In the early stage of the project, it was found that natural pentacyclic triterpenes liquidambaric acid regulates the NEDD8 modification of Cullin2. This study aims to find more triterpenoid natural active molecules targeting Cullin family members and reveal its mechanism of action. Western blot was used to detect natural products that can significantly change the total protein NEDD8 modification and specific Cullin protein NEDD8 modification in cells; microscale thermophoresis (MST) was used to detect the direct binding of candidate small molecule oleanonic acid to TRAF family proteins, and the binding at the level of living cells was verified by cellular thermal shift assay (CETSA). Proximity ligation assay (PLA) was used to investigate the regulatory effect of oleanonic acid on the protein interaction between TNF receptor-associated factor 1 (TRAF1) and Cullin1 NEDD8 modified complex. Three pentacyclic triterpenoids were found to significantly inhibit NEDD8 modification in cells, among which oleanonic acid had the strongest effect on blocking NEDD8 modification. Different from the previous identification that liquidambaric acid regulates Cullin2/5, oleanonic acid can also specifically induce NEDD8-modified Cullin1 to transform into its unmodified form. And binding experiments showed that oleanonic acid could directly bind to TRAF1 at the level of cell lysate and living cells. Further mechanism studies found that oleanonic acid significantly changed the protein interaction between TRAF1 and Cullin1 NEDD8 modified complex. The above results indicate that oleanonic acid targets TRAF1 and regulates its interaction with NEDD8 modification complex to inhibit NEDD8 modification of Cullin.

Exosomes are a kind of endosomal vesicles that are secreted by most if not all living cells. Due to their capability of delivering a variety of cargos, such as tissue- or cell-specific proteins, lipids, and genetic materials, and their broad biological activities, exosomes have gained substantial attention as emerging therapeutics. Exosomes derived from mesenchymal stem cells (MSCs) and dendritic cells (DCs) are two types of exosomes that are widely studied. Many preclinical and clinical studies have shown that they have a satisfactory treatment effect in lung diseases, liver diseases, nervous system diseases, tumors, and other diseases. In addition, exosomes from macrophages, tumor cells, plant cells, and many other cells are getting more attention due to their therapeutic potential. Besides natural exosomes, research on engineered exosomes has also made plenty of progress. There have been several engineering methods of exosomes, such as targeting modification and loading of active ingredients. In this review, we summarize the research progress of therapeutic exosomes from different sources, and further discusses the application prospects of exosomes and possible challenges in the future.

Hepatitis B virus (HBV) represents a significant global public health challenge. Despite the availability of several approved drugs for hepatitis B treatment, the persistence of covalently closed circular DNA (cccDNA) renders HBV eradication elusive, thereby leading to disease relapse after drug withdrawal. This paper reviews the regulatory mechanisms of cccDNA formation, transcription and replication, and summarizes the research progress of related small molecule regulators from the perspective of medicinal chemistry.

Gut microbiota is a complex and dynamic system, and is essential for the health of the body. As the "second genome" of the body, it can establish communication with the important organs by regulating intestinal nerves, gastrointestinal hormones, intestinal barrier, immunity and metabolism, thus affecting host′s physiological functions. Short chain fatty acid (SCFA), known as one important metabolite of intestinal microbiota, is regarded as a significant messenger of the gut-organ communication, due to its extensive regulation in the body′s immunity, metabolism, endocrine and signal transduction. In this review, we summarize the interaction between gut-liver/brain/kidney/lung axis and diseases, and focus on the role and mechanism of SCFA in the gut-organ communication, hoping to provide new ideas for the treatment of the related diseases.

Berberine is a naturally occurring benzylisoquinoline alkaloid with a wide range of pharmacological activities, such as antibacterial, anticancer, hypolipidemic, antidiabetic and antidiarrheal. Although berberine has a wide range of curative effects, the extremely low bioavailability (< 1%) limits its clinical application. Pure berberine preparations have not yet been approved for any specific disease. The low oral bioavailability of berberine is mainly due to poor solubility caused by self-aggregation under acidic conditions, low permeability, P-glycoprotein (P-gp)-mediated efflux, and liver and intestine metabolism. To improve the oral bioavailability of berberine, researchers have adopted a variety of strategies, including the application of various nano-delivery systems, penetration enhancers and P-gp inhibitors, structural modifications, and development of berberine derivatives. Improving the oral bioavailability of berberine can improve the pharmacological activity of berberine, reduce the dosage, and then reduce the toxic and side effects. This review summarized the various pharmacological activities, metabolism progress and pharmacokinetic characteristics of berberine, the newly discovered berberine target intestinal microbiota and focused on the strategies to improve the oral bioavailability of berberine by improving solubility and permeability, inhibiting P-gp efflux, and structural modification. The research on berberine was prospected, which provided guidance for the in-depth study of berberine.

To prepare a progesterone pressure-sensitive gel patch combined with a microneedle to enhance drug release, HPLC was used to determine the preparation's progesterone content. One-way and orthogonal experiments were used to optimize the patch's prescription. Adhesion, sensory evaluation, cumulative release, and cumulative penetration were used as evaluation indices. Three microneedles with varying needle heights were made using 3D printing, and the cumulative penetration of the patch and microneedles was calculated and compared with the patch alone. The orthogonal experiments showed that the optimal prescription for the patches was Duro-Tak 87-2677 pressure-sensitive adhesive (87.5%), tributyl citrate (2%), isopropyl myristate (5%), dibutylated hydroxytoluene (0.5%), and drug (5%). The patches were prepared according to the optimized prescription, resulting in good patch formability and adhesion. In the transdermal penetration test, the cumulative penetration of the patch was 52.35 ± 7.88 μg·cm^-2 at 24 h, and the cumulative penetration of the patch in combination with 500, 750, and 1 000 μm microneedles was 226.01 ± 7.46, 278.78 ± 6.59, 422.95 ± 16.81 μg·cm^-2, respectively. The experiment was approved by the Experimental Animal Ethics Committee of Xinjiang Medical University (IACUC-20220725-8). The optimal patch prescription was screened through one-way and orthogonal experiments, and the transdermal penetration effect of patch and microneedle combination preparation was better than that of single use, which can effectively increase the in vitro transdermal penetration of the drug, and the above study provides a theoretical basis for the application of transdermal patches of progesterone.

Natural deep eutectic solvent(NDES) is a kind of deep eutectic solvents(DESs) which is composed of natural substances with good biocompatibility. Those substances can function as hydrogen bond donor and acceptor, such as choline, amino acids, sugars, etc. NDES have been widely used in many fields due to their advantages of low cost, easy preparation and environmental friendliness. It is especially suitable for the pharmaceutical industry because of its good biocompatibility and safety for use. In this paper, we firstly review the molecular simulation methods for current design of DESs from the formation principle. And then, the materials and preparation of NDES are reviewed and the physicochemical properties are further described. Finally, we review the current application of NDES in pharmaceutics including increasing drug solubility, promoting drug permeability and enhancing oral drug absorption, and meanwhile their future applications in pharmaceutics were also prospected.

Autophagy is a lysosomal degradation pathway, and plays a crucial role in cellular homeostasis, development, immunity, tumor suppression, metabolism, prevention of neurodegeneration, and lifespan extension. Thus, pharmacological stimulation of autophagy may be an effective approach for preventing or treating certain human diseases and/or aging. Here, combined with allosteric site identification methods, high-throughput virtual screening, and in vitro activity evaluation, we found that compound 10 can activate autophagy and has good anti-MDA-MB-231 cell proliferation activity (the half maximal inhibitory concentration IC₅₀=8.25±1.53 μmol·L^-1). Subsequently, molecular docking, molecular dynamics simulation, and immunoblotting assay demonstrate that compound 10 can target and activate beclin-1. In vitro studies have shown that compound 10 can induce autophagy-associated cell death in MDA-MB-231 cells. In addition, it was found that compound 10 can induce apoptosis in MDA-MB-231 cells. Taken together, we identified the candidate compound 10 as an effective and selective targeting beclin-1 to activate autophagy as a lead compound, which provide a reference for further development and optimization of small molecule drugs targeting beclin-1 to activate autophagy for clinical treatment.

This study aimed to explore the effects of Ziziphi Spinosae Seme (ZSS) and Fried Ziziphi Spinosae Semen (FZSS) on metabolites and intestinal flora structure in insomnia mice by combining metabolomics analysis and 16S rRNA gene sequencing. Animal experimentation was approved according to the Committee on the Ethics of Animal Experiments of Shanxi University of Chinese Medicine (approval number: 2021DW172). The prediction targets were validated in para-chlorophenylalanine (PCPA) induced insomnia mice with administration of ZSS and FZSS for 5 days, respectively. Then pentobarbital sodium induced sleeping test were performed to evaluate the synergistic sleep-aiding effect of ZSS and FZSS. The metabolic profile of serum from insomnia mice was analyzed by UPLC-Q-Orbitrap MS. Different metabolites were screened combined with multivariate statistical analysis. The relevant metabolic pathways and networks were constructed by MetaboAnalyst 5.0. Intestinal flora changes were detected by 16S rRNA sequencing technology. Animal study indicated that, compared to PCPA-induced insomnia model, ZSS and FZSS shortened the sleeping latency and increased the sleeping duration. The serum metabolomics results showed that, there are 36 potential biomarkers associated with insomnia were identified. Compared with the model group, 25 and 27 differential metabolites were identified in the ZSS and FZSS groups, respectively. Both ZSS and FZSS groups could significantly adjust to the common 20 differential metabolites. A total of 10 pathways are closely related to insomnia, which including amino acid metabolism, energy metabolism and lipid metabolism. 5 metabolic pathways are shared in ZSS and FZSS, including phenylalanine, tyrosine and tryptophan biosynthesis, cysteine and methionine metabolism and so on. Arachidonic acid metabolism is the unique metabolic pathway for ZSS to improve sleep; 3 metabolic pathways including glutamate and glutamine are the unique regulatory pathway for FZSS to improve sleep. PCoA analysis showed that the structure of ZSS and FZSS recovered to blank group, ZSS and FZSS can decrease the abundance of Pvotella and increase the abundance of Lactobacillus. LEfSe and correlation analysis between metabolites and differential intestinal flora showed that kynurenic acid, L-glutamic acid, D-proline are significantly positively associated with Gammaproteobacteria in ZSS and 5-HT, acetylcholine, L-methionine are significantly positively associated with Lactobacillus in FZSS. In conclusion, both ZSS and FZSS can influence metabolic pathways such as amino acids, energy and lipids and regulate the structure of intestinal flora, and then play a part in treating insomnia. However, ZSS and FZSS improve the quality of sleeping by regulating different metabolic pathways and regulating gut microbiota structure. The results of this study provide an experimental basis for illustrating the action mechanism of ZSS and FZSS in treating insomnia and provide a scientific basis for clinical rational medication guidance.

As a Ginkgo biloba extract preparation, shuxuening injection has a unique advantage in the prevention and treatment of acute and subacute stroke, but its main active ingredient is still unclear. Using a subacute model of stroke in mice constructed earlier, we further explored the contribution and mechanism of the two main components of total ginkgo flavonol glycosides and total ginkgolides in facilitating the neurofunctional recovery in stroke-induced mice. The pharmacodynamics was mainly evaluated by neurobehavioral changes, cerebral infarction volume, blood-brain barrier permeability and brain edema. The pathway and targets were predicted by transcriptome and network pharmacology. Finally, the mechanism was verified at the mRNA and protein levels. The results showed that the beneficial effect of total ginkgolides was greater than that of total ginkgo flavonol glycosides in both the pharmacodynamics and the regulatory mechanism of granulocyte adhesion and diapedesis involving granulocyte colony-stimulating factor(G-CSF), macrophage-1 antigen(MAC-1) and E-selectin. These findings suggest that shuxuening injection may improve the prognosis for mice with subacute stroke by down-regulating GCSF-mediated granulocyte adhesion and diapedesis pathway mainly through the total ginkgolide components. This finding is expected to provide reference for optimizing prescription and searching for natural drugs for targeting the treatment of ischemic stroke prognosis. The animal experiments in this study followed the regulations of Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine.