraction efficiency of two extraction solvent systems for four different kinds of deer-derived medicinal materials was compared, and the optimum enzymatic hydrolysis conditions of the samples were optimized to form an efficient and simple sample pretreatment process. A specific analysis method, combining chemical markers and characteristic polypeptides was established to identify four different position of deer-derived medicinal materials by using liquid chromatography-triple quadrupole mass spectrometry and mass spectrometry multi-reaction monitoring technology (MRM). And methodological experiments were conducted as part of this analysis method. RESULTS The optimized sample pretreatment method with high extraction efficiency and good enzymatic effect was successfully applied, and the corresponding characteristic polypeptide ion pairs were successfully detected in 24 batches of samples. CONCLUSION The distinction and identification method with high-level specificity, sensitivity and robustness of deer-derived medicinal materials were established. The innovative analysis method can be effectively used to evaluate the quality of deer-derived medicinal materials, which provides a reference for ensuring and improving the quality of deer-derived medicinal materials.

OBJECTIVE To determine the antioxidant activities of Gossampim Flos waterextracts,establish UPLC fingerprints and study the spectral effect relationship. METHODS Based on the scavenging rates of 2, 2-diazo-di (3-ethyl-benzothiazolin-6-sulfonic acid) diamiammonium salt (ABTS) and 1, 1-diphenyl-2-picrohydrazyl radical (DPPH), the fingerprint of different parts (petal, calyx and stamen) of 12 batches of Gossampim Flos were established by UPLC method, and similarity analysis were carried out. Grey correlation analysis (GRA), Spearman analysis, stepwise regression analysis and partial least squares regression(PLSR) analysis were used to study the relationship between antioxidant spectrum effect. RESULTS Eight chromatographic peaks were identified by fingerprint identification of water extracts from different parts of 12 batches of Gossampim Flos. Mangiferin, quercetin, rutin, vitexin and other compounds were the main active components of Gossampim Flos and its different parts for antioxidant activities. CONCLUSION The antioxidant activity of Gossampim Flos and its different parts are the result of the combined effect of multiple components. This study can provide a comprehensive reference for the basis and quality control of antioxidant active substances of Gossampim Flos and its different parts.

OBJECTIVE To predict the potential quality marker (Q-Marker) components of Suanzaoren Decoction based on the prototype chemical composition of in vivo by using network pharmacology. METHODS Firstly, the plasma samples and urine samples of rats were collected within 4 h and 24 h after intragastric administration of Suanzaoren Decoction, respectively. Then, based on the chemical components found in vivo, the key action targets and key pathway were obtained by network pharmacology, and the network map of "component-target-pathway" was carried out to predict the quality markers of Suanzaoren Decoction against insomnia, anxiety and depression. RESULTS A total of 20 prototype compounds were detected in rats. The 23 core targets were predicted and 14 prototype components associated with pharmacodynamic efficacy were screened through network pharmacology, the 14 components including jujuboside A, enoxolone, ferulic acid, magnoflorine, liquiritigenin, isoliquiritigenin, poricoic acid A, spinosin, coclaurine, timosaponin BII, glycyrrhizic acid, jujuboside B, liquiritin and senkyunolide A played multiple targets and multiple pathways to regulate insomnia, anxiety, depression. CONCLUSION It is predicted that the 14 chemical components are considered as potential quality markers of Suanzaoren Decoction by analyzing prototype chemical composition in vivo combined with network pharmacology. The results can provide reference for the quality control of Suanzaoren Decoction, and lay a foundation for elucidating the mechanism of its pharmacodynamic substances.

OBJECTIVE To explore the antiepileptic activity of QO-83, a novel KCNQ channel opener, by establishing and optimizing a 6 Hz corneal kinked model, and to provide a scientific basis for the application of QO-83 in drug-resistant epilepsy. METHODS The mice were divided into control group, model group, positive drug levetiracetam (100 mg·kg^-1) group, control drug carbamazepine (50 mg·kg^-1) group, and QO-83 low (2 mg·kg^-1), medium (4 mg·kg^-1), and high (6 mg·kg^-1) dose groups. The 6 Hz kindled mice were established, and drugs was injected intraperitoneally. After 30 min, 6 Hz corneal stimulation was applied again to observe its protective effect on the animal. And set up a series of behavioral experiments: tail suspension test, T-maze test, and new object recognition test, observe the immobility time, selection accuracy, and recognition index of mice respectively, to evaluate its impact on animal depressive tendencies and memory abilities. Immunofluorescence staining was performed on 6 Hz model mice to observe the expression changes of c-Fos protein and evaluate the protective effect of QO-83 on specific brain regions. Observe the expression changes of its target KCNQ2 protein and evaluate its impact on target protein expression. RESULTS The established model mice were not sensitive to the action of carbamazepine (50 mg·kg^-1), but effective against levetiracetam (100 mg·kg^-1). When given 2, 4 and 6 mg·kg^-1 QO-83, dose-dependent anti-epileptic effects can be observed, which significantly reduced the incidence and duration of seizures in ignited mice. The 4 mg·kg^-1 of QO-83 can significantly reduce the immobility time of epileptic mice in tail suspension experiments, increase the accuracy of mice in T-maze test, and increase the recognition index (RI) of mice in new object recognition test. QO-83 can significantly downregulate the expression of c-fos in the hippocampus and piriform cortex of epileptic mice, and upregulate the expression of KCNQ2 protein in the CA3, DG regions and their surrounding fibers and PIR regions of late-stage model mice. CONCLUSION Compound QO-83 has good anti drug-resistant epilepsy activity, and its protective effect on epileptic animals is better than the positive drug levetiracetam. At the same time, it also has the effect of improving the behavior of epileptic animals, demonstrating the therapeutic potential for drug-resistant epilepsy. The mechanism of action may be related to upregulating the expression of the target protein KCNQ2.

OBJECTIVE To prepare puerarin/daidzein porous nanocrystals using polyethyleneglycol (PEG)and chitosan(CS)as carrier aterialsand evaluate their physicochemical properties and in vitro characteristics. METHODS The drug loading rate and entrapment rate were used as indicators to optimize the prescription,and the insoluble drug puerarin-daidzein was encapsulated in the form of inclusion compound by two-step method β-CD-puerarin-daidzein-PEG-CS nanocrystals. The inclusion degree of puerarin-daidzein nanocrystals was determined by Fourier transform infrared spectroscopy, thermogravimetry, X-ray diffraction, scanning electron microscope, antioxidant and bacteriostatic tests to verify the feasibility of the preparation method. And investigate its drug release behavior in simulated gastrointestinal fluid and gastric environment, and fit the drug release model. RESULTS The inclusion compound prepared by saturated aqueous solution method was the best when the volume fraction of inclusion compound was 1.000% acetic acid. Its cumulative release rate is about 90%, drug loading and inclusion rate are (26.13±0.74)% and (78.39±2.23)%, respectively. Puerarin-daidzein nanocrystals have many and dense pore diameters, and have good antioxidant performance. Obvious inhibition zone against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa can be observed, showing good antibacterial performance. CONCLUSION Puerarin-Daidzein porous nanocrystals can significantly improve the dissolution of insoluble drug daidzein, which lays a foundation for its application in the field of biomedicine.

OBJECTIVE To prepare osthole(OST) nanocrystallization and evaluate its characterization. METHODS OST-NSs were prepared by a media grinding method, and the average particle size and polydispersity index (PDI) were used as indicators to optimize the formulation and process parameters. The crystal morphology, crystal form, and interaction between OST-NCs and stabilizers were characterized. Their surface wettability, equilibrium solubility, and in vitro dissolution in various media were investigated, and the f₂ value was used to evaluate the similarity of the dissolution curves. RESULTS The formulation and process parameters for preparing OST-NCs were as follows: OST (3%): PVP-VA64, sodium deoxycholate 25∶5∶1, the volume ratio of grinding beads (0.4-0.6 mm) to OST suspension was 2∶1, and it was ground at 450 r·min^-1 for 70 min, and 9% mannitol was used as a lyophilized protective agent to prepare OST-NCs. The stability of OST-NCs decreases with increasing temperature. After reconstitution, the particle size of OST-NCs was (354.2±9.857) nm, the PDI was (0.259±0.023), and the Zeta potential was (-22.2±0.896) mV. OST-NCs basically maintain the rod-shaped crystal structure. There is no interaction between OST and the stabilizer. The surface wettability of OST-NCs is better than that of OST and its physical mixture. The equilibrium solubility of OST-NCs in pure water is 231% that of OST. Among the six dissolution media, the dissolution rate and cumulative dissolution of OST-NCs were significantly better than those of OST and its physical mixture. Additionally, the f₂ values for the physical mixtures were greater than 50, whereas those for OST-NCs were less than 50, as compared with the dissolution curves of OST. CONCLUSION The results show that the reproducibility of OST-NCs is good during their preparation. The grinding temperature should be reduced, and the grinding time should be optimized. The dissolution behavior of OST-NCs is significantly different from that of OST. The reduction of particle size is the main reason for the improved water solubility of OST-NCs. These results suggest that the nanocrystallization strategy based on media grinding can effectively improve the water solubility and hydrophobicity of OST and has a good application prospect in the development of new OST drugs.

OBJECTIVE To establish an UHPLC-MS/MS method for the determination of six alkaloids in Tianma capsule. METHODS Phenomenex Kinetex C₁₈ (2.1 mm×100 mm, 2.6 μm) column was used for separation. The mobile phase consisted of methanol and 0.1% formic acid (containing 2.5 mmol·L^-1 ammonium acetate) solution (gradient elution). The flow rate was 0.3 mL·min^-1 and the column temperature was maintained at 35 ℃. Electrospray positive ion source was used for mass spectrometry detection by multiple reaction monitoring mode. RESULTS The liner range of calibration curve was 79.38-3 969 ng·mL^-1 (r=0.999 9), 18.60-929.8 ng·mL^-1 (r=0.999 8), 38.20-1 910 ng·mL^-1 (r=0.999 9), 1.264-252.8 ng·mL^-1(r=0.999 3), 1.196-239.2 ng·mL^-1(r=1.000 0) and 1.268-253.6 ng·mL^-1(r=1.000 0)for benzoylmesaconine, benzoylaconine, benzoylhypaconine, mesaconitine, hypaconitine and aconitine. The RSD of repeatability test (content) was 0.5%-3.2%. The average recoveries was in the range of 98.3%-105.3%. RSD was 1.0%-3.4%. CONCLUSION The method has good sprcificity, sensitivity and accuracy, and it is suitable for the analysis of six alkaloids in Tianma capsule.

OBJECTIVE To analyze the common pharmaceutical issues in application of generic nifedipine controlled-release tablets, in order to provide reference for the research and manufacture of this formulation. METHODS Based on some references and review experience, the common issues in the application documents of generic nifedipine controlled-release tablets about prescription research, production process research, and quality control, etc., were analyzed. RESULTS and CONCLUSION Nifedipine controlled-release tablets have certain technical difficulties and individual drug characteristics. A comprehensive study should be conducted on the basis of osmotic pump controlled-release technology, combined with the characteristics of the reference formulation and nifedipine controlled-release tablets, to ensure that the quality of the generic formulation is equivalent to that of the reference formulation.

OBJECTIVE To understand the overall situation of Tibetan medicines on the market, and to explore the method of comprehensive evaluation of Tibetan medicines. METHODS Data were extracted from the drug information network of the State Drug Administration and analyzed from the dimensions of manufacturers and varieties of Tibetan medicines. RESULTS The 373 items of proprietary Tibetan medicine were included, 80 items (proprietary Chinese medicine) were excluded, 19 manufacturers were involved, and 182 different dosage forms of proprietary Tibetan medicine were involved. The protection level of Tibetan patent medicine protection varieties includes 16 varieties, and the provincial standard document number involves 16 varieties. The classification of systemic treatment diseases is mainly concentrated in the treatment of gastrointestinal diseases, the treatment of skin, yellow water and rheumatic drugs, the treatment of liver diseases. CONCLUSION The preliminary comprehensive evaluation catalogue of proprietary Tibetan medicines can be conducted in steps according to the existing evaluation methods of proprietary Chinese medicines in terms of protected varieties, multi-varieties of manufacturers and classification of major diseases.The first batch of varieties include 25 Flavor Song shi pills, 25 Flavor Coral pills, 25 Flavor Pearl pills, 25 Flavor Guijiu pills, Zhituo Jiebai pills, 15 Flavor Black pills, 70 Flavor Pearl pills, Renqing Changjue, Coral 70 Flavor pills, Pain Relieving patch, and Nodikon capsules.