Blockade of the programmed cell death-1 (PD-1)/programmed cell death ligand 1 (PD-L1) pathway is an attractive strategy for immunotherapy, but the clinical application of small molecule PD-1/PD-L1 inhibitors remains unclear. In this work, based on BMS-202 and our previous work YLW-106, a series of compounds with benzo[d]isothiazol structure as scaffold were designed and synthesized. Their inhibitory activity against PD-1/PD-L1 interaction was evaluated by a homogeneous time-resolved fluorescence (HTRF) assay. Among them, LLW-018 (27c) exhibited the most potent inhibitory activity with an IC50 value of 2.61 nM. The cellular level assays demonstrated that LLW-018 exhibited low cytotoxicity against Jurkat T and MDA-MB-231. Further cell-based PD-1/PD-L1 blockade bioassays based on PD-1 NFAT-Luc Jurkat cells and PD-L1 TCR Activator CHO cells indicated that LLW-018 could interrupt PD-1/PD-L1 interaction with an IC50 value of 0.88 μM. Multi-computational methods, including molecular docking, molecular dynamics, MM/GBSA, MM/PBSA, Metadynamics, and QM/MM MD were utilized on PD-L1 dimer complexes, which revealed the binding modes and dissociation process of LLW-018 and C2-symmetric small molecule inhibitor LCH1307. These results suggested that LLW-018 exhibited promising potency as a PD-1/PD-L1 inhibitor for further investigation.

The comprehensive evaluation of the quality of traditional Chinese medicines (TCM) is an important issue for the continuous progress and exploration of TCM. In this study, a "Yiqing" tablet (YQT) was taken as an example, and the sample quality was comprehensively investigated by multi-component quantification, multi-dimensional fingerprint construction, and antioxidant activity analysis. Based on high performance liquid chromatography (HPLC) and fourier transform infrared spectroscopy (FTIR) fingerprint, accurate and fast multi-component quantification is achieved by reliable Multi-markers assay by mono-linear method (MAML) method and verified partial least squares regression (PLSR) model. The basic HPLC fingerprint and the special FTIR quantitative fingerprint were evaluated by SQFM, and the rich fingerprint qualitative and quantitative information of the sample was obtained. The characteristic parameter (blocking rate (BR)) characterizing antioxidant activity in the electrochemical (EC) fingerprint was excavated for the first time, and the fingerprint-efficacy analysis results with HPLC and FTIR were obtained through bivariate correlation analysis (BCA). The results showed that 25 components in the HPLC fingerprint and had antioxidant activity, and most bands of FTIR showed antioxidant activity. Finally, by combining the evaluation results of HPLC and FTIR fingerprint using the mean method, all samples were classified as first level, except for S1, demonstrating the consistency of sample quality. Based on the comprehensive quality evaluation system combining vertical and horizontal combination, this study provides a new idea for achieving comprehensive quality evaluation of TCM.

We formerly reported that EZH2 inhibitors sensitized HIF-1 inhibitor-resistant cells and inhibited HIF-1α to promote SUZ12 transcription, leading to enhanced EZH2 enzyme activity and elevated H3K27me3 levels, and conversely, inhibition of EZH2 promoted HIF-1α transcription. HIF-1α and EZH2 interacted to form a negative feedback loop that reinforced each other's activity. In this paper, a series of 2,2- dimethylbenzopyran derivatives containing pyridone structural fragments were designed and synthesized with DYB-03, a HIF-1α inhibitor previously reported by our group, and Tazemetostat, an EZH2 inhibitor approved by FDA, as lead compounds. Among these compounds, D-01 had significant inhibitory activities on HIF-1α and EZH2. In vitro experiments showed that D-01 significantly inhibited the migration of A549 cells, clone, invasion and angiogenesis. Moreover, D-01 had good pharmacokinetic profiles. All the results about compound D-01 could lay a foundation for the research and development of HIF-1α and EZH2 dual-targeting compounds.

The development and utilization of new dienes and dienophiles for the controlled synthesis of isoquinuclidines is highly appealing. Herein, we describe a novel strategy for diastereoselective synthesis of indoline-fused isoquinuclidines via copper-catalyzed dearomative Diels-Alder reaction of cyclic amidines with indoles. This protocol avoids the use of unstable DHPs and activated alkenes, offering a more efficient and selective approach to synthesize isoquinuclidines.

Alkenyl oxindoles have been characterized as autophagosome-tethering compounds (ATTECs), which can target mutant huntingtin protein (mHTT) for lysosomal degradation. In order to expand the application of alkenyl oxindoles for targeted protein degradation, we designed and synthesized a series of hetero-bifunctional compounds by conjugating different alkenyl oxindoles with the BRD4 inhibitor JQ1. Through structure-activity relationship study, we successfully developed JQ1-alkenyl oxindole conjugates that potently degrade BRD4. Unexpectedly, we found that these molecules degrade BRD4 through the ubiquitin-proteasome system, rather than the autophagy-lysosomal pathway. Using pooled CRISPR interference (CRISPRi) screening, we revealed that JQ1-alkenyl oxindole conjugates recruit the E3 ubiquitin ligase complex CRL4DCAF11 for substrate degradation. Furthermore, we validated the most potent hetero-bifunctional molecule HL435 as a promising drug-like lead compound to exert antitumor activity both in vitro and in vivo . Our research provides new employable PROTAC moieties for targeted protein degradation, providing new possibilities for drug discovery.### Competing Interest StatementThe authors have declared no competing interest.

Epoxyeicosatrienoic acids with anti-inflammatory effects are inactivated by soluble epoxide hydrolase (sEH). Both sEH and histone deacetylase 6 (HDAC6) inhibitors are being developed as neuropathic pain relieving agents. Based on the structural similarity, we designed a new group of compounds with inhibition of both HDAC6 and sEH and obtained compound M9. M9 exhibits selective inhibition of HDAC6 over class I HDACs in cells. M9 shows good microsomal stability, moderate plasma protein binding rate, and oral bioavailability. M9 exhibited a strong analgesic effect in vivo, and its analgesic tolerance was better than gabapentin. M9 improved the survival time of mice treated with lipopolysaccharide (LPS) and reversed the levels of inflammatory factors induced by LPS in mouse plasma. M9 represents the first sEH/HDAC6 dual inhibitors with in vivo antineuropathic pain and anti-inflammation.

Based on the reported routes, a convergent eight-step approach to Gilteritinib fumarate was designed, which is suitable for industrial implementation. The first key intermediate, 3,5-dichloro-6-ethylpyrazine-2-carboxamide ( 6 ), was obtained from 2,6-dichloropyrazine in 49.3% yield by two subsequent one-pot Minisci reactions. The second key intermediate, 1-methyl-4-(piperidin-4-yl)piperazine ( 3 ), was synthesized from Boc-4-piperidone and N -methylpiperazine in 88.5% overall yield via two step reaction sequence that included reductive amination and the removal of Boc protective group. The target compound, Gilteritinib fumarate, was obtained in five steps from 1-fluoro-2-methoxy-4-nitrobenzene in 46.9% overall yield. Of special interest in this reaction sequence is the Ullmann-type coupling that was performed in the presence of the CuI- l -quebrachitol catalytic system providing an excellent catalytic effect.

Multiple studies have established the Pks13-TE domain as a promising target for anti-tuberculosis drug development. However, recent findings have revealed that the lead compound currently in the pipeline for Pks13-TE has significant cardiotoxicity issues. Given the pressing need for new chemical structures for Pks13-TE inhibitors, this study aims to provide a detailed understanding of the Pks13-TE domain binding site through the application of computational chemical biology techniques. Our results highlight the size and shape of the Pks13-TE domain binding pocket, key residues including Asp1644, Asn1640, Phe1670, and Tyr1674 within the pocket, and inhibitor pharmacophore characteristics such as aromatic ring sites, positively charged sites, and hydrogen bond donors. To our knowledge, these simulation results are novel and contribute to the discovery of next-generation Pks13-TE inhibitors without similar prior studies.

Reagent system NaBH 4 —ZnCl 2 was found to be suitable for the reduction of various substituted quinoxalines to obtain corresponding 1,2,3,4-tetrahydro derivatives. The reduction proceeds quickly under mild reaction conditions. The procedure is convenient and can be applied for a wide scope of substrates.

Hypoxia-inducible factor-1 (HIF-1) as a key mediator in tumor metastasis, angiogenesis and poor patient prognosis, has been recognized as an important cancer drug target. Up to now, some HIF-1 inhibitors with diverse skeletal structures were reported as anticancer agents, mostly natural product-derived compounds. In this study, we designed and synthesized a series of chalcone-based compounds with 2,2-dimethylbenzopyran using the combination principles to select benzopyrans and chalcones natural products. A novel series of chalcone-based compounds with 2,2-dimethylbenzopyran were evaluated as HIF-1 inhibitor. HRE luciferase reporter assay demonstrated compounds showed superior HIF-1 inhibitory activity. Among them, compound 16e exhibited the best features: the strongest HIF-1 inhibitory activity (IC50 = 2.38 μM, 3-fold higher than that of LXH-SYP-7). Meanwhile, it also significantly suppressed migration and VEGF-induced invasion of A549 cells in nontoxic concentrations. Additionally, tube formation assay demonstrated its anti-angiogenesis activity. Moreover, the in vivo study indicated that compound 16e could retard angiogenesis in the matrigel plug assay model, and almost no new blood vessels were formed in the suppository when it reached 20 μM. Finally, we also performed a subchronic toxicity test in which doses up to 50 mg/kg were administered orally for 10 days in Kunming mice with no toxic adverse effects and were well tolerated. These findings support the further investigation on the anti-invasive and anti-angiogenic potential of this class of compounds as HIF-1 inhibitor.

A photo and Cu-mediated radical-radical approach enabling the one-step synthesis of the phthalideisoquinoline skeleton has been reported. Under mild reaction conditions, a series of N-aryl phthalideisoquinolines containing various substituents were synthesized in moderate to good yields. Bioactivity data demonstrated that a new compound 4x can efficiently inhibit the growth of multiple tumor cell lines with enhancements of more than 10-fold by significantly increasing G2/M arrest compared with noscapine.

Research onβ3-AR, the new member of the adrenoceptor family, is in its infancy and few β3-AR agonists have been approved for marketing to date. Meanwhile, β3-AR exhibited obvious species differences in pharmacological properties, such as between human and animals, however, the 3D structure of human β3-AR has not been published, which makes it difficult to understand the interaction between human β3-AR and its agonists. Herein, binding patterns of β3-AR agonists are explored starting from the Alphafold predicted structural model, and the obtained model was optimized by using molecular dynamics simulations. Moreover, the human β3-AR and its agonists were subjected to molecular docking, dynamics simulations, binding free energy calculations and pharmacophore modeling to elucidate the characteristics of human β3-AR activity pockets and agonist conformational relationships, including a hydrophobic group, a positively charged group as well as two hydrogen-bonded donors, which provide comprehensive insights into the interactions between human β3-AR and its agonists.

Soluble epoxide hydrolase (sEH) has been identified as an attractive target for anti-inflammatory drug design in recent years. Picomolar level compound G1 against sEH was obtained by introducing the hydrophilic group homopiperazine and hydrophobic fragment propionyl onto the structure of lead compound A. G1 showed good microsomal stability, a moderate plasma protein binding rate, and good oral bioavailability and was well tolerated in rats. G1 has significant analgesic effects on CFA-induced AIA mice, ameliorated the pancreatic injury in acute pancreatitis induced by l-arginine, reversed pancreatic injury, edema, and neutrophil infiltration, and increased the survival time of C57BL/6 mice in a lipopolysaccharide (LPS)-induced sepsis model. Moreover the expression levels of sEH, COX-2, NOS-2, vascular cell adhesion molecule (VCAM), IL-6, MCP-5, and tumor necrosis factor α (TNF-α) were measured by Western blot or enzyme-linked immunosorbent assay (ELISA), with varying degrees of decrease. These results suggested that G1 is a drug candidate worthy of further evaluation for the treatment of inflammation-induced diseases such as arthritis, acute pancreatitis, and sepsis.

Antiviral oral liquid (AOL), which had antipyretic, antiviral, and anti-inflammatory pharmacological effects, was currently used to treat Coronavirus Disease 2019 (COVID-19). However, the mechanism of action was still unclear. Additionally, there had long been a lack of a comprehensive quality assessment and in vitro antioxidant activity study. The aim was to construct fingerprints of AOL and conduct the fingerprint-efficacy relationship study, as well as to investigate the mechanism of action of AOL in treating COVID-19. Firstly, four fingerprints of AOL were established using HPLC, GC, UV, and electrochemical analysis methods. Then, the quality consistency of AOL was evaluated qualitatively and quantitatively using the average linear quantitative fingerprint method and chemometric method. In addition, the fingerprint efficacy relationship between the fingerprint peaks of AOL and the antioxidant activity measured by DPPH was constructed using the OPLS model, while Pearson's correlation coefficient was used to explore the relationship between the fingerprint peaks and electrochemical analysis. Finally, network pharmacology and molecular docking were used to study AOL's potential active substances, targets, and mechanisms to treat COVID-19. The 22 samples were divided into different grades. AOL showed good antioxidant predictive power, providing a favorable direction for finding and predicting bioactive compounds and potential UV wavelengths. 100 compounds and 263 potential targets were identified, providing a theoretical basis for subsequent clinical studies. This paper investigated AOL in two dimensions of quality consistency and efficacy, providing a new way of thinking for other herbal compound preparations to achieve dual control of quality and effectiveness.

RARγ is a therapeutic target for many skin diseases and has potential in cancer treatment. In the current study, we put forward a comprehensive structure–activity relationship study of third and fourth generations of RARγ agonists, addressing multiple crystal structures of RARγ complexes and approved drugs. Adapalene and Trifarotene, through hybrid strategies including protein contacts Atlas analysis, molecular docking, dynamics simulations, MM-GBSA, ASM, and pharmacophore modeling. Our result revealed crucial amino acids Arg267, Ser278, Phe288, Phe230, Met272, Leu271, and Leu268 within the RARγ pocket, as well as pharmacophore features such as two hydrophobic groups, two aromatic rings, and negative ionic features, which are essential for the binding of RARγ agonists. Based on this study, the binding mechanism of RARγ agonists was elucidated, which will be helpful for the rational design of new RARγ agonists for skin diseases and cancer treatment. In this study, Schrödinger suite 2021–2 with OPLS_4 force field, Discovery Studio program 3.0, LigandScout 4.3, and PyMOL are utilized in the investigation.

In recent years, it has been proposed that G9a/EZH2 dual inhibition is a promising cancer treatment strategy. Herein, we present the discovery of G9a/EZH2 dual inhibitors that merge the pharmacophores of G9a and EZH2 inhibitors. Among them, the most promising compound 15h displayed potent inhibitory activities against G9a (IC50 = 2.90 ± 0.05 nM) and EZH2 (IC50 = 4.35 ± 0.02 nM), superior antiproliferative profiles against RD (CC50 = 19.63 ± 0.18 μM) and SW982 (CC50 = 19.91 ± 0.50 μM) cell lines. In vivo, 15h achieved significant antitumor efficacy in a xenograft mouse model of human rhabdoid tumor with a tumor growth inhibitory rate of 86.6% without causing observable toxic effects. The on-target activity assays illustrated that compound 15h can inhibit tumor growth by specifically inhibiting EZH2 and G9a. Therefore, 15h is a potential anticancer drug candidate for the treatment of malignant rhabdoid tumor.

Triple-negative breast cancer (TNBC) is an extremely aggressive tumor with limited treatment options and effectiveness. Dual-target inhibitors capable of simultaneously suppressing invasion may represent a promising therapeutic approach for TNBC. In this work, we developed a series of dual BRD4/Src inhibitors by connecting JQ1 and dasatinib using various linkers and evaluated their efficacy against TNBC both in vitro and in vivo. Among these compounds, HL403 demonstrated IC50 values of 133 nM for BRD4 inhibition and 4.5 nM for Src inhibition. Most importantly, HL403 not only exhibited potent anti-proliferative capabilities, but also effectively suppressed the invasion of MDA-MB-231 cells in vitro. Finally, the anti-tumor efficacy of HL403 was validated in a mouse MDA-MB-231 xenograft tumor model, achieving a tumor growth inhibition rate (TGI) of 70.7 %, which was superior to the combination of JQ1 and dasatinib (TGI = 54.0 %). Our research provides a promising and feasible new strategy for improving the treatment of TNBC.

Quality consistency evaluation of Paeonia Radix Alba combined with multidimensional quantitative fingerprinting and antioxidant analysis.

2,2'-dipyridylamine (DPA) is hereby firstly reported as a novel and efficient tridentate ligand for promoting copper(I)-catalyzed C-O/S coupling reactions of abundant aryl bromines with related phenols or thiophenols. The key to this very discovery is the identification of an N-donor tripod ligand (DPA), and such a reaction allows the conversion of simple starting materials to complex diaryl ethers or diaryl thioethers, which are important synthetic intermediates in the preparation of bioactive molecules.

Adenosine A1receptor (A1AR) and adenosine A2Areceptor (A2AAR) are AR isoforms that share high homology but play many different roles in terms of regulating arteriolar pressure and urine flow as well as relieving neurodegenerative disorders.