New drug research and development is a technology-intensive industry with high investment, high cycle and high risk. In recent years, with the rapid development of modern disciplines such as omics technology, bioinformatics, high-throughput and high-content screening, and artificial intelligence, the research and development of small-molecule drugs has presented a new paradigm characterized by "integrated medicinal chemistry". This review summarizes new enabling drug discovery technologies, the emergence of new subfields formed through integration innovations and practical chemistry toolbox in the field of medicinal chemistry.

At present, majority of the small molecular drugs used in clinics target proteins, they exert the efficacy through the binding to specific sites on the target protein. However, the "druggable" protein targets account for a small portion of the total number of proteins, and "non-druggable" proteins account for 80%, because of not having suitable drug binding sites. In the central rule, RNA is located in the upstream of proteins and controls the transcription of proteins. The research of small molecule drugs targeting RNA can solve the problem of protein "undruggable proteins" in some extent. This review summarizes the representative research achievements of small molecular drugs targeting RNA in recent years, and the screening methods applied to this field, with the focuses on the latest progress of small molecular drugs targeting novel coronavirus RNA.

The overexpression of doublecortin-like kinase 1 (DCLK1) is closely related to the occurrence and development of various malignant tumors. Discovery of novel anti-tumor agents targeting DCLK1 represents a hot spot in this field. So far, several DCLK1 small molecule inhibitors with excellent anti-tumor activity in vitro and in vivo have been designed and synthesized, which is expected to provide a new strategy for tumor therapy. This article reviews the research progress in the discovery, structure type, structural optimization, biological activity and mechanism of action of DCLK1 small molecule inhibitors, and provides research basis for the development of new anti-tumor small molecule inhibitors targeting DCLK1.

Heterotrimeric G proteins are classes of signal-transducing proteins that bind to guanine nucleotides and possess GTP hydrolase activity. G proteins are composed of three subunits α, β, and γ, and are considered as the "molecular switch" in the GPCR signaling pathway. The abnormal activation of G protein is strongly related to diseases such as uveal melanoma, asthma, et al., making directly targeting G protein as an promising strategy for combating diseases. In this review, the classification and physiological functions of G protein are briefly described, and the research progress of G proteins in diseases, G protein modulators are reviewed.

Oxidative stress is a redox imbalance in the body, which is one of the important factors leading to tissue damage and diseases. The nuclear factor E2-related factor 2 (Nrf2)-Kelch like ECH-associated protein 1 (Keap1) signaling pathway is not only an important defense system against oxidative damage, but also one of the key signaling pathways of the antioxidant capacity. Numerous studies have shown that targeting the Keap1-Nrf2 signaling pathway to activate Nrf2 has become an effective strategy for the treatment of oxidative stress and related diseases. Using small molecules to directly block the Keap1-Nrf2 protein-protein interaction (PPI) is one of the important directions for activating Nrf2 and exerting the cytoprotective effect, which can avoid the potential side effects of covalent modification of Nrf2. On the other hand, the Keap1 is an efficient E3 ubiquitin ligase that has been used in the design of proteolysis targeting chimeras (PROTACs). This review summarizes the research progresses of Keap1-Nrf2 protein interaction inhibitors and degraders based on the Keap1 E3 ubiquitination system in recent years.

Lysine-specific demethylase 1 (LSD1) plays vital roles in cell stemness, differentiation, cell motility, metabolic control and epithelial-mesenchymal transition, which is closely associated with tumorigenesis processes including cell proliferation, invasive, metastasis and poor prognosis. Besides, LSD1 also contributes to the occurrence of other diseases such as neurodegenerative diseases and viral infections. Since 2013, the irreversible inhibitors including tranylcypromine, ORY-1001, ORY-2001, GSK-2879552, IMG-7289, INCB059872, TAK-418, LH-1802 and reversible inhibitors including CC-90011 and SP-2577 have been approved for clinical assessment. This review comprehensively summarizes the clinical research of LSD1 drug candidates and briefly discusses the prospects, opportunities and challenges of LSD1-targeted drug discovery, aiming to provide a landscape for the related drug development.

The urate transporter 1 (URAT1) which controls urate reabsorption is a membrane transporter in the apical membrane of human renal proximal tubule epithelial cells. It was found that about 90% of patients suffer from hyperuricemia due to insufficient uric acid excretion. Therefore, the development of URAT1 inhibitors that can reduce the level of serum uric acid in vivo by enhancing renal urate excretion has been a hot spot in seeking anti-gout drugs in recent years. In this article, the representative URAT1 inhibitors with uric acid-lowering or anti-gout effects are reviewed, and related medicinal chemical strategies are analyzed, hoping to provide valuable insights into the discovery of new URAT1 inhibitors.

Hepatitis B virus (HBV) infection is a serious global public health problem. Chronic hepatitis B virus infection can cause health problems such as cirrhosis, liver metabolism disorders and hepatocellular carcinoma. Nucloes(t)ide analogues and interferon drugs used to treat chronic HBV infection do not completely eradicate covalently closed circular DNA (cccDNA) and integrated genome of HBV DNA, so that they cannot achieve the functional cure of chronic HBV infection. Currently, a series of drugs targeting the phases of HBV lifecycle and immunomodulators have entered clinical trials. Here, we review the current status of the therapeutic drugs as well as the recent advance of direct antiviral agents.

In this study, we investigated the effect of aspirin on tumor biological effects mediated by hepatocyte growth factor/cellular-mesenchymal-epithelial transition factor (HGF/c-Met) axis, and preliminarily explored the molecular mechanism of inhibiting tumor metastasis by aspirin. The binding of aspirin to c-Met was predicted by molecular docking; cellular thermal shift assay (CETSA) was used to verify the binding of aspirin to c-Met at the cellular level. The inhibitory effect of aspirin on c-Met kinase was detected by kinase activity; Western blot, cell scattering test, cell branching morphogenesis and Transwell test were used to evaluate the cell signal transduction, morphological changes and migration and invasion ability. The results showed that aspirin could effectively inhibit the kinase activity of c-Met with a half inhibitory concentration of 0.95 mmol·L^-1. The results of docking showed that aspirin could bind to the ATP pocket of c-Met protein, and the main binding sites were Tyr1230, Tyr1159 and Met1229. The CETSA test also showed that aspirin could form binding complex with c-Met protein. Western blot results showed that aspirin could inhibit the up-regulation of phosphorylated Met stimulated by HGF in a concentration-dependent manner. The results of cell scattering test showed that aspirin could block HGF/c-Met promoted cell scattering in a concentration dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. Similarly, the results of MDCK cell branching morphogenesis experiment showed that aspirin could inhibit HGF/c-Met mediated invasive growth in a concentration dependent manner. The results of Transwell test showed that aspirin could block HGF/c-Met mediated cell migration and invasion in a concentration-dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. The above results indicate that aspirin can bind to c-Met, thereby blocking the biological effects mediated by HGF/c-Met, and inhibiting tumor metastasis. This study revealed the new biological function of aspirin, and provided a new theoretical basis for a comprehensive understanding of the anti-metastatic effect of aspirin.

The epidemic of Zika virus (ZIKV) raises critical public health and safety problems. However, there are currently no vaccines or drugs that are clinically approved for ZIKV infections. Since RNA-dependent RNA polymerase (RdRp) plays an important role in replication and transcription of ZIKV and is absent in human beings, it is a potential drug screening target of anti-ZIKV agents. According to the fluorescence-based alkaline phosphatase-coupled polymerase assay method, we established the NS5 RdRp inhibitor screening model. Through screening from an anti-infection compound library, we found a compound octenidine dihydrochloride (OCT) that could inhibit ZIKV RdRp activity with a half maximal inhibitory concentration (IC₅₀) of 5.43 μmol·L^-1. Biolayer interferometry (BLI) assay showed that OCT could bind to ZIKV RdRp and had a strong affinity. Moreover, OCT exhibited an inhibitory effect on ZIKV replication with a half maximal effective concentration (EC₅₀) of 29.94 μmol·L^-1. All these results indicated that OCT had the anti-ZIKV activity by targeting ZIKV RdRp, and it is likely to be a promising lead compound against ZIKV.

Influenza virus is an RNA virus that classified into 4 types, A, B, C, and D, where influenza A and B virus infection may cause human acute respiratory tract infection and nearly 0.3 million deaths annually. The life cycle of influenza virus infection is highly dependent on the host response, demonstrating an important strategy of developing anti-influenza agents that target the host factors. This research utilized a transcriptome signature reversion (TSR) strategy to discover a list of multi-host-factor-target anti-influenza agents and determined their anti-influenza activities in vitro. BIX02189 was discovered and exhibited broad spectrum anti-influenza activity, with half maximal effective concentration (EC₅₀) of 17.1 μmol·L^-1 against influenza A virus H1N1 (A/Puerto Rico/8/1934) and 9.4 μmol·L^-1 for influenza B virus (B/Jiangxi Xinjian/BV/39/2008). The anti-influenza A virus activity of BIX01289 is stronger than the positive control ribavirin with EC₅₀ of 97.9 μmol·L^-1 for influenza A virus H1N1 (A/Puerto Rico/8/1934). According to the unsupervised transcriptomic profile similarity clustering analysis, BIX02189 was considered to inhibit viral protein synthesis and release of influenza virus mainly through inhibiting the Raf/MEK/ERK cascade, revealing its potential mechanism of inhibiting influenza virus infection.

Inosine 5′-monophosphate dehydrogenase (IMPDH) is a key enzyme catalyzing the rate-limiting step of de novo nucleotide synthesis in vivo. In recent years, it has become a therapeutic target for anti-virus, anti-bacterial, anti-cancer, anti-parasitic and other diseases. IMPDH inhibitors have been shown to inhibit viral prolife-ration in host cells by depleting guanosine 5′-monophosphate (GMP), the raw material required for viral replication in host cells, with broad-spectrum antiviral properties. In order to find novel anti-coronavirus drugs, this study screened 22 potential IMPDH inhibitors from 70 000 natural small molecule libraries based on IMPDH protein structure using molecular docking and ROC calculation for virtual screening. With ribavirin as the positive control drug, Huh7 cell and H460 cell models were used to verify the anti-coronavirus HCoV-229E and HCoV-OC43 activities of 22 selected target compounds. Among them, compounds 11, 12, 15 and 16 showed inhibitory activity against coronavirus HCoV-229E. The compounds 4, 12, 13 and 15 showed inhibitory activities against coronavirus HCoV-OC43. 12 and 15 showed significant inhibitory activity against both two coronaviruses, and their efficacy was similar to ribavirin at the same dose, which can be further studied as a lead compound for IMPDH inhibitors.

For rapid discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (M^pro) inhibitors from a natural product library, a novel colorimetric screening assay was developed. According to the colorimetric principle, the synthetic peptide TSAVLQ-para-nitroanilide (pNA) was used as the M^pro hydrolysis substrate. Subsequently, the working concentration of pNA substrate, M^pro working concentration, hydrolysis time and DMSO tolerance were optimized for the development of a simple and robust colorimetric screening assay. Through these systematic optimizations, we selected 0.4 μmol·L^-1 M^pro and 100 μmol·L^-1 pNA substrate as the optimal working concentrations in this colorimetric screening assay, and a high Z' factor of 0.9 was achieved. Using this screening assay, natural product ginkgolic acid C13:0 (GA13:0) was identified as a novel competitive M^pro inhibitor in vitro. Taken together, we have successfully developed a simple and optimized colorimetric screening assay, which will be vital for the discovery of novel SARS-CoV-2 M^pro inhibitors.

Arrhythmia is the abnormal heart-beat frequency and/or rhythm caused by the origin of cardiac activity and/or conduction disorder. Arrhythmia disease has various manifestations and complex etiology, which can occur alone or complicated with other cardiovascular diseases. A sudden arrhythmic onset may lead to sudden death, whereas a sustained onset may lead to heart failure. In cardiomyocytes, calcium overload induces apoptosis and leads to arrhythmia. Calcium channel blockers have been widely used in clinic as a routine cardiovascular drug to regulate calcium signal, but their efficacy on different arrhythmia complications vary, and they also have potential therapeutic risks. Therefore, it is of great significance to seek calcium signal modulators targeting new mechanisms from plants and other natural product resources and develop them into anti-arrhythmia drugs with higher safety and better curative effect. This review focuses on the calcium signal regulatory effects of plant-derived natural calcium channel antagonists in arrhythmia models, highlights the research progress in recent years, and summarizes the effects and mechanisms of various natural drugs such as alkaloids, saponins, quinones and flavonoids, which regulate Ca²⁺ homeostasis, to provide a theoretical basis for the drug development of natural calcium channel antagonists to prevent and treat arrhythmia in the future.

Depression is a psychiatric disease characterized by long-lasting low mood, accompanied by symptoms such as cognitive, sleep and social disturbances. In the theory of traditional Chinese medicine, depression is included in the categories of "depression syndrome" and "madness", and the main syndrome type is liver stagnation and spleen deficiency. Xiaoyao san, Sini san and Chaihushugan san are commonly used prescriptions for clinical treatment of depression, and flavonoids are the common active ingredients. Clinical and basic studies have shown that the flavonoids derived from antidepressant compound prescription have pharmacological effects such as anti-inflammatory, antioxidant, neuroprotective, and intestinal flora regulation which could prevent the occurrence and development of depression from different pathways. Based on this, this review focuses on the pathogenesis of depression, summarizes the common flavonoids in antidepressant prescriptions, and summarizes their regulatory mechanisms and effects in order to provide ideas for the prevention and treatment of depression.

Cerebrovascular diseases have the characteristics of high morbidity, high disability, high mortality and high recurrence rate, which seriously harm human health and increase the national health economic burden. 3-n-Butylphthalide (NBP) is a new drug commonly used in clinical treatment of cerebrovascular diseases, and it is also one of the main active components in traditional Chinese medicine such as Angelica sinensis and Chuanxiong. In this review, the pharmacological effects of NBP were systematically summarized. Studies have shown that NBP has pharmacological effects such as antiplatelet aggregation, anti-thrombosis, inhibiting neuronal apoptosis, anti-oxidation, anti-cerebral ischemia, anti-brain injury, and anti-vascular dementia. In clinical practice, it is often combined with edaravone, alteplase, fasudil, Xingnaojing injection, and compound Danshen injection to treat cerebral vascular diseases such as stroke, vascular dementia, and cerebral vasospasm, and plays a good synergistic effect. This summary could provide support for the rational clinical application of NBP, and also provide basis for the in-depth study of the interaction of its drug combination.

Dementia is a series of diseases with severe cognitive decline caused by brain diseases, that closely related to kidney deficiency in traditional Chinese medicine, including Alzheimer's disease (AD), dementia caused by cerebral stroke, vascular dementia (VAD) and so on. Dipsaci Radix is the dried root of Dipsacus asper Wall. ex Henry and its curative effects mainly focus on nourishing the liver and kidney, strengthening muscles and bones, as well as dredging blood vessels. The main chemical components of Dipsaci Radix are triterpenoid saponins and iridoid glycosides. In recent years, studies have found that Dipsaci Radix and its active compounds could ameliorate dementia symptoms via multiple targets and molecular mechanisms. In this review, we summarize the recent research progress of Dipsaci Radix in dementia prevention, which will provide reference for further exploration of its mechanism and application in the prevention and treatment of dementia.

Compared with the traditional two-dimensional (2D) monolayer culture, three-dimensional (3D) organoid can better simulate the physiological and pathological microenvironment of organs and tissues. In this study, 3D cardiac organoids were constructed using cardiac fibroblasts (CFs), cardiac myocytes (CMs) and endothelial cells (ECs) isolated from hearts of 1-3-day Sprague-Dawley (SD) neonatal rats. The experimental scheme was approved by the Experimental Animal Welfare and Ethics Committee of Tianjin University of Traditional Chinese Medicine and met the standards of experimental animal welfare and ethics. Optimal seeding cell density and culture time were determined by observing the sphere diameter and pulsation. The hierarchical structure and cardiac-like function were evaluated by fluorescence staining. The results showed that the cardiac-like microspheres constructed with cell number of 1×10⁴ still beated spontaneously even after 34 days in culture, and maintained characteristic cellular hierarchical structure. Then, based on these cardiac microspheres, a phenylephrine (PE)-induced cardiac hypertrophy model was established and evaluated by mitochondrial mass, intracellular Ca²⁺ concentration and mitochondrial membrane potential. Guanxinning Injection (GXNI) was tested to verify that the established model can be used for myocardial hypertrophy drug screen. The results showed that GXNI significantly reversed the enlargement of cardiac microsphere area and diameter, the increase of mitochondrial mass, intracellular Ca²⁺ concentration and the decrease of mitochondrial membrane potential caused by PE, and reduced upregulation of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC). In conclusion, this study successfully established a 3D in vitro model of cardiac remodeling induced by cardiac hypertrophy. In this new system, cardiac microspheres not only have cardiac-like morphology and extracellular matrix components, but also exhibit spontaneous and rhythmic systolic and diastolic function. Therefore, the cardiac microsphere is an effective model to investigate the pathological mechanism of cardiac hypertrophy and screen related drugs.

Doxorubicin (DOX) is an anthracycline antibiotic widely used in the treatment of certain types of tumors. However, DOX have some serious side effects in the body after long-term use, especially acute and chronic cardiotoxicity. This study explored the protective effect of tanshinone I (Tan I) on acute cardiotoxicity induced by DOX and its underlying molecular mechanisms. In vivo and in vitro acute cardiotoxicity models were established by injecting DOX (6 mg·kg^-1, twice per week) into the tail vein of C57 mice and stimulating H9C2 cardiomyocytes with DOX. In in vivo experiments, Tan I (10 mg·kg^-1) was administered daily by oral 5 days before the tail vein injection, till the end of the experiment. The effects of Tan I on mice heart function, myocardial tissue morphology and serological indicators were detected. Animal welfare and experimental procedures followed the regulations of the Animal Ethics Committee of Beijing University of Traditional Chinese Medicine. In in vitro experiments, the specific mechanism of Tan I against oxidative stress was further studied. Immunofluorescence was used to detect the expression of Nrf2 and its transcription into the nucleus. In addition, the levels of oxidative stress related proteins, protein kinase B (Akt), nuclear erythroid factor 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and NAD(P)H quinone dehydrogenase 1 (NQO1), were detected by Western blot. Finally, AutoDock software was used for molecular docking verification. The results showed that Tan I significantly improved cardiac function in mice. Meanwhile, the expression levels of creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) in serum were decreased. Immunofluorescence results indicated that Tan I could increase Nrf2 expression level in H9C2 cells and promote Nrf2 entry into the nucleus. Western blot results also indicated that the levels of oxidative stress related proteins, P-Akt, Nrf2, HO-1 and NQO1 in DOX plus Tan I group were significantly increased compared with DOX group. These results suggest that Tan I can alleviate DOX-induced acute cardiotoxicity by inhibiting oxidative stress through up-regulating the Akt-Nrf2 pathway, thereby alleviating DOX-induced acute myocardial injury.

The blood-brain barrier (BBB) plays an important role in maintaining the homeostasis of the central nervous system. BBB is disrupted in many neurological disorders such as ischemic stroke and Alzheimer's disease. Traditional Chinese medicine has great potential to prevent ischemic stroke, but the lack of clinically relevant models has been a challenge. We adapted a BBB organoid model formed by human brain microvascular endothelial cells (HBMEC), human astrocytes (HA), and human brain vascular pericytes (HBVP), and established conditions for oxygen-glucose deprivation/reoxygenation (OGD/R) on the cell vitality, barrier permeability, as well as BBB signature marker expression. The protective effect of Guanxinning injection (GXNI) on OGD/R-induced BBB dysfunction was then investigated in the organoid model. The results showed that OGD/R decreased BBB organoid cell viability, increased permeability (leakage), decreased the level of tight junction proteins zonula occludens-1 (ZO-1), claudin-5, occludin and P-glycoprotein (P-gp). GXNI significantly prevented OGD/R-induced BBB disruption such as the decreased cell viability and increased permeability. This study provides a new human cell-derived 3D ischemic brain disease model for central nervous system-targeted drug research and development and demonstrates that a Chinese injection medicine GXNI effectively protects BBB dysfunction in vitro.

In this study, a research strategy integrating network pharmacology analysis and animal experimental validation was applied to explore the molecular mechanism of Chuanxiong Qingnao Granules (CXQN) in improving migraine headache (MH). All animal experiments were followed the regulation of the Laboratory Animal Ethics Committee of the China Academy of Chinese Medical Sciences. Based on the network pharmacology analysis, the 27 active ingredients and their corresponding 940 targets were obtained, and 99 common targets of CXQN in the treatment of MH were obtained by intersection, and tumor necrosis factor-α (TNF-α), interleukin (IL)-6, vascular endothelial growth factor A (VEGFA), IL-1β, brain-derived neurotrophic factor (BDNF) were screened out as hub targets. Enrichment analysis showed that the targets of CXQN in the treatment of MH were mainly involved in cyclic adenosine monophosphate (cAMP), hypoxia inducible factor-1 (HIF-1), phosphoinositide 3-kinase-protein kinase B (PI3K-Akt) signaling pathways. In addition, the experimental verification in the MH rat induced by nitroglycerin showed that the CXQN administrated groups could significantly improve the behavioral symptoms and regulate the level of vasoactive substances, and reduce the expression of TNF-α, IL-6, VEGFA, IL-1β, and BDNF at gene and protein levels. This study revealed the multi-component, multi-target, and multi-pathway characteristics of CXQN in the treatment of MH, and elucidated the potential mechanism of CXQN in the treatment of MH, laying a theoretical foundation and scientific basis for its clinical application in the treatment of MH diseases.

In this study, we investigated the pharmacological effect and possible molecular mechanism of higenamine (HG) in isoproterenol (ISO)-induced myocardial infarction (MI). All procedures were approved by the Institutional Animal Care and Use Committee of the Guangzhou University of Chinese Medicine. ISO was used to induce MI model in rats and H9c2 cells. The effects of HG on biomarkers and cardiac function in MI rats were evaluated by enzyme linked immunosorbent assay (ELISA), echocardiography and hematoxylin-eosin staining (HE). The expression of apoptosis and autophagy related proteins were detected by Western blot in myocardial tissue and H9c2 cells, as well as methyltransferase-like 3 (METTL3) and transcription factor EB (TFEB) protein expression. Molecular docking was used to evaluate the interaction between HG and METTL3. The results showed that HG significantly improved cardiac function and pathologic changes in ISO-induced MI, and inhibited the levels of MI-related biomarkers such as creatine kinase Mb (CK-MB), creatine kinase (CK) and lactate dehydrogenase (LDH). Mechanism studies showed that HG inhibited the expression of apoptosis-related proteins (Bax/Bcl2, caspase3, cleaved-caspase3). Interestingly, HG up-regulated the expression of autophagy related protein Beclin1, promoted autophagy flux, and decreased the ratio of light chain 3B-I/light chain 3B-II (LC-3B-I/LC-3B-II). Further studies found that HG increased the autophagy regulator TFEB and inhibited METTL3 expression. Molecular docking results showed that HG had a good interaction with METTL3. Taken together, HG has a potential anti-MI effect via regulating METTL3/TFEB signaling pathway-mediated autophagy.

In this study, network pharmacology research and animal experiments were used to predict and validate the potential targets of ShengMaiSan (SMS) in the treatment of diabetic cardiomyopathy. The active components of SMS were obtained through TCMSP and BATMAN databases. The potential targets of the active components and diabetic cardiomyopathy were predicted by Swiss Target Prediction and GeneCards databases, respectively. The protein-protein interaction (PPI) network was constructed by String database. Cytoscape software was adopted to perform topological analysis to select the core action target. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were performed using Metascape platform. To validate the potential targets, a type I diabetic rat model which induced by intraperitoneal injection of streptozotocin (STZ) was prepared. Rats were divided into sham group, model group, SMS group and trimetazidine (TMZ) group. Left ventricular hemodynamics was detected after 4 weeks administrated of SMS or TMZ. Myocardial contraction and calcium transients were detected synchronously in cardiomyocytes, as well as sarcoplasmic reticulum calcium content, calcium leak level and ryanodine receptor 2 (RyR2) expression were detected. Based on network pharmacology, 1 288 targets of SMS, 1 066 targets of diabetic cardiomyopathy, and 180 overlapped targets were obtained. The 39 core targets were screened, and 159 pathways including calcium signaling pathway were screened by KEGG pathway analysis. According to the previous studies and focusing on the contractility of diabetic cardiomyopathy, this study was involved calcium signaling regulation pathway in the SMS protection mechanism. The results showed that, compared with sham group, the systolic function of left ventricular and myocardial cells were decreased, and the calcium transport was in disorder in model group; compared with model group, both SMS group and TMZ group increased the maximum systolic pressure of left ventricle and the maximum systolic rate of left ventricular contraction. In addition, SMS group and TMZ group increased the contraction amplitude of cardiomyocytes, decreased the diastolic calcium concentration, the sarcoplasmic reticulum calcium leak and decreased the phosphorylation level of RyR2. There was no significant difference between SMS and TMZ groups. In summary, SMS could reduce the calcium leak of the sarcoplasmic reticulum and enhance the myofilament sensitivity of calcium to increasing contractile function of diabetic rats. The animal welfare and experiment procedures of this study were in accordance with the regulations of the Experimental Animal Ethics Committee of Experimental Research Center, China Academy of Chinese Medical Sciences.

Autoimmune diseases (AID) are characterized by autoimmune disorder, as autologous tissue is attacked by the autoimmune system. It is reported that the imbalance of autoimmune tolerance and ingrained inflammatory response are the core events of AID undoubtedly. Peroxisome proliferator-activated receptor γ (PPARγ) which belongs to the nuclear hormone receptor superfamily is a ligand activated transcription factor. PPARγ combines with retinoid X receptor (RXR) to form heterodimer. When PPARγ is activated, the complex regulates gene expression by binding to a specific peroxisome proliferator response element (PPRE). In addition, PPARγ has diversified biological functions, playing important roles in regulating metabolism, controling inflammation, modulating glucose and lipid metabolism, ameliorating atherosclerosis, anti-tumor, and regulating immune response. However, recently researches indicate that PPARγ participates in the pathogenesis of AID. PPARγ plays key roles in regulating activation and polarization of macrophages, function of dendritic cells, proliferation and differentiation of T cells, and modulation of the function of related stromal cells. This article summarizes the biological functions and signal transduction pathways of PPARγ and the protective effects of agonists of PPARγ on AID, aiming to provide theoretical support for the research of mechanism and prevention and treatment of AID.

Leukotriene B₄ (LTB₄) is a proinflammatory lipid mediator that is synthesized by a number of inflammatory cells. Binding of LTB₄ to its receptor leukotriene B₄ receptor 1 (BLT1) can migrate neutrophils and macrophages to inflammatory sites through chemotaxis and up-regulation of adhesion molecules. Many researches have shown that LTB₄-BLT1 axis is related to the occurrence of autoimmune disorders and other inflammatory diseases. Receptor antagonists of LTB₄ are thus expected to be useful therapeutics for these diseases. In this review, we briefly describe the biological function of LTB₄ and summarize the preclinical and clinical developments of LTB₄ receptor antagonists.

Small interfering RNAs (siRNAs) are an emerging class of RNA interference (RNAi) therapeutics with unique pharmacokinetic properties. Five siRNA drugs based on two delivery systems have been approved, and an increasing number of siRNA drugs have already moved to the clinical study phase. Physiologically-based pharmacokinetic (PBPK) modeling is a useful tool and has been demonstrated to have wide ranging utility in drug development and regulatory review. However, PBPK modeling is still in its infancy in guiding the development of siRNA-based drugs in the context of its widespread use in small and large molecule areas. This article reviews the pharmacokinetic profiles of siRNA drugs, outlines the current state of PBPK model building in siRNA drug development, and describes the key parameters required for model building. This article provides insights into the future applications of PBPK models and for optimizing the key parameters when building the model for siRNA drug development.

Model-informed drug development (MIDD) in the development of pediatric drugs is drawing more and more attention due to the insufficiency of subjects, lack of research on ontogeny, and the limitation of ethic. The core of MIDD used for dose selection includes the population pharmacokinetic (PopPK) model and physiologically based pharmacokinetic (PBPK) model, as well as model-based simulation and prediction. PBPK model has the advantage of predicting the optimal pediatric dose before the clinical trials and has the ability of extrapolation from adult model to pediatric model. PopPK model characterizes the pediatric PK feature based on the analysis of clinical data and can be used to explore the significant covariates, which is a power tool for individualized medicine in children. With their own advantages and disadvantages, PBPK and PopPK model should be jointly used in the pediatric drug development to refine the dose regimen for children at different ages. In this study, the pediatric drug development of rivaroxaban was taken as an example to introduce the combined application of PBPK model and PopPK model in the design and validation of pediatric dose regimen in Phase Ⅰ, Ⅱ and Ⅲ trials, which may provide reference to MIDD in other pediatric drug development.

A new [7, 10∶1, 5]-patchoulane-type sesquiterpenoid was isolated and purified by silica gel column chromatography, medium pressure liquid chromatography, and semi-preparative high performance liquid chromatography. Based on IR, HR-ESI-MS, NMR, and X-single crystal diffraction data analyses, compound 1 was determined to be (-)-(3S, 4R, 5R, 7R, 10R)-[7, 10∶1, 5]patchoul-1(2)-en-3, 4-diol. It showed an inhibitory effect on LPS-induced NO production in RAW264.7 cells.

Nine compounds were isolated from the leaves of Chimonanthus nitens Oliv. by silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. They were identified as chimnitensene B (1), 1α-hydroxyisodauc-4-en-15-al (2), trans-4, 5-dihydroxycorocalane (3), trefoliol B (4), oplopanone (5), oplodiol (6), 3-(3'-hydroxybutyl)-2, 4, 4-trimethylcyclohexa-2, 5-dienone (7), 9(S)-4-oxo-7, 8-dihydro-β-ionol (8), and saniculamoid D (9) respectively, by MS, NMR and single crystal diffraction. Among them, compound 1 is a new guaiane-sesquiterpenoid, and compounds 2-9 were isolated from this plant for the first time.

We identified molecular mechanisms by which Isatidis Radix might prevent or mitigate influenza and corona virus disease 2019 (COVID-19) based on chemical composition and network pharmacology. High performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) was used to analyze the components of Isatidis Radix. Seventy compounds were identified, of which 33 prototype compounds entered the blood. Network pharmacological analysis of 41 potential active components demonstrated that Isatidis Radix can regulate protein kinase B1 (AKT1), serum albumin (ALB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), vascular endothelial growth factor A (VEGFA), tyrosine-protein kinase SRC (SRC), epidermal growth factor receptor (EGFR), intercellular adhesion molecule-1 (ICAM1) and other key genes, which have preventive effects on influenza and COVID-19 through hypoxia inducible factor-1 (HIF-1), vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF), influenza A, Toll-like receptor (TLR), phosphatidylinositol-3-kinase-protein kinase B (PI3K-AKT), COVID-19 and other signaling pathways. This study identifies mechanisms by which Isatidis Radix might act against influenza and COVID-19 that are related to the inflammatory response, immunomodulation and viral defense, and provides a basis for subsequent clinical research. All animal experiments were approved by the Ethics Committee of Shenyang Pharmaceutical University (SYPU-IACUC-S2020-12.23-201).

The UHPLC-LTQ-orbitrap-MS metabolomics technique was used to determine the effect of deapio-platycodin D (DPD) on endogenous metabolites in lung tissues of mice with ammonia-induced cough, and to identify the metabolic regulatory pathways of DPD in its antitussive and expectorant activities. This work was approved by the Animal Ethics Committee of Jiangxi University of Chinese Medicine (Approval No. JZLLSC-20190235). Metabolites were identified by UHPLC-LTQ-orbitrap-MS method and the metabolic pathways related to differentially-expressed metabolites were analyzed by the MetaboAnalyst platform. DPD significantly prolonged (P < 0.05) cough latency, reduced the number of coughs, and significantly increased (P < 0.05) phenol red excretion in ammonia-induced cough mice. Twenty-five metabolites related to cough and 38 metabolites related to sputum excretion were identified by UHPLC-LTQ-orbitrap-MS. Changes in the metabolism of linoleic acid, arachidonic acid, glycerophospholipid, alanine, aspartic acid and glutamic acid, pentose and glucuronate interconversions, and lysine degradation were evident with DPD treatment of ammonia-induced cough mice. Changes in the metabolism of linoleic acid, taurine and hypotaurine, glycerophospholipid, purines and pyrimidines, arachidonic acid and the biosynthesis of unsaturated fatty acids after DPD treatment were related to phenol red excretion. Linoleic acid metabolism, arachidonic acid metabolism and glycerophospholipid metabolism are common regulatory pathways by which DPD appears to exert its antitussive and expectorant activity. These metabolic pathways are closely related to anti-inflammatory pathways, immune function regulation, neurotransmitter release, cell signal transduction, energy metabolism and apoptosis. This study clarifies the antitussive and expectorant activity and mechanism of DPD.

Bitter almonds (Semen Armeniacae Amarum) are prone to oil deterioration during storage, so they often require mashing prior to clinical use. To confirm the medical value of bitter almonds "being mashed when used" and to determine the optimal storage conditions for bitter almonds, UPLC-MS/MS was used to perform a comparative study of the chemical composition of bitter almonds in different storage states (mashed and unmashed), storage times (0, 2 and 4 weeks), and storage temperatures (25 ℃ and 4 ℃). A total of 58 substances were identified in bitter almond extracts through literature review, this group's previous work, and a Compound Discoverer software search. Statistically significant differences were found in the chemical composition and content of bitter almonds in different storage states, storage times, and storage temperatures. The results show that the chemical composition of bitter almonds stored unmashed was more stable than that of bitter almonds stored mashed; the chemical composition of bitter almonds stored at 4 ℃ was more stable than that of bitter almonds stored at 25 ℃; and the shorter the storage time, the less the chemical composition changed. Amygdalin, the main medicinal component of bitter almonds, showed statistically significant differences in content under the above three storage conditions, which can be used as a potential quality marker for bitter almonds.

The enteric-hepatic axis plays an important role in the occurrence, progression and regression of nonalcoholic fatty liver disease (NAFLD). Obeticholic acid (OCA) is a farnesoid X receptor agonist. In this study, C57BL/6 mice were fed with methionine and choline deficient (MCD) diet for 8 weeks with OCA (6.5 mg·kg^-1·d^-1) administration by gavage at the same time. The effects of OCA on serum lipid and bile acid metabolomics and ileal gut microbiota (GM) of MCD mice were studied by UPLC-MS and 16S rDNA sequencing. The results were as follows: (1) OCA decreased the activities of alanine aminotransferase and aspartate aminotransferase in serum and the contents of triglyceride (TG) and malondialdehyde in liver, alleviated the accumulation of liver fat and inflammation of MCD mice. OCA down-regulated the contents of 2 eicosanoids (12, 13-EPOME, 9, 10-EPOME) and 4 free fatty acids (FFA16∶1, FFA18∶1, FFA16∶2, FFA18∶3) and TG (16∶1_16∶1_18∶2) in serum, and up-regulated the content of 1 eicosanoid thromboxanes B3. KEGG differential metabolite pathway analysis showed that fatty acid biosynthesis might be the main way that OCA ameliorated lipid metabolism disorder of MCD mice. OCA reduced the relative abundance of Christensenellaceae and Lachnospiraceae_UCG-006 in the GM of MCD mice; OCA decreased the serum levels of 23-deoxycholic acid, porcine deoxycholic acid, 3-deoxycholic acid, glycine deoxycholic acid, glycine cholic acid, taurine deoxycholic acid, taurocholic acid and taurine. These results suggest that the alleviating effect of OCA on NAFLD of MCD mice may be related to its above-mentioned regulation of the metabolism of the free fatty acids, oxidized lipids, 12α-hydroxylated bile acids and the abundance of GM. The animal experiments were approved by the Experimental Animal Ethics Committee of Hubei University (No. 20220036).

Ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) was applied to rapidly identify the phospholipids in human plasma and explore the mass spectrometric fragmentation pattern. An acquity UHPLC^TM BEH C18 column (50 mm × 2.1 mm, 1.7 μm) was utilized and eluted with a gradient system; the mobile phase consisted of 10 mmol·L^-1 ammonium formate aqueous solution-0.1% formic acid aqueous solution (A) and acetonitrile-isopropanol (1∶1) organic solution (B) containing 10 mmol·L^-1 ammonium formate-0.1% formic acid. The flow rate was 0.3 mL·min^-1 and the column temperature was set at 50 ℃. An electrospray ionization (ESI) source was used to collect mass spectra in positive and negative ion mode. Based on the precise relative molecular weight and elemental composition calculated by Masslynx 4.1 software, comparison with references, and secondary mass spectrometry fragment ions and lipid databases, a total of 82 plasma lipids were identified, including 14 lysophosphatidylcholines (LysoPCs), 39 phosphatidylcholines (PCs), 17 sphingomyelins (SMs), 7 ceramides (Cers), 4 phosphatidylethanolamines (PEs), and 1 phosphatidylinositol (PI). A simple, efficient, fast and stable analytical method was established in this study for the qualitative analysis of phospholipids in human plasma, and the fragmentation regularity of the main phospholipids was determined. This work provides a good foundation for further metabolomics studies of plasma phospholipids. This study was approved by the Second Affiliated Hospital of Guangzhou Medical University Clinical Research and Application Institutional Review Board Approval (No. 2020-hs-07).

We developed an in-vitro bioassay for determining the bioactivity of human insulin by homogeneous time-resolved fluorescence immunoassay. CHO-INSR B1284 transgenic cells were used as target cells. Key assay parameters, including the cell density, the range of working concentrations, and the stimulation time were optimized. The specificity, relative accuracy, intermediate precision, linearity, and range of the method were validated, as well as the passage stability of the CHO-INSR B1284 cell line. The national standard of recombinant human insulin was used as the benchmark to evaluate the relative potency of insulin analogues and drugs. The drugs and the reference human insulin showed a dose-response relationship, R² > 0.995, which conforms to the four-parameter equation: y = (A - D) / [1 + (x/C)^B] + D. Specificity of the method was good. The geometric mean, relative bias, and geometric coefficient of variation (GCV, %) of the five concentrations (n = 8, 64%, 80%, 100%, 125% and 156%) met the requirements of the General Rules of Chinese Pharmacopoeia, 2020 edition, Volume IV (9401). In summary, a bioassay for determining the in vitro bioactivity of human insulin based on a homogeneous time-resolved fluorescence technique was established; the method was simple, time-saving, accurate and precise, and could be used for the evaluation of biological activity and quality.

Alangium chinense is a commonly used medicinal plant of Alangiaceae Alangium, one of the characteristic Miao medicines in Guizhou. The genus is strongly differentiated and has complex morphological variation, with significant differences in active ingredients and potency among herbs. In this paper, we sequenced the chloroplast genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib using Illumina high-throughput sequencing technology. The genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplasts were sequenced, their assembly annotation and structural characterization were completed, and the genomes of the congeners Alangium chinense and Alangium alpinum were downloaded from NCBI for comparative analysis. Finally, the phylogenetic tree was constructed by selecting published species with close affinity to Alangium chinense family from NCBI. The results were as follows: Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplast genomes were 156 670, 156 672 and 156 871 bp in length, with a typical four-segment structure, all containing one long single copy region (LSC), one short single copy region (SSC) and two inverted repeat regions (IRa and IRb). All of them were annotated to 133 genes, including 88 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Three types of long repeat sequences and SSR loci, forward repeats, palindrome repeats and reverse repeats, were found in all chloroplast genomes. Comparative genomic analysis showed that there was diversity in the boundary transition regions of the five species, no rearrangements or inversions were found in the chloroplast genome, and there were significant differences in the coding regions of ndhA, ycf1, rpl16, ycf2, and petD genes, which provided new loci for molecular identification of Patagonia. In the phylogenetic analysis, Alangium kurzii var. kurzii clustered as a single species, and Alangium chinense subsp. Pauciflorum and Alangium chinense subsp. Strigosum clustered as a single species, with a support rate of 93 and close affinity, indicating that the phylogenetic tree can be used for the species identification. This paper can provide a basis for the taxonomic identification of Alangium, ensure the safety of clinical use of anise herbs and regulate the market of Alangium chinense herbs.

Gentiana rhodantha is a characteristic medicinal material of Miao Ethnomedicine. It has significant curative effect in the treatment of acute jaundice hepatitis, dysentery, pediatric pneumonia and bronchitis, etc. However, the evolutionary relationship and taxonomic identification of G. rhodantha are controversial. In this study, we sequenced the chloroplast genome of G. rhodantha using the second and third generation sequencing technology. Then, the structural characteristics and suitability evolution characteristics were analyzed. The results showed that the G. rhodantha chloroplast genome was 148 844 bp in length with 37.75% GC content, consisting of a large single copy region (LSC) of 80 076 bp, a small single copy region (SSC) of 17 596 bp and an inverted repeat region (IR) of 25 586 bp. A total of 124 genes were annotated, including 80 protein-coding genes, 36 tRNA genes, and 8 rRNA genes; the chloroplast genome of G. rhodantha has a weak codon preference, and the influencing factors are mainly natural selection. The optimal codons are CUU, UCU, UCA, CCA, and ACU. A total of 169 SSRs were found in MISA, of which the single nucleotide repeats were the most (114, 67.50%), followed by dinucleotide repeats (43, 25.44%). The phylogenetic analysis support that G. rhodantha belong to Sect. Stenogyne which can be clearly distinguished from other groups. Compared with other species, the Ka/Ks value of chloroplast genes of G. rhodantha is basically less than 1 except for psaI, rpl22 and rps11, indicating that they have been subjected to strong purification selection in the long-term evolutionary process. The photosynthesis gene psaI and the expression-related genes rpl22 and rps11 showed differences between groups, which supported the view that Sect. Stenogyne was an independent genus. This study will provide a reference for future researches on chloroplast genetic engineering and molecular breeding of G. rhodantha.