The previous virtual screening of ten million compounds yielded two novel nonlipopeptide-like chemotypes as TLR2 agonists. Herein, we present the chemical optimization of our initial hit, 1-phenyl-3-(thiophen-2-yl) urea, which resulted in the identification of SMU-C80 (EC₅₀ = 31.02 ± 1.01 nM) as a TLR2-specific agonist with a 370-fold improvement in bioactivity. Mechanistic studies revealed that SMU-C80, through TLR1/2, recruits the adaptor protein MyD88 and triggers the NF-κB pathway to release cytokines such as TNF-α and IL-1β from human, but not murine, cells. To the best of our knowledge, it is the first species-specific TLR1/2 agonist reported until now. Moreover, SMU-C80 increased the percentage of T, B, and NK cells ex vivo and activated the immune cells, which suppressed cancer cell growth in vitro. In summary, we obtained a highly efficient and specific human TLR1/2 agonist that acts through the MyD88 and NF-κB pathway, facilitating cytokine release and the simultaneous activation of immune cells that in turn affects the apoptosis of cancer cells. © 2021 American Chemical Society

BACKGROUND: Anti-malarial drug artesunate (ART), a semi-synthetic derivative of artemisnin, has immunosuppressive effects on several autoimmune diseases, including Systemic lupus erythematosus (SLE), Rheumatoid arthritis (RA), and Colitis. However, molecular mechanisms of ART, especially on follicular helper T cells (Tfh), central players in SLE pathology, are far from clear.

PURPOSE: The object for this work is to investigate the therapeutic effect of ART on lupus-prone MRL/lpr mice and its regulatory function on Tfh cells.

STUDY DESIGN AND METHODS: MRL/lpr mice were used to explore therapeutic effects of ART on lupus-prone MRL/lpr mice and its regulatory functions on Tfh cells. Then, experiments of renal function were accomplished using the biochemical kits. Effects of ART on histopathology of kidneys, inflammatory factors and autoantibodies were examined using H&E staining, ELISA and real-time PCR. Flow cytometry and western blot analysis were used to examine effects of ART on Tfh differentiation and Jak2-Stat3 signaling pathway.

RESULTS: Upon oral administration, ART significantly prolonged the survival of MRL/lpr mice, ameliorated the lupus nephritis symptoms, decreased the levels of anti-dsDNA antibodies deposited in the kidney, and the levels of pathogenic cytokines (IL-6, IFN-γ and IL-21). After ART treatment, T-cell compartment in the spleen of MRL/lpr mice was restored in terms of reduction in the number of Tfh cells and in the maintenance of the ratio of Tfr to follicular regulatory T cells (Tfh). In addition, ART has significantly inhibited the phosphorylation levels of Jak2 and Stat3 in the MRL/lpr mice.

CONCLUSION: ART showed therapeutic effects on lupus-prone MRL/lpr mice by inhibiting the differentiation of Tfh cells as well as altering the activation status of Jak2-Stat3 signaling cascade. Copyright © 2019 Elsevier GmbH

Mo-clay was used by German doctors to treat injured soldiers' wounds during the First World War. Today, there are anecdotal cases of mo-clay being beneficial for patients suffering from psoriasis, a chronic, inflammatory disease. There are several histological features in the psoriatic skin, including acanthosis, hyperkeratosis, pararkeratosis and a loss of granular layer. Mo-clay is a unique marine deposit, an Eocene clayed diatomite. It was formed 54 million years ago from deposits of single-celled algae along with clay minerals and volcanic ash. The major elements are silicon, aluminium and iron. It is found in Denmark and Germany. As mo-clay had been used to treat wounds, it was tested for antibacterial activity. Mo-clay did not show any anti-bacterial activity against a battery of Gram-positive and -negative bacteria. Mo-clay showed stimulation of cell proliferation at concentrations 39 – 78 µg/ml in splenic mouse lymphocytes, and at 156 µg/ml in HaCat cells, whereas an inhibition of proliferation was observed at 313 µg/ml. Mo-clay was tested for anti-psoriatic activity in vivo using the mouse tail test [1]. This model can be used to investigate agents for effect on psoriasis, since the adult mouse tail has regions of both orthokeratosis and parakeratosis. Mo-clay induced orthokeratosis and showed a significant increase in epidermis thickness. The results suggest that mo-clay may have anti-psoriatic effects.

Imidazole has an important five-membered aromatic heterocyclic ring, which is available widely in natural products and synthetic molecules. The special structural characteristics of imidazole ring enable it to bind with a variety of enzymes and receptors through hydrogen bonds, coordination, ion-dipole and cation-π interactions, hydrophobic effects, and Van der Waals forces. These interactions promote several biological activities involving anti-tumor, anti-inflammatory, anti-microbial, and anti-viral properties. Herein, we review and discuss recent developments in using imidazole derivatives and their special pharmacological activities for the treatment of various diseases. © 2021 Bentham Science Publishers.

Proteolysis-targeting chimeras (PROTACs), the hetero-bifunctional compounds containing a specific ligand to bind the target protein, a suitable linker, and an E3 ubiquitin ligase substrate, are being developed for therapeutic applications. PROTACs hijack the catalytic activity of ubiquitin E3 ligases to mediate proteasome dependent degradation of selected protein of interest (POI), by bringing the ligase and POI into close spatial proximity and initiating the poly-ubiquitination process. Compared to the traditional small-molecule drugs, PROTACs reduce the problems of dosage, drug resistance, side effects and undruggable targets that could not be targeted pharmacologically. In this review, all the POIs, and peptide to small-molecule based PROTACs developed during the past two decades are summarized and directions for future development are discussed. © 2020 Elsevier Inc.

Identification of new indications for existing drugs is crucial through the various stages of drug discovery. Computational methods are valuable in establishing meaningful associations between drugs and diseases. However, most methods predict the drug–disease associations based solely on similarity data, neglecting valuable biological and chemical information. These methods often use basic concatenation to integrate information from different modalities, limiting their ability to capture features from a comprehensive and in-depth perspective. Therefore, a novel multimodal framework called AMDGT was proposed to predict new drug associations based on dual-graph transformer modules. By combining similarity data and complex biochemical information, AMDGT understands the multimodal feature fusion of drugs and diseases effectively and comprehensively with an attention-aware modality interaction architecture. Extensive experimental results indicate that AMDGT surpasses state-of-the-art methods in real-world datasets. Moreover, case and molecular docking studies demonstrated that AMDGT is an effective tool for drug repositioning. Our code is available at GitHub: https://github.com/JK-Liu7/AMDGT. © 2023 The Author(s)

Ovarian cancer is one of the leading causes of death in women. It is also an important factor for an increased worldwide gynecological death observed recently. Here, we have elucidated the effect and mechanisms of Gancao Nurish-Yin Decoction (GNYD) on the viability of ovarian cancer cells. Network pharmacology analysis was performed to explore the putative therapeutic targets of GNYD on the ovarian cancer cells. Experiments at the cellular and molecular levels using ovarian cancer cells were performed to further verify the effect of GNYD. According to the network pharmacology analysis, 282 genes related to the targets of GNYD were identified. Among them, 123 genes were found to be overlapping as the potential targets for treating ovarian cancer. Subsequent experiments proved that GNYD medicated serum has significantly inhibited the survival, growth and migration of the ovarian cancer cells. The number of apoptotic cells has increased after the treatment with GNYD medicated serum, and a higher proportion of cells were arrested at G2/M phase. Importantly, genes present in the AMPK-p53/p21 pathway were found to be significantly promoted. Thus, this study has not only demonstrated the potential therapeutic value of GNYD in inhibiting the survival, growth and migration of ovarian cancer cells in vitro, but also provide clues for further exploring the pharmacological and molecular mechanisms of Chinese medicine preparations acting on various types of cancer. © 2022 The Authors. 

ETHNOPHARMACOLOGICAL RELEVANCE: Wang-bi tablet (WB) consists of 17 traditional Chinese medicines and has been used for treating rheumatoid arthritis (RA) in China for many years, however, its pharmacologic mechanism is not clear.

AIM OF STUDY: The aim of this study was to investigate the therapeutic effect of WB on collagen-induced mouse arthritis and explored the underlying mechanism.

MATERIALS AND METHODS: DBA/1 mice were used to establish a type II collagen-induced arthritis (CIA) model. From the day of arthritis onset, mice were treated daily by gavage with either total glucosides of paeony (TGP, 0.37  g/kg/d) or WB at a lower (1.11  g/kg/d, WBL) or higher dose of (3.33  g/kg/d, WBH) for 8 weeks. The severity of arthritis, levels of cytokines and the activation of signaling pathways were determined.

RESULTS: Our results revealed that WB treatment effectively alleviated inflammatory symptoms and prevented bone erosions and joint destructions. It obviously decreased the serum concentration of pro-inflammatory cytokines TNF-α, IL-6 and IL-17α, while increased the concentration of anti-inflammatory cytokine IL-10. Interestingly, the proportion of splenic Treg cells were increased significantly. In vitro experiments showed that WB inhibited the differentiation of osteoclasts. Consistently, the mRNA levels of tartrate-resistant acid phosphatase (TRAP) and cathepsin K (CtsK), and the activation of NF-κB and JAK-STAT3 signaling pathways in the paws of CIA mice were inhibited by WB treatment. On the other hand, up-regulation of osteogenic genes Runx2, Osterix mRNA, and activation of Wnt/β-catenin signaling pathway along with a decreased receptor activator of nuclear factor κB ligand (RANKL) expression were found in WB treated mice.

CONCLUSION: Our results suggest that the therapeutic effect of Wang-bi tablet could be attributed to its inhibitory activity on NF-κB and STAT3 signaling pathway-mediated osteoclast differentiation, and its enhancement on Wnt/β-catenin signaling pathway-mediated osteoblast functions.

OBJECTIVE: To present the prescribing patterns of general practitioners (GPs) at public primary care centres (PPCCs) in Halland, a county in the south-west of Sweden. GP share of the total prescribing of different drug groups 1988-1997 is presented, as well as changes in patterns. DESIGN: A descriptive prescription study performed 3 months each year in 10 consecutive years. SETTING: Medical service and pharmacies in Halland. SUBJECTS: Prescriptions from about 100 GPs of PPCCs and 550 physicians of various other specialties. MAIN OUTCOME MEASURES: Percentages and absolute numbers of GPs prescribing. RESULTS: GPs prescribed 45% and 51% of the prescriptions from physicians in 1988 and 1997, respectively, while the cost shares were 40% and 42%. An increase in prescriptions was seen both in relative and in absolute numbers (from 117414 in 3 months in 1988 to 161012 in 1995). The increase in cost per DDD (defined daily dose) during the study period was 47% for GPs and 72% for other doctors. CONCLUSIONS: GP prescribing increased in both absolute and relative numbers, while the cost increase per DDD was moderate compared to other physicians.

PURPOSE: To present and interpret drug prescription patterns, related to various groups of the population in a Swedish county, in order to estimate the prevalence of drug use in different age groups. METHODS: Data on prescriptions, dispensed March-May 1988-2002, were combined with population statistics of Halland, a county in the south of Sweden, and analysed. Number of defined daily doses (DDD) per 100 inhabitants and day and prescriptions per 100 inhabitants and 3 months were used as indicators of drug prevalence. RESULTS: The total drug exposure in the population of Halland nearly doubled during the 15-year period. The most frequently used drugs overall, in 2002, were psycholeptics (N05), analgesics (N02), antibacterials (J01) and sex hormones (G03). Nearly 30% of the women of 15-69 years were exposed to sex hormones. Multiplied drug prevalence among people above 60 was found for antithrombotic drugs (B01), agents acting on the renin-angiotensin system (C09), sex hormones (G03), serum lipid reducing agents (C10), antidepressants (N06) and drugs for peptic ulcer and GORD (A02B). CONCLUSIONS: The increase in drug prescribing over the 15 years concerned both symptom-related treatments, like hormone replacement therapy, analgesics, antidepressants and drugs for acid-related disorders, as well as preventive treatments, like antithrombotics, lipid-lowering drugs and antihypertensives. The unit DDD/100 inhabitants and day gives a fairly correct measure of the percentage treated for chronic disorders. However, for short-term treatment courses and especially for drug use in children, number of prescriptions/100 inhabitants and adequate period of time, is easier to interpret.

PURPOSE: To determine the prevalence of purchase of prescribed antihypertensive and/or serum lipid-reducing pharmaceutical preparations among different age groups, from the age of 45, in the Swedish population. Further, to calculate the percentage of the population, from the age of 60, who purchased these pharmaceuticals without having had a circulatory diagnosis in the Hospital Discharge Register the last 7 years, or having purchased nitrate vasodilators, as an attempt to estimate the proportion of primary preventive treatments. METHODS: A cross-sectional study, of individual data on prescriptions for antihypertensives (C02-C03, C07-C09) and serum lipid-reducing agents (C10), dispensed from July to December 2005 for the Swedish population. Data were obtained from the new Swedish Prescribed Drugs Register. The data were related to population statistics, and linked to data on diagnoses of cardiovascular disease (I00-I99), from the Swedish Hospital Discharge Register 1998-2004. Data on individuals with purchase of antihypertensive or serum lipid-reducing agents, but without a diagnosis of cardiovascular disease, were also linked to purchase of nitrate vasodilators (C01D). RESULTS: Among Swedes of 60 years and above, 53% purchased antihypertensive or serum lipid-reducing pharmaceuticals, and 30% purchased the pharmaceuticals without having been hospitalized for a coronary or cerebrovascular event during the previous 7 years, or having purchased prescribed nitrate vasodilators during 6 months. CONCLUSION: Over half of the Swedish senior population purchased prescribed antihypertensive or serum lipid-reducing drugs during 6 months in 2005. The magnitude of the prevalence points to the importance of intensified follow-up of both adverse effects and of effectiveness of these drugs.

BACKGROUND AND PURPOSE: Osteoclasts are unique cells to absorb bone. Targeting osteoclast differentiation is a therapeutic strategy for osteolytic diseases. Natural marine products have already become important sources of new drugs. The naturally occurring nitrobenzoyl sesquiterpenoids first identified from marine fungi in 1998 are bioactive compounds with a special structure, but their pharmacological functions are largely unknown. Here, we investigated six marine fungus-derived nitrobenzoyl sesquiterpenoids on osteoclastogenesis and elucidated the mechanisms.

EXPERIMENTAL APPROACH: Compounds were first tested by RANKL-induced NF-κB luciferase activity and osteoclastic TRAP assay, followed by molecular docking to characterize the structure-activity relationship. The effects and mechanisms of the most potent nitrobenzoyl sesquiterpenoid on RANKL-induced osteoclastogenesis and bone resorption were further evaluated in vitro. Micro-CT and histology analysis were used to assess the prevention of bone destruction by nitrobenzoyl sesquiterpenoids in vivo.

KEY RESULTS: Nitrobenzoyl sesquiterpenoid 4, with a nitrobenzoyl moiety at C-14 and a hydroxyl group at C-9, was the most active compound on NF-κB activity and osteoclastogenesis. Consequently, nitrobenzoyl sesquiterpenoid 4 exhibited suppression of RANKL-induced osteoclastogenesis and bone resorption from 0.5 μM. It blocked RANKL-induced IκBa phosphorylation, NF-κB p65 and RelB nuclear translocation, NFATc1 activation, reduced DC-STAMP but not c-Fos expression during osteoclastogenesis in vitro. Nitrobenzoyl sesquiterpenoid 4 also ameliorated LPS-induced osteolysis in vivo.

CONCLUSION AND IMPLICATIONS: These results highlighted nitrobenzoyl sesquiterpenoid 4 as a novel inhibitor of osteoclast differentiation. This marine-derived sesquiterpenoid is a promising lead compound for the treatment of osteolytic diseases.

Dengue fever is an acute infectious disease caused by dengue virus (DENV) and transmitted by Aedes mosquitoes. There is no effective vaccine or antiviral drug available to date to prevent or treat dengue disease. Recently, RNA-dependent RNA polymerase (RdRp), a class of polymerases involved in the synthesis of complementary RNA strands using single-stranded RNA, has been proposed as a promising drug target. Hence, we screened new molecules against DENV RdRp using our previously constructed virtual screening method. Mol-5, [1,2,4]triazolo[1,5-a]pyrimidine derivative, was screened out from an antiviral compound library (~8000 molecules). Using biophysical methods, we confirmed the direct interactions between mol-5 and purified DENV RdRp protein. In luciferase assay, mol-5 inhibited NS5-RdRp activity with an IC₅₀ value of 1.28 ± 0.2 μM. In the cell-based cytopathic effect (CPE) assay, mol-5 inhibited DENV2 infectivity with an EC₅₀ value of 4.5 ± 0.08 μM. Mol-5 also potently inhibited DENV2 RNA replication as observed in immunofluorescence assay and qRT-PCR. Both the viral structural (E) and non-structural (NS1) proteins of DENV2 were dose-dependently decreased by treatment with mol-5 (2.5-10 μM). Mol-5 treatment suppressed DENV2-induced inflammation in host cells, but had no direct effect on host defense (JAK/STAT-signaling pathway). These results demonstrate that mol-5 could be a novel RdRp inhibitor amenable for further research and development.

Dengue virus (DENV) annually infects 400 million people worldwide. Unfortunately, there is lack ofwidely protective vaccine or drugs against DENV. The viral RNA-dependent RNA polymerase (RdRp) ofNS5 protein is highly conserved among different DENV subtypes, thus presenting itself as an attractivetarget for drug design. In the current research, SPRi was performed to screen compounds against DENV2RdRp and 5(1H)-Quinazolinone,2-(4-bromophenyl)-2,3,4,6,7,8-hexahydro-7,7-dimethyl-1,3-diphenyl(Q63) was successfully screened out with a KD of 0.9 mM. Then, ITC and molecular docking assay wasperformed to access the binding mechanism between Q63 and DENV2 RdRp. Meanwhile, Q63 alsodecreased the intermediate dsRNA production, which was the product of RdRp. Further the antiviraleffects of Q63 were evaluated on mosquito C6/36 cells and mammalian BHK-21 cells. Q63 reduced CPEand cell toxicity effect after DENV2 infection on C6/36 and BHK-21 cells, with an EC50 of 2.08 mM. Time ofaddition assay revealed that Q63 affected the early genome RNA replication stage, including genome RNAreplication. In addition, Q63 down-regulated STAT1 phosphorylation, ISG15 and ISG54 after DENV2infection. In summary, Q63 was found to be a novel RdRp non-nucleoside inhibitor and a potential leadcompound for coping with DENV infectious disease in the future. © 2018 The Authors. 

Dengue virus (DENV) is the most prevalent mosquito borne viral pathogen worldwide. However, antiviral drugs against this infection are not available. To identify novel anti-DENV compound from traditional Chinese medicine, we discovered the ethanol extract of Acorus tatarinowii Schott containing potent anti-DENV activity and diasarone-I was isolated from this extract. Diasarone-I has antiviral effect with half maximal effective concentration (EC₅₀) of 4.5μM and half maximal cytotoxicity concentration (CC₅₀) of >80μM. Time of drug addition assay suggested that this compound inhibited at RNA replication step in the DENV life cycle. Further, in silico analysis indicated that diasarone-I might act as an inhibitor of 2'O Methyltransferase of NS5. Diasarone-I has also decreased the DENV2-induced STAT1 phosphorylation and ISGs. In summary, we suggest that diasarone-I may be a 2'O Methyltransferase inhibitor and might serve as a potential candidate for the treatment of DENV2 infections. © 2017 Elsevier B.V.

BACKGROUND: Acorus calamus l. (Acoraceae) is a well-known traditional Chinese medicinal plant, whose root are historically mainly used to treat neurodegenerative diseases, and for cholera treatment. This datum strongly indicates the antimicrobial activity of A. calamus.

PURPOSE: Our goal is to find the active constituents of A. calamus to treat dengue virus (DENV) infections, and to study the effects and mechanisms of these active substances.

METHODS: The root of A. calamus was extracted by ethanol. Mosquito larva C6/36 cells were used for DENV2 replication and transfection host. Mouse kidney fibroblast cells (BHK-21) were used as a host cell to study the infection ability of the virus. DENV2-induced cytopathic effect (CPE) and plaque assay were used to evaluate the inhibitory effect of A. calamus extracts on DENV2 infectivity inhibition. The levels of E and NS1 protein expression were measured by real-time PCR and western blot assays.

RESULTS: 12 compounds were isolated from ethanol extract of A. calamus root, tatanan A showed the best anti-DENV ability among these 12 compounds, which significantly alleviated DENV2-induced CPE and cytotoxicity effects, with an EC₅₀ of 3.9 µM. In addition, RNA replication assay further confirmed the antivirus ability of tatanan A. Time-addition assay showed that tatanan A affected the early stage of viral RNA replication, which in turn inhibited mRNA and protein levels of DENV2.

CONCLUSIONS: These results demonstrated the anti-DENV2 effect of tatanan A, in inhibiting DENV2 RNA replication and infections. In summary, tatanan A was found to be a novel natural DENV inhibitor and a potential candidate for the treatment of DENV infectious disease.

Introduction: Seasonal influenza vaccination, together with FDA-approved neuraminidase (NA) and polymerase acidic (PA) inhibitors, is the most effective way for prophylaxis and treatment of influenza infections. However, the low efficacy of prevailing vaccines to newly emerging influenza strains and increasing resistance to available drugs drives intense research to explore more effective inhibitors. Hemagglutinin (HA), one of the major surface proteins of influenza strains, represents an attractive therapeutic target to develop such new inhibitors. Areas covered: This review summarizes the current progress of HA-based influenza virus inhibitors and their mechanisms of action, which may facilitate further research in developing novel antiviral inhibitors for controlling influenza infections. Expert opinion: HA-mediated entry of influenza virus is an essential step for successful infection of the host, which makes HA a promising target for the development of antiviral drugs. Recent progress in delineating the crystal structures of HA, especially HA-inhibitors complexes, has revealed a number of key residues and conserved binding pockets within HA. This has opened up important insights for developing HA-based antiviral inhibitors that have a high resistance barrier and broad-spectrum activities.

Background: Human urate transporter 1 (hURAT1) is the most pivotal therapeutic target for treating hyperuricemia. However, the molecular interactions between uric acid and URAT1 are still unknown due to lack of structural details. Methods: In the present study, several methods (homology modeling, sequence alignment, docking, and mutagenesis) were used to explain the atomistic mechanisms of uric acid transport of hURAT1. Results: Residues W357-F365 in the TMD7 and P484-R487 in the TMD11 present in the hURAT1 have unique roles in both binding to the uric acid and causing subsequent structural changes. These residues, located in the transport tunnel, were found to be related to the structural changes, as demonstrated by the reduced V _max values and an unaltered expression of protein level. In addition, W357, G361, T363, F365, and R487 residues may confer high affinity for binding to uric acid. An outward-open homology model of hURAT1 revealed a crucial role for these two domains in the conformational changes of hURAT1. F241 and H245 in TMD5, and R477 and R487 in TMD11 may confer high affinity for uric acid, and as the docking analysis suggests, they may also enhance the affinity for the inhibitors. R477 relation to the structural changes was demonstrated by the V _max values of the mutants and the contribution of positive charge to the uric acid selectivity. Conclusions: W357-F365 in TMD7, P484-R487 in TMD11, and residues F241, H245, and R477 were found to be critical for the translocation and recognition of uric acid.

Pattern recognition receptors (PRRs) are key immune receptors of the innate immune system, which recognize the conserved pathogen-associated molecular patterns (PAMPs) of the invading pathogens. Compared to the adaptive immune receptors, PRRs have three distinguishing features, viz., universal expression, fast response and recognizing many kinds of microbes. Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), C-type lectin receptors (CLRs) and NOD-like receptors (NLRs) recognize viral nucleic acid/bacterial fragments and trigger anti-microbial innate immune responses. Upon recognition of their ligand species, PRRs recruit specific intracellular adaptor proteins to initiate signaling pathways culminating in the activation of nuclear factor-κB (NF-κB), mitogen-activated protein (MAP) kinases and interferon regulatory factors (IRFs) that control the transcription of genes encoding pro-inflammatory factors including type I interferon and other inflammatory cytokines, which are critical for eliminating the potential threat to the host. Here, we summarize the effects of small molecule regulators acting on signaling pathways initiated by TLR, RLR and NLR as well as their influence on innate and adaptive immune responses leading to therapy. © 2017 Elsevier Masson SAS