Parkinson's disease (PD) is the second most common neurodegenerative disease of the central nervous system. It is currently believed that PD is related to factors such as age, gender, family inheritance, gene mutation and environment. The pathogenesis of PD is complex and is related to dysfunction and loss of dopaminergic neurons, involving accumulation of α-synuclein, neuroinflammation, oxidative stress, mitochondrial dysfunction and excessive accumulation of neuromelanin. In many ways, various factors act both independently and through cross-promotion, resulting in an ongoing pattern of brain tissue damage and progressing PD pathology. This article reviews the recent research on the pathogenic factors and pathogenesis of PD and explores new ideas and potential targets for PD treatment and drug development.

Snake bite is a common acute and severe disease in tropical and subtropical regions, and its public health importance has been largely neglected. Snake venom is a complex mixture of active proteins, polypeptides, and other toxins. Many of these components can target multiple ion channels, cell receptors, and membrane transporters. Compared with traditional small molecule drugs, the proteins and polypeptides from snake venom have stronger specificity and affinity to targets and are especially suitable for novel drug design. The current studies show that snake venom and its components have great potential for development as leading compounds of new drugs. In this paper, the recent advances in main components, toxic effects, and detoxification strategies of snake venom, as well as its pharmacological activities and medical application are reviewed. The aim is to provide reference for clinical diagnosis and treatment of snake bite and development of new drugs based on snake venom.

Angiogenesis is the formation of new capillaries from pre-existing vasculature, which plays a critical role in several diseases. Under the normal physiological conditions, only about 0.5% of endothelial cells (ECs) undergo mitosis, while the most ECs are in a resting state. Angiogenesis is a dynamic process in which ECs shift from resting to activated state, including three basic steps:① excessive vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF), platelet-derived endothelial growth factor (PDGF) and other pro-angiogenic factors secreted by ECs can promote the germination of ECs in the original blood vessels; ② the sprouts are continuously elongated through the proliferation of ECs and the degradation and migration of basement membrane. At this time, the ECs present two phenotypes with filamentous feet and strong proliferation ability; ③ the buds are continuously elongated to form a tubular structure and connect with adjacent blood vessels, and the junction is wrapped by wall cells and basement membrane to form new blood vessels. Nowadays, angiogenesis has become a target for clinical treatment of multifarious diseases. On one hand, anti-angiogenesis is used to treat various diseases with excessive angiogenesis, such as cancer, atherosclerosis, and diabetic retinopathy, etc. On the other hand, the diseases caused by insufficient angiogenesis, including myocardial infarction, myocardial ischemia/reperfusion injury, stroke, wound long-term healing and other ischemic diseases can be improved by pro-angiogenesis therapy. Large numbers of researches have shown that many active ingredients of traditional Chinese medicine can effectively treat the previously mentioned diseases by regulating angiogenesis in different ways. Therefore, the anti-and pro-angiogenesis effects of some active ingredients derived from traditional Chinese medicine and their mechanism were summarized in this manuscript, arming to provide theoretical basis for the development of new drugs for the treatment of angiogenesis-related diseases.

Ursodeoxycholic acid (UDCA) is an essential drug for the treatment of cholestatic liver diseases. As the most important representative of endogenous drugs, the metabolism and disposition of UDCA in human is characterized by both host-gut microbial co-metabolism and hepato-billilary-intestinal circulation. These distinct metabolic and pharmacokinetic features have brought great challenges into the bioequivalence (BE) evaluation of UDCA generic formulations. These challenges include not only biopharmaceutical problems derived from the unique physiochemical properties of amphiphilic molecules and the large single dose, but also the drug metabolism and pharmacokinetic problems associated with endogenous metabolism, long terminal half-life, high inter-and intra-individual variations, as well as accurate determination of UDCA and its metabolites. This review summarized academic and industrial literatures about the clinical pharmacokinetics and endogenous metabolism of UDCA. Current guidelines and technical challenges of UDCA BE studies were extensively discussed. Knowledge summarized in this review is expected to provide valuable reference for the development of UDCA generic formulations.

The 2-oxoglutarate-dependent dioxygenase (2-ODD) gene is regarded as the key enzyme gene involved with aryl naphthalene lignan-podophyllotoxin synthesis. To study the expression pattern and function of the Sc2-ODD gene, a full-length cDNA of the gene was cloned. Bioinformatic analysis, the expression pattern, and prokaryotic expression and purification were implemented. The open reading frame of Sc2-ODD gene was 1 077 bp and encoded 358 amino acids with a molecular weight of 40.16 kD. The Sc2-ODD protein contained the conserved 2OG-FeII-oxy sequence of the 2-ODD protein. The results of phylogenetic analysis revealed that Sc2-ODD is most closely related to Corchorus olitorius 2-ODD. qRT-PCR results showed that Sc2-ODD expression displayed obvious up-regulation at the fruit-swelling stage, then down-regulation in the fruit-coloring period. The Sc2-ODD gene was cloned into the bacterial expression vector pGS21T, the recombinant Sc2-ODD protein was expressed in Escherichia coli Rosetta (DE3) cells and the fusion protein was obtained and purified by GST fusion protein purification technology. This study will lay a foundation for further research on the function and expressional regulation of the Sc2-ODD gene in the aryl naphthalene lignans biosynthesis pathway, and also provides a scientific basis for improving the lignan content and the medicinal quality of Schisandra chinensis using plant genetic engineering.

The pharmacodynamic material basis and mechanisms of Ju-Hong Tan-Ke liquid (JHTKL) for its anti-tussive, anti-asthmatic and expectorant effects were investigated by using network pharmacology. We collected, screened, and predicted potential targets and signaling pathways for 24 compounds in the 8 herbs of JHTKL and grouped them according to their efficacy. Combined with the evidence analysis in the literature database, we explored molecular mechanisms of the components of this formula in different diseases and analyzed their compatibility laws. To verify the network analysis results, we used software to perform molecular docking of a part of the pivotal targets with their corresponding compounds. The results show that the main active ingredients in JHTKL may be naringin, L-ephedrine, glaucogenin C, amygdalin, deoxyschizandrin, neotuberostemonine, pachymic acid and glycyrrhizic acid. Moreover, efficacy groups of JHTKL may play a role by acting on pivotal gene targets such as the muscarinic acetylcholine receptor M1, acetylcholinesterase, beta-2 adrenergic receptor, prostaglandin G/H synthase 2, tumor necrosis factor, epidermal growth factor receptor and biological pathways such as the neuroactive ligand-receptor interaction, cholinergic synapses, calcium signaling pathway, NF-kappa B signaling pathway, MAPK signaling pathway, and PI3K-Akt signaling pathway. In this study, we have confirmed the pharmacodynamic material basis and mechanisms of JHTKL by using network pharmacology, laying a foundation for improving the quality standards of JHTKL and providing a reference basis for its potential expansion in clinical applications.

Eighteen dihydroartemisinin-fluoroquinolone molecules conjugated with L-homoserine were designed and synthesized using fragmented drug splicing approaches. The in vitro activities of the synthesized conjugates against Mycobacterium tuberculosis (MTB) and the lipid-lowering target PCSK9 were evaluated. The bioassay test results showed that most of the synthesized molecules had anti-tuberculosis (anti-TB) activity. Five compounds showed greater than 80% inhibitory activity against MTB in the replication state and three compounds exhibited more than 50% inhibitory activity against H₃₇Rv in the non-replication state. A structure-activity relationship analysis demonstrated that TM2 series compounds (Boc protection) have better anti-TB activity than TM1 series compounds (Cbz protection). There were 13 compounds with strong inhibitory activity toward PCSK9 (>73%) and TM1-3 compounds reached 92%. The determination of physical parameters showed that all the molecules are largely nontoxic. The structure-toxicity relationship analysis showed that the safety of TM2 is higher than that of TM1 in all parameters, perhaps related to the protecting group of the amino acid in the target molecule, and provides new ideas for the design and structural modification of subsequent molecules. This study sets a precedent for L-homoserine as a linking structural unit in multi-target drug molecules.

Chalcone synthase (CHS) is the rate-limiting enzyme involved in the biosynthetic pathway of flavonoids in Glycyrrhiza uralensis. It plays an important role in the regulation and control of flavonoids biosynthesis. In this study, X-ray irradiated G. uralensis samples with high or low content of flavonoids were studied. The CHS gene polymorphism in these samples were analyzed, the specific haplotypes were identified, and CHS function was parsed. 109 CHS cDNA sequences with a length of 1 170 bp were cloned, 220 variable sites (116 missense mutation sites) were found and 85 haplotypes were identified, which encoded 65 amino acid sequences with 96 variable sites. aa-20 and aa-45 were the most common amino acid sequences in samples with high flavonoid content, while aa-11 was the most sequence in samples with low flavonoid content. Molecular docking results showed that the mutation sites at 383 in aa-20 and 229 in aa-45 were related to substrate binding, while the mutation sites at 383 and 229 in aa-11 were not involved. Therefore, we speculate that the two mutation sites have significant influence on the function of CHS. We analyzed a large number of CHS cDNA sequences and identified the important functional sites, which will provide a basis for further functional studies. This paper will provide ideas for further research of the molecular regulation of the flavonoid biosynthetic pathway in G. uralensis.

Drug metabolites in the systemic circulation can be closely related to the safety or efficacy of drugs, so it is necessary to evaluate the pharmacokinetics of both the parent drug and its major metabolites in plasma. Bisthianostat, a novel histone deacetylase (HDAC) inhibitor, is currently under development. An LC-MS/MS method was developed and validated for the simultaneous determination of bisthianostat and its hydrolyzed N-hydroxyamide metabolite M351 in human plasma to evaluate their pharmacokinetic characteristics in humans. After extraction from the plasma by acetonitrile-induced protein precipitation, the analytes and endogenous substances were separated on a Waters BEH C₁₈ column (2.1 mm×50 mm, 1.7 μm). The mobile phase consisted of acetonitrile and 5 mmol·L^-1 ammonium acetate (containing 0.2% formic acid, v/v) for gradient elution. Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 367.1→235.0 for bisthianostat, m/z 352.1→207.0 for M351, m/z 371.1→235.0 for d₄-bisthianostat, and m/z 357.1→208.0 for d₅-M351. The method was linear over a concentration range of 2.00-2000 ng·mL^-1 for bisthianostat and 4.00-4 000 ng·mL^-1 for M351. The results of quality control samples showed that the intra-and inter-day precision were no more than 6.2% for bisthianostat and 6.8% for M351. The accuracy ranged from -1.1% to 4.3% for bisthianostat and -0.5% to 4.9% for M351. The pharmacokinetic results show that after a single oral administration of 100 mg bisthianostat, the time to peak (t_max) of M351 in the plasma of three patients with tumors was significantly longer than that of the parent drug (t_max was 4.00 h and 0.67 h, respectively), and the C_max and plasma exposure of M351 were about 1.7 times and 11 times higher, respectively, than that of the parent drug. This clinical trial was approved by the society of ethics and conducted in Renji Hospital, Shanghai Jiaotong University School of Medicine.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen that caused the global COVID-19 outbreak. The 3C-like protease (3CL^pro) of SARS-CoV-2 plays a key role in virus replication and has become an ideal target for antiviral drug design. In this paper, we report the validation and use of bioluminescence resonance energy transfer (BRET) technology to establish a cell-based assay for screening for SARS-CoV-2 virus 3CL protease inhibitors. The results show that the method is able to monitor the cleavage efficiency of 3CL protease with good reproducibility (Z' factor is 0.59), and is consistent with antiviral activity analysis in cell culture. This work demonstrates that this method can be applied to the screening and evaluation of 3CL protease inhibitors, providing a powerful tool for the development of new drugs.